INSTRUMENT HANDBOOK. Biacore 2000

Transcription

1 INSTRUMENT HANDBOOK Biacore 2000

2 BIACORE 2000 Instrument Handbook.ERYEV]

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

4 Contents M Contents 1. Introduction &MSQSPIGYPEVMRXIVEGXMSREREP]WMW 7EJIX]MRJSVQEXMSR 'SRJSVQERGI[MXLWXERHEVHW 2. Description &-%'36)TVSGIWWMRKYRMX 4YQTW %YXSWEQTPIV 'SRRIGXSVFPSGO -RXIKVEXIH *PYMHMG'EVXVMHKI-*' 7IRWSVGLMTW 3TXMGEPW]WXIQ 8IQTIVEXYVIGSRXVSP 0)(WXEXYWMRHMGEXSVW &-%'36)W]WXIQGSRXVSP &-%'36)'SRXVSP7SJX[EVI &-%IZEPYEXMSRWSJX[EVI &-%WMQYPEXMSRWSJX[EVI 3. Installation -RWXVYQIRXTPEGMRK 1SZMRK&-%'36) ;MXLMRXLIPEFSVEXSV] 0SRKIVHMWXERGIW 'SRRIGXMSRW 7SJX[EVIMRWXEPPEXMSR 4S[IVYT 0MUYMHW]WXIQWXEVXYT 4. Operation 9WMRKXLIWSJX[EVI +IRIVEP;MRHS[WXIGLRMUYIW 7XEVXMRK&-%'36)'SRXVSP7SJX[EVI &-%'36)WSJX[EVIWGVIIRVIKMSRW 'SQQERHWERHXSSPFEVFYXXSRW 4VITEVMRKXLIW]WXIQ 4VITEVMRKWSPYXMSRW

5 Contents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

6 Contents MMM 5. Application Wizards 9WMRKETTPMGEXMSR[M^EVHW 7XEVXMRKETTPMGEXMSR[M^EVHW ;M^EVHXIQTPEXIW +IXXMRKLIPTSR[M^EVHW 8LIWYVJEGITVITEVEXMSR[M^EVH 7YVJEGITVITEVEXMSR[M^EVHJYRGXMSRW -QQSFMPM^EXMSRT,WGSYXMRK -QQSFMPM^EXMSR 6IKIRIVEXMSRWGSYXMRK 7YVJEGITIVJSVQERGIXIWX 8VSYFPIWLSSXMRKWYVJEGITVITEVEXMSR 8LIFMRHMRKEREP]WMW[M^EVH 8LIOMRIXMGEREP]WMW[M^EVH /MRIXMGEREP]WMW[M^EVHJYRGXMSRW 2SXIWSRXLIOMRIXMGEREP]WMW[M^EVH -RXITVIXMRKVIWYPXWJVSQXLIOMRIXMGEREP]WMW[M^EVH 6. Customized Applications 3ZIVZMI[SJXLIGYWXSQM^IHETTPMGEXMSR[M^EVH (IJMRMRKXLIGSQQERHWIUYIRGI %WWMKRMRKZEPYIWXSZEVMEFPIW %WWMKRMRKVEGOTSWMXMSRW 4VITEVMRKXLIVYR 'SRWXERXERHZEVMEFPIGSQQERHTEVEQIXIVW 'YWXSQM^IHETTPMGEXMSRGSQQERHW 9WMRKHIXIGXMSRQSHIERHJPS[TEXL (IXIGXMSRQSHI *PS[TEXL *PS[GIPPREQIW 9WMRKOI][SVHW 9WMRKIQTX]QM\MRKZMEPW 9WMRK1-< 9WMRKVITSVXTSMRXW 9WMRKGSRHMXMSREPGSQQERHW 9WMRK;%-8 'YWXSQM^IHETTPMGEXMSRI\EQTPIW -RLMFMXMSREWWE] 'SRHMXMSREPVIKIRIVEXMSR 'SRHMXMSREPGSRXVSPI\TIVMQIRXW

7 Contents MZ 7. Maintenance 6IKYPEVQEMRXIRERGI %XXLIIRHSJXLIHE] 7]WXIQGPIERMRK 3XLIVVSYXMRIW 9WIVXSSPW ;SVOMRKXSSPW 8IWXXSSPW 7IVZMGIXSSPW 8. Troubleshooting 7IRWSVKVEQTVSFPIQW 7MKREPRSMWI 0S[HEXEUYEPMX] 7TMOIW (VMJX 6ITVSHYGMFPIHMWXYVFERGIW 7LMJXW 2SVIWTSRWISRMRNIGXMSR 4SSVVITVSHYGMFMPMX] 7EQTPIHMWTIVWMSR 3XLIVETTPMGEXMSRTVSFPIQW -RWXVYQIRXTVSFPIQW +IRIVEPJEMPYVI 0MUYMHHIPMZIV] %YXSWEQTPIV 'SRRIGXSVFPSGOERH-*' 7IRWSVGLMT 8IQTIVEXYVIGSRXVSP 7SJX[EVITVSFPIQW 9. BIACORE Control Software reference 7XEVXMRK&-%'36)'SRXVSP7SJX[EVI 7XEVXMRKJVSQXLI;MRHS[W7XEVXQIRY 7XEVXMRKJVSQ;MRHS[W)\TPSVIV 1YPXMTPIWSJX[EVIMRWXERGIW 6IGSZIVMRKPSWXHEXE 7XEVXYTVSYXMRIW 7GVIIRHMWTPE] 1IRYFEV 8SSPFEV 7IRWSVKVEQ[MRHS[ 6ITSVXTSMRXXEFPI )ZIRXPSK[MRHS[ 7XEXYW[MRHS[ 1IXLSHIHMXSV[MRHS[ 1IRYWERHFYXXSRW

8 Contents *MPIQIRY )HMXQIRY 7IRWSVKVEQ[MRHS[ 1IXLSHIHMXSV :MI[QIRY 'SQQERHQIRY 6YRQIRY 8SSPWQIRY ;MRHS[QIRY,IPTQIRY A. System specifications %7]WXIQTIVJSVQERGI %8IGLRMGEPWTIGMJMGEXMSRW %8IGLRMGEPHIWGVMTXMSR % % % B. Application guidelines &+IRIVEPGSRWMHIVEXMSRW &8]TIWSJETTPMGEXMSR &1ERYEPZIVWYWQIXLSHGSRXVSPPIHSTIVEXMSR &7TIGMEPJIEXYVIWSJ&-%'36) &+VEHMIRXWYVJEGIW &-RPMRIVIJIVIRGIGIPP &)\TIVMQIRXEPGSRWMHIVEXMSRW &7EQTPITVITEVEXMSR &0MKERHMQQSFMPM^EXMSR &*PS[VEXI &-RNIGXMSRXIGLRMUYIW &9WMRKEVIJIVIRGIGIPP &6IHYGMRKGEVV]SZIV &8IQTIVEXYVIGLERKIW &6IKIRIVEXMSR &;EWLMRKXLIPMUYMHLERHPMRKW]WXIQ & & & & & & & & & & & & & & & & C. Chemical resistance and recommended solutions '+IRIVEPGLIQMGEPVIWMWXERGI ' '&YJJIVWSPYXMSRW ' '6IKIRIVEXMSRWSPYXMSRW ' '(MWMRJIGXERXW ' D. Status and event log messages Z

9 Contents E. Writing BIACORE 2000 methods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ethod language reference Index ZM

10 Introduction 1. Introduction 1.1 Biomolecular interaction analysis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

11 Introduction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

12 Introduction 1.2 Safety information &-%'36)4VSGIWWMRK9RMXGSRXEMRWQEMRWZSPXEKIERHLERHPIW PMUYMHW[LMGLQE]FILE^EVHSYW&IJSVISTIVEXMRKSVQEMRXEMRMRKXLI W]WXIQ]SYQYWXVIEHXLIQERYEPERHFIE[EVISJXLILE^EVHW HIWGVMFIHFIPS[*SPPS[XLIMRWXVYGXMSRWTVSZMHIHXSEZSMHTIVWSREP MRNYVMIWSVHEQEKIXSXLIIUYMTQIRX(SRSXYWIXLIIUYMTQIRXMRER] SXLIV[E]XLERHIWGVMFIHMRXLMWQERYEP ;%62-2+ *SV]SYVS[RWEJIX]EP[E]W[IEVETTVSTVMEXITVSXIGXMZIGPSXLMRK HYVMRKSTIVEXMSRERHQEMRXIRERGISJ&-%'36) ;% WIVIUYMVIHWEJIX]IUYMTQIRX[LIRLERHPMRKLE^EVHSYWWYFWXERGIW ;% JXLIMRWXVYQIRXMWGSRXEQMREXIH[MXLFMSLE^EVHWTPIEWIGSRXEGX ]SYVPSGEP&MEGSVI9WIV7YTTSVXJSVJYVXLIVMRJSVQEXMSREFSYX HIGSRXEQMREXMSRTVSGIHYVIW ;%62-2+ &-%'36)4VSGIWWMRK9RMXGSRXEMRWQEMRWZSPXEKISJYTXS :EG (SRSXSTIRGSZIVW(MWGSRRIGXQEMRWGSVHFIJSVIVITPEGMRKJYWIW

13 Introduction 1.3 Conformance with standards &-%'36)4VSGIWWMRK9RMXQIIXWXLIJSPPS[MRKWEJIX]WXERHEVHW ERHHMVIGXMZIW -)')27EJIX]VIUYMVIQIRXWJSVIPIGXVMGEP IUYMTQIRXJSVQIEWYVIQIRXGSRXVSPERHPEFSVEXSV]YWI ))')YVSTIERHMVIGXMZIGSRGIVRMRKIPIGXVSQEKRIXMG GSQTEXMFMPMX] ))')YVSTIERHMVIGXMZIGSRGIVRMRKPS[ZSPXEKIHIZMGIW

14 Description 2. Description &-%'36)GSRWMWXWSJETVSGIWWMRKYRMX[MXLPMUYMHLERHPMRKERH STXMGEPW]WXIQWERHE4'VYRRMRK&-%'36)'SRXVSP7SJX[EVI7IRWSV GLMTWERHSXLIVGSRWYQEFPIWEVIEZEMPEFPIJVSQ&MEGSVI%& Figure 2-1. BIACORE 2000 system processing unit and controlling computer. 2.1 BIACORE 2000 processing unit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

15 Description &-%'36)[LMGLLEWERMRNIGXMSRTSVXJSVPSEHMRKWEQTPIWMRXS XLI-*' IFC Connector block Autosampler Sensor Chip Eluent pump Autosampler pump Waste bottle Buffer bottle Figure 2-2. The liquid delivery system with two pumps and an autosampler transports liquid to the connector block where samples and buffers are injected to the IFC. 8LIHSSVXSXLIEYXSWEQTPIVERHFYJJIVGSQTEVXQIRXMWPSGOIHHYVMRK MRWXVYQIRXSTIVEXMSRXSTVSXIGXXLISTIVEXSVJVSQMRNYV]F]XLI QSZMRKRIIHPISJXLIEYXSWEQTPIV8LIMRWXVYQIRXGERRSXFISTIVEXIH [MXLXLIHSSVSTIR Pumps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

16 Description HMWTIRWMRKWEQTPIWMRXLIEYXSWEQTPIV8LIEYXSWEQTPIVTYQTGER EPWSFITVSKVEQQIHXSHMWTIRWIFYJJIVJVSQXLIFYJJIVZIWWIPMRXS EYXSWEQTPIVZMEPW7EQTPIWEVITVIZIRXIHJVSQQM\MRK[MXLTVMQMRK FYJJIVF]WQEPPEMVWIKQIRXWQEMRXEMRIHMRXLITYQTXYFMRK 8LIFYJJIVGSQTEVXQIRXGERLSPHX[SQPGSRXEMRIVWSRIJSV FYJJIVERHSRIJSVGSPPIGXMRK[EWXIIJJPYIRXJVSQXLIJPS[GIPPW Autosampler 8LIEYXSWEQTPIVMWYWIHJSVXVERWJIVVMRKHMPYXMRKERHQM\MRKWEQTPIW MRNIGXMRKWEQTPIWMRXSXLI-*'ERHJSVWEQTPIVIGSZIV]ERHJVEGXMSR GSPPIGXMSR-XMWWMXYEXIHHMVIGXP]EFSZIXLIFYJJIVGSQTEVXQIRX Figure 2-3. The autosampler compartment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

17 Description Connector block 8LIGSRRIGXSVFPSGOLEWX[SMRPIXTSVXWETYQTTSVXJSVFYJJIVJPS[ [MXLEWGVI[RMTTPIJMXXMRKJSVXLITYQTXYFMRKERHERMRNIGXMSRTSVX JSVXLIEYXSWEQTPIVRIIHPIXSMRNIGXWEQTPIERHVIEKIRXW8LIWIMRPIXW GSRRIGXHMVIGXP][MXLJPS[GLERRIPWMRXLI-*'8LIVIEVISYXPIXTSVXW JSV[EWXIERHJSVVIGSZIVIHWEQTPI 1HHGOH ZDVK 7R ZDVWH 5HFRYHU,QMHFW Figure 2-4. Side view of the connector block, showing the injection port, wash station and recovery cup Integrated µ-fluidic Cartridge (IFC) 8LI-*'GSRWMWXWSJEWIVMIWSJGLERRIPWERHTRIYQEXMGZEPZIWIRGEWIH MRETPEWXMGLSYWMRKERHWIVZIWXSGSRXVSPHIPMZIV]SJPMUYMHXSXLI WIRWSVGLMTWYVJEGI7EQTPIWEVIMRNIGXIHJVSQXLIEYXSWEQTPIVMRXS XLI-*'[LMGLGSRRIGXWHMVIGXP][MXLXLIHIXIGXSVJPS[GIPPW Figure 2-5. Diagram of the channels and valves in the IFC as seen from above. The inset shows a side view of how the flow cells are formed by pressing the sensor chip against the IFC. Not to scale.

18 Description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

19 Description Figure 2-6a. Schematic illustration showing the flow paths during direct injection. The symbols o and represent open and closed valves respectively. Not to scale. During preparation, the leading sample is flushed to waste to ensure minimum dispersion at the front of the injected sample plug. The continuous flow pump maintains a constant flow of buffer through the flow cell. The enlargement shows two air segments separating the sample from buffer which prevent mixing in the autosampler tubing and IFC channels. During sample injection, the sample is driven directly through the flow cell by the autosampler pump.

20 Description Figure 2-6b. Schematic illustration showing the flow paths during loop injection. The symbols o and represent open and closed valves respectively. Not to scale. During loop loading, the continuous flow pump maintains a constant flow of buffer through the flow cell. During sample injection, buffer from the continuous flow pump pushes the sample out of the sample loop over the sensor surface in the opposite direction from loading.

21 Description Prism Opto-interface Sensor chip Prism Integrated µ-fluidic Cartridge with flow cell block Flow cells Figure 2-7. Precision-cast channels in the surface of the IFC define four flow cells on the sensor chip surface. These are matched by ridges of transparent polymer on the opto-interface, ensuring good optical contact between the components of the system. Not to scale. 8LI-*'EPPS[WWMRKPISVQYPXMGLERRIPEREP]WMWMRYTXSJSYVJPS[GIPPW *PS[GIPPWERHGERFIYWIHEWSRPMRIVIJIVIRGIGIPPWEPPS[MRK FPEROWYFXVEGXIHHEXEXSFITVIWIRXIHHMVIGXP]SRXLIWGVIIRHYVMRK EREP]WMW Figure 2-8. Simplified diagrammatic illustration of sample flow through two flow cells. The symbols o and represent open and closed valves respectively. Valves and channels not used in other flow path arrangements are not included. Not to scale. 7EQTPIMRNIGXMSRGSQQERHWEGXMZEXIEWIUYIRGISJSTIVEXMSRWGSZIVMRK EWTMVEXMSRSJWEQTPIJVSQXLIEYXSWEQTPIVZMEPHIPMZIVMRKXLIVIUYMVIH EQSYRXSJWEQTPIXSXLIJPS[GIPPWERHJPYWLMRKXLI-*'[MXLFYJJIV VIEH]JSVXLIRI\XMRNIGXMSR

22 Description Sensor chips 7IRWSV'LMT'1MWEKIRIVEPTYVTSWIGLMTGSEXIH[MXLGEVFS\] QIXL]PEXIHHI\XVERXS[LMGLFMSQSPIGYPIWGERFIPMROIH[MXLEZEVMIX] SJIWXEFPMWLIHGLIQMGEPQIXLSHWWII&-%XIGLRSPSK],ERHFSSOJSVE HIWGVMTXMSRSJMQQSFMPM^EXMSRGLIQMWXV]3XLIVWIRWSVGLMTWEVIEPWS EZEMPEFPIJSVWTIGMJMGETTPMGEXMSREVIEW Surface matrix Linker layer Gold film Glass support Figure 2-9. Sensor Chip CM5 is a glass slide coated with a thin gold film to which the carboxymethyl dextran surface matrix is covalently bonded. The matrix surface forms one wall of the detector flow cell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ptical system 8LIKPEWWWMHISJXLIWIRWSVGLMTMWTVIWWIHMRXSGSRXEGX[MXLEKPEWW TVMWQMRXLISTXMGEPYRMX%WMPMGSRISTXSMRXIVJEGIFIX[IIRXLIWIRWSV GLMTERHXLITVMWQQEXGLIHMRVIJVEGXMZIMRHI\XSXLIKPEWWIRWYVIW KSSHSTXMGEPGSYTPMRKFIX[IIRXLITVMWQERHXLIWIRWSVGLMT

23 Description 0MKLXJVSQERIEVMRJVEVIHPMKLXIQMXXMRKHMSHI0)(MWJSGYWIH XLVSYKLXLITVMWQSRXSXLIWIRWSVGLMTWYVJEGIMRE[IHKIWLETIH FIEQKMZMRKEJM\IHVERKISJMRGMHIRXPMKLXERKPIW0MKLXVIJPIGXIH JVSQXLIWIRWSVGLMTMWQSRMXSVIHF]EPMRIEVEVVE]SJPMKLXWIRWMXMZI HMSHIWGSZIVMRKXLIVERKISJMRGMHIRXPMKLXERKPIWXSEVIWSPYXMSRSJ ETTVS\MQEXIP] &]YWMRKE[IHKISJMRGMHIRXPMKLXERHEJM\IHEVVE]SJHIXIGXSVWXLI 746ERKPIMWQSRMXSVIHEGGYVEXIP]MRVIEPXMQI[MXLRSTL]WMGEP QSZIQIRXSJPMKLXWSYVGIWIRWSVGLMTSVHIXIGXSV8LI746ERKPIMW HIXIVQMRIHEXEJVIUYIRG]SJ,^QIEWYVIQIRXWWERHXLIR EZIVEKIHF]XLIQMGVSTVSGIWWSVMR&-%'36)ERHXVERWQMXXIHXS &-%'36)'SRXVSP7SJX[EVIEXVEXIWYTXS,^ Figure The detection unit houses the optical unit with light source and detector, the opto-interface and the removable sensor chip. Flow cells on the sensor chip are connected to the liquid handling system through the Integrated µ-fluidic Cartridge. The broken line shows the boundaries of the thermally insulated box Temperature control 746QIEWYVIQIRXWEVIWIRWMXMZIXSGLERKIWMRXIQTIVEXYVI-XMW MQTSVXERXXLEXEGSRWXERXXIQTIVEXYVIMWQEMRXEMRIHEXXLIWIRWSVGLMT WYVJEGIXLVSYKLSYXXLIVYR 8LIXIQTIVEXYVIGSRXVSPYWIW4IPXMIVIPIQIRXWXSQEMRXEMREGSRWXERX TVIWIXXIQTIVEXYVIEXXLIWIRWSVGLMTWYVJEGI-JXLIXIQTIVEXYVIMWRSX WXEFPIXLI]IPPS[0)(SRXLITVSGIWWMRKYRMXJPEWLIWERHXLI XIQTIVEXYVIMRHMGEXMSRMR&-%'36)'SRXVSP7SJX[EVIWLS[W EPXIVREXIP]XLITVIWIXERHEGXYEPXIQTIVEXYVI8LIXIQTIVEXYVIGSRXVSP GERQEMRXEMRETVIWIXXIQTIVEXYVIMRXLIVERKI 'EWPSRKEWXLI STIVEXMRKXIQTIVEXYVIMWRSXQSVIXLER 'FIPS[XLIEQFMIRX XIQTIVEXYVI

24 Description 8LIXIQTIVEXYVIEXXLIWIRWSVGLMTWYVJEGIMWVIGSVHIHEYXSQEXMGEPP] F]&-%'36)'SRXVSP7SJX[EVIEXXLIFIKMRRMRKERHIRHSJIZIV]VYR LED status indicators 8LIVIEVIJMZI0)(WXEXYWMRHMGEXSVWSRXLIJVSRXTERIPSJXLI MRWXVYQIRX 6IEH]KVIIR 0MKLXW[LIRTS[IVMWSRERHXLIMRWXVYQIRXMWVIEH]JSVYWI% JPEWLMRKPMKLXMRHMGEXIWXLEXGSQQYRMGEXMSR[MXLXLI4'MWFVSOIR )VVSVVIH 0MKLXWXSMRHMGEXIEWIVMSYWIVVSVMRXLIMRWXVYQIRX 8LMW0)(MWPMXJSVEFSYXWIGSRHWEJXIVXLITS[IVMWW[MXGLIHSR-J XLI0)(PMKLXWMRSXLIVGMVGYQWXERGIWXYVRSJJXLIMRWXVYQIRXERHGEPP ]SYV&MEGSVIWIVZMGIVITVIWIRXEXMZI 8IQTIVEXYVI]IPPS[ 0MKLXWWXIEHMP][LIRXLIXIQTIVEXYVIEXXLIJPS[GIPPMWWXEFPIEXXLI TVIWIXXIQTIVEXYVI*PEWLIW[LIRXLIXIQTIVEXYVIMWRSXWXEFPI 7IRWSVGLMTKVIIR 0MKLXW[LIREWIRWSVGLMTMWHSGOIHERHVIEH]9RPMX[LIRXLIVIMWRS WIRWSVGLMTERHJPEWLMRKMJEWIRWSVGLMTMWMRWIVXIHFYXRSXHSGOIH 6YRKVIIR 0MKLXW[LIREQIXLSHGSRXVSPPIHSVQERYEPWIRWSVKVEQMWVYRRMRK

25 Description 2.2 BIACORE 2000 system control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kWTVMQEV] QIQSV]ERHEZIVEKIHZEPYIWEVITEWWIHXSXLIW]WXIQGSRXVSPPIV-R XLIW]WXIQGSRXVSPPIVXLIQIEWYVIQIRXWEVIHMWTPE]IHSRXLIWGVIIREW EWIRWSVKVEQERHWXSVIHTIVQERIRXP]SRXLIGSQTYXIVLEVHHMWO &IGEYWIGSPPIGXMSRSJVE[HEXEMWQEREKIHF]EWITEVEXIQMGVS GSQTYXIVMR&-%'36)XLIXMQIVIWSPYXMSRSJQIEWYVIQIRXWMW RSXEJJIGXIHF]GSQTYXMRKEGXMZMX]SRXLI4'VYRRMRK&-%'36) 'SRXVSP7SJX[EVI7MQYPXERISYWYWISJXLI4'JSV&-%'36)GSRXVSP ERHJSVSXLIVTVSKVEQWMWLS[IZIVRSXVIGSQQIRHIH 2.3 BIACORE Control Software &-%'36)'SRXVSP7SJX[EVIVYRWYRHIV1MGVSWSJX;MRHS[WSV ;MRHS[W28ZIVWMSRSRER-&1GSQTEXMFPI4'8LIWSJX[EVI MRGPYHIWXLIJSPPS[MRKJYRGXMSRW XSSPFEVERHQIRYHVMZIRMRXIVJEGIJSVMRXIVEGXMZIQERYEPGSRXVSP ETTPMGEXMSR[M^EVHWJSVKYMHERGIMRWXERHEVHEREP]WMWTVSGIHYVIW QIXLSHIHMXSVJSVTVITVSKVEQQIHSTIVEXMSR HMVIGXVIEPXMQIHMWTPE]SJWIRWSVKVEQW GSRXMRYSYWHMWTPE]SJWIRWSVKVEQGSSVHMREXIWIPETWIHXMQIERH VIWTSRWIJPS[VEXIERHTEXLQIEWYVMRKXIQTIVEXYVIERHHIXIGXMSR QSHI EYXSQEXMGIZIRXQEVOIVWJSVIKWXEVXERHIRHSJWEQTPIMRNIGXMSR TYQTVIJMPPERHI\XVE[EWL

26 Description VIJIVIRGIPMRIJSVHMWTPE]SJIPETWIHXMQIERHEFWSPYXIVIWTSRWIEX ER]TSMRXSRXLIWIRWSVKVEQ IZIRXPSK[LMGLVIGSVHWEPPIZIRXWMRXLIGYVVIRXVYR VITSVXTSMRXXEFPI[LMGLVIGSVHWTVIHIJMRIHWIRWSVKVEQMRJSVQEXMSR EXER]VIUYIWXIHTSMRXSRXLIWIRWSVKVEQ RSXIFSSOJSVI\TIVMQIRXEPHIXEMPW YWIVXSSPWJSVWXERHEVHSTIVEXMSRWERHQEMRXIRERGI 2.4 BIAevaluation software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

27 Description 2.5 BIAsimulation software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

28 Installation 3. Installation &-%'36)[MPPFIMRWXEPPIHF]E&MEGSVIVITVIWIRXEXMZI 3.1 Instrument placing 4PEGIXLIMRWXVYQIRXSREJMVQFIRGLGPIEVSJWXVSRKIPIGXVMGEPSV QEKRIXMGJMIPHW(SRSXTPEGIZMFVEXMRKIUYMTQIRXIKFIRGL GIRXVMJYKIWSRXLIWEQIFIRGL-JTSWWMFPIEZSMHTPEGMRKXLI MRWXVYQIRXGPSWIXSE[MRHS[[LIVIHMVIGXWYRPMKLXSVZIRXMPEXMSRQE] GEYWIWYHHIRXIQTIVEXYVIGLERKIW *SVGSVVIGXJYRGXMSRMRKSJXLIXIQTIVEXYVIGSRXVSPW]WXIQQEOIWYVI XLEXXLIJERMRPIXYRHIVRIEXLXLIMRWXVYQIRXEXXLIJVSRXPIJXLERH WMHIERHSYXPIXSRXLIFEGOTERIPEXXLIPIJXLERHWMHIEVIRSX SFWXVYGXIH8LIVIWLSYPHFIEXPIEWXGQGPIEVERGIFILMRHERHEFSZI XLIMRWXVYQIRXXSEPPS[EHIUYEXIEMVGMVGYPEXMSR 3.2 Moving BIACORE Within the laboratory -J&-%'36)MWXSFIQSZIHPSGEPP][MXLMRXLIPEFSVEXSV] YRHSGOXLIWIRWSVGLMTERHWLYXHS[RXLIW]WXIQ7II7IGXMSRW ERHJSVEJYPPHIWGVMTXMSRSJXLIWISTIVEXMSRW 0MJXXLIMRWXVYQIRXF]XLILERHPIWSRIEXIEGLWMHI/IITXLI MRWXVYQIRXYTVMKLXHYVMRKQSZMRK 'EYXMSR 8LIMRWXVYQIRXMWLIEZ]OKERHQE]RIIHX[STISTPIXSPMJXMX Longer distances 'SRXEGX]SYV&MEGSVIVITVIWIRXEXMZIJSVMRWXVYGXMSRW

29 Installation 3.3 Connections 8LIMAINS INLETXLIMAINS POWERW[MXGLERHXLIQEMRWJYWIW EVIPSGEXIHSRXLIQEMRWMRTYXTERIPSRXLIPIJXWMHISJXLI MRWXVYQIRX-RTYXZSPXEKIVERKIQE\MQYQTS[IVGSRWYQTXMSR ERHJYWIVEXMRKWEVITVMRXIHSRXLIQEMRWMRTYXTERIP 'SRRIGXXLITS[IVGSVHXSXLIQEMRWMRPIX -RWXEPPGSQTYXIVTIVMTLIVEPIUYMTQIRXEGGSVHMRKXSXLIVIWTIGXMZI MRWXVYGXMSRQERYEPW9WIETEVEPPIPTSVXJSVTVMRXIVGSRRIGXMSR RSVQEPP]048ERHWIVMEPTSVX'31JSV&-%'36) VMKLXLERHWMHISJXLIMRWXVYQIRX7IVMEPTSVX'31MWJSV WIVZMGITYVTSWIW 2SXI 3R'SQTEUTIVWSREPGSQTYXIVWXLI'31WIVMEPTSVXMW QEVOIHl-3-3-%z3RSXLIVQEOIWSJGSQTYXIVXLITSVX QE]FIQEVOIHl'31zl7IVMEP4SVXzSVWMQMPEVSVRSX QEVOIHEXEPP6IJIVXS]SYVGSQTYXIVHSGYQIRXEXMSRMJ]SY EVIYRGIVXEMREFSYXXLIPSGEXMSRSJXLIWIVMEPTSVX 'LIGOXLEXXLIZSPXEKIWIPIGXSVSRXLI4'MWWIXGSVVIGXP] 3.4 Software installation &-%'36)7SJX[EVIMWWYTTPMIHSR'(6318LIWSJX[EVI[MPPFI TVIMRWXEPPIHSRHIPMZIV]-JXLIWSJX[EVIRIIHWXSFIVIMRWXEPPIHJSV ER]VIEWSRJSPPS[XLIWXITWFIPS[ 8YVRSRXLI4'ERHWXEVX1MGVSWSJX;MRHS[W -J]SYLEZIEQIQSV]VIWMHIRXZMVYWTVSXIGXMSRTVSKVEQMRWXEPPIH ]SYQE]RIIHXSHMWEFPIXLMWTVSKVEQFIJSVIMRWXEPPMRK&-%'36) 7SJX[EVI -RWIVXXLI&-%'36)7SJX[EVI'(631MRXLI'(HVMZI -JXLIWIXYTTVSKVEQHSIWRSXWXEVXEYXSQEXMGEPP]GLSSWI Add/Remove ProgramsJVSQXLI;MRHS[W'SRXVSP4ERIPERH GPMGOSRXLIInstallFYXXSR8LIMRWXEPPEXMSRTVSKVEQMWsetup.exe SRXLI'(631HMWO

30 Installation 6IEHXLIMRWXVYGXMSRWSRXLIWGVIIRXLIRGPMGONext>)RXIVXLI YWIVVIKMWXVEXMSRMRJSVQEXMSR8LITVSHYGXOI]RYQFIVMWTVMRXIH SRXLI'(FS\ 8LITVSHYGXOI]MWZEPMHJSVXLIWSJX[EVITVSKVEQW]SYLEZI TYVGLEWIH7SJX[EVIGERRSXFIMRWXEPPIH[MXLSYXEZEPMHTVSHYGX OI]'LIGOXLIFS\IWJSV&-%'36)'SRXVSP7SJX[EVI &-%IZEPYEXMSR7SJX[EVIERH&-%WMQYPEXMSRERHGPMGOSRInstall 8LIWSJX[EVI[MPPFIMRWXEPPIHMRHIJEYPXJSPHIVWYRHIV C:\Program Files\ 2SXI -J]SY[ERXXSMRWXEPPXLIWSJX[EVIMRHMJJIVIRXJSPHIVW GPMGOSRXLIETTVSTVMEXIWSJX[EVIMXIQ[MXLXLIVMKLX QSYWIFYXXSRERHGLSSWICustomized installation8lmw [MPPEPPS[]SYXSWTIGMJ]XLIHIWXMREXMSRJSPHIV -RWXEPPEXMSRMRXLIHIJEYPXJSPHIVWMWLS[IZIV VIGSQQIRHIH

31 Installation 'PMGOSRFinish[LIRXLIMRWXEPPEXMSRMWGSQTPIXI 6IIREFPI]SYVZMVYWTVSXIGXMSRTVSKVEQMJ]SYXYVRIHMXSJJMR WXIT-J]SYEVIYTHEXMRK&-%'36)7SJX[EVIWSQIZMVYW TVSXIGXMSRTVSKVEQWQE]HIXIGXXLEXI\IGYXEFPIJMPIWLEZIFIIR EPXIVIHERH[EVR]SYXLEX]SYQE]LEZIEZMVYW-KRSVIXLMW [EVRMRKXLIJMPIWLEZIFIIRlEPXIVIHzFIGEYWIXLI]EVIYTHEXIH RSXFIGEYWIXLI]EVIMRJIGXIH(SRSXLS[IZIVMKRSVIZMVYW [EVRMRKWXLEXETTIEVEXXMQIW[LIRXLIWSJX[EVILEWRSXNYWX FIIRYTHEXIH

32 Installation 3.5 Power up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iquid system start-up 4YXFYJJIVERH[EWXIVIWIVZSMVWMRTPEGI-RWIVXXLIMRPIX XYFMRK MRXSXLIFYJJIVVIWIVZSMV,&7FYJJIVMWVIGSQQIRHIHEWE KIRIVEPTYVTSWIVYRRMRKFYJJIVERHMWWYTTPMIHF]&MEGSVI%&MR HMWTSWEFPIVIEH]XSYWITEGOW %QPJPEWOSVFSXXPIMWVIGSQQIRHIHEWE[EWXI VIWIVZSMV -RWIVXERI[WIRWSVGLMT'PSWIXLIMRWXVYQIRXHSSVERHGLSSWI Command:DockMRXLI&-%'36)'SRXVSP7SJX[EVIQIRY6YR Working Tools:PrimeEJXIVHSGOMRK 2SXI 8LIMRWXVYQIRXHSSV[MPPFIPSGOIHHYVMRKSTIVEXMSR 8LMWMWEWEJIX]QIEWYVIXSTVIZIRXMRNYV]F]XLIWLEVT EYXSWEQTPIVRIIHPI 6YRWorking Tools:RinseXSJPYWLXLIPMUYMHLERHPMRKW]WXIQE WIGSRHXMQI'LIGOXLEXXLIVIEVIRSPIEOWEXXYFMRKGSRRIGXSVW 6YR Test Tools:System checkxsgligoxliw]wxiqtivjsvqergi 7II7IGXMSRJSVHIXEMPWSJXLMWTVSGIHYVI

33 Installation

34 Operation 4. Operation 8LMWGLETXIVHIWGVMFIWXLISTIVEXMSRWMRZSPZIHMRYWMRK&-%'36) 8LIGLETXIVMWSVKERM^IHMRXSXLIJSPPS[MRKWIGXMSRW 9WMRKXLIWSJX[EVI 4VITEVMRKXLIW]WXIQ 9WMRKETTPMGEXMSR[M^EVHW 1ERYEPSTIVEXMSR 5YIYMRKQERYEPGSQQERHW 1IXLSHGSRXVSPPIHSTIVEXMSR :MI[MRKERHIZEPYEXMRKVIWYPXW 4.1 Using the software General Windows techniques &-%'36)'SRXVSP7SJX[EVIVYRWYRHIV1MGVSWSJX;MRHS[WERH JSPPS[WXLIKIRIVEP;MRHS[WWXERHEVHWJSVLERHPMRKWGVIIR[MRHS[W QIRYWERHHMEPSKYIFS\IW Starting BIACORE Control Software %WLSVXGYXXSXLI&-%'36)'SRXVSP7SJX[EVIMWMRWXEPPIHYRHIVBIA ProgramsSRXLIStartQIRY -XHSIWRSXQEXXIV[LIXLIV]SYW[MXGLSRXLIMRWXVYQIRXSVWXEVXXLI WSJX[EVIJMVWX-J]SYWXEVXXLIWSJX[EVIJMVWXXLIWXEXYW[MRHS[[MPP HMWTPE]XLIQIWWEKIInstrument Offline'SRRIGXMSR[MPPFIIWXEFPMWLIH [LIRXLIMRWXVYQIRXMWW[MXGLIHSR (ITIRHMRKSRXLIGYVVIRXMRWXVYQIRXWXEXYW[LIRXLIWSJX[EVIMW WXEVXIH]SY[MPPFITVSQTXIHJSVXLIETTVSTVMEXIWIXYTEGXMSRIK HSGOEWIRWSVGLMTTVMQIXLIW]WXIQ7II7IGXMSRJSVJYVXLIV HIXEMPWSJXLIWIEGXMSRW

35 Operation BIACORE 2000 software screen regions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

36 Operation 7II'LETXIV&-%'36)'SRXVSP7SJX[EVIJSVEQSVIHIXEMPIH HIWGVMTXMSRSJXLIWGVIIRVIKMSRW Commands and toolbar buttons -RXLIHIWGVMTXMSRWMRXLMWGLETXIVGSQQERHWEVIVIJIVVIHXSF]REQI [MXLXLIQIRYREQIWITEVEXIHJVSQXLIGSQQERHREQIF]EGSPSR IKCommand:Dock1ER]SJXLIGSQQSRP]YWIHGSQQERHWEVI EWWSGMEXIH[MXLXSSPFEVFYXXSRW1ER]GSQQERHWEPWSLEZIWLSVXGYX OI]W%PMWXSJXSSPFEVFYXXSRWERHWLSVXGYXOI]WMWKMZIRSRXLIRI\X TEKI'SQQERHWRSXMRGPYHIHMRXLMWPMWXEVIEZEMPEFPISRP]JVSQXLI QIRYFEV8SSPFEVFYXXSRWEVIEPWSWLS[RMRXLIQEVKMR[LIVI ETTVSTVMEXIMRXLMWGLETXIV

37 Operation Menu/Command Toolbar button Shortcut key File Open Save Print Edit (sensorgram) Copy Graph Add Report Point Edit (method) Undo Cut Copy Paste Find Replace View Unzoom Reference Line Baseline Notebook Wizard Results Command Inject/Stop Inject Ctrl-O Ctrl-S Ctrl-P Ctrl-H Ctrl-Z Ctrl-X Ctrl-C Ctrl-V Ctrl-F Ctrl-R Ctrl-U F8 F9 Ctrl-R Transfer Mix Flow/Stop Flow Run Run/Stop Sensorgram Stop Sensorgram Tools Evaluation Ctrl-E

38 Operation 4.2 Preparing the system (SRSXTYXTETIVWYRHIVXLIMRWXVYQIRXTEVXMGYPEVP]EXXLIPIJXLERH WMHI4ETIVMWIEWMP]WYGOIHEKEMRWXXLIJERMRPIXFPSGOMRKXLIEMV MRXEOI1EOIWYVIXLEXXLIWTEGIFIX[IIRXLIFSXXSQSJXLIMRWXVYQIRX ERHXLIFIRGLMWRSXSFWXVYGXIHF]PEFSVEXSV]IUYMTQIRXRSXIFSSOW IXG Preparing solutions,&7fyjjivwevivieh]xsywi,)4)7fyjjivihwepmriwspyxmsrwwyttpmih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ocking the sensor chip &IJSVI&-%'36)GERFIYWIHEWIRWSVGLMTQYWXFIHSGOIHMR XLIMRWXVYQIRXERHXLIJPS[W]WXIQTVMQIH[MXLFYJJIV 8SHSGOXLIWIRWSVGLMTSTIRXLIWIRWSVGLMT TSVXGSZIVTYPPSYXXLI WPMHIERHMRWIVXEWIRWSVGLMTGEWWIXXIMRXLIHSGOMRKWXEXMSR[MXLXLI PEFIPJEGMRKYT[EVHWERHXLIEVVS[TSMRXMRKMRXSXLIMRWXVYQIRX(S RSXJSVGIXLIGEWWIXXIMRXSXLIWXEXMSR(SRSXWPMHIXLIWIRWSVGLMTSYX SJXLITVSXIGXMZIGEWWIXXI 7YVJEGXERX4MWWYTTPMIHEWEEWXSGOWSPYXMSR8STVITEVIFYJJIV GSRXEMRMRKWYVJEGXERXEHH PWXSGOWSPYXMSRXSQPFYJJIV

39 Operation 1. Place the sensor chip cassette on the slide with the label facing upwards and the arrow pointing into the instrument. 2. Push the slide into the instrument. 4YWLMRXLIWPMHIERHGPSWIXLIWIRWSVGLMTTSVXGSZIVXLIRGLSSWI Command:DockMRXLI&-%'36)'SRXVSP7SJX[EVIXSHSGOXLIGLMT Initiating the liquid system %JXIVHSGOMRKERI[WIRWSVGLMTVYRXLI[SVOMRKXSSPPrimeXSTVMQI XLIJPS[W]WXIQ[MXLFYJJIV8LI[SVOMRKXSSPWHMEPSKYIFS\MW EYXSQEXMGEPP]STIRIHEJXIVEHSGOMRKSTIVEXMSR 4PEGIVYRRMRKFYJJIVERHE[EWXIFSXXPIMRXLIFYJJIVGSQTEVXQIRX FIWMHIXLITYQTW4PEGIXLIFYJJIVXSXLIVMKLXERHXLI[EWXIFSXXPIXS XLIPIJX-RWIVXXLITYQTXYFMRKWQEOIWYVIXLIWIEVIGPIERJVSQ FSXLTYQTWMRXSXLIFYJJIVGEVXSRERHXLI[EWXISYXPIXXYFMRKMRXSXLI [EWXIFSXXPI -RXLI[SVOMRKXSSPWHMEPSKYIFS\QEOIWYVIPrimeMWWIPIGXIHERH GPMGOSRStart*SPPS[XLIMRWXVYGXMSRWSRXLIWGVIIR

40 Operation -J]SYHSRSX[ERXXSTVMQIXLIW]WXIQMQQIHMEXIP]EJXIVHSGOMRK GPMGOSRCancelMRXLI;SVOMRK8SSPWHMEPSKYIFS\2SXILS[IZIV XLEX]SYWLSYPHVYRPrimeFIJSVIYWMRKXLIWIRWSVGLMTJSVPMKERH MQQSFMPM^EXMSRSVMRXIVEGXMSREREP]WMW Setting autosampler rack types 8LIEYXSWEQTPIVGERLSPHX[SWEQTPIVEGOW[LMGLQE]FIIMXLIV QMGVSXMXVITPEXIWSVSRISJXLVIIXLIVQSVEGOWWII7IGXMSR% 8STPEGIVEGOWSRXLIEYXSWEQTPIVVEGOFEWITYPPFEGOXLIWPMHIVPSGO EXXLIJVSRXSJXLIFEWIERHTPEGIXLIVEGOSRXLIFEWI[MXLXLIXI\XEX XLIJVSRX8LIVEGO[MPPFIPSGOIHMRTPEGI[LIR]SYVIPIEWIXLIWPMHIV 1EOIWYVIXLEXXLIVEGOMWTVSTIVP]WIEXIHSRXLIFEWI Pull the slider lock towards you to insert or remove autosampler racks.

41 Operation 'LSSWICommand:Define Rack BaseXSXIPPXLIGSRXVSPWSJX[EVI[LMGL VEGOX]TIWEVIFIMRKYWIH8LMWWIXXMRKHIXIVQMRIW[LMGLVEGOTSWMXMSRW EVIZEPMHJSVGSQQERHWPMOITransfer Mix DiluteERHInject 7IPIGXETTVSTVMEXIVEGOX]TIWJVSQXLITYPPHS[RPMWXWERHGPMGOSROK 2SXI 8LIVIMWRSEYXSQEXMGWIRWMRKSJVEGOX]TIMRXLIWSJX[EVI-J ]SYWIXXLI[VSRKVEGOX]TIXLIEYXSWEQTPIVRIIHPIQE]FI HMVIGXIHXSXLI[VSRKSVRSRI\MWXIRXTSWMXMSRW6IQIQFIVXS VIHIJMRIXLIVEGOFEWIYWMRKXLMWGSQQERHMJ]SYGLERKIXLI X]TISJVEGOMRXLIEYXSWEQTPIV Normalizing the signal response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

42 Operation The autosampler compartment viewed from the front, showing the position R2F2 used for BIAnormalizing solution. Thermo-rack type B is shown in both base positions. 2SXI 8LIKP]GIVSPGSRGIRXVEXMSR[[MWGVMXMGEPJSVGSVVIGX RSVQEPM^EXMSR 'PSWIXLIHSSVXSXLIEYXSWEQTPIVGSQTEVXQIRX7IPIGXTools: Working Tools.7IPIGXNormalizeMRXLIPMWXERHGPMGOSRStart*SPPS[XLI MRWXVYGXMSRWSRXLIWGVIIR

43 Operation Setting the temperature 7IPIGXCommand:Set TemperatureERHIRXIVXLIVIUYMVIHEREP]WMW XIQTIVEXYVI 8LIXIQTIVEXYVIHMWTPE]MRXLIWXEXYW[MRHS[XSKKPIWFIX[IIRXLIWIX XIQTIVEXYVIERHXLIEGXYEPXIQTIVEXYVIERHXLIXIQTIVEXYVI0)(SR XLITVSGIWWMRKYRMXJPEWLIWYRXMPXLIQIEWYVIHXIQTIVEXYVIMWWXEFPI ;EMXYRXMPXLIWIRWSVGLMTWYVJEGIXIQTIVEXYVIMWWXEFPIFIJSVIWXEVXMRK EWIRWSVKVEQ%XIQTIVEXYVIGLERKISJ 'XEOIWEFSYXQMRYXIW 2SXI 8LISet TemperatureGSQQERHGSRXVSPWXLIHIXIGXMSR XIQTIVEXYVIEXXLIWIRWSVGLMTWYVJEGI7EQTPIWERHFYJJIV IUYMPMFVEXIXSXLIHIXIGXMSRXIQTIVEXYVIHYVMRKTEWWEKI XLVSYKLXLI-*'IZIREXLMKLJPS[VEXIW PQMR=SY QE][ERXXSQEMRXEMRXLIWEQTPIWMRXLIEYXSWEQTPIV GSQTEVXQIRXEXEHMJJIVIRXXIQTIVEXYVIJSVWXEFMPMX]VIEWSRW 8LIVQSWXEXXMRKPMUYMHGERFIGMVGYPEXIHXLVSYKLXLIVEGOFEWI MRXLIEYXSWEQTPIVYWMRKERI\XIVREPTYQT 4.3 Using application wizards %TTPMGEXMSR[M^EVHWLIPTFSXLRI[ERHI\TIVMIRGIHYWIVWXSTIVJSVQ XLIMVEREP]WIWUYMGOP]ERHGSRJMHIRXP]%RETTPMGEXMSR[M^EVHMWE WSJX[EVIXSSPXLEXKYMHIW]SYXLVSYKLXLIWXITWMRERI\TIVMQIRXJVSQ WXEVXXSJMREPEREP]WMW4VSGIHYVIWERHVIGSQQIRHEXMSRWMRXLI ETTPMGEXMSR[M^EVHWEVIFEWIHSREGGYQYPEXIHI\TIVXMWIJVSQ &-%'36)EREP]WIW(IXEMPIHSRPMRILIPTGERFIEGGIWWIHJVSQXLI HMEPSKYIFS\IWMRXLI[M^EVHW %TTPMGEXMSR[M^EVHWTVSZMHIH[MXL&-%'36)EVIHIWGVMFIHMR QSVIHIXEMPMR'LETXIV

44 Operation 4.4 Manual operation 8LMWWIGXMSRHIWGVMFIWLS[XSSTIVEXI&-%'36)QERYEPP] XLVSYKLXLIWSJX[EVIMRXIVJEGI7II7IGXMSRJSVEHIWGVMTXMSRSJ QIXLSHGSRXVSPPIHSTIVEXMSR 2SXI 8LIMRWXVYQIRXHSSVMWPSGOIHJSVWEJIX]VIEWSRWHYVMRK STIVEXMSRW[LMGLYWIXLIEYXSWEQTPIVIKWEQTPIQM\MRKERH MRNIGXMSR1EOIWYVIEPPWEQTPIWERHZMEPWEVIMRTPEGIFIJSVI MRMXMEXMRKWYGLSTIVEXMSRW8LIWISTIVEXMSRWGERRSXFI TIVJSVQIHMJXLIHSSVMWRSXGPSWIH Transferring, mixing and diluting samples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redip Needle STXMSREPPS[MRK]SYXSWIPIGXETSWMXMSR EX[LMGLXLIRIIHPI[MPPFIHMTTIHFIJSVIXLIXVERWJIVSTIVEXMSR8LMW STIVEXMSREPWSEWTMVEXIW PJVSQXLITVIHMTTSWMXMSRMRXSXLI EYXSWEQTPIVXYFMRKXSTVIZIRXXLIWEQTPIJVSQGSQMRKMRXSGSRXEGX [MXLVYRRMRKFYJJIV9WIXLMWJIEXYVIXSQMRMQM^IGEVV]SZIVFIX[IIR WEQTPIZMEPWIK[LIRWSPYXMSRWSJ[MHIP]HMJJIVIRXGSQTSWMXMSREVI YWIHMRWYGGIWWMSRERHEPWS[LIRMXMWMQTSVXERXXLEXXLIWEQTPIMRXLI ZMEPHSIWRSXFIGSQIGSRXEQMREXIH[MXLVYRRMRKFYJJIV

45 Operation 8VERWJIVVMRKWEQTPIW 8SXVERWJIVWEQTPIJVSQSRIVEGOTSWMXMSRXSERSXLIVWIPIGX Command:Transfer )RXIVXLIVIUYMVIHVEGOTSWMXMSRWERHZSPYQIXLIRGPMGOSROK 1M\MRKWEQTPIW 8SQM\WEQTPIMRSRITSWMXMSRWIPIGXCommand:Mix9WIXLMW GSQQERHJSVI\EQTPIEJXIVXVERWJIVVMRKX[SHMJJIVIRXWSPYXMSRWXSE GSQQSRZMEPXSTVITEVIELSQSKIRISYWQM\XYVI )RXIVXLIVIUYMVIHVEGOTSWMXMSRERHETTVSTVMEXIZSPYQIXSFIQM\IH QE\MQYQ PXLIRGPMGOSROK%WEKIRIVEPKYMHIPMRIQM\ YWMRKSJXLIXSXEPZSPYQIMRXLIZMEP8LIQM\MRKTVSGIHYVI EWTMVEXIWXLIWTIGMJMIHZSPYQISJWEQTPIERHXLIRHMWTIRWIWMXXSXLI WEQIZMEP8LMWMWTIVJSVQIHXLVIIXMQIW

46 Operation (MPYXMRKWEQTPIW 8SHMPYXIEWEQTPIWIPIGXCommand:Dilute8LMWGSQQERHGEREPWSFI YWIHMRWXIEHSJX[SWYGGIWWMZITransferSTIVEXMSRWXSTVITEVIE QM\XYVISJX[SWEQTPIW 8LMWGSQQERHTVITEVIWEHMPYXMSRSJWEQTPIMRWEQTPIEXXLI WTIGMJMIHQM\MRKTSWMXMSR8LIJMREPZSPYQISJXLIHMPYXIHWEQTPIMW EP[E]W PJSVI\EQTPIVIUYIWXMRKEHMPYXMSRXS[MPPXEOI PWEQTPIERH PWEQTPI8LIHMPYXMSRZEPYIQYWXFIMRXLI VERKI*SVFIWXTVIGMWMSRYWIZEPYIWMRXLIVERKI 3RIDiluteGSQQERHTIVJSVQWXLIIUYMZEPIRXSJX[S Transfer GSQQERHWJSPPS[IHF]EMix*SVI\EQTPI (MPYXI WEQTPI6% WEQTPI6% QM\TSWMXMSR6% HMPYXMSR MWIUYMZEPIRXXS 8VERWJIV PJVSQ6% XS6% 8VERWJIV PJVSQ6% XS6% 1M\MR6% 2SXI DiluteMWSTXMQM^IHJSVFIWXTIVJSVQERGIERHWLSYPHFIYWIH [LIVITSWWMFPIMRTVIJIVIRGIXSTransferERHMix *SVFIWXVITIEXEFMPMX]FIX[IIRHMPYXMSRSTIVEXMSRWEP[E]W WTIGMJ]XLIWSPYXMSRWMRXLIWEQISVHIVMRXLIHMEPSKYIFS\

47 Operation Data collection rate 8LIHEXEGSPPIGXMSRVEXIHIXIVQMRIWXLIRYQFIVSJTSMRXWGSPPIGXIHTIV YRMXXMQIHYVMRKXLIWIRWSVKVEQMIXLIXMQIVIWSPYXMSRSJXLI EREP]WMW(EXEGSPPIGXMSRVEXIGERFIWIXXSLowMediumSVHigh[MXL XLIRun:Data Collection RateGSQQERH%GXYEPGSPPIGXMSRVEXIWJSV ZEVMSYWWIXXMRKWEVIPMWXIHMRXLIXEFPIFIPS[ (IXIGXMSRQSHI (EXEGSPPIGXMSRVEXI 7MRKPI 1YPXMGLERRIP 0S[ TSMRXW TSMRXW 1IHMYQ TSMRXW TSMRXW,MKL TSMRXWW TSMRXWW 2SXI :EPYIWMRQYPXMGLERRIPQSHIVIJIVXSXLIGSPPIGXMSRVEXITIV GLERRIP 8LIHIJEYPXWIXXMRKSJMedium MWEHIUYEXIJSVQSWXTYVTSWIW7IXXLI VEXIXSHighJSVETTPMGEXMSRWVIUYMVMRKZIV]LMKLVIWSPYXMSR7IXXLI VEXIXSLowXSVIHYGIXLIWM^ISJXLIVIWYPXJMPIJSVPSRKVYRW %VIQMRHIVMWMWWYIHMJ]SYWXEVXEVYR[MXL Data Collection RateWIXXS HighSVLow Starting a manual sensorgram ;MXLEWIRWSVGLMTHSGOIHGLSSWIRun:Run SensorgramXSWXEVX VIGSVHMRKHEXE ;LIRIZIV]SYWXEVXEWIRWSVKVEQQERYEPP]]SY[MPPFIEWOIHXSWIPIGX XLIHIXIGXMSRQSHIERHJPS[VEXI7II7IGXMSRWERHJSVE HIWGVMTXMSRSJXLIHMEPSKYIFS\IW8LIQSWXVIGIRXP]YWIHZEPYIWEVI WYKKIWXIHJSVXLIWIWIXXMRKW -JAutosaveMWIREFPIHJSVQERYEPWIRWSVKVEQWWII7IGXMSR]SY [MPPEPWSFIEWOIHXSWTIGMJ]XLIJMPIREQIJSVWEZMRKXLIVIWYPXW 8LIWIRWSVKVEQ[MPPWXEVXEJI[WIGSRHWEJXIV]SYLEZIEGGITXIHXLI PEWXHMEPSKYIFS\(YVMRKXLMWHIPE]XLITYQTWEVIJMPPIHERHXLI RIIHPIMW[EWLIHFVMIJP]MRTVITEVEXMSRJSVXLIVYR:EVMSYWJPS[VEXI ERHJPS[TEXLWIXXMRKWETTIEVMRXLIWXEXYW[MRHS[EXXLIWIRWSVKVEQ WXEVXVIKEVHPIWWSJXLIWIXXMRKW]SYLEZIVIUYIWXIH8LMWMWERSVQEP TEVXSJXLI[EWLMRKTVSGIHYVI=SYVVIUYIWXIHJPS[WIXXMRKW[MPPFI IWXEFPMWLIHFIJSVIXLIWIRWSVKVEQEGXYEPP]WXEVXW

48 Operation 3RGIXLIWIRWSVKVEQMWWXEVXIHHEXE[MPPFIVIGSVHIHYRXMP]SYGLSSWI Run:Stop Sensorgram)ZIRXWWYGLEWWEQTPIMRNIGXMSRWHYVMRKXLI WIRWSVKVEQEVIVIGSVHIHMRXLIIZIRXPSK=SYGERWEZIXLIWIRWSVKVEQ [MXLFile:SaveSVFile:Save AsEXER]XMQI[MXLSYXMRXIVVYTXMRK VIGSVHMRKWII7IGXMSR,S[IZIVSRGI]SYLEZIWXSTTIHE WIRWSVKVEQ]SYGERRSXVIWYQIVIGSVHMRKI\GITXEWERI[WIRWSVKVEQ IMXLIVMRERI[JMPISVETTIRHIHXSXLITVIZMSYWJMPI 7II7IGXMSRJSVEHIWGVMTXMSRSJWIRWSVKVEQHMWTPE]JEGMPMXMIW Setting the detection mode and flow path =SYWIXXLIHIXIGXMSRQSHIERHJPS[TEXLEXXLIWXEVXSJXLI WIRWSVKVEQJVSQXLIDetectionHMEPSKYIFS\ 8LIGLSMGIMRXLMWHMEPSKYIFS\HIXIVQMRIWFSXLXLIJPS[TEXLERH HIXIGXMSRQSHIEXXLIWXEVXSJXLIVYR7IRWSVKVEQWEVIVIGSVHIHSRP] JVSQXLIJPS[GIPPWMRXLIGLSWIRJPS[TEXL=SYGERGLERKIXLIJPS[ TEXLHYVMRKEVYR[MXLCommand:Flow pathxlihixigxmsrqshi[mpp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etectionHMEPSKYIFS\EXXLIWXEVXSJXLIWIRWSVKVEQERHGERRSXFI GLERKIHHYVMRKEWIRWSVKVEQ6IJIVIRGIGYVZIWYFXVEGXMSR EYXSQEXMGEPP]GSVVIGXWJSVXLIWQEPPXMQIHIPE]FIX[IIRXLIJPS[GIPPW

49 Operation 8LIEZEMPEFPIWIXXMRKWJSVVIJIVIRGIGYVZIWYFXVEGXMSREVIWYQQEVM^IH FIPS[ *PS[TEXL 6IJIVIRGIWYFXVEGXMSR *G *G ERH SV ERH DQIERWXLEXXLIVIWTSRWIMRJPS[GIPPMWWYFXVEGXIHJVSQXLEXMR JPS[GIPPMI*GMWXLIFPEROJSV*G 'LSSWINoneMRXLIReference cellfs\mj]syhsrsx[erxvijivirgi WYFXVEGXMSR 2SXI -J]SYVYREWIRWSVKVEQ[MXLSYXVIJIVIRGIGYVZIWYFXVEGXMSR ERH[ERXXSWYFXVEGXXLIVIJIVIRGIGYVZISREPEXIVSGGEWMSR YWI&-%IZEPYEXMSRWSJX[EVI 1YPXMTPIWIRWSVKVEQHMWTPE] =SYGERYWIXLIView:Overlay, View:Plot Curve ClassesERH Window:New WindowGSQQERHWXSGSRXVSPLS[WIRWSVKVEQWEVI HMWTPE]IH7II7IGXMSRJSVHIXEMPW'VIEXMRKQYPXMTPI[MRHS[W[MXL Window:New WindowMWTEVXMGYPEVP]YWIJYP[LIRVIJIVIRGIGYVZI WYFXVEGXMSRMWYWIHXSHMWTPE]SVMKMREPERHWYFXVEGXIHGYVZIW WMQYPXERISYWP]SRHMJJIVIRXWGEPIW Setting the flow rate 8SGLERKIXLIJPS[VEXIGLSSWICommand:Flow ERHIRXIVXLI VIUYMVIHJPS[VEXIMRXLIHMEPSKYIFS\8LIRI[JPS[VEXI[MPPETTP] [LIR]SYGPMGOSROK 8LIJPS[[MPPGSRXMRYIEXXLIWIXVEXIYRXMP]SYWXSTSVGLERKIXLI JPS[8SWXSTXLIJPS[IMXLIVGLSSWICommand:Stop FlowSVWIXXLI JPS[VEXIXS^IVS=SYGERWXSTXLIJPS[[LMPIEWIRWSVKVEQMW VYRRMRK2SXILS[IZIVXLEXMRNIGXMSRWGERRSXFITIVJSVQIH[LMPIXLI JPS[MWWXSTTIH

50 Operation *MPPMRKXLIGSRXMRYSYWJPS[TYQT 8LITYQTWEVIJMPPIH[LIREWIRWSVKVEQMWWXEVXIHERHEVIVIJMPPIH EYXSQEXMGEPP]EXXLIIRHSJIZIV]TYQTWXVSOISVEXXLIWXEVXERHIRH SJIEGLG]GPIMRQIXLSHGSRXVSP8LIWXVSOIZSPYQIMW PERHXLI VIJMPPSTIVEXMSRXEOIWEFSYXWIGSRHW8SEZSMHHMWXYVFERGIWEVMWMRK JVSQXLIVIJMPPSTIVEXMSRHYVMRKEGVMXMGEPEREP]WMW]SYGERVIJMPPXLI TYQTEXER]XMQIMREQERYEPVYR'LSSWICommand:Flow ERHGLIGO XLIRefillFS\8LITYQT[MPPFIVIJMPPIH[LIR]SYGPMGOSROKIZIRMJ ]SYLEZIRSXGLERKIHXLIJPS[VEXI 8LIWXEVXERHIRHSJXLIVIJMPPSTIVEXMSRFSXLEYXSQEXMGERHQERYEPP] VIUYIWXIHEVIVIGSVHIHMRXLIIZIRXPSKMJEWIRWSVKVEQMWVYRRMRKWS XLEX]SYGERMHIRXMJ]TSWWMFPIWMKREPHMWXYVFERGIWEVMWMRKJVSQXLMW WSYVGI Injecting sample 8SMRNIGXWEQTPISZIVXLIWIRWSVGLMTWYVJEGIGLSSWICommand:Inject =SYGERRSXGLSSWIXLMWGSQQERHYRPIWWEWIRWSVKVEQMWVYRRMRK -RNIGXGSQQERHWJSVHMJJIVIRXETTPMGEXMSRTYVTSWIWEVIWYQQEVM^IHFIPS[

51 Operation Command Comments Injected volume Sample consumption Max flow rate INJECT General purpose inject µl Injected vol +30 µl 100 µl/min KINJECT QUICKINJECT COINJECT BIGINJECT MANUAL INJECT Low dispersion inject for kinetics and for samples sensitive to dilution with running buffer. Controlled dissociation time after sample injection. Low sample consumption. Less preparation time than INJECT but more sample dispersion. Injection of second sample immediately following the first (e.g. for dissociation in the presence of ligand). Variable contact times for first sample only. Recovery of either first or second sample, but not both. Large volume inject. Direct inject up to 325 µl, sample loop between 326 and 425 µl, direct between 426 and 750 µl. Manually controlled inject. See Section µl Injected vol +40 µl 100 µl/min µl Injected vol +10 µl 100 µl/min Sample 1: µl Sample 2: µl Sample 1: injected vol +40 µl Sample 2: injected vol +40 µl 100 µl/min µl Injected vol +52 µl 50 µl/min* µl / loading Loaded vol +30 µl *BIGINJECT cannot be performed at higher flow rates because the autosampler tubing must be refilled during injection from the sample loop for samples larger than 425 µl. 8LIHMEPSKYIFS\IWJSVXLIZEVMSYWMRNIGXGSQQERHWI\GITX 1%29%0-2.)'8WII7IGXMSRLEZIERYQFIVSJGSQQSR JIEXYVIW PositionJMIPHWLEZIETYPPHS[RPMWXSJZEPMHTSWMXMSRWEGGSVHMRKXS XLIVEGOWIXXMRKW 100 µl/min =SYIRXIVXLIZSPYQISJWEQTPIXSFIMRNIGXIHMRXLIVolumeJMIPHW 8LIHMEPSKYIFS\EPWSHMWTPE]WGSRXEGXXMQISJXLIWEQTPI[MXLXLI WIRWSVWYVJEGIGSRXEGXXMQI!MRNIGXIHZSPYQIJPS[VEXIERHXLI WEQTPIGSRWYQTXMSRJSVXLIMRNIGXMSRWIIXEFPIEFSZI -JXLIVariable Contact TimesFS\MWGLIGOIHZSPYQIJMIPHWETTIEV JSVXLIJPS[GIPPWMRXLIGYVVIRXJPS[TEXLIKMJ]SYLEZIWIXJPS[ TEXLXLIVI[MPPFIJSYVZSPYQIJMIPHWJSVER-2.)'8 GSQQERH)EGLZSPYQIQYWXFIPIWWXLERSVIUYEPXSXLIZSPYQIMR

52 Operation XLITVIGIHMRKJPS[GIPP*SVI\EQTPI[MXLJSYVJPS[GIPPW 4IVQMXXIH 2SXTIVQMXXIH :EVMEFPIGSRXEGXXMQIMRNIGXMSRWEVIMQTPIQIRXIHF]W[MXGLMRK EHHMXMSREPJPS[GIPPWMRXSXLIJPS[TEXLEWXLIMRNIGXMSRTVSGIIHW -RNIGXMSRWMRXLIWITEVEXIJPS[GIPPWXLYWWXEVXEXHMJJIVIRXXMQIWERH IRHEXXLIWEQIXMQI8LMWMWMPPYWXVEXIHF]XLIZEVMEFPIGSRXEGXXMQI WIRWSVKVEQWWLS[RFIPS[ Variable Contact TimesMWSRP]EZEMPEFPIMJ]SYLEZIGLSWIRE QYPXMGLERRIPJPS[TEXLERHEVIRSXYWMRKVIJIVIRGIGYVZIWYFXVEGXMSR

53 Operation 8LIMore>>FYXXSRSTIRWEHHMXMSREPSTXMSRWJSVXLIMRNIGXMSR 'LIGOPredip NeedleERHIRXIVEVEGOTSWMXMSRXSHMTXLIRIIHPIMRXLI WTIGMJMIHTSWMXMSRFIJSVIEWTMVEXMRKWEQTPI8LMWSTIVEXMSREPWS EWTMVEXIW PJVSQXLITVIHMTTSWMXMSRMRXSXLIEYXSWEQTPIVXYFMRKXS TVIZIRXXLIWEQTPIJVSQGSQMRKMRXSGSRXEGX[MXLVYRRMRKFYJJIV9WI XLMWJIEXYVIXSQMRMQM^IGEVV]SZIVFIX[IIRWEQTPIZMEPWIK[LIR WSPYXMSRWSJ[MHIP]HMJJIVIRXGSQTSWMXMSREVIYWIHMRWYGGIWWMSR9WISJ Predip NeedleMRERI\XVEZMEPSJWEQTPIGEREPWSLIPTXSQMRMQM^I WEQTPIHMWTIVWMSRMRXLIMRNIGXMSRF]MQTVSZMRKXLIWITEVEXMSR FIX[IIRMRNIGXIHWEQTPIERHVYRRMRKFYJJIV 'LIGOExtra CleanupXSTIVJSVQEREHHMXMSREP[EWLSJXLIRIIHPIERH -*'EJXIVXLIMRNIGXMSR9WIXLMWJIEXYVIXSQMRMQM^IGEVV]SZIV[LIR XLIRI\XMRNIGXMSRYWIWWEQTPISJ[MHIP]HMJJIVIRXGSQTSWMXMSR8LI -*'MWVSYXMRIP][EWLIHEWTEVXSJXLIWXERHEVHMRNIGXMSRWIUYIRGI Extra CleanupWLSYPHFIYWIHMJTEVXMGYPEVGEVIMWRIGIWWEV]IKEJXIV MRNIGXMSRSJVIKIRIVEXMSRWSPYXMSR 8LIWEQTPIVIGSZIV]ERHJVEGXMSRGSPPIGXMSRSTXMSRWEVIHIWGVMFIH FIPS[7IGXMSRWERH 'PMGOSRStart InjectionXSWXEVXXLIMRNIGXMSR8LIW]WXIQXEOIWE[LMPI XSTVITEVIXLIMRNIGXMSRZEV]MRKJVSQEFSYXYTXSQMRYXIW HITIRHMRKSRXLIOMRHSJMRNIGXMSRXLIWEQTPIZSPYQIWERHXLI STXMSRWGLSWIRWIIXEFPIFIPS[8LIIZIRXPSKVIGSVHWFSXLXLIXMQI [LIRXLIMRNIGXMSR[EWVIUYIWXIHERHXLIXMQIEXXLIEGXYEPWXEVXERH IRHSJWEQTPIMRNIGXMSR

54 Operation Inject command Injected volume (µl) Preparation time (s) Finishing time (s) No. of air segments INJECT QUICKINJECT KINJECT COINJECT 20, , 100 BIGINJECT MANUAL INJECT Preparation time for loop loading 100 µl: 80 s Approximate times for preparation and completion of various inject commands at a flow rate of 20 µl/min. Preparation time is the time between the inject command and the actual start of sample injection. Finishing time is the time between the end of sample injection and completion of the command, during which the flow system is washed. Finishing times for KINJECT do not include the dissociation time specified in the command. Predip Needle adds about 8 s to the preparation time. Extra Cleanup adds about s to the finishing times for INJECT, QUICKINJECT and KINJECT and about 70 s to the finishing times for COINJECT and BIGINJECT.. Recover of 25 µl adds about 210 s to the finishing times Stopping an injection 8SWXSTERMRNIGXMSRGLSSWICommand:Stop InjectSVGPMGOSRXLIStop Inject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

55 Operation 2SXIW -J]SYWXSTERMRNIGXMSRJSV[LMGLWEQTPIVIGSZIV]MWVIUYIWXIH WII7IGXMSRXLIZSPYQISJVIGSZIVIHWEQTPI[MPPFI VIHYGIHMREGGSVHERGI[MXLXLIPS[IVZSPYQIMRNIGXIH-J]SY WXSTERMRNIGXMSRJSV[LMGLJVEGXMSRGSPPIGXMSRMWVIUYIWXIHWII 7IGXMSRJVEGXMSRW[MPPRSXFIGSPPIGXIH (YVMRKXLIJMVWXMRNIGXMSRJSV'3-2.)'8XLIStop Inject GSQQERHGLERKIWXSSkip Inject'LSSWMRKXLMWGSQQERH WXEVXWXLIWIGSRHMRNIGXMSRMQQIHMEXIP]7EQTPIVIQEMRMRKMR XLIWEQTPIPSSTJVSQXLIJMVWXMRNIGXMSRMWJPYWLIHXS[EWXI EJXIVXLIMRNIGXMSRERHGERRSXFIVIGSZIVIH8LIWIGSRH MRNIGXMSRGERFIWXSTTIHMRXLIWEQI[E]EWSXLIVMRNIGXMSR GSQQERHW *SV&-+-2.)'8]SYGERVIGSZIVWEQTPI[LMGLMWGYVVIRXP]MR XLIEYXSWEQTPIVXYFMRK[LIRXLIMRNIGXMSRMWWXSTTIHFYXRSX XLEX[LMGLMWMRXLIWEQTPIPSSTMRXLI-*' 7II7IGXMSRJSVIQIVKIRG]WXSTTVSGIHYVIW Manual injections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njectHMEPSKYIFS\ERHGPMGOSRContinue>> 8LIHMEPSKYIFS\WLS[WXLIEGGYQYPEXIHMRNIGXIHZSPYQIERHXLI ZSPYQISJFYJJIVVIQEMRMRKMRXLIGSRXMRYSYWJPS[TYQT8LMWPEXXIV JMKYVIKMZIW]SYERMRHMGEXMSRSJ[LIRXLITYQT[MPPFIVIJMPPIHWS XLEX]SYGEREZSMHVIJMPPSTIVEXMSRWHYVMRKMRNIGXMSRW

56 Operation *SPPS[XLIWXITWFIPS[XSTIVJSVQEQERYEPMRNIGXMSR 'PMGOSRLoad Loop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ommand:FlowJVSQXLI QIRYSVGPMGOSRXLIFlowFYXXSRSRXLIQEMRXSSPFEVGLIGOXLI RefillFS\ERHGPMGOSROKWII7IGXMSR 'LSSWIIMXLIVVolumeSVManualQSHI Volume 8LIMRNIGXMSR[MPPWXSTEYXSQEXMGEPP]EJXIVXLIWTIGMJMIH MRNIGXIHZSPYQI9WIXLMWQSHIJSVMRNIGXMSRSJ TVIHIXIVQMRIHZSPYQIW Manual 8LIMRNIGXMSR[MPPGSRXMRYIYRXMP]SYGPMGOSRStop9WI XLMWQSHIMJ]SY[ERXXSWXSTXLIMRNIGXMSRMRXIVEGXMZIP] EGGSVHMRKXSXLIETTIEVERGISJXLIWIRWSVKVEQ 8LIMRNIGXMSRGERGSRXMRYIMRHIJMRMXIP]MRManual QSHI&IGEVIJYPRSXXSMRNIGXQSVIXLERXLIZSPYQI PSEHIHMRXLIPSST =SYGERGLERKIXLIJPS[VEXIHYVMRKEQERYEPQSHIMRNIGXMSR FYXRSXHYVMRKEZSPYQIQSHIMRNIGXMSR=SYGERGLERKIXLIJPS[ TEXLMRQYPXMGLERRIPQSHIERHWXSTXLIMRNIGXMSREXER]XMQIMR FSXLQSHIW 'PMGOSRStart8LIHMEPSKYIFS\HMWTPE]WFSXLXLIEGGYQYPEXIH MRNIGXIHZSPYQIMIXLIWYQSJEPPZSPYQIWMRNIGXIHWMRGIXLI WXEVXSJQERYEPMRNIGXMSRERHXLIZSPYQIVIQEMRMRKMRXLIWEQTPI

57 Operation MRNIGXMSRTYQT8LIZSPYQIMRNIGXIHWMRGIXLIPEWXStartMWWLS[R MRXLIWIRWSVKVEQWXEXYW[MRHS[ %WEVYPISJXLYQFHSRSXMRNIGXQSVIXLERSJXLI PSEHIHZSPYQIQSVIEXLMKLIVJPS[VEXIWXSQMRMQM^IXLIVMWOSJ WEQTPIHMWTIVWMSRIJJIGXWEXXLIIRHSJXLIMRNIGXIHWEQTPI 8SMRXIVVYTXXLIMRNIGXMSREXER]XMQIGPMGOSRStop8LMW[MPP W[MXGLJVSQWEQTPIXSFYJJIVJPS[SZIVXLIWIRWSVWYVJEGIFYX [MPPRSXIQTX]XLIWEQTPIPSSTSV[EWLXLI-*'=SYGERVIPSEH XLIWEQTPIPSST[LMPIXLIMRNIGXMSRMWWXSTTIH=SYGEREPWSYWI XLIEYXSWEQTPIVXSTVITEVIWEQTPIWYWMRKMixTransferERHDilute [LMPIXLIMRNIGXMSRMWWXSTTIHYWIXLIGSQQERHWMRXLIQIRYSV XLIFYXXSRWSRXLIQEMRXSSPFEVJSVXLIWISTIVEXMSRW'PMGOSR StartXSVIWYQIXLIMRNIGXMSR 8SIRHXLIQERYEPMRNIGXMSRGPMGOSRExit8LMW[MPPIQTX]XLI WEQTPIPSSTXS[EWXIERH[EWLXLI-*' Recovering sample 8LIWEQTPIVIGSZIV]JIEXYVIGERFIYWIHIMXLIV XSVIGSZIVXLIMRNIGXIHWEQTPISV XSVIGSZIVFSYRHEREP]XIXLEXMWVIPIEWIHHYVMRKVIKIRIVEXMSRXLMWMW VIEPP]VIGSZIVMRKlWEQTPIzHYVMRKMRNIGXMSRSJVIKIRIVEXMSR WSPYXMSR %28 &IJSVIWXEVXMRKERMRNIGXMSR[MXLWEQTPIVIGSZIV]TIVJSVQEWash [MXLRecoverySTXMSRXS[EWLXLIVIGSZIV]GYTERH-*'VIGSZIV] GLERRIPERHIQTX]XLIVIGSZIV]GLERRIPWII7IGXMSR-J]SY HSRSXHSXLMWXLIVIGSZIVIHWEQTPIQE]FIHMPYXIH[MXLYTXS P FYJJIVJVSQXLIVIGSZIV]GLERRIP ;LIRWEQTPIVIGSZIV]MWEGXMZEXIHIJJPYIRXJVSQXLI-*'MWHMZIVXIHXS XLIVIGSZIV]GYTSRXLIGSRRIGXSVFPSGO*VSQXLIVIMXMWGSPPIGXIHF] XLIEYXSWEQTPIVERHXVERWJIVVIHXSEWTIGMJMIHVEGOTSWMXMSR 7IPIGXVIGSZIV]MRXLIMore>>TERIPSJXLIInjectionHMEPSKYIFS\8LMW JIEXYVIGERRSXFIYWIHMRGSQFMREXMSR[MXL&-+-2.)'8SV1%29%0-2.)'8SV[MXLJVEGXMSRGSPPIGXMSRWIIFIPS[

58 Operation )RXIVXLIZMEPTSWMXMSRXS[LMGLXLIVIGSZIVIHWEQTPIMWXSFI XVERWJIVVIHERHXLIZSPYQIXSFIVIGSZIVIHTIVQMXXIHVERKI P VIGSQQIRHIH" PJSVFIWXTVIGMWMSR8LIZSPYQIHIPMZIVIHXSXLI VIGSZIV]GYTMWXLIWEQIEWXLIMRNIGXIHZSPYQILS[IZIVMJ]SY[ERX XSFIWYVISJVIGSZIVMRKEPPXLIMRNIGXIHWEQTPIWTIGMJ] PQSVI JSVVIGSZIV]XLERXLIZSPYQISJWEQTPIMRNIGXIHWSXLEXEWQEPPZSPYQI SJEMVMWlVIGSZIVIHzEJXIVXLIWEQTPI =SYGERYWIXLITransferGSQQERHXSTVMQIXLIVIGSZIV]TSWMXMSR [MXLRIYXVEPM^MRKFYJJIVFIJSVIXLIMRNIGXMSR8LMWMWTEVXMGYPEVP] VIPIZERX[LIR]SYYWIVIGSZIV]JSPPS[MRKEVIKIRIVEXMSRMRNIGXMSRXS GSPPIGXQEXIVMEPVIPIEWIHJVSQXLIWYVJEGI Collecting fractions 8LIJVEGXMSRGSPPIGXMSRJIEXYVIGSPPIGXWIJJPYIRXJVSQXLI-*' HYVMRK FYJJIVJPS[MQQIHMEXIP]JSPPS[MRKERMRNIGXMSR9WIXLMWJIEXYVIXS GSPPIGXERHSVJVEGXMSREXIQEXIVMEP[LMGLMWVIPIEWIHJVSQXLIWYVJEGI HYVMRKXLIHMWWSGMEXMSRTLEWI %28 &IJSVIWXEVXMRKERMRNIGXMSR[MXLJVEGXMSRGSPPIGXMSRTIVJSVQEWash [MXLRecoverySTXMSRXS[EWLXLIVIGSZIV]GYTERH-*'VIGSZIV] GLERRIPERHIQTX]XLIVIGSZIV]GLERRIPWII7IGXMSR-J]SY HSRSXHSXLMWXLIJMVWXJVEGXMSRWQE]FIHMPYXIH[MXLYTXS P FYJJIVJVSQXLIVIGSZIV]GLERRIP 7IPIGXJVEGXMSRGSPPIGXMSRMRXLIMore>>TERIPSJXLIInjectionHMEPSKYI FS\8LMWJIEXYVIGERRSXFIYWIHMRGSQFMREXMSR[MXL&-+-2.)'8 1%29%0-2.)'8SV'3-2.)'8MRNIGXMSRWSV[MXLWEQTPIVIGSZIV] WIIEFSZI

59 Operation )RXIVXLIZMEPTSWMXMSRJSVXLIJMVWXJVEGXMSRXSFIGSPPIGXIHXLIJVEGXMSR ZSPYQIERHXLIRYQFIVSJJVEGXMSRW*VEGXMSRW[MPPFIGSPPIGXIHMR WIUYIRGIJVSQXLIWTIGMJMIHJMVWXTSWMXMSR8LIRI\XJVEGXMSREJXIVXLI IRHSJEVS[MWXLIFIKMRRMRKSJXLIRI\XVS[IKVE Æ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ashing liquid handling system components 8LIPMUYMHLERHPMRKW]WXIQMW[EWLIHEYXSQEXMGEPP]EWVIUYMVIHHYVMRK STIVEXMSR 8LIEYXSWEQTPIVRIIHPIMW[EWLIHEJXIVIEGLWEQTPILERHPMRK STIVEXMSRInjectTransferMixDilute 8LI-*'GLERRIPWEVI[EWLIHEJXIVIEGLMRNIGXMSR 8LIVIGSZIV]GYTMW[EWLIHEJXIVIEGLMRNIGXMSRXLEXYWIWWEQTPI VIGSZIV]SVJVEGXMSRGSPPIGXMSR 8LIWash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

60 Operation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orking Tools:Flush WII7IGXMSR8LMWQE]FIRIGIWWEV]MJ]SYEVIYWMRKEZMWGSYW VIKIRIVEXMSRWSPYXMSRWYGLEWIXL]PIRIKP]GSP Saving the sensorgram 8SWEZIEWIRWSVKVEQEWE&-%'36)VIWYPXJMPIGLSSWIFile:SaveSV File:Save As8LIJMVWXXMQIERI[WIRWSVKVEQMWWEZIHXLIWI GSQQERHWEVIIUYMZEPIRX8LIVIEJXIVSaveWEZIWXLIWIRWSVKVEQ YRHIVXLIWEQIREQISZIV[VMXMRKXLIIEVPMIVZIVWMSR[LMPISave As EPPS[W]SYXSWEZIXLIWIRWSVKVEQYRHIVERI[REQIPIEZMRKER] TVIZMSYWZIVWMSRYRGLERKIH 6IWYPXJMPIWEVIWEZIH[MXLXLIHIJEYPXI\XIRWMSRFPV -JXLIAutosaveJYRGXMSRYRHIVTools:PreferencesMWXYVRIHSRXLI WIRWSVKVEQ[MPPFIWEZIHEYXSQEXMGEPP]EXXLIWTIGMJMIHMRXIVZEPWII 7IGXMSR;IVIGSQQIRHXLMWJIEXYVIEWETVIGEYXMSREKEMRWXPSWMRK HEXEMJXLIGSQTYXIVWLSYPHPSGOYTSVPSWITS[IVHYVMRKEVYR-J

61 Operation AutosaveMWXYVRIHSR]SYQYWXWTIGMJ]XLIJMPIREQIJSVXLI WIRWSVKVEQEXXLIWXEVXSJXLIVYR 8SWEZIEWIRWSVKVEQERHEWWSGMEXIHHEXESVXLIVITSVXTSMRXXEFPIMRE JSVQEX[LMGLGERFISTIRIHF]SXLIVTVSKVEQWGLSSWIFile:Export ERHWIPIGXIMXLIVCurvesSVReport Point TableJVSQXLIWYFQIRY 8LMWGVIEXIWEXI\XJMPI[MXLXEFWITEVEXIHHEXEWYMXEFPIJSVYWIMR WTVIEHWLIIXWERHSXLIVETTPMGEXMSRW =SYGERWEZISVI\TSVXWIRWSVKVEQHEXEEXER]XMQIHYVMRKEVYR [MXLSYXMRXIVVYTXMRKHEXEGSPPIGXMSR Stopping a sensorgram 8SWXSTHEXEGSPPIGXMSRGLSSWISensorgram:Stop SensorgramSVGPMGO SRXLIStop SensorgramXSSPFEVFYXXSR8LI-*'JPS[GLERRIPWEVI [EWLIHEYXSQEXMGEPP]MJERMRNIGXMSRLEWFIIRTIVJSVQIHERHXLI FYJJIVJPS[MWXYVRIHSJJ 2SXI 3RGI]SYLEZIWXSTTIHEWIRWSVKVEQ]SYGERRSXVIWYQIHEXE GSPPIGXMSRMRXLIWEQIWIRWSVKVEQ-J]SYRIIHXSGSQFMRI HEXEJVSQX[SWITEVEXIVYRW]SYQYWXYWIGYVZITVIWIRXEXMSR JIEXYVIWMR&-%IZEPYEXMSRSVIUYMZEPIRXWSJX[EVI Emergency stop procedure 8SEFSVXEVYREXER]XMQITVIWWCtrl-Break8LIBreakOI]MWEPWS QEVOIHPauseSRWXERHEVHOI]FSEVHW8LMW[MPPMQQIHMEXIP]WXSTEPP GYVVIRXSTIVEXMSRWMRGPYHMRKFYJJIVJPS[ERHHEXEGSPPIGXMSR=SY WLSYPHRSVQEPP][EWLXLIPMUYMHLERHPMRKW]WXIQMRGSRRIGXMSR[MXL XLIIQIVKIRG]WXSTWMRGIRSEYXSQEXMG[EWLMRKTVSGIHYVIWEVI TIVJSVQIH8LIVIEVIRSJEGMPMXMIWJSVVIGSZIVMRKWEQTPIMRXLMW WMXYEXMSR (SRSXYWI'XVP&VIEOYRPIWW]SYRIIHXSWXSTEPPSTIVEXMSRW MQQIHMEXIP]IKMJGSRXMRYIHSTIVEXMSRGSRWXMXYXIWEVMWOSJHEQEKMRK XLIMRWXVYQIRXSVPSWMRKZEPYEFPIQEXIVMEP%GSRXVSPPIHWXSTYWMRK Stop InjectERHStop SensorgramMWEP[E]WTVIJIVEFPIXS'XVP&VIEO Undocking the sensor chip =SYGERPIEZIXLIWIRWSVGLMTHSGOIHMRXLIMRWXVYQIRXJSVWIZIVEP HE]WTVSZMHIHXLEXPMUYMHMWOITXJPS[MRKSZIVXLIWYVJEGI=SY[MPP LS[IZIVRIIHXSYRHSGOXLIWIRWSVGLMT [LIREPPJSYVJPS[GIPPWEVIYWIHERH]SYRIIHERI[GLMT

62 Operation [LIRXLIWXEFMPMX]SJ]SYVMQQSFMPM^IHPMKERHVIUYMVIWXLEXXLIGLMT WLSYPHFIWXSVIHYRHIVWTIGMEPGSRHMXMSRW [LIR]SYEVIWLYXXMRKHS[RXLIW]WXIQJSVQSVIXLERHE]W 8SYRHSGOXLIWIRWSVGLMTGLSSWICommand:Undock.(SRSXEXXIQTX XSVIQSZIXLIGLMT[MXLSYXJMVWXYRHSGOMRK -JXLIEmpty flow cellsfs\mwgligoihxliw]wxiq[mppiqtx]xlijps[ GIPPWFIJSVIYRHSGOMRK%P[E]WGLIGOXLMWSTXMSRYRPIWWMXMWIWWIRXMEP XSOIITXLIWIRWSVWYVJEGI[IXWSQIPMKERHWERHWTIGMEPWYVJEGIWQE] HIXIVMSVEXISRGSRXEGX[MXLEMV0MUYMH[MPPRSXPIEOMRXSXLISTXMGEP W]WXIQIZIRMJ]SYYRHSGOXLIGLMT[MXLSYXIQTX]MRKXLIJPS[GIPPW ;LIRXLIYRHSGOMRKMWGSQTPIXIXLIWIRWSVGLMT0)(SRXLI MRWXVYQIRXJVSRXTERIPJPEWLIWSTIRXLIWIRWSVGLMTTSVXGSZIVERH VIQSZIXLIWIRWSVGLMTGEWWIXXI8LIWSJX[EVI[MPPEYXSQEXMGEPP] TVSQTX]SYXSMRWIVXERHHSGOERI[GLMT 4.5 Queuing manual commands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ommandQIRYEVIKVI]ERHGERRSXFIYWIHEWPSRKEWXLIMRWXVYQIRX MWFYW]'SQQERHWMWWYIH[LIRXLIUYIYIMWRSXEGXMZIGERRSXFIYWIH XSKIRIVEXIEQIXLSH Starting the command queue (ITIRHMRKSRXLIWIXXMRKYRHIVTools:PreferencesXLIGSQQERH UYIYIQE]FIWXEVXIHEYXSQEXMGEPP][LIREWIRWSVKVEQMWWXEVXIH-J XLMWMWRSXXLIGEWIWXEVXXLIUYIYIF]GLSSWMRK Command:Command Queue=SYGERSRP]WXEVXXLIUYIYI[LMPIEWIRWSVKVEQMWVYRRMRK

63 Operation ;LIRXLIGSQQERHUYIYIMWEGXMZIXLIUYIYI[MRHS[MWWLS[RIMXLIV QMRMQM^IHSRXLI;MRHS[WXEWOFEVSVI\TERHIHMRXLIWIRWSVKVEQ [MRHS[EVIE8LII\TERHIH[MRHS[WLS[WEPPGSQQERHWMRXLI UYIYIMRGPYHMRKXLSWI[LMGLLEZIFIIRGSQTPIXIH]SYQE]LEZIXS WGVSPPXLI[MRHS[XSWIIEPPGSQQERHWMJXLIUYIYIMWPSRK The command queue window. A KINJECT command is in progress. 8SI\TERHXLIQMRMQM^IHUYIYI[MRHS[WMQTP]GPMGOSRXLIFYXXSR SRXLIXEWOFEV=SYGERVIWM^IXLII\TERHIH[MRHS[F]HVEKKMRKSR XLI[MRHS[FSVHIVWXLIWM^ISJXLIQMRMQM^IH[MRHS[MWJM\IH8S QMRMQM^IXLII\TERHIH[MRHS[GPMGOSRXLIMinimizeFYXXSRSRXLI [MRHS[XSSPFEVWXERHEVH;MRHS[WJYRGXMSREPMX]8LII\TERHIH [MRHS[GERFIQSZIHER][LIVISRXLIWGVIIRF]HVEKKMRK8LI QMRMQM^IH[MRHS[GEREPWSFIQSZIHFYX[MPPEP[E]WWRETXSXLI RIEVIWXGSVRIVSJXLIWIRWSVKVEQ[MRHS[ Command queue window display 'SQQERHWEVIHMWTPE]IHMRXLIGSQQERHUYIYI[MRHS[EWJSPPS[W 8LIQMRMQM^IH[MRHS[HMWTPE]WERMGSRJSVXLIGSQQERHGYVVIRXP] FIMRKI\IGYXIH8LIMGSRMWXLIWEQIEWXLEX[LMGLETTIEVWMRXLI HMEPSKYIFS\JSVXLIGSQQERHERHSRXLIXSSPFEVJSVGSQQERHW[MXL XSSPFEVFYXXSRW8LI[MRHS[WLS[WEStopMGSRMJXLIUYIYIMW WXSTTIHSVRSMGSRMJI\IGYXMSRSJXLIUYIYIMWGSQTPIXI 8LII\TERHIH[MRHS[WLS[WGSPYQRWJSVIEGLGSQQERHWII MPPYWXVEXMSREFSZI[MXLGSQQERHWXEXYWGSQQERHMGSRERHQIXLSH GSQQERHW'SQQERHW[LMGLLEZIFIIRI\IGYXIHEVIKVI]

64 Operation 'SQQERHWXEXYW 8LIGSQQERHWXEXYWMWMRHMGEXIHF]ERMGSR MRHMGEXIWXLEXXLIGSQQERHLEWFIIRGSQTPIXIHXLI GSQQERHXI\XMWEPWSKVI] MRHMGEXIWXLIGSQQERHGYVVIRXP]FIMRKI\IGYXIH MRHMGEXIWXLEXXLIUYIYI[MPPWXSTEXXLMWTSMRX MRHMGEXIWXLEXXLIUYIYILEWWXSTTIHEXXLMWTSMRX MRHMGEXIWXLEXXLMWGSQQERHLEWJEMPIHIMXLIVXLVSYKL MRWXVYQIRXJEMPYVISVFIGEYWIXLIYWIVLEWTVIWWIH Ctrl-Break 'SQQERHW[LMGLEVI[EMXMRKXSFITVSGIWWIHLEZIRSWXEXYWMGSR 'SQQERHMGSR 8LMWMGSRMWXLIWEQIEWXLEX[LMGLETTIEVWMRXLIHMEPSKYIFS\JSVXLI GSQQERHERHSRXLIXSSPFEVJSVGSQQERHW[MXLXSSPFEVFYXXSRW 8LIJSPPS[MRKMGSRWEVIRSXMRGPYHIHMRXLIXSSPFEVFYXEVIYWIHMRXLI GSQQERHUYIYI Dilute Wash 1IXLSHGSQQERHW 8LIQIXLSHGSQQERHWGSPYQRWLS[WXLIGSQQERHEWMX[SYPH ETTIEVMREQIXLSH*SVWSQIGSQQERHWXLIVIQE]FIQSVIXLERSRI QIXLSHGSQQERHJSVEWMRKPIGSQQERHMGSRJSVI\EQTPIERInject GSQQERH[MXLSTXMSRWPredip needle ERHExtracleanETTIEVWEW JSPPS[W Managing the command queue %HHMRKGSQQERHWXSXLIUYIYI 8SEHHEGSQQERHXSXLIUYIYIWMQTP]GLSSWIXLIGSQQERHJVSQ XLIXSSPFEVSVXLICommandQIRY8LIGSQQERH[MPPFITPEGIHEX

65 Operation XLIIRHSJXLIUYIYIEWWSSREW]SYGPMGOSROKMRXLIHMEPSKYIFS\ JSVXLIGSQQERH 2SXI 8LIManual injectgsqqerhgerrsxfiuyiyih -JXLIUYIYI[MRHS[MWQMRMQM^IH]SY[MPPRSXWIIGSQQERHWEHHIH EXXLIIRHSJXLIUYIYI)\TERHXLI[MRHS[XSHMWTPE]EPPUYIYIH GSQQERHW 2SVQEPP]GSQQERHWMRXLIUYIYIEVII\IGYXIHEWWSSREWTSWWMFPI-J XLIUYIYIMWIQTX]EVIUYIWXIHGSQQERH[MPPFII\IGYXIH MQQIHMEXIP]-J]SY[ERXXSLSPHXLIUYIYI[LMPI]SYEHHGSQQERHW GLSSWIRun:Stop QueueJVSQXLIUYIYI[MRHS[QIRYFIJSVI]SYEHH MRWXVYQIRXGSQQERHWWIIFIPS[ (IPIXMRKGSQQERHWJVSQXLIUYIYI 8SHIPIXIGSQQERHWJVSQXLIUYIYIWIPIGXSRISVQSVIGSQQERHWMR XLIUYIYI[MRHS[ERHGLSSWIEdit:RemoveJVSQXLIGSQQERHUYIYI [MRHS[QIRY=SY[MPPFIEWOIHXSGSRJMVQHIPIXMSRSJXLIWIPIGXIH GSQQERHW%XIQTSVEV]StopMGSR[MPPFITPEGIHEXXLIJMVWXWIPIGXIH GSQQERHERHI\IGYXMSR[MPPWXSTMJXLIUYIYIVIEGLIWXLMWGSQQERH [LMPIXLIGSRJMVQEXMSRHMEPSKYIFS\MWSTIR=SYGERHIPIXI GSQQERHWXLEXLEZIFIIRI\IGYXIHlKVI]zGSQQERHWERH GSQQERHWXLEXEVI[EMXMRKXSFII\IGYXIHFYX]SYGERRSXHIPIXIXLI GSQQERHXLEXMWGYVVIRXP]FIMRKI\IGYXIHMRXLIMRWXVYQIRX 8SHIPIXIEPPGSQQERHWMRXLIUYIYIMRGPYHMRKlKVI]zGSQQERHW GLSSWIEdit:Clear Queue8LIUYIYI[MPPFIGPIEVIHMQQIHMEXIP]FYX I\IGYXMSRSJXLIGYVVIRXGSQQERH[MPPGSRXMRYIXSGSQTPIXMSR )HMXMRKGSQQERHWMRXLIUYIYI =SYGERIHMXGSQQERHWMRXLIUYIYITVSZMHIHXLEXXLI]LEZIRSXFIIR I\IGYXIHSVEVIGYVVIRXP]FIMRKI\IGYXIH'SQQERHWXLEXEVI GSQTPIXIHlKVI]zGSQQERHWERHXLIGYVVIRXGSQQERHGERRSXFI IHMXIH 8SIHMXEGSQQERHHSYFPIGPMGOSRXLIGSQQERHSVWIPIGXXLI GSQQERHERHGLSSWIEdit:Edit Command8LMW[MPPTPEGIEXIQTSVEV] StopMGSREXXLIGSQQERHERHSTIRXLIETTVSTVMEXIHMEPSKYIFS\JSV XLIGSQQERH)\IGYXMSR[MPPWXSTMJXLIUYIYIVIEGLIWXLMWGSQQERH [LMPIXLIEditHMEPSKYIFS\MWSTIR'LERKIXLIWIXXMRKWEWVIUYMVIHMR XLIHMEPSKYIFS\8LIStopMGSR[MPPFIVIQSZIH[LIR]SYGPMGOSR OKXSMQTPIQIRXXLIGLERKIHGSQQERH =SYGERSRP]GLERKIXLIWIXXMRKW[MXLMRXLIWEQIGSQQERHMRXLMW [E]8SGLERKISRIGSQQERHXSERSXLIVIKXSGLERKITransferXS Dilute]SYQYWXHIPIXIXLISPHGSQQERHERHMRWIVXXLIRI[SRI

66 Operation -RWIVXMRKGSQQERHWMRXLIUYIYI 8SMRWIVXEGSQQERHEXEKMZIRTPEGIMR XLIUYIYIWIPIGXXLI GSQQERHHMVIGXP]EJXIVXLIMRWIVXMSRTSWMXMSRERHGLSSWIEdit:Insert Command8LMW[MPPMRWIVXEFPEROPMRIMRXLIUYIYI[MXLEXIQTSVEV] StopMGSRSRXLIFPEROPMRI)\IGYXMSR[MPPWXSTMJXLIUYIYIVIEGLIW XLMWTSMRX[LMPIXLIGSQQERHHMEPSKYIFS\MWSTIR;MXLXLIFPERO PMRIWIPIGXIHGLSSWIXLIGSQQERH]SY[MWLXSMRWIVX)\IGYXMSR[MPP WXSTMJXLIUYIYIVIEGLIWXLMWTSMRX[LMPIXLIGSQQERHHMEPSKYIFS\MW STIR8LIStop MGSR[MPPFIVIQSZIH[LIR]SYGPMGOSROKXS MQTPIQIRXXLIMRWIVXIHGSQQERH =SYGERSRP]MRWIVXGSQQERHWMRXLIUYIYIEJXIVXLIGSQQERH GYVVIRXP]FIMRKI\IGYXIH 7XSTTMRKXLIUYIYI 8SWXSTXLIUYIYIFIJSVIEWIPIGXIHGSQQERHGPMGOSRXLIGSQQERH ERHGLSSWIRun:Stop at CommandJVSQXLIGSQQERHUYIYI[MRHS[ QIRY8LMW[MPPMRWIVXEStop at CommandPMRIMRXLIUYIYI MQQIHMEXIP]FIJSVIXLIWIPIGXIHGSQQERH;LIRI\IGYXMSRVIEGLIWE Stop at commandxliuyiyiwxstwerhxlipmriwxexywglerkiwxse StopMGSR8SGSRXMRYII\IGYXMSREJXIVEStopGLSSWIRun:Start queue JVSQXLIGSQQERHUYIYI[MRHS[QIRY 8SVIQSZIEStop at CommandPMRI[LMGLLEWRSX]IXFIIRVIEGLIHMR XLIUYIYIWIPIGXXLIPMRIERHGLSSWIEdit:Remove 8SWXSTXLIUYIYIEJXIVI\IGYXMSRSJXLIGYVVIRXGSQQERHLEW GSQTPIXIHGLSSWIRun:Stop QueueJVSQXLIGSQQERHUYIYI[MRHS[ QIRY8LMWTPEGIWEStop at CommandPMRIHMVIGXP]EJXIVXLIGYVVIRX GSQQERH=SYGERLSPHXLIUYIYIFIJSVIMRWIVXMRKER]MRWXVYQIRX GSQQERHWF]GLSSWMRKStop Queue[LIRXLIUYIYIMWIQTX] 8SWXSTXLIUYIYIMQQIHMEXIP]TVIWWCtrl-BreakWII7IGXMSR 8LMW[MPPWXSTXLIWIRWSVKVEQERHQE]EFSVXXLIGYVVIRXGSQQERH [LMGL[MPPXLIRFIQEVOIH[MXLEFailedWXEXYWMGSRMRXLIUYIYI 2SXI 8LIUYIYI[MRHS[VIQEMRWSTIREJXIVECtrl-BreakWXSTERH QYWXFIGPSWIHFIJSVIERI[WIRWSVKVEQGERFIWXEVXIH=SY GERKIRIVEXIEQIXLSHJVSQXLIUYIYIFIJSVIGPSWMRKXLI UYIYI[MRHS[ 'PSWMRKXLIUYIYI 8SGPSWIXLIGSQQERHUYIYIGLSSWIFile:CloseJVSQXLIUYIYI [MRHS[QIRYSVCommand:Command QueueJVSQXLIQEMR[MRHS[ QIRY'PSWMRKXLIUYIYI[MPPGPIEVER]UYIYIHGSQQERHW8LI WIRWSVKVEQ[MPPLS[IZIVGSRXMRYIXSVYR

67 Operation 7XSTTMRKXLIWIRWSVKVEQ =SYGERSRP]WXSTXLIWIRWSVKVEQ[LMPIGSQQERHWEVI[EMXMRKJSV I\IGYXMSRMJXLIGSQQERHUYIYIMW WXSTTIH-JXLIVIEVIGSQQERHWMR XLIUYIYI]SY[MPPFI[EVRIHXLEXWXSTTMRKXLIWIRWSVKVEQ[MPPGPIEV XLIUYIYI-J]SY[ERXXSKIRIVEXIEQIXLSHJVSQXLIUYIYIH GSQQERHWHSXLMWFIJSVIWXSTTMRKXLIWIRWSVKVEQ7XSTTMRKXLI WIRWSVKVEQGPSWIWXLIGSQQERHUYIYI[MRHS[ )QIVKIRG]WXSTTVSGIHYVI'XVP&VIEO -J]SYTVIWWCtrl-Break[LMPIGSQQERHWEVIUYIYIHXLIVYR[MPPFI EFSVXIHFYXXLIGSQQERHW[MPPVIQEMRMRXLIUYIYI[MRHS[ERHGER FIWEZIHEWEQIXLSH=SYQYWXGPSWIXLIUYIYI[MRHS[FIJSVI]SY GERWXEVXERSXLIVWIRWSVKVEQ Generating methods from the command queue 8SKIRIVEXIEQIXLSHJVSQXLIGSQQERHUYIYIGLSSWIFile:Generate MethodJVSQXLIGSQQERHUYIYI[MRHS[QIRY8LMW[MPPKIRIVEXIE QIXLSHMREQIXLSHIHMXSV[MRHS[GSRXEMRMRK MRXVSHYGXSV]GSQQIRXWMRGPYHMRKXLIGSRXVSPWSJX[EVIZIVWMSRJVSQ [LMGLXLIQIXLSH[EWKIRIVEXIHERHEHEXIERHXMQIWXEQT 3RIAPROGl'SQQERH5YIYIzGSRXEMRMRKXLIGSQQERHWJVSQXLI UYIYI[MRHS[-JXLIUYIYIGSRXEMRWEGSQQERH[MXLFailedWXEXYW EWEVIWYPXSJIMXLIVXIGLRMGEPJEMPYVISVYWIVMRXIVZIRXMSR[MXL'XVP &VIEOXLMW[MPPFIQEVOIH[MXLEGSQQIRXMRXLIQIXLSH EMAINFPSGO[LMGLWIXWXLIXLIVQSVEGOWJPS[TEXLERHJPS[VEXI GEPPWXLIAPROG'SQQERH5YIYIERHJMRMWLIW[MXLERAPPEND standbymrwxvygxmsr Stop at commandpmriwhsrsxkirivexier]gsqqerhmrxliqixlsh 8LIQIXLSHMWGSQTPIXIMRMXWIPJERHGERFIWEZIHXSHMWO6YRRMRKMX [MPPVITIEXXLIEGXMSRWMRXLIGSQQERHUYIYI-J]SY[ERXXSYWIE KIRIVEXIHQIXLSHEWXLIWXEVXMRKTSMRXJSV]SYVS[RQIXLSH HIZIPSTQIRXFIEVXLIJSPPS[MRKTSMRXWMRQMRH )EGLQIXLSHKIRIVEXMSRGVIEXIWEWIPJGSRXEMRIHQIXLSH[MXLER APROG[MXLXLIWEQIREQI-J]SY[ERXXSGSQFMRIAPROGWJVSQ HMJJIVIRXKIRIVEXIHQIXLSHWYWMRKGYXERHTEWXIMRXLIQIXLSH IHMXSVVIQIQFIVXSGLERKIXLIREQIWMRXLIDEFINE APROGFPSGOW ERHXLIGSVVIWTSRHMRKGEPPWMRXLIMAINFPSGO %KIRIVEXIHQIXLSHMRGPYHIWEPPGSQQERHWMRXLIUYIYIEXXLIXMQI SJKIRIVEXMSR=SYQE][ERXXSHIPIXIWYTIVJPYSYWGSQQERHWJVSQ XLIUYIYIFIJSVIKIRIVEXMRKXLIQIXLSH

68 Operation 'SQQERHWMRXLIKIRIVEXIHQIXLSHEVIRSXXMQIGSRXVSPPIHERH VITSVXTSMRXWEHHIHHYVMRKXLIQERYEPUYIYIGSRXVSPPIHVYREVI RSXMRGPYHIHMRXLIQIXLSH=SYQE][MWLXSEHHXMQIWTIGMJMGEXMSRW ERHVITSVXTSMRXWXSXLIKIRIVEXIHQIXLSH 2SXI 1IXLSHGSQQERHWEVIKIRIVEXIHI\EGXP]EWXLI]ETTIEVMRXLI UYIYI[MRHS[-J]SYGLSSWIStop injecthyvmrkeuyiyih MRNIGXMSRXLIKIRIVEXIHQIXLSH[MPPMRGPYHIERINJECT GSQQERHJSVXLIZSPYQISVMKMREPP]VIUYIWXIHRSXJSVXLI ZSPYQIEGXYEPP]HIPMZIVIH-J]SYKIRIVEXIEQIXLSHMR GSRRIGXMSR[MXLHIZIPSTQIRX[SVO[LIVI]SYLEZIWXSTTIH MRNIGXMSRWMRXIVEGXMZIP]]SYQE]RIIHXSQSHMJ]XLIKIRIVEXIH QIXLSHMREGGSVHERGI[MXLXLIIZIRXPSKSJXLIEGXYEPVYR Command queue window menu -RXLII\TERHIHJSVQXLIGSQQERHUYIYI[MRHS[LEWEQIRYFEV [MXLXLIJSPPS[MRKJYRGXMSRW File:Generate Method File:Close Edit:Copy Edit:Insert Edit:Delete +IRIVEXIWEQIXLSHJVSQXLIGSQQERHWMR XLIUYIYI8LIQIXLSH[MPPETTIEVMRERI[ YRXMXPIHQIXLSHIHMXSV[MRHS[7II7IGXMSR JSVHIXEMPW 'PSWIWXLIGSQQERHUYIYI8LMWSTIVEXMSR GPIEVWEPPGSQQERHWJVSQXLIUYIYI;LIR XLIUYIYIMWGPSWIHGSQQERHWVIUYIWXIH JVSQXLICommandQIRYEVII\IGYXIH HMVIGXP][MXLSYXFIMRKUYIYIH 'STMIWQEVOIHGSQQERHWJVSQXLIUYIYI XSXLIQIXLSHIHMXSVGPMTFSEVH'STMIH GSQQERHWGERFITEWXIHMREQIXLSHIHMXSV [MRHS[8LI]GERRSXLS[IZIVFITEWXIHMR ERSXLIVTSWMXMSRMRXLIUYIYI -RWIVXWERI[GSQQERHMRXLIUYIYI MQQIHMEXIP]FIJSVIXLIQEVOIHGSQQERH 'SQQERHWGERRSXFIMRWIVXIHFIJSVIXLI GSQQERHGYVVIRXP]FIMRKI\IGYXIH (IPIXIWXLIQEVOIHGSQQERHWJVSQXLI UYIYI

69 Operation Edit:Clear Queue Edit:Edit Command Run:Start queue Run:Stop Queue Run:Stop at Command (IPIXIWEPPGSQQERHWJVSQXLIUYIYI MRGPYHMRKER][LMGLLEZIRSX]IXFIIR I\IGYXIH(SIWRSXGPSWIXLIGSQQERH UYIYI[MRHS[ 3TIRWXLIETTVSTVMEXIHMEPSKYIFS\JSV IHMXMRKTEVEQIXIVWJSVXLIQEVOIH GSQQERH3RP]GSQQERHWXLEXLEZIRSX ]IXFIIRI\IGYXIHGERFIIHMXIH 7XEVXWUYIYII\IGYXMSRMJMXMWGYVVIRXP] WXSTTIHWII7IGXMSR8LMWGSQQERH GLERKIWXSStop queue[lirxliuyiyimw VYRRMRK 7XSTWXLIUYIYIEJXIVI\IGYXMSRSJXLI GYVVIRXGSQQERH8LMWGSQQERHGLERKIW XSStart queue[lirxliuyiyimwwxsttih 2SXIXLEXWXSTTMRKXLIUYIYIMWRSXXLI WEQIEWGPSWMRKMXGSQQERHWMWWYIH[LMPI XLIUYIYIMWWXSTTIH[MPPFITPEGIHMRXLI UYIYIERHI\IGYXIH[LIRXLIUYIYIMW WXEVXIH 4PEGIWEStop queuemrwxvygxmsrmqqihmexip] FIJSVIXLIQEVOIHGSQQERH8LIUYIYI [MPPWXST[LIRXLMWTSMRXMWVIEGLIH 4.6 Method-controlled operation &-%'36)GERFIGSRXVSPPIHJVSQE TVITVSKVEQQIHQIXLSHJSV YREXXIRHIHSTIVEXMSR%QIXLSHGERMRGPYHIER]RYQFIVSJEREP]WMW G]GPIWIEGLVITVIWIRXIHF]EWIRWSVKVEQMRXLIVIWYPXJMPI8LMWWIGXMSR HIWGVMFIWLS[XSSTIVEXI&-%'36)YRHIVQIXLSHGSRXVSP 2SXI 8LIMRWXVYQIRXHSSVMWPSGOIHHYVMRKQIXLSHSTIVEXMSR I\GITX[LIRSTIVEXSVEGGIWWXSXLIEYXSWEQTPIVMW VIUYMVIH 1IXLSHMRWXVYGXMSRWXLEXVIUYMVIXLMWEGGIWWEVITPEGIHMRE WTIGMEPDEFINE ALERTFPSGOMRXLIQIXLSHWII%TTIRHM\) Creating a method 8SGVIEXIERI[QIXLSHGLSSWIFile:New MethodJVSQXLI&-%'36) 'SRXVSP7SJX[EVIQIRY*SPPS[XLIWXITWFIPS[XSIRXIVERIPIQIRXEV] QIXLSH[LMGLGERFIYWIHXSMPPYWXVEXIQIXLSHGSRXVSPPIHSTIVEXMSR 6IJIVXS%TTIRHMGIW)ERH*JSVHIXEMPWSJQIXLSHTVSKVEQQMRK 2SXI =SYGERRSXYWIXLIQIXLSHIHMXSV[LMPIEWIRWSVKVEQMW VYRRMRK

70 Operation 8]TIXLIJSPPS[MRKPMRIWMRXLIQIXLSHIHMXSV[MRHS[ define aprog practice flow 30 dilute r1a1 r1a2 r1a3 50 inject r1a3 25 end main detection 1 aprog practice end 8LMWMWEGSQTPIXIQIXLSH[LMGL[MPPTVITEVIMRTSWMXMSRVE P SJEQM\XYVISJWEQTPIWJVSQVEERHVEXLIRMRNIGX PSJ XLMWQM\XYVISZIVXLIWIRWSVGLMTWYVJEGIMRJPS[GIPPEXEJPS[VEXISJ PQMR 'LSSWIEdit:Adjust Method:Minutes8LMW[MPPEHNYWXXLIQIXLSH ETTIEVERGIWSXLEXOI][SVHWEVIWIXMRGETMXEPWTEVEQIXIVREQIWEVI MRPS[IVGEWIERHGSQQERHWEVIMRHIRXIHXSWLS[XLIFPSGOWXVYGXYVI SJXLIQIXLSH DEFINE APROG practice FLOW 30 DILUTE r1a1 r1a2 r1a3 50 INJECT r1a3 25 END MAIN DETECTION 1 APROG practice END 8LISecondsERHMinutesSTXMSRWEHNYWXXMQIVIJIVIRGIWMRXLI QIXLSHXSXLIVIWTIGXMZIXMQIFEWI8LIVIEVIRSXMQIVIJIVIRGIWMR XLMWI\EQTPIWSXLIGLSMGISJSTXMSRMWMVVIPIZERXMRXLMWGEWI7II 7IGXMSRJSVEHIWGVMTXMSRSJXMQIFEWIEHNYWXQIRX Checking and saving the method -XMWMQTSVXERXXLEXQIXLSHMRWXVYGXMSRWEVIIRXIVIHGSVVIGXP]WMRGIE QIXLSH[MXLW]RXE\IVVSVWGERRSXFIVYR8LITVIVYRJEGMPMX]GLIGOW XLIQIXLSHW]RXE\ERHMHIRXMJMIWMRGSVVIGXMRWXVYGXMSRW 2SXI 8LITVIVYRJEGMPMX]GERSRP]GLIGOXLIQIXLSHW]RXE\RSXXLI MRXIRXMSRSJXLIQIXLSH-XMW]SYVVIWTSRWMFMPMX]XSIRWYVIXLEX XLIQIXLSHMRWXVYGXMSRWTIVJSVQXLIVMKLXSTIVEXMSRWIKXLEX

71 Operation INJECTGSQQERHWEHHVIWWXLIGSVVIGXVEGOTSWMXMSRERH WEQTPIZSPYQI%QIXLSH[MXLW]RXE\IVVSVWGERFIWEZIH FYXGERRSXFIVYR 'LSSWIRun:Prerun Method6IJIVXS7IGXMSR)MJ]SYVIGIMZIER] W]RXE\IVVSVQIWWEKIW'SVVIGXW]RXE\IVVSVWERHVITIEXXLITVIVYR TVSGIHYVI 'LSSWIFile:Save AsXSWEZIXLIQIXLSH1IXLSHJMPIWEVIWEZIH[MXL XLII\XIRWMSRFPQ Running a method 1IXLSHWEVII\IGYXIH[MXLXLIGSQQERH Run:Run Method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un MethodGSQQERHTVSQTXWJSV EQIXLSHXSSTIR

72 Operation %TVIVYRMWTIVJSVQIHEYXSQEXMGEPP][LIRIZIV]SYI\IGYXIEQIXLSH -JXLIVIEVIW]RXE\IVVSVWMRXLIQIXLSHXLIWI[MPPFIMHIRXMJMIHMRXLI QIXLSHIHMXSV[MRHS[=SYQYWXGSVVIGXXLIIVVSVWERHGLSSWI Run:Run MethodEKEMRXSWXEVXXLIQIXLSH -JXLIAutosaveJYRGXMSRMWEGXMZEXIHJSVQIXLSHVYRWYRHIV Tools:Preferences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ethod readyqiwweki FS\ 'LSSWIFile:Save AsXSWEZIXLIWIRWSVKVEQWJVSQXLIQIXLSHVYRMRE VIWYPXJMPI8LIVIWYPXJMPI[MPPGSRXEMRSRISVJSYVWIRWSVKVEQWJSVIEGL G]GPIMRXLIQIXLSHEGGSVHMRKXSXLIHIXIGXMSRQSHI Tracing method operation (YVMRKEQIXLSHGSRXVSPPIHVYRXLIViewQIRYMRGPYHIWEMethod TraceGSQQERH'LSSWMRKXLMWGSQQERHHMWTPE]WEQIXLSHXVEGI [MRHS[[LIVIXLITVSKVIWWSJXLIQIXLSHGERFIJSPPS[IH

73 Operation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topping a method 8SWXSTEQIXLSHGLSSWIRun:Stop Method8LIQIXLSH[MPPGSRXMRYI XSXLIIRHSJXLIGYVVIRXGSQQERHMRXLIMAINFPSGOYWYEPP]XLIIRH SJXLIGYVVIRXAPROGERHXLIRWXST8LIJPS[W]WXIQMW[EWLIH EYXSQEXMGEPP][LIRXLIQIXLSHWXSTW =SYGERWXSTEQIXLSHMQQIHMEXIP]YWMRKXLIIQIVKIRG]WXST'XVP &VIEOWII7IGXMSR2SXIXLEXMJXLIGYVVIRXGSQQERHMRXLI MAINFPSGOGERRSXFIGSQTPIXIHIKMJXLIXIQTIVEXYVIVIUYIWXIHF] ETEMPGSQQERHGERRSXFIEGLMIZIH]SYQYWXYWIXLIIQIVKIRG] WXST'XVP&VIEOXSWXSTXLIQIXLSH

74 Operation 4.7 Closing down the system BIACORE 2000 Processing Unit *SPPS[SRISJXLITVSGIHYVIWFIPS[XSGPSWIHS[RXLI&-%'36) 4VSGIWWMRK9RMXEGGSVHMRKXSLS[PSRKXLIW]WXIQ[MPPFIMHPI 3ZIVRMKLX 8LIMRWXVYQIRXGERFIPIJXSZIVRMKLX[MXLXLIWIRWSVGLMTHSGOIHERH XLI-*'ERHJPS[GIPPWJMPPIH[MXLFYJJIV;IVIGSQQIRHXLEXXLI TYQTWEVIEPPS[IHXSVYREXEPS[JPS[VEXI PQMRF]QIERWSJ XLIStandbyTVSGIHYVIWII7IGXMSR1EOIWYVIXLEXXLIVIMW WYJJMGMIRXFYJJIVMRXLIFYJJIVVIWIVZSMVXLITVSGIHYVIYWIWEFSYXQP QSVIFYJJIVXLERVIUYMVIHJSVGSRXMRYSYWJPS[MILSTIVEXMSR VIUYMVIWEFSYXQPFYJJIV%WERI\XVETVIGEYXMSRTEVXMGYPEVP]MJ ]SYVFYJJIVLEWEXIRHIRG]XSTVIGMTMXEXI]SYQE]VITPEGIXLIVYRRMRK FYJJIV[MXL[EXIV9WIXLI[SVOMRKXSSPRinseXSJPYWLXLI-*'ERH JPS[GIPPW[MXL[EXIVERHXLIRVYRStandbyXSPIEZI[EXIVJPS[MRK XLVSYKLXLIW]WXIQ =SYGERETTIRHStandbyXSEQIXLSHXSWXEVXXLITVSGIHYVI EYXSQEXMGEPP][LIRXLIQIXLSHMWGSQTPIXIHWII%TTIRHM\* 9TXSJMZIHE]W 8SGPSWIHS[RXLIW]WXIQJSVYTXSJMZIHE]WVYRWorking Tools:Close WII7IGXMSR8LMWJPYWLIWXLIW]WXIQ[MXLPMUYMH [EXIVMW VIGSQQIRHIHJVSQETVIHIJMRIHEYXSWEQTPIVVEGOTSWMXMSRXLIR IQTXMIWXLI-*'ERHJPS[GIPPW 1SVIXLERJMZIHE]W 8SGPSWIHS[RXLIW]WXIQJSVQSVIXLERJMZIHE]WVYRWorking Tools:ShutdownWII7IGXMSR*SPPS[XLIMRWXVYGXMSRWSRXLI WGVIIR=SY[MPPRIIH[EXIVERHIXLERSPJSVXLMWTVSGIHYVI Control Software 8SGPSWI&-%'36)'SRXVSP7SJX[EVIGLSSWIFile:ExitSVGPSWIXLI [MRHS[YWMRKWXERHEVH;MRHS[WXIGLRMUYIW-JXLIGYVVIRX WIRWSVKVEQSVQIXLSHLEWRSXFIIRWEZIH]SY[MPPFITVSQTXIHXSHS WSFIJSVIGPSWMRKXLIWSJX[EVI=SYGERRSXGPSWI&-%'36)'SRXVSP 7SJX[EVI[LMPIEWIRWSVKVEQMWVYRRMRK -J]SYEWOXSGPSWI&-%'36)'SRXVSP7SJX[EVI[MXLEWIRWSVGLMT HSGOIH]SYEVIKMZIRERSTTSVXYRMX]XSTIVJSVQXLIWLYXHS[R TVSGIHYVI

75 Operation 'LSSWIYesXSWLYXHS[RXLIMRWXVYQIRXERHGPSWIXLIWSJX[EVINoXS GPSWIXLIWSJX[EVI[MXLSYXWLYXXMRKHS[RSVCancelXSVIXYVRXSXLI WSJX[EVI 2SXI 'PSWMRKXLI&-%'36)'SRXVSP7SJX[EVI[MPPRSXGPSWIXLI &-%IZEPYEXMSRERH&-%WMQYPEXMSRETTPMGEXMSRWIZIRMJXLIWI [IVIWXEVXIHJVSQXLIGSRXVSPWSJX[EVI 4.8 Viewing and evaluating results 8LIWIRWSVKVEQ[MRHS[HMWTPE]WXLIVIWYPXWSJXLIGYVVIRXVYRSVXLI GSRXIRXWSJEWEZIHVIWYPXJMPI7IRWSVKVEQWJVSQFSXLWSYVGIWEVI I\EQMRIHMRXLIWEQI[E]8SSTIREVIWYPXJMPIGLSSWIFile:OpenERH WIPIGXXLIVIUYMVIHJMPI Using multiple windows =SYGERHMWTPE]QYPXMTPIWIRWSVKVEQ[MRHS[WSRXLIWGVIIRXSWLS[ XLIWIRWSVKVEQEXHMJJIVIRXWGEPIWHMJJIVIRXTEVXWSJXLIWEQI WIRWSVKVEQSVHMJJIVIRXGYVZIWMRXLIWEQIVYR 8LIHMWTPE]TVSTIVXMIWSJWITEVEXI[MRHS[WEVIMRHITIRHIRXSJIEGL SXLIV[MXLXLII\GITXMSRSJG]GPIRYQFIVXLIG]GPIGERFIGLSWIR MRHITIRHIRXP]MREWEZIHVIWYPXJMPIFYXEPP[MRHS[WWLS[XLIGYVVIRX G]GPIHYVMRKEVYR 'LSSWIWindow:New WindowXSGVIEXIERI[[MRHS[%VVERKI [MRHS[WIMXLIVF]HVEKKMRKXLIETTVSTVMEXI[MRHS[FSVHIVWSV GLSSWMRKSRISJXLITileGSQQERHWJVSQXLIWindowsQIRY

76 Operation 2SXI 1YPXMTPIWIRWSVKVEQ[MRHS[WGERSRP]WLS[HMJJIVIRXZMI[W SJEWMRKPIVIWYPXJMPI8SGSQFMRIWIRWSVKVEQWJVSQWIZIVEP VIWYPXJMPIWSTIRXLIJMPIWMR&-%IZEPYEXMSRERHGSQFMRIXLI WITEVEXIVIWYPXWMRE&-%IZEPYEXMSRTVSNIGX[MRHS[ Selecting sensorgrams %G]GPIMWHIJMRIHEWXLIVIWYPXSJSRIVYRIMXLIVEQERYEPVYRSVER APROGMREQIXLSHWII7IGXMSR)%QERYEPVYRKIRIVEXIWSRI G]GPISRP]%QIXLSHVYRQE]KIRIVEXIER]RYQFIVSJG]GPIW)EGL G]GPIGSRXEMRWIMXLIVSRISVJSYVWIRWSVKVEQWWMRKPIERHQYPXMGLERRIP HIXIGXMSRQSHIWVIWTIGXMZIP]ERHQE]EPWSGSRXEMREHHMXMSREPGYVZIW IKXIQTIVEXYVISVFYJJIVZSPYQIMJXLIWIEVIVIUYIWXIHMRXLI QIXLSHWII7IGXMSR) 'LSSWIXLIG]GPIXSHMWTPE]MRXLICycle[MRHS[SRXLIXSSPFEV8LI GYVVIRXG]GPIMWEP[E]WHMWTPE]IHHYVMRKEQIXLSHVYRFYX]SYGER GLERKIXLIHMWTPE]IHG]GPI[LIRXLIQIXLSHMWGSQTPIXI'LSSWIXLI EGXMZIGYVZIMRXLICurve[MRHS[SRXLIXSSPFEV8LIEGXMZIGYVZIMW XLISRIXS[LMGLXLIVIJIVIRGIPMRIERHFEWIPMRIJYRGXMSRWETTP]WII 7IGXMSRERHXLISRIJSV[LMGLGSSVHMREXIZEPYIWEVIHMWTPE]IH MRXLIWXEXYW[MRHS[ -J]SYLEZIWIZIVEPGYVZIWMRXLIWIRWSVKVEQ[MRHS[XLI View:OverlayGSQQERH[MPPXSKKPIFIX[IIRHMWTPE]SJEPPGYVZIWERH HMWTPE]SJXLIEGXMZIGYVZISRP] Viewing curve classes -J]SYLEZIYWIHSRPMRIVIJIVIRGIWYFXVEGXMSRWIPIGXMRKXLIView:Plot Curve ClassesGSQQERH[MPPTPSXEPPGYVZIWMRXLIWEQIGPEWWSVMKMREP SVVIJIVIRGIWYFXVEGXIHEWXLIGYVVIRXP]WIPIGXIHGYVZI8LMWWMQTPMJMIW EYXSQEXMGWGEPMRKWMRGIXLISVMKMREPERHVIJIVIRGIWYFXVEGXIHGYVZIWEVI YWYEPP][MHIP]WITEVEXIHMRXIVQWSJVIWTSRWIZEPYIW

77 Operation Scaling the display 8SIRPEVKI^SSQETEVXSJXLIWIRWSVKVEQ[MRHS[HVEKXLIGYVWSV SZIVXLIEVIEXSFIIRPEVKIHLSPHHS[RXLIPIJXQSYWIFYXXSREXSRI GSVRIVSJXLIEVIEQSZIXLIQSYWIXSXLISTTSWMXIGSVRIVXLIRVIPIEWI XLIQSYWIFYXXSR-J]SYHSXLMW[LMPIEWIRWSVKVEQMWVYRRMRKERH WGEPMRKMWWIXXSAutoXLIWGEPIW[MPPFIEHNYWXIHEWWSSREWXLI WIRWSVKVEQKSIWSYXWMHIXLIHMWTPE] 8SVIZIVXXSXLITVIZMSYWHMWTPE]EJXIV^SSQMRKWIPIGX View:Unzoom SVHSYFPIGPMGOMRXLI[MRHS[[MXLXLIPIJXQSYWIFYXXSR9TXS PIZIPWSJ^SSQMRKGERFIVIWXSVIHMRXLMW[E] 7IPIGXView:ScaleXSSTIREHMEPSKYIFS\[LIVI JM\IHPMQMXWGERFIWIX JSVXLI\ERH]E\IW'LSSWIAutoXSEHNYWXXLIWGEPIWXSMRGPYHIXLI [LSPIWIRWSVKVEQ'LSSWIMethodXSHMWTPE]XLIWIRWSVKVEQ EGGSVHMRKXSXLIWGEPIHIJMRIHMRXLIQIXLSHXLMWSTXMSRMWSRP] EZEMPEFPIMJXLIQIXLSHMRGPYHIWESCALEMRWXVYGXMSRWII%TTIRHM\* 'LSSWIManualXSWIXXLIWGEPIPMQMXWQERYEPP]=SYGEREPWSGLSSWI XSHMWTPE]XLIWIRWSVKVEQSREPSKEVMXLQMGWGEPI 7IPIGXView:Adjust ScaleXSEHNYWXXLIWGEPIWSXLEXXLIIRXMVI WIRWSVKVEQMWHMWTPE]IHMRXLI[MRHS[VIKEVHPIWWSJER]TVIZMSYW JM\IHSVEYXSQEXMGWGEPMRK Reading coordinate values =SYGERVIEHVIWTSRWIERHXMQIZEPYIWSRXLIEGXMZIGYVZIYWMRKXLI VIJIVIRGIPMRIERHFEWIPMRI 7IPIGXView:Reference LineXSHMWTPE]XLIVIJIVIRGIPMRI=SYGERHVEK XLIZIVXMGEPVIJIVIRGIPMRI[MXLXLIGYVWSVXLILSVM^SRXEPVIJIVIRGI PMRIJSPPS[WXLI]GSSVHMREXISJXLIEGXMZIGYVZIERHXLIEFWSPYXI GSSVHMREXIWSJXLIVIJIVIRGIPMRIEVIHMWTPE]IHMRXLIWXEXYW[MRHS[ =SYGEREPWSQSZIXLIVIJIVIRGIPMRI[MXLXLIEVVS[OI]WSRXLI OI]FSEVH

78 Operation ;MXLXLIVIJIVIRGIPMRIHMWTPE]IHWIPIGXView:BaselineXSJM\EFEWIPMRI EXXLIGYVVIRXTSWMXMSRSJXLIVIJIVIRGIPMRI8LMWFEWIPMRI[MPPVIQEMR MRTSWMXMSREW]SYQSZIXLIVIJIVIRGIPMRIERHXLIWXEXYW[MRHS[[MPP RS[HMWTPE]XLI]GSSVHMREXIEXXLIVIJIVIRGIPMRIVIPEXMZIXSXLIJM\IH FEWIPMRI 8LYWXSVIEHEVIPEXMZIVIWTSRWIZEPYIJSVI\EQTPIEXXLIIRHSJER MRNIGXMSR 'LSSWIView:Reference Line 1SZIXLIVIJIVIRGIPMRIXSETSWMXMSRFIJSVIXLIWXEVXSJXLI MRNIGXMSR 'LSSWIView:Baseline 1SZIXLIVIJIVIRGIPMRIXSXLIIRHSJXLIMRNIGXMSRERHVIEHXLI VIPEXMZIVIWTSRWIJVSQXLIWXEXYW[MRHS[ 8SWIXERI[FEWIPMRIZEPYIGLSSWIView:BaselineXSXYVRXLIFEWIPMRI SJJERHVITIEXWXITWERHEFSZI 2SXI 8LIVIJIVIRGIPMRIGSSVHMREXIWWLS[RMRXLIWXEXYW[MRHS[EVI ZEPYIWJVSQWMRKPIHEXETSMRXW9WIVITSVXTSMRXWWII7IGXMSR XSSFXEMRZEPYIWEZIVEKIHSZIVWIZIVEPHEXETSMRXWXS IPMQMREXIRSMWIZEVMEXMSRW

79 Operation Setting report points %VITSVXTSMRXVIGSVHWXLIXMQIVIWTSRWIERHSXLIVTEVEQIXIVW HIJMRIH[MXLXLIView:KeywordsGSQQERHWII7IGXMSREXE WTIGMJMIHTSMRXMRXLIWIRWSVKVEQ-RQYPXMGLERRIPQSHIVITSVXTSMRXW GERFIWIXIMXLIVJSVMRHMZMHYEPWIRWSVKVEQWSVJSVEPPJSYVWIRWSVKVEQW XSKIXLIV6ITSVXTSMRXWEVIHMWTPE]IHMRXLIVITSVXTSMRXXEFPI =SYWIXVITSVXTSMRXW MRXIVEGXMZIP]MRQERYEPVYRWGSQTPIXIHQIXLSHVYRWERHWEZIH WIRWSVKVEQW[MXLXLIEdit:Add Report PointGSQQERHSVXLIVITSVX TSMRXXSSPFEVFYXXSR MRQIXLSHVYRW[MXLXLIRPOINTMRWXVYGXMSR=SYGERRSXWIXVITSVX TSMRXWMRXIVEGXMZIP]HYVMRKEQIXLSHVYR 8LIEdit:Add Report PointGSQQERHSTIRWEHMEPSKYIFS\JSV WTIGMJ]MRKXLIVITSVXTSMRXGLEVEGXIVMWXMGW )RXIVXLIXMQIJSVXLIVITSVXTSMRX-JXLIVIJIVIRGIPMRIMWHMWTPE]IH WII7IGXMSRXLITSWMXMSRSJXLIVIJIVIRGIPMRIMWWYKKIWXIHEWXLI XMQIJSVXLIVITSVXTSMRX9WIXLMWJIEXYVIEWEREMHMRTPEGMRKVITSVX TSMRXWMRXIVEGXMZIP] 8LIWindowTEVEQIXIVHIXIVQMRIWXLI[MRHS[SZIV[LMGLXLI VIWTSRWIZEPYIW[MPPFIEZIVEKIHJSVXLIVITSVXTSMRX8LIHIJEYPX ZEPYISJWMWWYMXEFPIJSVQSWXTYVTSWIW=SYQE][ERXXSMRGVIEWI XLI[MRHS[MJ]SYVWIRWSVKVEQMWRSMW] 8LIIdTEVEQIXIVETTIEVWEWERMHIRXMJMGEXMSRSJXLIVITSVXTSMRXMRXLI VITSVXTSMRXXEFPI=SYGERIRXIVYTXSGLEVEGXIVWMRXLMWJMIPH 'LIGOXLIBaselineFS\MJ]SY[ERXXLMWVITSVXTSMRXXSFIYWIHEWE FEWIPMRIJSVGEPGYPEXMRKVIPEXMZIVIWTSRWIZEPYIWMRWYFWIUYIRXVITSVX TSMRXW 'LIGOApply to all curvesmj]sy[erxxlivitsvxtsmrxxsfiwixjsvepp WIRWSVKVEQWMRQYPXMGLERRIPQSHI-JXLMWFS\MWRSXGLIGOIHXLI VITSVXTSMRX[MPPFIWIXSRXLIEGXMZIWIRWSVKVEQSRP]8LMWWIXXMRKMW MVVIPIZERXMRWMRKPIGLERRIPQSHI

80 Operation )HMXMRKVITSVXTSMRXW 8SGLERKIXLIWIXXMRKWJSVERI\MWXMRKVITSVXTSMRXHSYFPIGPMGOSRXLI ETTVSTVMEXIPMRIMRXLIVITSVXTSMRXXEFPI8LMW[MPPSTIRXLIEdit Report PointHMEPSKYIFS\[LMGLLEWXLIWEQIJMIPHWEWAdd Report PointHIWGVMFIHEFSZI'LERKIXLIWIXXMRKWEWVIUYMVIHERHGPMGOSR OK Report point and event log markers 6ITSVXTSMRXWEVIQEVOIH[MXLERxSRXLIGYVZIERHERMRHMGEXMSRSJ XLIVITSVXTSMRX[MRHS[)ZIRXWVIGSVHIHMRXLIIZIRXPSKWII7IGXMSR EVIQEVOIHEWWQEPPXVMERKPIWSRXLI\E\MWSJXLIWIRWSVKVEQ 'LSSWIView:Markers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ow TSMRXW

81 Operation Event log 8LIIZIRXPSKVIGSVHWEPPIZIRXWHYVMRKEVYRMRGPYHMRKE XMQIWXEQT EXXLIFIKMRRMRKSJXLIVYRERHXLIJPS[GIPPXIQTIVEXYVIEXXLI FIKMRRMRKERHIRHSJXLIVYR'LSSWIView:Extended Event LogXS HMWTPE]EJYPPZIVWMSRSJXLIIZIRXPSKMRGPYHMRKPS[HEXEUYEPMX] IZIRXW8LMWGERFIYWIJYPMRMRWXVYQIRXXVSYFPIWLSSXMRKWMXYEXMSRW Printing sensorgrams 8STVMRXWIRWSVKVEQWERHEWWSGMEXIHHEXEGLSSWIFile:Print7IPIGXXLI MXIQWXSTVMRXJVSQXLIHMEPSKYIFS\ 'LIGOXLIFit To PageFS\XSJMXIEGLMXIQXSEWITEVEXITEKI'PMGOSR XLIMore>>FYXXSRJSVTEKIWIXYTSTXMSRW*SVQSVIHIXEMPWWII File:Print7IGXMSR 2SXI 7IRWSVKVEQWERHVITSVXTSMRXXEFPIW[MPPFITVMRXIHEWXLI]EVI HMWTPE]IHSRXLIWGVIIR[MXLVIWTIGXXSWGEPMRKSZIVPE]ERH EGXMZIGYVZI -J]SY[ERXXSYWIWIRWSVKVEQWEWMPPYWXVEXMSRWMRSXLIV;MRHS[W TVSKVEQWIK[SVHTVSGIWWMRKTVSKVEQWGLSSWIEdit:Copy Graph 8LMW[MPPGST]XLIGYVVIRXWIRWSVKVEQ[MRHS[XSXLI;MRHS[W GPMTFSEVHJVSQ[LIVIMXGERFITEWXIHMRXSSXLIVETTPMGEXMSRW

82 Operation Evaluating sensorgrams =SYGERYWI&-%'36)'SRXVSP7SJX[EVIXSHMWTPE]ERHTVMRX WIRWSVKVEQWERHXSWIXVITSVXTSMRXWEWHIWGVMFIHEFSZI*SVQSVI EHZERGIHIZEPYEXMSRGLSSWITools:Evaluation8LMWKMZIWEGGIWWXS &-%IZEPYEXMSRWSJX[EVI[LMGLTVSZMHIWI\XIRWMZIJEGMPMXMIWJSV IZEPYEXMRKWIRWSVKVEQHEXE[MXLQSHIPJMXXMRKTVSGIHYVIWJSV GEPGYPEXMRKOMRIXMGGSRWXERXWJVSQXLIWIRWSVKVEQW&-%IZEPYEXMSREPWS MRGPYHIWJYRGXMSRWJSVWIRWSVKVEQTVIWIRXEXMSRERHIZEPYEXMSRSJ EREP]XIGSRGIRXVEXMSRJVSQVEXISVFMRHMRKPIZIPQIEWYVIQIRXW8LIWI JYRGXMSRWEVIHIWGVMFIHWITEVEXIP]MRXLI&-%IZEPYEXMSR7SJX[EVI,ERHFSSO =SYGEREPWSGLSSWIXSI\TSVXWIRWSVKVEQHEXESVVITSVXTSMRXXEFPIW XSXI\XJMPIWJSVYWIMRSXLIVWTVIEHWLIIXSVHEXEIZEPYEXMSRTVSKVEQW

83 Operation

84 Application Wizards 5. Application Wizards %TTPMGEXMSR;M^EVHWLIPTFSXLRI[ERHI\TIVMIRGIHYWIVWTIVJSVQ XLIMVEREP]WIWUYMGOP]ERHGSRJMHIRXP]%RETTPMGEXMSR[M^EVHMWE WSJX[EVIXSSPXLEXKYMHIW]SYXLVSYKLXLIWXITWMRERI\TIVMQIRXJVSQ WXEVXXSJMREPEREP]WMW;LIR]SYYWIERETTPMGEXMSR[M^EVH]SYIRXIV EPPVIUYMVIHWIXXMRKWJSVXLIVYREW]SYWXITXLVSYKLXLIHMEPSKYI FS\IW3TIVEXMSRSJXLIMRWXVYQIRXHSIWRSXWXEVXYRXMP]SYGPMGOSR Start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

85 Application Wizards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sing application wizards Starting application wizards 8SWXEVXERETTPMGEXMSR[M^EVHGLSSWIRun:Application WizardsJVSQ XLIQEMRQIRY'LSSWIXLI[M^EVH]SY[ERXXSVYRERHGPMGOSRStart 8SYWIEWEZIH[M^EVHXIQTPEXIWIIFIPS[GPMGOSRUse TemplateMR XLI[M^EVHWIPIGXMSRHMEPSKYIFS\ERHWIPIGXXLIXIQTPEXIJMPIXSYWI ;LIR]SYYWIEXIQTPEXIXLIWEZIHWIXXMRKWJSVEPPXLI[M^EVHHMEPSKYI FS\IWETTIEVEWHIJEYPX]SYGERQEOIER]GLERKIW]SYVIUYMVIXSXLI WIXXMRKWEW]SYWXITXLVSYKLXLI[M^EVH9WMRKEXIQTPEXIXLYWTVSZMHIW EGSRZIRMIRX[E]SJVITIEXMRKEREREP]WMWIMXLIVI\EGXP]SV[MXLQMRSV GLERKIW2SXIXLEXMJ]SYYWIEXIQTPEXIXLIGLSMGISJ[M^EVHMW

86 Application Wizards HIXIVQMRIHF]XLIGSRXIRXSJXLIXIQTPEXIJMPIXLI[M^EVHWIPIGXIHMR XLIHMEPSKYIFS\MWMVVIPIZERX =SYGEREPWSWXEVXERETTPMGEXMSR[M^EVHJVSQ XLIVIWYPXJMPIWEZIH JVSQE[M^EVHWYTTSVXIHVYR8LMW[MPPSTIRXLIETTVSTVMEXI ETTPMGEXMSR[M^EVH[MXLWIXXMRKWEWYWIHMRXLIVIWYPXJMPI8SHSXLMW STIRXLIVIWYPXJMPIGLSSWIView:Wizard TemplateERHGPMGOSREditSV RunMRXLIHMEPSKYIFS\ 8LIEditSTXMSRSTIRWXLI[M^EVHXIQTPEXIEXXLIJMVWXWGVIIRWSXLEX ]SYGERWXITJSV[EVHWXLVSYKLXLI[M^EVHQEOMRKGLERKIWXSXLI WIXXMRKWEWVIUYMVIH8LIRunSTXMSRSTIRWXLI[M^EVHXIQTPEXIEXXLI IRHSJXLI[M^EVHWSXLEX]SYWMQTP]RIIHXSGPMGOStartMRXLI[M^EVH HMEPSKYIMRSVHIVXSVITIEXXLIVYRI\EGXP]=SYQE]SJGSYVWIWXIT FEGO[EVHWXLVSYKLXLI[M^EVHXSQEOIER]GLERKIW]SYRIIH ;LIR]SYGPSWIE[M^EVH[LIVI]SYLEZIIRXIVIHWEQTPIHIXEMPW]SY EVIKMZIRXLISTXMSRSJWEZMRKXLI[M^EVHXIQTPEXIJSVJYXYVIYWI Wizard templates 8LIWIXXMRKWYWIHMRERETTPMGEXMSR[M^EVHGERFIWEZIHMRE XIQTPEXI JSVPEXIVYWI8LMWQEOIWMXIEW]XSVITIEXE[M^EVHFEWIHEREP]WMW -J]SYGPSWIE[M^EVHEJXIVLEZMRKQEHIGLERKIWXSXLIXIQTPEXI]SY [MPPFIKMZIRERSTTSVXYRMX]XSWEZIMX

87 Application Wizards =SYGEREPWSWEZIXLIXIQTPEXIJVSQXLIMenuSTXMSRXLEXMWEZEMPEFPI MRWSQI[M^EVHHMEPSKYIFS\IWWII7IGXMSR;M^EVHXIQTPEXIWEVI WEZIHF]HIJEYPX[MXLJMPIX]TIFP[MRXLIBIA Users\TemplatesJSPHIV 8SGVIEXIE[M^EVHXIQTPEXIERHWEZIMXJSVPEXIVYWI]SYGEREPWS GLSSWINew:Application WizardJVSQXLIFileQIRY8LMW[MPPEPPS[ ]SYXSWIPIGXE[M^EVHERHWXITXLVSYKLMXXLIRWEZIXLIXIQTPEXI [LIREPPTEVEQIXIVWEVIHIJMRIH %WEZIH[M^EVHXIQTPEXIGSRXEMRWEPPWIXXMRKWYTXSERHMRGPYHMRKXLI TSMRXEX[LMGLMX[EWWEZIHERHXLIWIWIXXMRKWEVITVIWIRXIHEWHIJEYPX [LIRXLIXIQTPEXIMWYWIH Common wizard functions 8LIJYRGXMSRWFIPS[EVIGSQQSRXSQSWXETTPMGEXMSR[M^EVHW %YXSWEQTPIVVEGOTSWMXMSRW 8LI[M^EVHWYKKIWXWEYXSWEQTPIVVEGOTSWMXMSRWJSVWEQTPIWVIEKIRXW ERHIQTX]ZMEPWMJRIGIWWEV]FEWIHSRXLIVIUYMVIQIRXWSJ]SYV ETTPMGEXMSR=SYGERGLERKIXLITPEGMRKSJZMEPWMRXLIVEGOWWMQTP]F] HVEKKMRKXLIETTVSTVMEXIJMPPIHZMEPTSWMXMSR=SYGERRSXQSZIEZMEP XSETSWMXMSR[LMGLHSIWRSXLEZIWYJJMGMIRXZSPYQIGETEGMX]

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108 Application Wizards ;LIR]SYWXEVXXLIOMRIXMGEREP]WMW[M^EVH]SYGERGLSSWIXSVYRER] SVEPPSJXLIGSRGIRXVEXMSRWIVMIWERHGSRXVSPI\TIVMQIRXWMREWMRKPI lfexglzvyr'ligoxlii\tivmqirxw]sy[mwlxstivjsvq %WEVIGSQQIRHEXMSRVYRXLIGSRGIRXVEXMSRWIVMIWJMVWX[MXLSYX GLIGOMRKER]GSRXVSPI\TIVMQIRXW;LIRXLIEREP]WMWMWJMRMWLIHGPMGO Next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

109 Application Wizards 6YRRMRKWIZIVEPEREP]XIWMRSRIVYR =SYGERIRXIVWIZIVEPHMJJIVIRXEREP]XIWMRXLIWEQTPIXEFPIJSVXLI OMRIXMGEREP]WMW[M^EVH 2SXILS[IZIVXLEXXLIWEQIVIKIRIVEXMSRGSRHMXMSRW[MPPFIYWIHJSV EPPG]GPIWMRXLIWEQIVYR-J]SY[ERXXSYWIHMJJIVIRXVIKIRIVEXMSR GSRHMXMSRWJSVHMJJIVIRXEREP]XIWVYRXLIEREP]XIWMRWITEVEXIWIWWMSRW Notes on the kinetic analysis wizard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

110 Application Wizards 'SRGIRXVEXMSRWIVMIW ;LIRIRXIVMRKHIXEMPWSJXLIGSRGIRXVEXMSRWIVMIWMXMWQSWXGSRZIRMIRX XSIRXIVXLILMKLIWXEREP]XIGSRGIRXVEXMSRJMVWX8LI[M^EVH[MPP EYXSQEXMGEPP]WYKKIWXZEPYIWJSVXLIRI\XGIPPVS[[LIR]SYTVIWW Enter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extXSGSRXMRYI[MXLXLI[M^EVH -XMWEPWSEHZMWEFPIXSMRGPYHIEWEQTPI[MXL^IVSEREP]XIGSRGIRXVEXMSR XLMWQIERWERMRNIGXMSRSJEREP]XIFYJJIV-HIEPP]XLMWWLSYPHFIE WXVEMKLXFEWIPMRIEJXIVWYFXVEGXMSRSJXLIVIJIVIRGIVIWTSRWILS[IZIV MRGPYWMSRSJWYGLEWEQTPIMRXLIGSRGIRXVEXMSRWIVMIWGERJEGMPMXEXI EWWIWWQIRXSJFEWIPMRIHIZMEXMSRWMRXLIJMREPHEXE

111 Application Wizards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ntepreting results from the kinetic analysis wizard 6IWYPXWJVSQXLIOMRIXMGEREP]WMW[M^EVHEVITVIWIRXIHMREXEFFIH HMEPSKYIFS\[MXLWITEVEXIHXEFWJSVXLIGSRGIRXVEXMSRWIVMIWERH GSRXVSPI\TIVMQIRXW-JQSVIXLERSRIEREP]XILEWFIIRVYRXLI EREP]XIGERFIWIPIGXIHJVSQXLITYPPHS[RPMWXEXXLIXSTSJXLI HMWTPE] 2SXI -J]SYVYRXLIGSRGIRXVEXMSRWIVMIW ERHGSRXVSPI\TIVMQIRXWMR WITEVEXI[M^EVHWIWWMSRWGPSWMRKXLI[M^EVHHMEPSKYIFS\ FIX[IIRVYRWXLIVIWYPXWSJIEGLWIWWMSR[MPPFIWEZIHMR WITEVEXIJMPIW8LIXEFWMRXLI[M^EVHVIWYPXTVIWIRXEXMSR VITVIWIRXI\TIVMQIRXWVYR[MXLMRXLIWEQI[M^EVHWIWWMSR 'SRGIRXVEXMSRWIVMIW 8LIVIWYPXWJVSQXLIGSRGIRXVEXMSRWIVMIWEVIWLS[REWERSZIVPE]TPSX [MXLXLIWIRWSVKVEQWEPMKRIHEXXLIXMQISJMRNIGXMSRERHXLIFEWIPMRI EHNYWXIHXS^IVSFIJSVIMRNIGXMSR8LIVIWYPXWEVIIZEPYEXIHEGGSVHMRK XSEWMQTPIMRXIVEGXMSRQSHIPI\TIVMQIRXEPGYVZIWEVIWLS[RMR VIHERHJMXXIHGYVZIWMRFPYI%TTEVIRXOMRIXMGGSRWXERXWEGGSVHMRKXS XLIJMXXMRKEVIPMWXIHXSKIXLIV[MXLXLIIWXMQEXIHQE\MQYQEREP]XI FMRHMRKGETEGMX]6 QE\

112 Application Wizards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aseline LevelXEFHMWTPE]WXLIFEWIPMRIVIWTSRWI MQQIHMEXIP] EJXIVVIKIRIVEXMSREWEJYRGXMSRSJG]GPIRYQFIV6IWYPXWEVITVIWIRXIH WITEVEXIP]JSVXLIJPS[GIPPWYWIHMRXLIEREP]WMWEGGIWWIHJVSQXLI TYPPHS[RPMWXERHJSVXLIVIJIVIRGIWYFXVEGXIHWMKREPMJERMRPMRI VIJIVIRGIGIPPMWYWIH%PPG]GPIWEVITVIWIRXIHMRGPYHMRKER]GSRXVSP I\TIVMQIRXWXLIZEPYIWEVIRSXHMWXMRKYMWLIHEGGSVHMRKXSEREP]XI [LIRXLIVYRYWIWWIZIVEPEREP]XIW

113 Application Wizards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

114 Application Wizards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r"%&r"%& [LIVIHMWWSGMEXMSRSJ%&TVSGIIHWEXEWMKRMJMGERXP]HMJJIVIRXVEXI JVSQHMWWSGMEXMSRSJ%&%WWYQMRKXLEXXLIVEXISJGSRZIVWMSRSJ%&XS %&MWWYGLXLEXXLITVSTSVXMSRSJ%&GLERKIWFIX[IIRERH QMRYXIWXLMWVIEGXMSRWGLIQI[MPPKMZIHMJJIVIRXP]WLETIHHMWWSGMEXMSR TVSJMPIWEJXIVHMJJIVIRXMRGYFEXMSRXMQIW8LII\EQTPIFIPS[HSIWRSX MRHMGEXIPMROIHVIEGXMSRWWMRGIXLIHMWWSGMEXMSRTLEWIWEVITVEGXMGEPP] MHIRXMGEP7IIXLISRPMRILIPTJSVQSVII\EQTPIW

115 Application Wizards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

116 Customized applications 6. Customized applications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lMJXLIRzGSQQERHWXLEXGSRXVSPMRWXVYQIRXSTIVEXMSR SRXLIFEWMWSJZEVMEFPIWSVVITSVXTSMRXZEPYIW 6.1 Overview of the customized application wizard

117 Customized applications Defining the command sequence 7XEVXMRKXLIGYWXSQM^IHETTPMGEXMSR[M^EVHSTIRWE[MRHS[[LIVI]SY HIJMRIXLIWIUYIRGISJGSQQERHWXLEXGSRWXMXYXISRIG]GPIMRXLI ETTPMGEXMSR ;LIRJMVWXSTIRIHXLI[MRHS[WMWIQTX]I\GITXJSVE*03; GSQQERHXLEXWIXWXLIFYJJIVJPS[XS PQMREXXLIFIKMRRMRKSJXLI VYR=SYGERIHMXXLMWGSQQERHXSGLERKIXLIJPS[VEXIFYX]SY GERRSXHIPIXIMX8LII\EQTPIWLS[REFSZIMWEWMQTPIEQMRIGSYTPMRK WIUYIRGI8LITEVEQIXIVWDetectionERHLigandIRGMVGPIHMRXLI MPPYWXVEXMSREFSZIWLS[RMRFPYIXI\XMRXLIHMWTPE]EVIZEVMEFPIW [LMGLEPPS[XLIMQQSFMPM^EXMSRWIUYIRGIXSFIYWIHJSVHMJJIVIRX PMKERHWMRHMJJIVIRXJPS[GIPPW[MXLMREWMRKPIVYR 8SEHHGSQQERHWXSXLIIRHSJXLIWIUYIRGIGPMGOSRXLIXSSPFEV FYXXSRJSVXLIGSQQERH]SY[ERXXSEHH%PXIVREXMZIP]]SYGER GLSSWIXLIGSQQERHJVSQXLIMenuFYXXSRSVVMKLXGPMGOMRXLI [MRHS[EVIEERHGLSSWIXLIGSQQERHJVSQXLIGSRXI\XQIRY 7LSVXGYXOI]GSQFMREXMSRWJSVXLIGSQQERHWEVIPMWXIHMRXLI QIRYW%HHMRKEGSQQERHSTIRWEHMEPSKYIFS\[LIVI]SYWIX TEVEQIXIVWJSVXLIGSQQERH 8SMRWIVXEGSQQERHMRERI\MWXMRKWIUYIRGIWIPIGXXLIGSQQERH MQQIHMEXIP]EJXIVXLIMRWIVXMSRTSMRXERHJSPPS[XLITVSGIHYVIJSV EHHMRKEGSQQERH

118 Customized applications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lXMIHzMRXLIHMWTPE]XSXLIVIWTIGXMZI GSQQERH[MXLEZIVXMGEPPMRI6ITSVXTSMRXWQSZI[MXLEGSQQERH [LIRXLIGSQQERHMWQSZIHERHEVIHIPIXIHMJXLIGSQQERHMW HIPIXIH 8LI-2.)'8GSQQERHMWEP[E]WHMWTPE]IHXSKIXLIV[MXLE GSVVIWTSRHMRK-2.)'8)2(8LMWLEWRSSTIVEXMSREPJYRGXMSRFYX EPPS[WVITSVXTSMRXWXSFIWIXMRVIPEXMSRXSXLIIRHSJEWEQTPI MRNIGXMSR[MXLSYXLEZMRKXSGEPGYPEXIXLIGSRXEGXXMQIVITVIWIRXIHF] XLIMRNIGXMSR

119 Customized applications Assigning values to variables ;LIR]SYLEZIHIJMRIHXLIWIUYIRGISJGSQQERHWGPMGOSRNextXS KSXSXLICyclesHMEPSKYIFS\,IVI]SYWIXXLIRYQFIVSJVITPMGEXIVYRWJSVIEGLG]GPIERHZEPYIWJSV ER]ZEVMEFPIWYWIHMRXLIGSQQERHWIUYIRGIZEVMEFPIWEPPS[]SYXS VYRQYPXMTPIG]GPIWYWMRKXLIWEQIWIUYIRGISJGSQQERHW=SYGER EPWSGLSSWI[LIXLIVXLIG]GPIW[MPPFIVYRMRVERHSQSVHIVSVMRXLI SVHIVIRXIVIHMRXLIXEFPI 8LIXEFPIMRXLMWHMEPSKYIFS\GSRXEMRWSRIGSPYQRJSVXLIRYQFIVSJ VITPMGEXIWERHSRIJSVIEGLZEVMEFPIYWIHMRXLIGSQQERHWIUYIRGI 8LII\EQTPILIVIHIJMRIWJSYVG]GPIWXSMQQSFMPM^IJSYVHMJJIVIRX PMKERHWMRJPS[GIPPWVIWTIGXMZIP] -J]SY[ERXXSGLERKIXLISVHIVMR[LMGLXLIGSPYQRWEVIHMWTPE]IH GPMGOSRXLIMenuFYXXSRERHWIPIGXSet column order=sygerrsx GLERKIXLISVHIVJSVXLIReplERHDetectionGSPYQRW -JXLISTXMSRUse Separate Vials for Different Solution VariablesMW GLIGOIHIRXVMIWXLEXLEZIXLIWEQIZEPYIWMRHMJJIVIRXZEVMEFPI GSPYQRW[MPPYWIWITEVEXIZMEPWMRXLIEYXSWEQTPIVVEGOIRXVMIW[MXL XLIWEQIZEPYI[MXLMRSRIGSPYQRYWIXLIWEQIZMEP-JXLMWSTXMSRMW RSXGLIGOIHIRXVMIW[MXLXLIWEQIZEPYIMRHMJJIVIRXGSPYQRW[MPPFI QETTIHXSXLIWEQIZMEPTSWMXMSR 'LIGOXLIUse Separate Vials for Different Solution VariablesFS\MJ ]SY[ERXXSYWITSWMXMSRREQIWXLEXQMKLXRSXYRMUYIP]MHIRXMJ]E WEQTPIJSVI\EQTPIMJ]SYYWIXLIWEQTPIREQIEWXLIZEVMEFPIREQI ERHEGSRGIRXVEXMSREWXLITSWMXMSRREQIMRXLIXEFPI9RGLIGOXLI FS\MJ]SYYWIXLIWEQIWSPYXMSRMRHMJJIVIRXGSRXI\XWMREREREP]WMW WYGLEWITMXSTIQETTMRK[LIVIXLIWEQIERXMFSHMIWQE]FIYWIHEW

120 Customized applications JMVWXSVWIGSRHERXMFSHMIWMPPYWXVEXIHMRXLIJSPPS[MRKI\EQTPI*SVER ETTPMGEXMSR[MXLX[SMRNIGXMSRWYWMRKZEVMEFPIWPrimary_AbERH Secondary_AbXLIXEFPI [MXLUse Separate Vials for Different Solution VariablesGLIGOIH[MPP KMZIJSYVWEQTPIWMRXLIRack PositionsHMEPSKYI,S[IZIV[MXLUse Separate Vials for Different Solution VariablesRSX GLIGOIH SRP]X[SWEQTPITSWMXMSRWEVIKIRIVEXIHWMRGIXLIZEVMEFPIREQIWMab- 123ERHMab-456IEGLQETXSXLIWEQITSWMXMSRVIKEVHPIWWSJXLI ZEVMEFPIGSPYQRMR[LMGLXLI]EVIYWIH Assigning rack positions ;LIRZEPYIWLEZIFIIREWWMKRIHXSEPPZEVMEFPIWGPMGOSRNextXSKSXS XLIRack PositionsHMEPSKYIFS\7II7IGXMSRJSVEHIWGVMTXMSRSJ XLMWHMEPSKYIFS\

121 Customized applications Preparing the run 'PMGOSRNextMRXLIRack positionshmepskyifs\xsksxsxliprepare runwyqqev]8lmwhmepskyiwyqqevm^iwxliwspyxmsrwerhzmep TSWMXMSRWVIUYMVIHMRXLIEYXSWEQTPIVWII7IGXMSR-RWSQI GEWIWMJ]SYLEZIYWIH86%27*)6GSQQERHWMR]SYVWIUYIRGIIQTX] ZMEPW[MPPFIRIIHIHMRWSQIEYXSWEQTPIVTSWMXMSRW 2SXI -RWSQIGMVGYQWXERGIWTEVXMGYPEVP]MRWIUYIRGIWMRZSPZMRK GSRHMXMSREPGSQQERHWXLIVIUYMVIHZSPYQISJWSPYXMSR GERRSXFIEGGYVEXIP]HIXIVQMRIHMREHZERGI8LIZSPYQIW WLS[RMRXLIPrepare runhmepskyifs\evil[svwxgewiz ZSPYQIWGEPGYPEXIHEWXLSYKLEPPGSQQERHWEVII\IGYXIHMR EPPG]GPIW 6.2 Constant and variable command parameters 8LIGYWXSQM^IHETTPMGEXMSR[M^EVHEPPS[W]SYXSYWIIMXLIVGSRWXERX SVZEVMEFPITEVEQIXIVWJSVQSWXSJXLIGSQQERHWMRXLIWIUYIRGI 9WIEGSRWXERXTEVEQIXIVMJXLIZEPYIMWXLIWEQIMRIZIV]G]GPISJER EREP]WMW*SVI\EQTPIMJ]SYEVIKSMRKXSMRNIGXXLIWEQIZSPYQISJ WEQTPIMRIEGLG]GPI]SY[SYPHYWIEGSRWXERXZEPYIJSVXLIWEQTPI ZSPYQI 1SWXGSRWXERXTEVEQIXIVWEVIIRXIVIHEWWXVEMKLXJSV[EVHZEPYIWIK lzjsveweqtpizspyqisj P:EPYIWXLEXGSVVIWTSRHXS TSWMXMSRWMRXLIEYXSWEQTPIVVEGOLS[IZIVWYGLEWWEQTPI MHIRXMJMGEXMSRJSVER-2.)'8GSQQERHEVIIRXIVIHEWEREQI[LMGL MWEWWSGMEXIH[MXLEVEGOTSWMXMSRMRXLIRack PositionsHMEPSKYIFS\ PEXIVMRXLI[M^EVH 9WIEZEVMEFPITEVEQIXIVGLIGOVary by CycleMJ]SY[ERXXSZEV] XLIZEPYIFIX[IIRG]GPIW*SVI\EQTPIMJ]SYEVIVYRRMRKE WGVIIRMRKETTPMGEXMSRERH[ERXXSMRNIGXEHMJJIVIRXWEQTPIMRIEGL G]GPIYWIEZEVMEFPIJSVXLIWEQTPIMHIRXMJMGEXMSR=SYWIXXLI RYQFIVSJG]GPIWERHEWWMKREZEPYIXSXLIZEVMEFPIWJSVIEGLG]GPIMR XLICyclesHMEPSKYIFS\PEXIVMRXLI[M^EVH -RKIRIVEPYWIZEVMEFPITEVEQIXIVWEWWTEVMRKP]EWTSWWMFPI7MRGI ZEVMEFPIWEVIEWWMKRIHEZEPYIJSVIEGLG]GPIMRXLIEREP]WMW]SY[MPP GVIEXIEPSXSJHEXEIRXV][SVOMJ]SYYWIZEVMEFPIWYRRIGIWWEVMP] 2SXI 8LIToTSWMXMSRJSV86%27*)6MWVITVIWIRXIHF]EWTIGMEPOMRH SJZEVMEFPI[LMGLGVIEXIWERI[IQTX]ZMEPJSVIEGLG]GPIMRE VYRXLITVMQEV]YWISJ86%27*)6MWXSTVITEVIQM\IHWEQTPIW JSVMRNIGXMSRERHXLMWJIEXYVITVIZIRXW]SYJVSQTVITEVMRKE JVIWLQM\XYVIMREYWIHWEQTPIZMEP8LIIQTX]ZMEPZEVMEFPI HSIWRSXETTIEVMRXLICyclesXEFPIWMRGITSWMXMSRWEVI EWWMKRIHEYXSQEXMGEPP]=SYHSRSXRIIHXSGLIGOVary by CycleMRHMEPSKYIFS\IWJSVGSQQERHWPMOI-2.)'8MRSVHIVXS EHHVIWWXLMWOMRHSJZEVMEFPI

122 Customized applications 6.3 Customized application commands 'SQQERHWWYTTSVXIHMRXLIGYWXSQM^IHETTPMGEXMSR[M^EVHEVIEPP EGGIWWMFPIJVSQXLIXSSPFEV1SWXSJXLIGSQQERHWLEZIGPIEV GSYRXIVTEVXWMRXLIQERYEPSTIVEXMSRQIRYWWII'LETXIVWERH Keyword Flow path Report point Mix Wait Conditional (if-then) Cycle settings Flow Inject Transfer :DVK Comment Cycle settings 7IXWXLIXIQTIVEXYVIEXXLIJPS[GIPPERHXLIHIXIGXMSRQSHIJSVXLI GYVVIRXG]GPI 2SXI7IXXMRKXLIHIXIGXMSRQSHIEPWSWIXWXLIJPS[TEXL7II 7IGXMSRJSVLMRXWSRYWMRKXLIHIXIGXMSRQSHIXSFIWX EHZERXEKI Keywords (IJMRIWOI][SVHWXLEX[MPPETTIEVMRXLIVITSVXTSMRXXEFPIJSVXLI VYR=SYWIXXLIOI][SVHZEPYIEWEZEVMEFPIXLIZEPYISJXLI ZEVMEFPIJSVIEGLG]GPI[MPPETTIEVMRXLIOI][SVHGSPYQRMRXLI VITSVXTSMRXXEFPIWII7IGXMSR Flow 7IXWXLIJPS[VEXI 2SXI;LIR]SYGVIEXIERI[WIUYIRGIEWMRKPI*03;GSQQERH XLEXWIXWXLIJPS[VEXIXS PQMRMWEYXSQEXMGEPP]MRWIVXIH EXXLIFIKMRRMRKSJXLIWIUYIRGI=SYGERGLERKIXLI TEVEQIXIVWJSVXLMWGSQQERHFYX]SYGERRSXHIPIXIMXERH ]SYGERRSXTPEGISXLIVGSQQERHWI\GITXG]GPIWIXXMRKWERH /)=;36(FIJSVIXLIJMVWX*03;8LMWIRWYVIWXLEXXLIJPS[ VEXIMWWIXFIJSVIER]SXLIVGSQQERHWEVII\IGYXIHMRER EREP]WMW Flow path 7IXWXLIJPS[TEXL Inject -RNIGXWWEQTPISZIVXLIWIRWSVWYVJEGI Report point 7IXWVITSVXTSMRXWEXEJM\IHXMQIVIPEXMZIXSEGLSWIRGSQQERH

123 Customized applications Transfer 8VERWJIVWWSPYXMSRJVSQSRIEYXSWEQTPIVZMEPXSERSXLIVSVJVSQ XLIFYJJIVVIWIVZSMVXSEZMEP Mix 1M\IWXLIGSRXIRXWSJEWTIGMJMIHZMEP Wash ;EWLIWIMXLIVXLIEYXSWEQTPIVRIIHPISVXLI-*'GLERRIPW Wait -RWIVXWEHIPE]TIVMSHFIX[IIRGSQQERHW Comment -RWIVXWEGSQQIRXMRXLIGSQQERHWIUYIRGIJSVHSGYQIRXEV] TYVTSWIW'SQQIRXWHSRSXEJJIGXI\IGYXMSRSJXLIGSQQERHWMR XLIWIUYIRGI Conditional commands (if-then) %PPS[WGSRHMXMSREPI\IGYXMSRSJGSQQERHWSVXIVQMREXMSRSJXLI G]GPISVVYRHITIRHMRKSRXLIZEPYISJEVITSVXTSMRXSVZEVMEFPI 8LIWIGXMSRWFIPS[SJJIVWSQILMRXWSRLS[XSGSRWXVYGXETTPMGEXMSRW YWMRKXLIGSQQERHWMRXLIGYWXSQM^IHETTPMGEXMSR[M^EVH*SVQSVI HIXEMPIHHIWGVMTXMSRWSJXLIGSQQERHWXLIQWIPZIWWIIXLISRPMRILIPT 6.4 Using detection mode and flow path Detection mode 8LI()8)'8-32GSQQERHMRCycle settingswixwfsxlxlihixigxmsr QSHIERHXLIJPS[TEXL8LIHIXIGXMSRQSHIGERFIWIXXSEGSRWXERX ZEPYIMRXIVQWSJJPS[GIPPWERHVIJIVIRGIWYFXVEGXMSRSTXMSRWGLSWIR JVSQXLITYPPHS[RPMWXSVXSEZEVMEFPIREQIHDetection8LMW ZEVMEFPIREQIMWTVIHIJMRIHERHGERRSXFIGLERKIH 7IXXMRKXLIHIXIGXMSRQSHIXSEZEVMEFPIEPPS[W]SYXSGLERKI HIXIGXMSRQSHIERHGSVVIWTSRHMRKJPS[TEXLFIX[IIRG]GPIWMREVYR 8LII\EQTPIMPPYWXVEXIHMR7IGXMSRWLS[WLS[XLMWGERFIYWIHXS EHHVIWWEHMJJIVIRXJPS[GIPPMRIEGLG]GPI2SXIXLEXXLIHIXIGXMSR QSHIHIXIVQMRIWFSXLXLIMRMXMEPJPS[TEXLERHXLIRYQFIVSJ WIRWSVKVEQGYVZIWHMWTPE]IH -J]SYVGYWXSQM^IHGSQQERHWIUYIRGIHSIWRSXMRGPYHIE()8)'8-32 GSQQERH]SY[MPPFIEWOIHXSWIXXLIHIXIGXMSRQSHIEXXLI FIKMRRMRKSJXLIVYR=SYGERSRP]GLSSWIEHIXIGXMSRQSHIXLEXMW GSQTEXMFPI[MXLXLIGSQQERHWMRXLIGSQQERHWIUYIRGI*SV I\EQTPIMJ]SYVWIUYIRGIMRGPYHIWE*03;4%8,GSQQERH]SYGER SRP]GLSSWIEHIXIGXMSRQSHIXLEXMRGPYHIW*G

124 Customized applications Flow path 8LI*03;4%8,GSQQERHEPPS[W]SYXSGLERKIXLIJPS[TEXL[MXLMR XLIPMQMXEXMSRWWIXF]XLIHIXIGXMSRQSHI*SVI\EQTPIMJXLIHIXIGXMSR QSHIMWWIXXS*G]SYGERSRP]GLSSWIJPS[GIPPSVMRXLI *03; 4%8,GSQQERH =SYGERRSXYWIEZEVMEFPITEVEQIXIV[MXLXLI*03;4%8,GSQQERH [LMGLQIERWXLEX]SYGERRSXYWIXLMWGSQQERHXSGLERKIXLIJPS[ TEXLFIX[IIRG]GPIW9WI*03;4%8,SRP]MJ]SY[ERXXSGLERKIXLI JPS[TEXL[MXLMREG]GPI9WIEZEVMEFPIHIXIGXMSRQSHIMJ]SY[ERXXS GLERKIXLIJPS[TEXLFIX[IIRG]GPIW Flow cell names %TEVXJVSQXLI()8)'8-32GSQQERH]SYGERVIJIVXSJPS[GIPPWMR *03;4%8,GSQQERHWXSHMVIGXXLIJPS[ERHMR -*8,)2GSQQERHWXS WIXEGSRHMXMSRFEWIHSRXLIVIWTSRWIMREWTIGMJMIHJPS[GIPP -J]SYLEZIYWIHESRISVX[SGLERRIPHIXIGXMSRQSHISVMJXLI HIXIGXMSRQSHIMWRSXWTIGMJMIH]SYGERGLSSWIXLIPSKMGEPJPS[GIPP REQIWSampleReferenceERHSubtractedJSV*03;4%8,ERH-*8,)2 GSQQERHW8LIXEFPIFIPS[PMWXWXLIGSVVIWTSRHIRGIFIX[IIRPSKMGEP JPS[GIPPREQIWERHEGXYEPJPS[GIPPWJSVXLIHMJJIVIRXHIXIGXMSRQSHIW (IXIGXMSRQSHI Sample Reference Subtracted *G *G RE RE *G *G RE RE *G *G RE RE *G *G RE RE *G6IJ *G *G *G *G6IJ *G *G *G *G *G RE RE *G *G RE RE *G6IJ *G *G *G *G6IJ *G *G *G 8LMWSTXMSRMWTEVXMGYPEVP]YWIJYPJSV-*8,)2GSQQERHWERHEPPS[W ]SYXSWIXEGSRHMXMSRFEWIHSREVIWTSRWI[MXLSYXORS[MRKMR EHZERGI[LMGLJPS[GIPPMWMRYWIWII7IGXMSRJSVI\EQTPIW

125 Customized applications 6.5 Using keywords /I][SVHWKIRIVEXIYWIVHIJMRIHGSPYQRWMRXLIVITSVXTSMRXXEFPIJSV XLIVYR[MXLXLIOI][SVHREQIEWGSPYQRLIEHMRK8LIREQIMW HIJMRIHEWEZEVMEFPIMRXLIGYWXSQM^IHWIUYIRGI8LIGSPYQRGSRXIRX MWXLIZEVMEFPIZEPYIJSVIEGLG]GPIEWIRXIVIHMRXLICyclesXEFPI -J]SYEHHOI][SVHWEJXIVIRXIVMRKXLISXLIVGSQQERHWMRXLI WIUYIRGIXLIKeyword valuejmiphtviwirxwetypphs[rpmwxsjepp ZEVMEFPIWYWIHMRXLIGSQQERHWQEOMRKMXIEW]XSWIPIGXXLI ETTVSTVMEXIZEVMEFPIREQI8LI/)=;36(GSQQERHMXWIPJMWLS[IZIV EP[E]WTPEGIHEXXLIFIKMRRMRKSJXLIWIUYIRGI

126 Customized applications 6.6 Using empty mixing vials %TTPMGEXMSRWXLEXYWIXLI86%27*)6ERH1-<GSQQERHWXSTVITEVI WSPYXMSRWMRXLIEYXSWEQTPIVZMEPWKIRIVEPP]VIUYMVIERI[ZMEPJSV IZIV]G]GPIXSEZSMHGEVV]SZIVIJJIGXWFIX[IIRG]GPIW8LIHMEPSKYI FS\JSV86%27*)6EYXSQEXMGEPP]LERHPIWXLMWVIUYMVIQIRX 8LIToTSWMXMSRKIRIVEXIWERI[IQTX]ZMEPJSVIEGLG]GPIMRXLIVYR MRXLMWI\EQTPIJSVTVITEVMRKJVIWL2,7)('JSVIEGLG]GPI8LI TSWMXMSRREQIJSVXLIToTSWMXMSRGERFIEHHVIWWIHJSVQM\MRK MRNIGXMRKIXGFYXHSIWRSXETTIEVMRXLICyclesXEFPI)QTX]ZMEP TSWMXMSRWEVIEWWMKRIHEYXSQEXMGEPP]JSVIEGLG]GPIERHEVIWLS[RMR KVIIRMRXLIGSQQERHWIUYIRGI8LIWIUYIRGIFIPS[WLS[WLS[ 86%27*)61-<ERH-2.)'8GSQQERHWQE]FIYWIHXSTVITEVIERH MRNIGX2,7)(' 2SXI 8LIJMVWXSGGYVVIRGISJXLITSWMXMSRREQIJSVEToTSWMXMSRMR XLIWIUYIRGIQYWXFIMRE86%27*)6GSQQERHWMRGIXLI TSWMXMSRVITVIWIRXWERMRMXMEPP]IQTX]ZMEP8LI[M^EVH[MPPRSX EGGITXYWEKIMRER]IEVPMIVGSQQERHMRXLIWIUYIRGI

127 Customized applications 6.7 Using MIX 8LI1-<GSQQERHQM\IWXLIWSPYXMSRMREWTIGMJMIHVEGOTSWMXMSRWII 7IGXMSR -J]SYGLIGOXLIAutomixSTXMSRXLI[M^EVH[MPPHIXIVQMRIXLI STXMQYQQM\MRKZSPYQIEGGSVHMRKXSXLIZSPYQISJPMUYMHGYVVIRXP]MR XLIZMEP;MXLXLMWSTXMSRYRGLIGOIH]SYWTIGMJ]XLIQM\MRKZSPYQIMR PEWEKIRIVEPKYMHIPMRIXLMWWLSYPHFISJXLIZSPYQISJPMUYMH MRXLIZMEP 6.8 Using report points 6IVTSVXTSMRXWEVIWIXEXEJM\IHXMQIFIJSVISVEJXIVSXLIVGSQQERHW 8SWIXEVITSVXTSMRXWIPIGXXLIGSQQERHXS[LMGLXLIVITSVXTSMRX VIJIVWXLIRGPMGOSRXLIReport PointFYXXSRSRXLIXSSPFEV =SYGEREPWSWIXVITSVXTSMRXWEWMWSPEXIHGSQQERHW[MXLMRER -*8,)2 GSRWXVYGXMSRWII7IGXMSR8SHSXLMWIRXIVER -*8,)2GSQQERH YWMRKXLIGSQQERHFPSGOEPXIVREXMZIWIPIGXXLI)2(-*8,)2GSQQERH ERHMRWIVXEVITSVXTSMRX

128 Customized applications 6.9 Using conditional commands 'SRHMXMSREPGSQQERHWEPPS[]SYXSGSRXVSPXLIWIUYIRGISJ GSQQERHWXLEXEVIEGXYEPP]I\IGYXIHHITIRHMRKSRXLISYXGSQISJXLI EREP]WMW8LIFEWMGWXVYGXYVISJEGSRHMXMSREPGSQQERHMWEWJSPPS[W -*GSRHMXMSR8,)2 GSRWIUYIRGI )2(-* GSQQERHW -JXLIGSRHMXMSRMWJYPJMPPIHXLIGSQQERHWVITVIWIRXIHF] lgsrwiuyirgizevii\igyxihjspps[ihf]xligsqqerhwejxivxli )2(-*PMRI-JXLIGSRHMXMSRMWRSXJYPJMPPIHI\IGYXMSRGSRXMRYIWHMVIGXP] [MXLXLIGSQQERHWXLEXJSPPS[)2(-* 8LIGSRHMXMSRMWSRISJXLIJSPPS[MRK RelResp 6IPEXMZIVIWTSRWIJSVEWTIGMJMIHVITSVXTSMRXSREWTIGMJMIH WIRWSVKVEQGYVZI8LIVITSVXTSMRXQYWXFIE6)74327)WIX FIJSVIXLIGSRHMXMSREPGSQQERHMWMRWIVXIHERHQYWXFI TVIGIHIHF]E&%7)0-2)GSQQERHJVSQ[LMGLXSGEPGYPEXIE VIPEXMZIVIWTSRWI AbsResp %FWSPYXIVIWTSRWIJSVEWTIGMJMIHVITSVXTSMRXSREWTIGMJMIH WIRWSVKVEQGYVZI8LIVITSVXTSMRXQYWXFIWIXFIJSVIXLI GSRHMXMSREPGSQQERHMWMRWIVXIH Slope Variable 7PSTIJSVEWTIGMJMIHVITSVXTSMRXSREWTIGMJMIHWIRWSVKVEQ GYVZI8LIVITSVXTSMRXQYWXFIWIXFIJSVIXLIGSRHMXMSREP GSQQERHMWMRWIVXIH 8LIZEPYIJSVEWTIGMJMIHZEVMEFPIDetectionERHWSPYXMSR ZEVMEFPIWGERRSXFIYWIHMRGSRHMXMSREPGSQQERHW8LI ZEVMEFPIQYWXFIHIJMRIHFIJSVIXLIGSRHMXMSREPGSQQERHMW MRWIVXIH -JVIUYMVIHEREHHMXMSREPGSRHMXMSRGERFIEHHIH[MXLXLIPSKMGEP STIVEXSVANDFSXLGSRHMXMSRWQYWXFIJYPJMPPIHSVORSRP]SRI GSRHMXMSRRIIHWXSFIJYPJMPPIH

129 Customized applications 8LIGSRWIUYIRGIQE]FISRISJXLIJSPPS[MRK Exit cycle Exit method Command block Command block ELSE Command block )RHXLIGYVVIRXG]GPIERHGSRXMRYI[MXLXLIRI\X G]GPI )RHXLIVYRMKRSVMRKER]G]GPIWXLEXQE]RSXLEZI FIIRTIVJSVQIH %YWIVHIJMRIHWIUYIRGISJGSQQERHW 8[SGSQQERHWIUYIRGIWWITEVEXIHF])07)-RXLMW GSRWXVYGXMSRXLIJMVWXWIUYIRGIMWI\IGYXIHMJXLI GSRHMXMSRMWJYPJMPPIHERHXLIWIGSRHMJXLIGSRHMXMSR MWRSXJYPJMPPIH-RIMXLIVGEWII\IGYXMSRXLIR GSRXMRYIWEJXIVXLI)2(-*PMRI )HMXMRKHIPIXMRKERHQSZMRKGSRHMXMSREPGSQQERHW &IGEYWISJXLIPSKMGEPGSQTPI\MX]XLEXGERFIMRXVSHYGIHF]XLIYWISJ GSRHMXMSREPGSQQERHWXLIVIEVIWSQIVIWXVMGXMSRWSRLS[XLIWI GSQQERHWGERFILERHPIHMRXLIGYWXSQM^IHGSQQERHWIUYIRGI =SYGERRSXEHHGSRHMXMSREPGSQQERHWWSXLEXEPVIEH]I\MWXMRK GSQQERHWEVIHMVIGXP]MRGPYHIHMRXLIGSRHMXMSREPFPSGO8STPEGI I\MWXMRKGSQQERHWMREGSRHMXMSREPFPSGOEHHXLIGSRHMXMSREP GSQQERHJMVWXERHXLIRHVEKXLISXLIVGSQQERHWMRXSXLIFPSGO =SYGERRSXQSZIEGSRHMXMSREPGSQQERH=SYGERLS[IZIVHVEK GSQQERHWSYXSJEGSRHMXMSREPGSQQERH3RI[E]XSIJJIGXEQSZI MWXSIRXIVERI[GSRHMXMSREPGSQQERHMRXLIVIUYMVIHRI[TSWMXMSR ERHHVEKER]GSQQERHWJVSQXLISPHGSRHMXMSRXSXLIRI[SRI =SYGERRSXHIPIXIEGSRHMXMSREPGSQQERH[MXLSYXHIPIXMRKXLI GSQQERHFPSGOWMRGPYHIHMRMX8SHIPIXIEGSRHMXMSREPGSQQERH FYXVIXEMRXLIMRGPYHIHGSQQERHWJMVWXHVEKXLIGSQQERHWSYXWMHI XLIGSRHMXMSRERHXLIRHIPIXIXLIGSRHMXMSR 3RGI]SYLEZIIRXIVIHEGSRHMXMSR]SYGERRSXGLERKIXLIX]TISJ GSRWIUYIRGI-J]SY[ERXXSGLERKIXLIGSRWIUYIRGIIRXIVERI[ GSRHMXMSREPWXEXIQIRXERHHIPIXIXLISPHSRI6IQIQFIVXSHVEKER] GSQQERHW]SY[ERXXSOIITSYXSJXLISPHGSRHMXMSRFIJSVI]SY HIPIXIMX

130 Customized applications 6.10 Using WAIT 8LI;%-8GSQQERHMRWIVXWEHIPE]TIVMSHFIX[IIRGSQQERHWMRXLI WIUYIRGI8LIHIPE]WXEVXW[LIRXLIGSQQERHTVIGIHMRKXLI;%-8 MRGPYHMRKWIXXMRKER]VITSVXTSMRXWEWWSGMEXIH[MXLXLIGSQQERHMW GSQTPIXIHERHIRHW[LIRXLIRI\XGSQQERHI\GPYHMRKER]VITSVX TSMRXWXLEXEVIWIXFIJSVIXLIGSQQERHMWMRMXMEXIH*SV-2.)'8 STIVEXMSRWXLIWXEVXSJXLIGSQQERHMWGSYRXIHEWXLIXMQI[LIRXLI STIVEXMSREGXYEPP]FIKMRWI\GPYHMRKER]TVITEVEXMSRXMQIXLEXQMKLX FIRIIHIH2SXIXLEX;%-8GSQQERHMQQIHMEXIP]TVIGIHMRKER-2.)'8 GSQQERH[MPPLEZIRSIJJIGXMJXLIHIPE]TIVMSHMWWLSVXIVXLERXLI EYXSWEQTPIVTVITEVEXMSRXMQIWII7IGXMSR 8LMWMWMPPYWXVEXIHMRXLIHMEKVEQFIPS[ Diagram showing the time relationships of WAIT to other commands, using a WAIT between the end of one injection and the beginning of the next for illustration purposes. The arrow at the left indicates the duration of the delay specified in the WAIT command Customized application examples 8LMWWIGXMSRWLS[WWSQIWMQTPII\EQTPIWSJLS[ETTPMGEXMSRWGER ETTIEVMRXLIGYWXSQM^IHETTPMGEXMSR[M^EVH8LII\EQTPIWWLSYPHFI WIIREWMPPYWXVEXMSRWSJXLITVMRGMTPIWSJGYWXSQM^IHWIUYIRGIWVEXLIV XLEREW[SVOMRKI\TIVMQIRXEPXIQTPEXIW Inhibition assay 8LMWI\EQTPIMPPYWXVEXIWERMRLMFMXMSREWWE]JSVGSRGIRXVEXMSR HIXIVQMREXMSRSJEPS[QSPIGYPEV[IMKLXEREP]XI8LIWIUYIRGIQM\IW WEQTPI[MXLEORS[REQSYRXSJERXMFSH]ERHMRNIGXWXLIQM\XYVISZIV EWYVJEGISR[LMGLTYVMJMIHEREP]XIMWMQQSFMPM^IH8LIVIWTSRWI

131 Customized applications QIEWYVIWXLIEQSYRXSJJVIIERXMFSH]VIQEMRMRKMRXLIWEQTPIERHMW MRZIVWIP]VIPEXIHXSXLIEQSYRXSJEREP]XI 8LIWIUYIRGIHSIWRSXMRGPYHIE()8)'8-32GSQQERHWSXLEXXLI HIXIGXMSRQSHIERHJPS[TEXLEVIWIXEXXLIFIKMRRMRKSJIEGLVYR 8LMWEPPS[WXLIWIUYIRGIXSFIYWIH[MXLEJVIIGLSMGISJJPS[GIPP 8LIOI][SVHWSampleERHConcIRWYVIXLEXVIWTSRWIWVIGSVHIHMRXLI VITSVXTSMRXXEFPIEVIEHIUYEXIP]MHIRXMJMIHJSVPEXIVIZEPYEXMSR8LI ZEVMEFPISampleMRXLI/)=;36(GSQQERHMWXLIWEQIEWXLEXMRXLI -2.)'8GSQQERHXLIZEVMEFPIConcMWYWIHSRP]EWEOI][SVHXS VIGSVHGSRGIRXVEXMSRWMRXLIVITSVXTSMRXXEFPI Conditional regeneration 8LMWI\EQTPIMPPYWXVEXIWXLIYWISJER-*8,)2GSQQERHXSVIKIRIVEXI XLIWYVJEGISRP]MJEREP]XILEWFSYRH8LMWOMRHSJGSRWXVYGXMSRGER GSRWMHIVEFP]VIHYGIXLIXSXEPXMQIVIUYMVIHJSVEWIVMIWSJEREP]WIW 2SXIXLEXXLIWIUYIRGIHSIWRSXGSRXEMRE()8)'8-32GSQQERH EPPS[MRKJVIIGLSMGISJJPS[GIPPEXXLIFIKMRRMRKSJXLIVYR 'SVVIWTSRHMRKP]XLI-*8,)2GSQQERHVIJIVWXSXLIVIPEXMZIVIWTSRWI MRJPS[GIPPSample[LMGL[MPPQETXSXLIETTVSTVMEXIJPS[GIPP EGGSVHMRKXSXLIHIXIGXMSRQSHIYWIHMRER]KMZIRVYRWII7IGXMSR

132 Customized applications Conditional control experiments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

133 Customized applications

134 Maintenance. Maintenance *SV]SYVS[RWEJIX] %P[E]W[IEVKPSZIWERHTVSXIGXMZIGPSXLMRKHYVMRKSTIVEXMSRERH QEMRXIRERGI 9WIVIUYMVIHWEJIX]IUYMTQIRX[LIRLERHPMRKLE^EVHSYW WYFWXERGIW -JXLIMRWXVYQIRXMWGSRXEQMREXIH[MXLFMSLE^EVHWTPIEWIGSRXEGX ]SYVPSGEP&MEGSVI7IVZMGI7YTTSVXJSVJYVXLIVMRJSVQEXMSREFSYX HIGSRXEQMREXMSRTVSGIHYVIW 6IKYPEVQEMRXIRERGITVSGIHYVIWEVIHIWGVMFIHMRXLMWGLETXIV-JQSVI I\XIRWMZIQEMRXIRERGISVWIVZMGISJXLIMRWXVYQIRXMWVIUYMVIHTPIEWI GSRXEGX]SYVPSGEP&MEGSVI7IVZMGI7YTTSVX 7.1 Regular maintenance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

135 Maintenance At the end of the day %XXLIIRHSJEHE]kW[SVOVYRXLI[SVOMRKXSSPStandbyXSOIIT FYJJIVJPS[MRKEXEPS[VEXIXLVSYKLXLI-*'(SRSXPIEZIXLI MRWXVYQIRXSZIVRMKLXSVPSRKIV[MXLWXEXMSREV]FYJJIVMRXLIJPS[GIPPW 8LIStandbyXSSPQE]FIETTIRHIHXSEQIXLSHJSVEYXSQEXMG STIVEXMSR[LIRXLIQIXLSHJMRMWLIW -J]SYMRXIRHXSVYRStandbyJSVERI\XIRHIHTIVMSHQEOIWYVIXLEX XLIVIMWWYJJMGMIRXFYJJIVMRXLIFYJJIVTEGOStandby[MPPVYRJSVYTXS HE]W[LMGLVIUYMVIWEFSYXQPFYJJIV-JXLIW]WXIQMWXSFIPIJX JSVQSVIXLERHE]WVYRCloseSVShutdown MRWXIEHSJStandbyWII 7IGXMSR 6ITPEGIXLIVYRRMRKFYJJIV[MXL[EXIVMJ]SYVFYJJIVXIRHWXS TVIGMTMXEXISRWXERHMRK-RXLMWGSRXI\XVIQIQFIVXLEXXLIEQFMIRX XIQTIVEXYVIQE]HVSTHYVMRKXLIRMKLX System cleaning 6IQSZMRKEHWSVFIHTVSXIMRW -RGPYWMSRSJ7YVJEGXERX4XSWEQTPIERH VYRRMRKFYJJIVMW VIGSQQIRHIHXSQMRMQM^IRSRWTIGMJMGEHWSVTXMSRSJQEXIVMEPXSXLI TYQTXYFMRKERH-*'GLERRIPW6YRXLI[SVOMRKXSSPDesorbXSGPIER XLIXYFMRKERH-*'GLERRIPWEXPIEWXSRGIE[IIOSVQSVISJXIRMJ]SY HSRSXYWIWYVJEGXERXMRWEQTPIERHVYRRMRKFYJJIV *SVWSQIETTPMGEXMSRWEGSEXMRKSJTVSXIMRSRXLIXYFMRKERH-*' GLERRIP[EPPWLIPTWXSKMZIVITVSHYGMFPIVIWYPXWERHXLIJMVWXJI[VYRW EJXIVDesorbQE]FIYRVIPMEFPI-J]SYSFWIVZIXLMWFILEZMSYVVYR Desorb[LIR]SYGLERKI]SYVTVSXIMRW]WXIQVEXLIVXLERSRGIE[IIO 8LIVIGSQQIRHIHGPIERMRKEKIRXJSVDesorbMW&-%HIWSVF7SPYXMSR 7(7JVSQ&-%QEMRXIRERGI/MX-J]SY[MWLXSYWISXLIV GPIERMRKEKIRXWGSRWYPXXLIGLIQMGEPVIWMWXERGIXEFPIWMR%TTIRHM\' 6IQSZMRKWEPXVIWMHYIW 1ER]FYJJIVWMRGPYHMRK,&7YWITL]WMSPSKMGEPWEPXGSRGIRXVEXMSRW X]TMGEPP]Q12E'P8STVIZIRXWEPXHITSWMXWEGGYQYPEXMRKMRXLI PMUYMHLERHPMRKW]WXIQ[EWLXLIVIGSZIV]FPSGOERHXLIRIIHPI[MXL HIMSRM^IH[EXIVSRGIE[IIO;MTIXLISYXWMHISJXLIRIIHPI[MXLE [IXXMWWYIERHVYRCommand:Wash[MXLWIPIGXMSRNeedleXS[EWLXLI MRWMHI 4VIZIRXMRKQMGVSFMEPKVS[XL 8STVIZIRXQMGVSFMEPKVS[XLMRXLIMRWXVYQIRXOIITELMKLWXERHEVH SJL]KMIRIMRXLIWSPYXMSRWYWIH7XSVIFYJJIVWJVS^IRSVMRE VIJVMKIVEXSVERHJMPXIVXLVSYKLE QJMPXIVFIJSVIYWI

136 Maintenance 6YRXLI[SVOMRKXSSPSanitizeEXPIEWXSRGIEQSRXLXSVIQSZI QMGVSFMEPGSRXEQMRERXW*SVFIWXVIWYPXWVYRDesorbFIJSVISanitize 2SXI 8LIWIRWSVGLMTWYVJEGIQE]FIHEQEKIHF]XLIDesorbERH SanitizeTVSGIHYVIW=SYQE]OIITEWTIGMEPGLMTJSVVYRRMRK XLIWIXSSPWFYX]SYWLSYPHRSXYWIMXJSVER]SXLIVTYVTSWI %WGLIHYPIV[MPPVIQMRH]SYXLEXMXMWXMQIXSVYRXLI[SVOMRKXSSPW SanitizeERHSVDesorb[LIRQSVIXLERXLIVIGSQQIRHIHMRXIVZEPLEW IPETWIHWMRGIXLIVIWTIGXMZIQEMRXIRERGIXSSP[EWPEWXYWIH=SYQE] GLSSWIXSVYRXLIQEMRXIRERGIXSSPSVMKRSVIXLIVIQMRHIVERHVYRXLI XSSPEXEPEXIVHEXI-J]SYMKRSVIXLIVIQMRHIVMX[MPPFIVITIEXIHEJXIV SRI[IIO Other routines -RWTIGXEPPZMWMFPIPMUYMHGSRRIGXMSRWTYQTW]VMRKIWERHZEPZIW MRNIGXMSRTSVXERHGSRRIGXSVFPSGOJSVPIEOWIEGLHE]-JER]PIEOWEVI HMWGSZIVIHGPIERERHXMKLXIRXLIGSRRIGXMSRWSVMJRIGIWWEV]VITPEGI WIEPW -JHIXIVMSVEXMSRMRTIVJSVQERGIMWWYWTIGXIHVYRTest Tools:System checkxsgligoxliw]wxiqtivjsvqergiewsyxpmrihmr7igxmsr

137 Maintenance 7.2 User tools %PPYWIVXSSPWEVIEZEMPEFPIJVSQXLIToolsQIRY8LI]EVISVKERM^IH YRHIVXLVIIGSQQERHWJSV[SVOMRKXSSPWXIWXXSSPWERHWIVZMGIXSSPW VIWTIGXMZIP]8SVYREYWIVXSSPGLSSWIXLIETTVSTVMEXIGSQQERH WIPIGXXLIYWIVXSSPERHJSPPS[XLIMRWXVYGXMSRWSRXLIWGVIIR 8LIXSSPGLSMGIHMEPSKYIFS\MRGPYHIWWLSVXMRJSVQEXMSRXSMHIRXMJ]XLI LMKLPMKLXIHXSSP8LIHEXISR[LMGLXLIWIPIGXIHXSSP[EWPEWXVYRMW WLS[REXXLIFSXXSQSJXLIHMEPSKYIFS\ Working tools 8LI[SVOMRKXSSPWEVIEWIXSJVSYXMRIWJSVWXEVXMRKYTERHWLYXXMRK HS[RXLIW]WXIQERHJSVWTIGMEPGPIERMRKTVSGIHYVIW Close %TVSGIHYVIJSVWLYXXMRKHS[RXLIW]WXIQJSVYTXSHE]W8LI TVSGIHYVIJPYWLIWXLI-*'ERHJPS[GIPPW[MXLPMUYMHJVSQEJM\IH EYXSWEQTPIVTSWMXMSRVJXLIRIQTXMIWXLI-*'SJPMUYMH %TTVS\MQEXIP]QPPMUYMHMWVIUYMVIH(IMSRM^IHSVHMWXMPPIH[EXIV MWVIGSQQIRHIHEWJPYWLMRKPMUYMH8SXEPVYRXMQIMWEFSYXQMRYXIW 6IUYMVIHWSPYXMSRW (IMSRM^IH[EXIV

138 Maintenance Desorb 6IQSZIWEHWSVFIHTVSXIMRWJVSQXLIEYXSWEQTPIVERHXLI-*'F] JPYWLMRK[MXL7(7%WEKIRIVEPVIGSQQIRHEXMSRVYRXLMWTVSGIHYVI SRGIE[IIO*SVWSQIETTPMGEXMSRWEGSEXMRKSJTVSXIMRSRXLIXYFMRK ERH-*'GLERRIP[EPPWLIPTWXSKMZIVITVSHYGMFPIVIWYPXWERHXLIJMVWX JI[VYRWEJXIVDesorbQE]FIYRVIPMEFPI-J]SYSFWIVZIXLMW FILEZMSYVVYRDesorb[LIR]SYGLERKI]SYVTVSXIMRW]WXIQVEXLIV XLERSRGIE[IIO8SXEPVYRXMQIMWEFSYXQMRYXIW %WGLIHYPIV[MPPMWWYIEVIQMRHIVEXXLIFIKMRRMRKSJXLI[IIOMJQSVI XLERSRI[IIOLEWIPETWIHWMRGIDesorb[EWPEWXVYR-JXLIW]WXIQMW FYW]SVW[MXGLIHSJJEXXLIWGLIHYPIHVIQMRHIVXMQIXLIVIQMRHIV[MPP FIHMWTPE]IHEXXLIRI\XTSWWMFPISTTSVXYRMX][LIRXLIW]WXIQMW GSRRIGXIHXSXLI4'ERHMWRSXFYW] 2SXIW 8LIWIRWSVGLMTWYVJEGIQE]FIHEQEKIHF]XLIDesorb TVSGIHYVI=SYQE]OIITEWTIGMEPGLMTJSVVYRRMRKDesorb FYX]SYWLSYPHRSXYWIMXJSVER]SXLIVTYVTSWI6MRWIXLIGLMT EJXIVIEGLYWEKIERHQEOIWYVIMXMWGPIERFIJSVIVIMRWIVXMRKMX WII&-%XIGLRSPSK]2SXI &-%HIWSVFWSPYXMSRWEVIEZEMPEFPIJVSQ&MEGSVI%&EWTEVXSJXLI &-%QEMRXIRERGI/MX(SRSXVYRXLITVSGIHYVIFIPS[ 'WMRGIXLMW WSPYXMSRGSRXEMRW7(7[LMGLQE]TVIGMTMXEXIEXPS[IVXIQTIVEXYVIW 6IUYMVIHWSPYXMSRW &-%HIWSVFWSPYXMSR &-%HIWSVFWSPYXMSR Flush *PYWLIWXLI-*'ERHJPS[GIPPW[MXLFYJJIV9WIXLMWTVSGIHYVIJSVJEWX GPIERMRKSJXLI-*'ERHJPS[GIPPW8SXEPVYRXMQIMWEFSYX QMRYXIWFlushQE]FIVYRFIX[IIRG]GPIWMREQIXLSHQIXLSH GSQQERHFLUSH 6IUYMVIHWSPYXMSRW 6YRRMRKFYJJIV Normalize 2SVQEPM^IWXLIVIWSRERGIWMKREPMREPPJPS[GIPPWXSGSQTIRWEXIJSV WQEPPHMJJIVIRGIWFIX[IIRMRHMZMHYEPWIRWSVGLMTW9WIXLMWTVSGIHYVI E[LIRXLISTXSMRXIVJEGIMWGLERKIHF[LIRXLIXIQTIVEXYVIMW GLERKIHERHG[LIRERI[WIRWSVGLMTMWHSGOIHERH QE\MQYQ WIRWMXMZMX]MWVIUYMVIH8SXEPVYRXMQIMWEFSYXQMRYXIW Normalize QE]FIVYRFIX[IIRG]GPIWMREQIXLSHQIXLSHGSQQERHNORMALIZE [MXLETEVEQIXIVJSVXLIRSVQEPM^EXMSRWSPYXMSRVEGOTSWMXMSR 2SXI -JNormalizeMWMRGPYHIHMREQIXLSHERHXLIRSVQEPM^EXMSR JEMPWXLIQIXLSH[MPPXIVQMREXI

139 Maintenance 8LIGSRGIRXVEXMSRSJKP]GIVSPYWIHJSVRSVQEPM^EXMSRMWGVMXMGEP &-%RSVQEPM^MRKWSPYXMSRMWEZEMPEFPIJVSQ&MEGSVI%&EWTEVXSJXLI &-%QEMRXIRERGI/MX-J]SYGERRSXTIVJSVQEWYGGIWWJYP RSVQEPM^EXMSRXLISTXSMRXIVJEGISVWIRWSVGLMTQE]FIHIJIGXMZI 6IUYMVIHWSPYXMSRW &-%RSVQEPM^MRKWSPYXMSR Prime 4VMQIWXLIPMUYMHW]WXIQF]JPYWLMRKTYQTW-*'ERHEYXSWEQTPIV[MXL FYJJIV9WIXLMWTVSGIHYVIEXWXEVXYTERH[LIRXLIFYJJIVMWGLERKIH ERHEPWSXSVIQSZIWQEPPEMVFYFFPIW[LMGLQE]EGGYQYPEXIHYVMRK STIVEXMSREXPS[JPS[VEXIWJSVPSRKTIVMSHW8SXEPVYRXMQIMWEFSYX QMRYXIW PrimeQE]FIVYRFIX[IIRG]GPIWMREQIXLSHQIXLSH GSQQERHPRIME 6IUYMVIHWSPYXMSRW 6YRRMRKFYJJIV Rinse ;EWLIWXLIMRNIGXMSRTSVXXLSVSYKLP]XLIRVMRWIW XLI-*'ERHJPS[ GIPPW[MXLFYJJIV9WIXLMWTVSGIHYVIXSGPIERXLI-*'JPS[GIPPWERH VIGSZIV]GYT8SXEPVYRXMQIMWEFSYXQMRYXIW RinseQE]FIVYR FIX[IIRG]GPIWMREQIXLSHQIXLSHGSQQERHRINSE 6IUYMVIHWSPYXMSRW 6YRRMRKFYJJIV Sanitize 'PIERWTYQTW-*'ERHEYXSWEQTPIVJVSQQMGVSSVKERMWQW6YRXLMW TVSGIHYVISRGIEQSRXL*SVFIWXVIWYPXWVYRDesorbFIJSVILERH 8SXEPVYRXMQIMWEFSYXQMRYXIW %WGLIHYPIV[MPPMWWYIEVIQMRHIVEXXLIFIKMRRMRKSJXLI[IIOMJQSVI XLERJSYV[IIOWLEZIIPETWIHWMRGISanitize[EWPEWXVYR-JXLIW]WXIQ MWFYW]SVW[MXGLIHSJJEXXLIWGLIHYPIHVIQMRHIVXMQIXLIVIQMRHIV [MPPFIHMWTPE]IHEXXLIRI\XTSWWMFPISTTSVXYRMX][LIRXLIW]WXIQMW GSRRIGXIHXSXLI4'ERHMWRSXFYW] 2SXI 8LIWIRWSVGLMTWYVJEGIQE]FIHEQEKIHF]XLISanitize TVSGIWW=SYQE]OIITEWTIGMEPGLMTJSVVYRRMRKSanitizeFYX ]SYWLSYPHRSXYWIMXJSVER]SXLIVTYVTSWI6MRWIXLIGLMT EJXIVIEGLYWEKIERHQEOIWYVIMXMWGPIERFIJSVIVIMRWIVXMRKMX WII&-%XIGLRSPSK]2SXI 6IUYMVIHWSPYXMSRW &-%HMWMRJIGXERXWSPYXMSR &-%TVIWIVZEXMSRWSPYXMSR SV7YVJEGXERX4MRHIMSRM^IH[EXIV

140 Maintenance Shutdown %TVSGIHYVIJSVWLYXXMRKHS[RXLIW]WXIQJSVQSVIXLERHE]W8LI TVSGIHYVIJPYWLIWXLI-*'ERHJPS[GIPPW[MXL[EXIVIQTXMIWXLI-*' SJPMUYMHERHXLIRJPYWLIWXLITYQTWERHXYFMRK[MXLIXLERSP 8SXEPVYRXMQIMWEFSYXQMRYXIW %28 *SPPS[XLIMRWXVYGXMSRWSRXLIWGVIIRGEVIJYPP]8LI-*'HSIWRSX [MXLWXERHI\TSWYVIXSIXLERSPERHQYWXFIHMWGSRRIGXIH FIJSVIXLITYQTWEVIJPYWLIH[MXLIXLERSP 6IUYMVIHWSPYXMSRW (IMSRM^IH[EXIV ZZIXLERSP Standby 8LMWTVSGIHYVIQEMRXEMRWEPS[JPS[SJFYJJIVXLVSYKLXLI-*'ERH JPS[GIPPWJSVYTXSHE]W&YJJIVGSRWYQTXMSRMWE TTVS\MQEXIP] QPHE]8LITVSGIHYVIYWIWFSXLGSRXMRYSYWJPS[ERHEYXSWEQTPIV TYQTWXSIRWYVIEGPIERW]WXIQStandbyQE]FIETTIRHIHXSE QIXLSHQIXLSHGSQQERHAPPEND STANDBY 6IUYMVIHWSPYXMSRW 6YRRMRKFYJJIV 2SXI 8LIVIEVIXLVIIXSSPWJSVJPYWLMRKXLIW]WXIQ[MXLFYJJIV8LI IWWIRXMEPHMJJIVIRGIWFIX[IIRXLIWIEVIWYQQEVM^IHFIPS[ 8SSP (IWGVMTXMSR 8MQI Flush *PYWLIWXLIGSRXMRYSYWJPS[TYQTERH-*' FVMIJP]EXEQSHIVEXIJPS[VEXI9WIFlushMJ ]SY[ERXXSGPIERXLI-*'VETMHP]ERHHS RSXLEZITVSFPIQW[MXLEMVFYFFPIW QMR Rinse *PYWLIWTYQTWERH-*'GLERRIPWEXELMKL JPS[VEXIERH[EWLIWXLIVIGSZIV]GYT8LI LMKLJPS[VEXILIPTWXSVIQSZIEMVFYFFPIW XLEXQE]FIXVETTIHMRXLIW]WXIQ9WI RinseVIKYPEVP]MJ]SYVYREXEPS[JPS[VEXI SVIPIZEXIHXIQTIVEXYVI[LIVIEMVFYFFPIW QE]FIETVSFPIQ QMR Prime ;EWLIWXLITYQTWEYXSWEQTPIVXYFMRKERH -*'[MXLFYJJIV9WIPrime[LIR]SY[ERX XSGLERKIXLIFYJJIVMRXLITYQTW QMR

141 Maintenance Test tools 8LIXIWXXSSPWEVIEWIXSJVSYXMRIWJSVXIWXMRKXLIW]WXIQTIVJSVQERGI Signal quality check 8LMWTVSGIHYVIGLIGOWXLIWMKREPJVSQXLIJPS[GIPPW8LIXIWXJMRMWLIW F]HMWTPE]MRKXLIJPS[GIPPWXEXYW -JWMKREPUYEPMX]MWEGGITXEFPIXLIWXEXYWMWl3/z-JXLIWXEXYWMWl3YX SJH]REQMGVERKIzSVl4SSVWMKREPUYEPMX]zXLIVIQE]FIEMVTEVXMGPIW SVELMKLVIJVEGXMZIMRHI\WSPYXMSRIKKP]GIVSPMRXLI-*'GLERRIPW SVXLISTXSMRXIVJEGIQE]FIFEH -JXLIWMKREPUYEPMX]MWRSXEGGITXEFPIXV]VYRRMRK Working ToolsPrime SRGIXSVYRFYJJIVXLVSYKLXLI-*'-JXLMWHSIWRXLIPTVYR Normalize SRGI-JWMKREPUYEPMX]WXMPPLEWRXMQTVSZIHGSRXEGX&MEGSVIPSGEP WIVZMGIWYTTSVX System check 8LMWTVSGIHYVITIVJSVQWEGSQTVILIRWMZIGLIGOSJW]WXIQ TIVJSVQERGIYWMRKEWXERHEVHWYGVSWIWSPYXMSR[LMGLMWTVSZMHIHMR XLI&-%QEMRXIRERGIOMXJVSQ&MEGSVI%&8SXEPVYRXMQIMWEFSYX QMRYXIW

142 Maintenance =SYWLSYPHVYRXLITVSGIHYVIEX '[MXLE7IRWSV'LMT'1 VIWIVZIHJSVXLMWTYVTSWISR[LMGLRSPMKERHLEWFIIRMQQSFMPM^IH *SVQE\MQYQVIPMEFMPMX][IVIGSQQIRHXLEX]SYYWIERI[7IRWSV 'LMT'1 6IUYMVIHWSPYXMSRW &-%XIWXWSPYXMSR,&7FYJJIV&-%'IVXMJMIH 8LIWIRWSVKVEQVIWYPXMRKJVSQW]WXIQGLIGOMWHMWTPE]IHMRERYRXMXPIH WIRWSVKVEQ[MRHS[=SYQE]IZEPYEXIXLIYRWEZIHWIRWSVKVEQ HMVIGXP]ERHSVWEZIXLIWIRWSVKVEQ[MXLFile:SaveJSVPEXIVMRWTIGXMSR System check evaluation 8LMWTVSGIHYVIKIRIVEXIWEVITSVXSRXLIWEZIHVIWYPXWSJW]WXIQGLIGO 'LSSWIXLIW]WXIQGLIGOVIWYPXJMPIMRXLIOpenHMEPSKYIFS\8LI IZEPYEXMSRTVSXSGSPTEKIWMWHMWTPE]IHSRXLIWGVIIR[MXLWGVSPPMRK JEGMPMXMIWERHGERFITVMRXIH[MXLXLIPrintFYXXSR8LITVSXSGSP MRGPYHIWMRWXVYGXMSRWJSVEWWIWWMRKXLIW]WXIQGLIGOVIWYPXW 2SXI 9RWEZIHW]WXIQGLIGOVIWYPXWGERFIIZEPYEXIH[MXLXLIWEQI TVSGIHYVIF]GLSSWMRKEvaluate[LIRXLIW]WXIQGLIGO TVSGIHYVIMWGSQTPIXI Service tools 8LIWIVZMGIXSSPWEVIEWIXSJVSYXMRIWJSVW]WXIQQEMRXIRERGI

143 Maintenance Injection port %KYMHIXSXLIQEMRXIRERGISVVITPEGIQIRXSJXLIMRNIGXMSRTSVXERH GSRRIGXSVFPSGO 2SXI 8LMWTVSGIHYVIQYWXFIJSPPS[IHF]ERIIHPITSWMXMSRMRKXIWX Needle replacement/positioning Needle replacement/positioning %KYMHIXSVITPEGIQIRXSJXLIEYXSWEQTPIVRIIHPIERHRIIHPI TSWMXMSRMRKXIWX2IIHPITSWMXMSRMRKGERFIGLIGOIH[MXLSYXRIIHPI VITPEGIQIRX Software problem report %YXMPMX]XSLIPTMRJSVQYPEXMRKETVSFPIQVITSVXMRXLIIZIRXSJ QEPJYRGXMSR-JTSWWMFPIVYRXLMWYXMPMX]SRXLI4'GSRXVSPPMRKXLI W]WXIQERHEPPS[XLIYXMPMX]XSGSPPIGXGSQTYXIVW]WXIQMRJSVQEXMSR EYXSQEXMGEPP]8LIVITSVXKIRIVEXIHF]XLMWYXMPMX]QE]FIWYFQMXXIH F]IQEMPSVTVMRXIHSYXERHJE\IHXS&MEGSVI Syringe/tip (left) %KYMHIXSVITPEGMRKXLIW]VMRKIXMTSRXLIPIJXLERHTYQT Syringe/tip (right) %KYMHIXSVITPEGMRKXLIW]VMRKIXMTSRXLIVMKLXLERHTYQT Unclogging 'PIEVWTEVXMEPP]FPSGOIH-*'JPS[GIPPWERHPSSTWF]JPYWLMRK[MXL FYJJIVEXELMKLJPS[VEXI8SXEPVYRXMQIMWEFSYXQMRYXIW UncloggingQE]FIVYRFIX[IIRG]GPIWMREQIXLSHQIXLSHGSQQERH UNCLOG 6IUYMVIHWSPYXMSRW 6YRRMRKFYJJIV

144 Troubleshooting 8. Troubleshooting -J]SYLEZITVSFPIQW[MXL&-%'36)SV[MXL]SYVVIWYPXW %WEJMVWXQIEWYVI -J]SYSFXEMRYRI\TIGXIHSV YREGGITXEFPIWIRWSVKVEQVIWYPXW -J]SYWYWTIGXEWTIGMJMG MRWXVYQIRXQEPJYRGXMSR -J]SYLEZITVSFPIQW[MXL &-%'36)'SRXVSP7SJX[EVI -J]SYGERRSXMHIRXMJ]SVGSVVIGX TVSFPIQW[MXLXLILIPTSJXLI MRJSVQEXMSRLIVI 6YRTest Tools:System checkxs ZIVMJ]W]WXIQTIVJSVQERGI 6IQIQFIVXSHSGOERYRYWIH WIRWSVGLMTFIJSVIVYRRMRKXLMW TVSGIHYVI-JXLIW]WXIQGLIGO VIWYPXWHSRSXMRHMGEXIER] MRWXVYQIRXQEPJYRGXMSRGLIGO ]SYVQIXLSHERHVIEKIRXW GEVIJYPP]JSVTSWWMFPIWSYVGIWSJ IVVSV 6IEH7IGXMSR 6IEHXLIVIPIZERXTEVXWSJ 7IGXMSR 6IEH7IGXMSR 'EPP]SYV&MEGSVIWIVZMGI VITVIWIRXEXMZI 8.1 Sensorgram problems Signal noise 2SVQEPWMKREPRSMWIPIZIPWEVI TIEOXSTIEOEX HEXEGSPPIGXMSRVEXI,^)\GIWWMZIRSMWIQE]FIGEYWIHF] EMVSVTEVXMGPIWMRXLIWEQTPISVFYJJIV(IKEWERHJMPXIVWSPYXMSRW 6YRWorking Tools:PrimeXSVIQSZIEMV[LMGLQE]EGGYQYPEXIMR XLIJPS[GIPPGLERRIPW QIGLERMGEPZMFVEXMSR8LIMRWXVYQIRXWLSYPHFITPEGIHSREJMVQ FIRGL(SRSXTPEGILIEZMP]ZMFVEXMRKIUYMTQIRXIKERYRFEPERGIH FIRGLGIRXVMJYKISRXLIWEQIFIRGLEW&-%'36) IPIGXVMGEPRSMWI(SRSXTPEGIXLIMRWXVYQIRXMREREVIEWYFNIGXXS WXVSRKIPIGXVMGEPSVQEKRIXMGJMIPHW

145 Troubleshooting Low data quality 0S[HEXEUYEPMX]MWMRHMGEXIHF]EXLMGOPMRISRXLIWIRWSVKVEQERHER IRXV]MRXLII\XIRHIHIZIRXPSKGLSSWIView:Extended Event Log 0S[HEXEUYEPMX]MWRSVQEPMJXLIWMKREPVMWIWEFSZIEFSYX69 GSVVIWTSRHMRKXSEFYPOVIJVEGXMZIMRHI\SJ"8LMWQE]FI GEYWIHF]MRNIGXMSRSJEMV[LMGLWLMJXWXLIFEWIPMRIWSXLEXXLIWMKREPMW SYXWMHIXLIH]REQMGVERKISJXLIMRWXVYQIRX0S[HEXEUYEPMX]FIPS[ 69QE]FIGEYWIHF]EKKVIKEXMSRSJQEXIVMEPSRXLIWIRWSV GLMTWYVJEGI-JPS[HEXEUYEPMX]MWVITSVXIHMRSXLIVGMVGYQWXERGIW HSGOERI[WIRWSVGLMTERHVYRWorking Tools:Normalize-JXLI TVSFPIQTIVWMWXWGEPP&MEGSVIWIVZMGI 2SXI 8LIQEVOMRKJSVPS[HEXEUYEPMX]SRXLIWIRWSVKVEQI\XIRHW JVSQXLIJMVWXPS[UYEPMX]TSMRXXSXLIJMVWXRSVQEPUYEPMX] TSMRX-JXLIVIWTSRWIGYVZIMWZIV]WXIITXLMWQE]WSQIXMQIW KMZIXLIMQTVIWWMSRXLEXXLIHEXEUYEPMX]MWPS[IZIREXRSVQEP VIWTSRWIPIZIPWWIIXLIMPPYWXVEXMSREFSZI8LIIZIRXPSK VIGSVHWWXEVXSJPS[HEXEUYEPMX]ERHXLIVIXYVRXSRSVQEPHEXE UYEPMX]GSVVIGXP] Spikes 7TMOIWMRXLIWIRWSVKVEQQE]FIGEYWIHF] WQEPPEMVFYFFPIWMRXLIJPS[GIPP8LIWIQE]ETTIEVER][LIVIMRXLI WIRWSVKVEQ XVERWMIRXQEXVM\IJJIGXWWII7IGXMSR&SVGSRJSVQEXMSREPGLERKIW MRXLIMQQSFMPM^IHPMKERH8LIWIETTIEVEXXLIFIKMRRMRKERHSVIRH SJWEQTPIMRNIGXMSR 2SXI %WEVIWYPXSJXLI[E]HEXEMWHMWTPE]IHSRXLIWGVIIRWTMOIW XLEXEVIVITVIWIRXIHF]SRP]SRIHEXETSMRXQE] HMWETTIEVEX GIVXEMR^SSQJEGXSVW8LIWEQIIJJIGXQE]SGGEWMSREPP]FI WIIRMJXLIWIRWSVKVEQMWTVMRXIHEXHVEJXUYEPMX]

146 Troubleshooting Drift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orking Tools:Prime 9WIERMRPMRIVIJIVIRGIGIPP[LIVITSWWMFPIXSGSVVIGXJSVW]WXIQEXMG HVMJXIJJIGXW Reproducible disturbances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

147 Troubleshooting Shifts 7LMJXWMRWMKREPFEWIPMRIQE]FIGEYWIHF]WQEPPEMVFYFFPIWHIZIPSTMRK MRXLIJPS[GIPPERHXLIRFVIEOMRKJVIIMRXSXLIFYJJIVJPS[7II7IGXMSR JSVEMVFYFFPIGSRWMHIVEXMSRW 1IGLERMGEPMRWXEFMPMX]GEREPWSGEYWIWLMJXWMRFEWIPMRI No response on injection 'LIGOXLIHIXIGXSVF]MRNIGXMSRSJWYGVSWISVKP]GIVSPMR[EXIVMR QYPXMGLERRIPQSHI'LSSWIView:ScaleERHWIXXLIVIWTSRWIWGEPI] E\MWXSAuto-JXLIVIMWRSVIWTSRWIGSRXEGX&MEGSVIWIVZMGI 2SXI -J]SYSFXEMRRSVIWTSRWISRMRNIGXMSRMREQIXLSHMRWMRKPI GLERRIPQSHIGLIGOXLEX]SYLEZIRSXYWIHE()8)'8-32 GSQQERHMR1%-2ERHE*03;4%8,GSQQERHMR%463+ [MXLHMJJIVIRXJPS[GIPPRYQFIVW7II7IGXMSR) Poor reproducibility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orking Tools:PrimeERHEPPS[FYJJIVJPS[JSV QMRYXIWFIJSVIWXEVXMRKXLIJMVWXG]GPI-JXLMWHSIWRSXLIPTVYRE JI[HYQQ]G]GPIW[MXLWEQTPIMRNIGXMSRERHVIKIRIVEXMSRXS GSRHMXMSRXLIWYVJEGI -JXLIZEVMEFMPMX]MWVERHSQXLIVIQE]FIEJEYPXMRXLIPMUYMH LERHPMRKW]WXIQWII7IGXMSR6ITIEXIHMRNIGXMSRWTIVJSVQIH YRHIVQERYEPGSRXVSPQE]WLS[QSVIZEVMEXMSRXLERVITIEXIH

148 Troubleshooting QIXLSHG]GPIWYRPIWWGEVIMWXEOIRXSYWII\EGXP]XLIWEQI GSRHMXMSRWJSVIEGLMRNIGXMSR Sample dispersion 7II%TTIRHM\&JSVEHMWGYWWMSRSJWEQTPIHMWTIVWMSREXXLIFIKMRRMRK ERHIRHSJMRNIGXMSRW2SVQEPHMWTIVWMSRPIZIPWEVIVITVIWIRXIHF]E VIWTSRWIGLERKISJ[MXLMREFSYX PEJXIVXLIWXEVXERHIRH SJXLIMRNIGXMSRHITIRHMRKSRXLIRYQFIVSJJPS[GIPPWMRXLIJPS[ TEXL-JHMWTIVWMSRMWI\GIWWMZIGLIGOXLIEMVWIKQIRXWWITEVEXMRK WEQTPIJVSQFYJJIVMRXLIEYXSWEQTPIVXYFMRK 9RHYIHMWTIVWMSRMRQIXLSHGSRXVSPPIHVYRWQE]FIGEYWIHF]IEVP] TVITEVEXMSRSJEXMQIHMRNIGXMSR7II%TTIRHM\& Other application problems 8SEZSMHTVSFPIQW[MXL]SYVETTPMGEXMSRWMIXLIW]WXIQJYRGXMSRW GSVVIGXP]FYX]SYVETTPMGEXMSRlHSIWRX[SVOzVIEHXLI &-%ETTPMGEXMSRW,ERHFSSOJSVKIRIVEPEWTIGXWERH%TTIRHM\&SJXLMW QERYEPJSVEWTIGXWWTIGMJMGXS&-%'36) 8.2 Instrument problems General failure -JXLIVIH)6636MRHMGEXSVPMKLXWSRXLIJVSRXTERIPSJXLIMRWXVYQIRX W[MXGLXLIMRWXVYQIRXERH4'SJJ[EMXQMRYXIXLIRVIWXEVXXLI W]WXIQ-JXLITVSFPIQTIVWMWXWGEPP&MEGSVIWIVZMGI Liquid delivery 4YQTJYRGXMSRW *EYPX]PMUYMHLERHPMRKQE]GEYWIEZEVMIX]SJTVSFPIQWMRGPYHMRK MRGSVVIGXMRNIGXMSRZSPYQIWHIPE]IHMRNIGXMSRWFVSOIREMVWIKQIRXW HYVMRKMRNIGXMSRMRGSVVIGXHMPYXMSRWERHXVERWJIVVIHZSPYQIWIXG8IWX PMUYMHLERHPMRK[MXLMRNIGXMSRWSJWYGVSWISVKP]GIVSPWSPYXMSRW[LMGL KMZIE[IPPHIJMRIHVIJVEGXMZIMRHI\TYPWI8IWXEYXSWEQTPIVJYRGXMSRW HMPYXIXVERWJIVERHQM\F]TVITEVMRKHIJMRIHQM\XYVIWSJWYGVSWISV KP]GIVSPWSPYXMSRERHFYJJIV8LITPEXIEYVIWTSRWIHYVMRKMRNIGXMSR WLSYPHFIHMVIGXP]TVSTSVXMSREPXSXLIGSRGIRXVEXMSR

149 Troubleshooting *EYPX]PMUYMHLERHPMRKGEREJJIGXDiluteTransferMixDipneedle InjectERHSXLIVMRNIGXMSRGSQQERHWFractionsERHRecoverERH QE]FIGEYWIHF] MRGSVVIGXZSPYQIWMRGSQQERH'LIGOXLIZEPYIWMRXLIQERYEP HMEPSKYIFS\SVXLIQIXLSH TVSFPIQW[MXLEMVWIKQIRXWWIIFIPS[ MRGSVVIGXRIIHPITSWMXMSRMRKWII7IGXMSR PIEOEKIMRXYFMRKGSRRIGXMSRW*SVGSVVIGXMRNIGXMSRWXLIVIWLSYPHFI EHIPE]FIX[IIRXLIWXEVXSJXLIMRNIGXMSREWVIGSVHIHMRXLIIZIRXPSK ERHXLIFIKMRRMRKSJXLIVIWTSRWIMRXLIJMVWXJPS[GIPPGSVVIWTSRHMRK XSRSXQSVIXLER PPMUYMHIKWIGSRHEX PQMR8LIHIPE] FIX[IIRVIWTSRWIWMRQYPXMGLERRIPQSHIWLSYPHGSVVIWTSRHXSEFSYX PPMUYMH0SRKIVHIPE]WTVSFEFP]MRHMGEXIPIEOEKI Illustration of the normal delay between sensorgrams in multichannel mode. Note the enlarged time axis. TYQTJEMPYVI'SRXEGX&MEGSVIWIVZMGIMJXLITYQTWHSRSXSTIVEXI JSVQEXMSRSJEMVFYFFPIWMRWSPYXMSRW JPS[SFWXVYGXMSR'LIGOXLEXXLIXYFMRKMWRSXWLEVTP]FIRXSV XVETTIHWSEWXSSFWXVYGXPMUYMHJPS[6ITPEGIER]HEQEKIHXYFMRK EMVMRTYQTW]VMRKI)\EQMRIXLITYQTJSVEMVFYFFPIWMRXLI W]VMRKI8SVIQSZIEMVFYFFPIWXETXLIW]VMRKIKIRXP]HYVMRKEPrime STIVEXMSR

150 Troubleshooting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orking Tools:PrimeEXVIKYPEVMRXIVZEPWFIX[IIRIEGLG]GPIMR EQIXLSH MRGVIEWIXLIJPS[VEXI *SVEMVFYFFPIWMRNIGXIHXSKIXLIV[MXLXLIWEQTPIWII7IGXMSR Autosampler -VVITVSHYGMFPIEMVWIKQIRXWMRPMUYMHLERHPMRK -JXLIEMVWIKQIRXWWITEVEXMRKWEQTPIJVSQFYJJIVMRXLIEYXSWEQTPIV XYFMRKEVIXSSPEVKISVETTIEVXSFVIEOYTEMVQE]FIMRNIGXIHSZIV XLIWIRWSVWYVJEGIKMZMRKELMKLWMKREPSYXWMHIXLIH]REQMGVERKISJXLI MRWXVYQIRX-RSXLIVEYXSWEQTPIVSTIVEXMSRWWYGLEWDilute Fractions RecoverJEYPX]EMVWIKQIRXWGERPIEHXSHIPMZIV]SJMRGSVVIGXZSPYQIW Sensorgram from amine coupling with air spike in the NHS/EDC injection and air bubble at the beginning of the sample injection.

151 Troubleshooting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ervice Tools:Needle replacement/positioning -JXLIRIIHPIQSZIWXSERMRZEPMHVEGOTSWMXMSRGLIGOXLEXXLIVEGO YWIHMRXLIEYXSWEQTPIVMWGSVVIGXP]HIJMRIHMRXLIGSRXVSPWSJX[EVI Command:Define Rack Base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

152 Troubleshooting Connector block and IFC 0IEOEKIEXXLIMRNIGXMSRTSVX 'PIERSVVITPEGIXLIMRNIGXMSRTSVXSRXLIGSRRIGXSVFPSGOService Tools:Injection Port -*'ZEPZIQEPJYRGXMSR 8LI-*'ZEPZIWQE]WXMGOMJXLI]EVIPIJXGPSWIHJSVPSRKTIVMSHWSJ XMQI6YRService Tools:UncloggingJSPPS[IHF]Working Tools:Prime MJ]SYWYWTIGXXLEXZEPZIWEVIRSXJYRGXMSRMRKTVSTIVP] 8LIZEPZIWEVIPIJXSTIRMRXLIRSVQEPWXERHF]WXEXI[LIRRS WIRWSVKVEQMWVYRRMRK8LI[SVOMRKXSSPStandbyW[MXGLIWZEPZIWEX MRXIVZEPWXSTVIZIRXWXMGOMRK Sensor chip -J]SYGERRSXHSGOSVYRHSGOEWIRWSVGLMTW[MXGLXLIMRWXVYQIRXSJJ ERHVIWXEVX-JXLITVSFPIQTIVWMWXWGSRXEGX&MEGSVIWIVZMGI 6IQIQFIVXLEXXLIGSQQERHWDockERHUndockEVIRSXEZEMPEFPI HYVMRKEVYRMRGPYHMRKHYVMRKSTIVEXMSRSJStandby Temperature control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

153 Troubleshooting WYHHIRGLERKIWMREQFMIRXXIQTIVEXYVI" 'SZIVPIWWXLEREFSYX QMRYXIW %RMRGVIEWIMRXIQTIVEXYVI[MPPKMZIEHIGVIEWIMREFWSPYXIVIWTSRWI 8SGLIGOXLIXIQTIVEXYVIHYVMRKEVYR GLIGOXLIXIQTIVEXYVIEXXLIFIKMRRMRKERHIRHSJIEGLG]GPIEW VIGSVHIHMRXLIIZIRXPSK YWIXLICURVE tempgsqqerhmrxliqixlshxsvigsvhxli XIQTIVEXYVIXLVSYKLSYXXLIVYR -JEGSRWXERXXIQTIVEXYVIMWRSXQEMRXEMRIHMRWTMXISJVIEWSREFP] GSRWXERXEQFMIRXGSRHMXMSRWXLIXLIVQSWXEXXMRKYRMXQE]FIHIJIGXMZI 'SRXEGX&MEGSVIWIVZMGI 8.3 Software problems 2SGSRXEGX[MXLMRWXVYQIRX 8LIMRWXERGISJ&-%'36)'SRXVSP7SJX[EVI[LMGLLEWGSRXEGX[MXL XLIMRWXVYQIRXMWMHIRXMJMIHEWBIACORE 2000 on COM1:MRXLIXMXPIFEV SJXLI[MRHS[-J]SYGERRSXIWXEFPMWLGSRXEGX[MXLXLIMRWXVYQIRX 'LIGOXLEXXLIGEFPIGSRRIGXMRKXLI4'XSXLIMRWXVYQIRXMWTVSTIVP] WIEXIH'LIGOEPWSXLEXXLIGEFPIMWGSRRIGXIHXSXLIVMKLXTSVXSR XLI4'ERHXLIMRWXVYQIRXWII'LETXIV 'LIGOXLEX]SYHSRSXLEZIERSXLIVMRWXERGISJ&-%'36)'SRXVSP 7SJX[EVIVYRRMRK'PSWIEPPMRWXERGIWSJXLIGSRXVSPWSJX[EVIERH VIWXEVXMXJVSQXLI;MRHS[WStart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

154 Troubleshooting 3XLIVTVSFPIQW *SVTVSFPIQWVIPEXMRKXSXLIGSQTYXIVLEVH[EVISV;MRHS[W STIVEXMSRVIJIVXSXLIVIWTIGXMZITVSHYGXHSGYQIRXEXMSR *SVTVSFPIQW[MXL&-%'36)'SRXVSP7SJX[EVI[LMGLHSRSXLEZIE VIEHMP]ETTEVIRXWSPYXMSR -JTSWWMFPITIVJSVQXIWXWXSHIXIVQMRI[LIXLIVXLITVSFPIQMW VITVSHYGMFPI 6YRXLISoftware Problem ReportMRService Tools7IGXMSR XSGVIEXIEVITSVXMRGPYHMRKW]WXIQWTIGMJMGMRJSVQEXMSR-RXLI VITSVXXLIREXYVISJXLITVSFPIQERHLS[]SYGERVITVSHYGIMX SVEPXIVREXMZIP][LEX]SY[IVIHSMRK[LIRMXEVSWIMREWQYGL HIXEMPEWTSWWMFPIERHWYFQMXXLIVITSVXF]IQEMPSVJE\XS &MEGSVIXLIEHHVIWWMWTVSZMHIHMRXLI Software Problem Report YXMPMX] -JXLIWSJX[EVIQEPJYRGXMSRTVIZIRXW]SYJVSQVYRRMRKXLI Software Problem ReportJVSQService ToolsMRWIVXXLI&MEGSVI 7SJX[EVI'(MR]SYV'(HVMZIERHVMKLXGPMGOSRXLIHVMZIMGSRMR ;MRHS[W)\TPSVIV8LIWSJX[EVITVSFPIQVITSVXGERFIEGGIWWIH JVSQXLIGSRXI\XQIRYJSVXLI'( )VVSVW[LMGLEVMWIHYVMRKEVYREVIVIGSVHIHMRXLIIZIRXPSKJSVXLI VYR

155 Troubleshooting

156 BIACORE Control Software reference 9. BIACORE Control Software reference 8LMWGLETXIVTVSZMHIWEJYPPVIJIVIRGIXSXLI&-%'36)'SRXVSP 7SJX[EVIYWIVMRXIVJEGIJSV&-%'36)SVKERM^IHMREGGSVHERGI [MXLXLIWSJX[EVIQIRYWXVYGXYVI7II'LETXIVJSVERSTIVEXMSREP HIWGVMTXMSRSJLS[XSYWIXLIWSJX[EVI 9.1 Starting BIACORE Control Software Starting from the Windows Start menu %WLSVXGYXXSXLI&-%'36)'SRXVSP7SJX[EVIMWMRWXEPPIHYRHIV&-% ProgramsSRXLIStartQIRY -XHSIWRSXQEXXIV[LIXLIV]SYW[MXGLSRXLIMRWXVYQIRXSVWXEVXXLI WSJX[EVIJMVWX-J]SYWXEVXXLIWSJX[EVIJMVWXXLIWXEXYW[MRHS[[MPP HMWTPE]XLIQIWWEKIInstrument Offline'SRRIGXMSR[MPPFIIWXEFPMWLIH [LIRXLIMRWXVYQIRXMWW[MXGLIHSR (ITIRHMRKSRXLIGYVVIRXMRWXVYQIRXWXEXYW[LIRXLIWSJX[EVIMW WXEVXIH]SY[MPPFITVSQTXIHJSVXLIETTVSTVMEXIWIXYTEGXMSRIK HSGOEWIRWSVGLMTTVMQIXLIW]WXIQ7II7IGXMSRJSVJYVXLIV HIXEMPWSJXLIWIEGXMSRW Starting from Windows Explorer &-%'36)'SRXVSP7SJX[EVIMWEWWSGMEXIHMR;MRHS[W)\TPSVIV[MXL JMPII\XIRWMSRWFPVFPQERHFP[WSXLEXHSYFPIGPMGOMRKSRJMPIWSJ XLIWIX]TIWMR)\TPSVIV[MPPWXEVXXLI'SRXVSP7SJX[EVI&-%'36) 'SRXVSP7SJX[EVIWXEVXIHMRXLMW[E]LS[IZIV[MPPRSXFIGSRRIGXIHXS XLIMRWXVYQIRXERHGERSRP]FIYWIHJSV[VMXMRKERHIHMXMRKQIXLSHW ERHI\EQMRMRKVIWYPXJMPIW

157 BIACORE Control Software reference Multiple software instances =SYGERWXEVXWIZIVEPMRWXERGIWSJ&-%'36)'SRXVSP7SJX[EVIFYX SRP]XLIJMVWXMRWXERGIGERFIGSRRIGXIHXSXLIMRWXVYQIRX-J]SYWXEVX QYPXMTPIMRWXERGIW]SY[MPPFI[EVRIHXLEXXLIWSJX[EVIMWEPVIEH] VYRRMRK 8LIGSRXVSPWSJX[EVIMRWXERGIXLEXMWGSRRIGXIHXSXLIMRWXVYQIRXMW MHIRXMJMIHEWBIACORE Control Software on COM1MRXLI[MRHS[XMXPI FEVERHMRXLIMGSRXMXPIMJXLI[MRHS[MWVIHYGIHXSERMGSRSRXLI HIWOXST3XLIVMRWXERGIWEVIMHIRXMJMIHWMQTP]EWBIACORE Control Software 2SXIXLEXSRP]SRIMRWXERGISJXLIWSJX[EVIGERFIGSRRIGXIHXSXLI MRWXVYQIRX 8LIRYQFIVSJMRWXERGIW]SYGERSTIRHITIRHWSRXLI;MRHS[W W]WXIQVIWSYVGIWEZEMPEFPIMR]SYVGSQTYXIV8LIJSPPS[MRKIVVSV QIWWEKIWETTIEVMRKEXWXEVXYTQE]MRHMGEXIXLEXW]WXIQVIWSYVGIWEVI MRWYJJMGMIRX Out of memory ConnectToBIACORE, unexpected error=20100 Could not start communication server -J]SYVIGIMZISRISJXLIWIQIWWEKIWGPSWIWSQISXLIVETTPMGEXMSRWERH XV]EKEMR Recovering lost data %QIWWEKILost data foundhmwtpe]ih[lir]sywxevxxligsrxvsp WSJX[EVIMRHMGEXIWXLEXETVIZMSYWVYR[EWRSXGSVVIGXP]WEZIHXLMWGER LETTIHJSVI\EQTPIMJXLIVIMWETS[IVJEMPYVIHYVMRKEVYR-J]SY GLSSWIXSVIGSZIVXLIPSWXHEXEXLIHEXE[MPPETTIEVMREWIRWSVKVEQ [MRHS[ERHGERFIWEZIHEWERSVQEPVIWYPXJMPI-J]SYGLSSWIRSXXS VIGSZIVXLIHEXE[LIRXLIGSRXVSPWSJX[EVIWXEVXWXLIPSWXHEXE[MPPFI HIPIXIHTIVQERIRXP]ERHGERRSXFIVIGSZIVIHEXEPEXIVHEXI Start-up routines %XWXEVXYTXLIW]WXIQQE]HMWTPE]EVIQMRHIVXLEXMXMWXMQIXSVYR XLI[SVOMRKXSSPWSanitizeERHSVDesorbWII7IGXMSR8LMW VIQMRHIVETTIEVW[LIRQSVIXLERXLIVIGSQQIRHIHMRXIVZEPLEW IPETWIHWMRGIXLIVIWTIGXMZIXSSP[EWPEWXYWIH=SYQE]GLSSWIXSVYR XLIQEMRXIRERGIXSSPSVMKRSVIXLIVIQMRHIVERHVYRXLIXSSPEXEPEXIV HEXI-J]SYMKRSVIXLIVIQMRHIVMX[MPPFIVITIEXIHEJXIVSRI[IIO

158 BIACORE Control Software reference 9.2 Screen display 8LI&-%'36)WGVIIRMWHMZMHIHMRXSWM\QEMREVIEW 8LIQIRYFEVTVSZMHIWEGGIWWXS&-%'36)QIRYGSQQERHW 1ER]SJXLIGSQQERHWGEREPWSFIEGGIWWIHXLVSYKLWLSVXGYXOI]W WII7IGXMSR &YXXSRWSRXLIXSSPFEVHYTPMGEXIJVIUYIRXP]YWIHQIRYGSQQERHW JSVQSVIGSRZIRMIRXEGGIWW

159 BIACORE Control Software reference 8LIWIRWSVKVEQ[MRHS[HMWTPE]WXLITVSKVIWWSJXLIMRXIVEGXMSREWE WIRWSVKVEQ8LMWEVIEEPWSHMWTPE]WXLIQIXLSHIHMXSV[MRHS[ 8LIVITSVXTSMRXXEFPIVIGSVHWWIRWSVKVEQMRJSVQEXMSREXIEGL VITSVXTSMRX 8LIIZIRXPSK[MRHS[VIGSVHWMQTSVXERXTEVEQIXIVWEXXLIWXEVXERH IRHSJIEGLVYRERHEXIZIRXWHYVMRKXLIVYR 8LIWXEXYW[MRHS[HMWTPE]WXLIGYVVIRXMRWXVYQIRXWXEXYWMRGPYHMRK GYVVIRXVIWTSRWIGSRXMRYSYWJPS[VEXIXIQTIVEXYVIHEXEGSPPIGXMSR VEXI7]WXIQWXEXYWQIWWEKIWEVIHMWTPE]IHEXXLIFSXXSQSJXLMW [MRHS[ Menu bar 1IRYGSQQERHWJSPPS[;MRHS[WWXERHEVHYWEKI&VMIJP] 8SWIPIGXEQIRYGPMGOSRXLIQIRYREQIMRXLIQIRYFEVSVTVIWW AltJSPPS[IHF]XLIYRHIVPMRIHPIXXIVMRXLIREQI 8SGPSWIEQIRYGPMGOER][LIVISYXWMHIXLIQIRYSVTVIWWEsc 8SGLSSWIEGSQQERHWIPIGXXLIQIRYXLIRGPMGOSRXLIGSQQERH SVTVIWWXLIYRHIVPMRIHPIXXIVMRXLIGSQQERH%PXIVREXMZIP]TVIWW XLIWLSVXGYXOI]SVOI]GSQFMREXMSRWII7IGXMSR=SYHSRSX RIIHXSWIPIGXXLIQIRYMRSVHIVXSYWIXLIWLSVXGYXOI] 'SQQERHW[LMGLEVIHMQQIHGERRSXFIYWIHMRXLIGYVVIRX GMVGYQWXERGIWIK]SYGERRSXWXEVXEWIRWSVKVEQYRPIWW]SYLEZI HSGOIHEWIRWSVGLMT 7SQIGSQQERHWGLERKIEGGSVHMRKXSXLIGYVVIRXGMVGYQWXERGIWIK Run SensorgramGLERKIWXSStop Sensorgram[LIREWIRWSVKVEQMW WXEVXIH

160 BIACORE Control Software reference Toolbar 8LIXSSPFEVGSRXEMRWFYXXSRWJSVGSQQSRP]YWIHGSQQERHW'PMGOSR EXSSPFEVFYXXSRXSGLSSWIXLIGSVVIWTSRHMRKGSQQERH%PP GSQQERHWEVIEPWSEZEMPEFPIJVSQXLIQIRYW &YXXSRW[LMGLEVIHMQQIHGERRSXFIYWIHMRXLIGYVVIRX GMVGYQWXERGIWIK]SYGERRSXWXEVXEWIRWSVKVEQYRPIWW]SYLEZI HSGOIHEWIRWSVGLMT 8LICyclePMWXFS\WLS[WXLIGYVVIRXG]GPIHYVMRKEQIXLSHVYRERH EPPS[W]SYXSWIPIGX[LMGLG]GPIXSHMWTPE])EGLG]GPIGSVVIWTSRHWXS ERAPROGMRXLIQIXLSHWII7IGXMSRERHGSRXEMRWSRIX[SSV JSYVWIRWSVKVEQWHITIRHMRKSRXLIHIXIGXMSRQSHI 8LICurvePMWXFS\MHIRXMJMIWERHEPPS[W]SYXSWIPIGXXLIlEGXMZIz GYVZIMRQYPXMGLERRIPQSHI'SSVHMREXIWJSVXLIEGXMZIGYVZIEVI HMWTPE]IHMRXLIWXEXYW[MRHS['LERKMRKXLIEGXMZIGYVZIHSIWRSX EJJIGXHEXEGSPPIGXMSR Sensorgram window 8LIWIRWSVKVEQ[MRHS[HMWTPE]WXLITVSKVIWWSJXLIGYVVIRXEREP]WMW 8LIHMWTPE]MWEYXSQEXMGEPP]WGEPIHF]HIJEYPXXSEGGSQQSHEXIXLI WIRWSVKVEQW7GEPMRKGERFIWIXQERYEPP][MXLView:ScaleSV EHNYWXIHXSJMXXLIGYVVIRXWIRWSVKVEQW[MXL View:Adjust Scale7II XLIGSQQERHHIWGVMTXMSRWMR7IGXMSRJSVQSVIHIXEMPW 8S^SSQXLIHMWTPE]HVEKXLIQSYWIQEVOIVSZIVXLIEVIEXSFI IRPEVKIH8SVIXYVRXLIWXERHEVHHMWTPE]GLSSWIView:UnzoomSV HSYFPIGPMGO[MXLXLIPIJXQSYWIFYXXSRMRXLIWIRWSVKVEQ[MRHS[ 8LIWIRWSVKVEQWQE]WLS[IZIRXQEVOIVWXVMERKPIWSR\E\MW VITSVXTSMRXWGVSWWIWSRWIRWSVKVEQ[MXLXLIVITSVX[MRHS[ MRHMGEXIHERHFEWIPMRIERHSVVIJIVIRGIPMRILSVM^SRXEPERHZIVXMGEP PMRIWVIWTIGXMZIP]7II7IGXMSRJSVQSVIHIXEMPW *M\IHGSPSYVWEVIYWIHXSHMWXMRKYMWLXLIJPS[GIPPJSVIEGLWIRWSVKVEQ *G VIH *G KVIIR *G FPYI *G QEKIRXE VIJIVIRGIWYFXVEGXIH FPEGO *SVFIWXHMWTPE]SJXLIWIRWSVKVEQ[MRHS[QEOIWYVIXLEXXLI [MRHS[FEGOKVSYRHGSPSYVMWWIXXS[LMXIXLMWMWXLIRSVQEP ;MRHS[WWIXXMRK

161 BIACORE Control Software reference Report point table 8LIVITSVXXEFPIVIGSVHWWIRWSVKVEQMRJSVQEXMSREXIEGLVITSVXTSMRX *SVHIXEMPWSRXLIX]TISJMRJSVQEXMSRVIGSVHIHWII7IGXMSR Event log window 8LIIZIRXPSK[MRHS[VIGSVHWEPPIZIRXWMRXLIGYVVIRXG]GPIl)ZIRXWz QE]FIJYRGXMSREPFVIEOTSMRXWWYGLEWMRNIGXMSRWSVGLERKIWMRXLI JPS[VEXISVWXEXYWMRJSVQEXMSRWYGLEWXMQIWXEQTERHHIXIGXSV XIQTIVEXYVI2SXIXLEXIZIRXWSYXWMHIG]GPIWMRQIXLSHWMIMRXLI MAINFPSGO[MPPRSXFIVIGSVHIHMRXLIIZIRXPSK 8LIIZIRXPSKQE]FIHMWTPE]IHMRFVMIJSVI\XIRHIHQSHI'LSSWI View:Extended Event Log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

162 BIACORE Control Software reference Status window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urvePMWXSRXSSPFEV -JFSXLVIJIVIRGIPMRIERHFEWIPMRIEVIHMWTPE]IHXLIIPETWIHXMQI ERHVIPEXMZIVIWTSRWIFEWIPMRI!EXXLIVIJIVIRGIPMRIEVIWLS[R JSVXLIEGXMZIWIRWSVKVEQ Method editor window 8LIQIXLSHIHMXSVMWYWIHJSV[VMXMRKERHIHMXMRKQIXLSHW3TIRXLI QIXLSHIHMXSV[MRHS[[MXLIMXLIVFile:New MethodJSVERI[QIXLSH SVFile:OpenJSVEWEZIHQIXLSHJMPI1YPXMTPIQIXLSHIHMXSV[MRHS[W GERFISTIRWMQYPXERISYWP]

163 BIACORE Control Software reference 2SXI 8LIQIXLSHIHMXSVGERRSXFIYWIHMJEWIRWSVKVEQMWVYRRMRK 7XEVXERSXLIVMRWXERGISJXLIGSRXVSPWSJX[EVIMJ]SY[ERXXS IHMXQIXLSHW[LMPIEWIRWSVKVEQMWVYRRMRK *SVEHIWGVMTXMSRSJLS[XS[VMXIQIXLSHW[MXLXLIQIXLSHIHMXSV VIJIVXS%TTIRHM\) 9.3 Menus and buttons 7IZIVEPGSQQERHWMR&-%'36)LEZIGSVVIWTSRHMRKFYXXSRWMR XLIXSSPFEVJSVUYMGOEGGIWW%PPGSQQERHWEVIEZEMPEFPIJVSQXLI QIRYWQER]LEZIMREHHMXMSRWLSVXGYXOI]WEWEREPXIVREXMZIWLSVXGYX OI]WEVIWLS[REPSRKWMHIXLIGSQQERHWMRXLIQIRYW[LIVI ETTVSTVMEXI8LIXEFPISRXLIRI\XTEKIPMWXWXSSPFEVFYXXSRWERH WLSVXGYXOI]WJSVQIRYGSQQERHW 8SSPFEVFYXXSRMGSRWEVIHMWTPE]IHMRWSQIHMEPSKYIFS\IWIKInject XSLIPTXSMHIRXMJ]XLIHMEPSKYIFS\%WTIGMEPMGSRMRWSQIFS\IWIK DockMRHMGEXIWXLEXEQERYEPSTIVEXMSRMWVIUYMVIH -RHMGEXIWQERYEPSTIVEXMSR

164 BIACORE Control Software reference Menu/Command Toolbar button Shortcut key File Open Save Print Edit (sensorgram) Copy Graph Add Report Point Edit (method) Undo Cut Copy Paste Find Replace View Unzoom Reference Line Baseline Notebook Wizard Results Command Inject/Stop Inject Ctrl-O Ctrl-S Ctrl-P Ctrl-H Ctrl-Z Ctrl-X Ctrl-C Ctrl-V Ctrl-F Ctrl-R Ctrl-U F8 F9 Ctrl-R Transfer Mix Flow/Stop Flow Run Run/Stop Sensorgram Stop Sensorgram Tools Evaluation Ctrl-E

165 BIACORE Control Software reference 9.4 File menu New Method 3TIRWERI[IQTX]QIXLSHIHMXSV[MRHS[ 7II%TTIRHM\)JSVEHIWGVMTXMSRSJLS[XS[VMXIQIXLSHWMRXLI QIXLSHIHMXSV File:New MethodMWRSXEZEMPEFPI[LMPIEWIRWSVKVEQMWVYRRMRK New Application Wizard 3TIRWXLI%TTPMGEXMSR;M^EVHHMEPSKYIWII'LETXIVJVSQ[LMGL ]SYGERHIJMRIERI[[M^EVHXIQTPEXI9WIXLMWSTXMSRXSHIJMRI [M^EVHXIQTPEXIW[LIR]SYHSRSXLEZIEGSRRIGXMSRXSXLI MRWXVYQIRXIKMREWIGSRHMRWXERGISJXLIGSRXVSPWSJX[EVI Open (ctrl-o) 3TIRWERI\MWXMRKQIXLSHSVVIWYPXJMPI 'LERKIXLIJSPHIVMJRIGIWWEV]MRXLILook injmiph7ipigxxlijmpixs STIRSVIRXIVXLIJMPIREQIMRXLIFile NameJMIPH 3TIRMRKEVIWYPXJMPIHIJEYPXI\XIRWMSR&06STIRWEWIRWSVKVEQ [MRHS[GSRXEMRMRKXLIVIWYPXW3RP]SRIWIRWSVKVEQ[MRHS[GERFI STIREXSRGI 3TIRMRKEQIXLSHJMPIHIJEYPXI\XIRWMSR&01STIRWEQIXLSH IHMXSV[MRHS[GSRXEMRMRKXLIQIXLSH7IZIVEPQIXLSHIHMXSV[MRHS[W GERFISTIRIHWMQYPXERISYWP]XLIQE\MQYQRYQFIVHITIRHWSRXLI EZEMPEFPI;MRHS[WW]WXIQVIWSYVGIW File:OpenMWRSXEZEMPEFPI[LMPIEWIRWSVKVEQMWVYRRMRK

166 BIACORE Control Software reference Close 'PSWIWXLIEGXMZI[MRHS[WIRWSVKVEQSVQIXLSHIHMXSV-JXLIGYVVIRX GSRXIRXWSJXLI[MRHS[LEZIRSXFIIRWEZIH]SY[MPPFIKMZIRER STTSVXYRMX]XSWEZIXLIVIWYPXWSVQIXLSH File:CloseMWRSXEZEMPEFPI[LMPIEWIRWSVKVEQMWVYRRMRK Save (ctrl-s) 7EZIWXLIGSRXIRXWSJXLIGYVVIRXP]EGXMZI[MRHS[EWEJMPI8LMWSTXMSR MWIUYMZEPIRXXSFile:Save As[LIREJMPIMWWEZIHJSVXLIJMVWXXMQI 8LIJMPIMWWEZIHEWEVIWYPXSVQIXLSHJMPIEGGSVHMRKXS[LIXLIVXLI GYVVIRX[MRHS[MWEWIRWSVKVEQSVQIXLSHIHMXSV[MRHS[ VIWTIGXMZIP]%WEZIHVIWYPXJMPIMRGPYHIWWIRWSVKVEQHEXEVITSVXTSMRX XEFPIIZIRXPSKQIXLSHHIJMRMXMSRERHRSXIFSSO%WEZIHQIXLSHJMPI GSRXEMRWEXI\XVITVIWIRXEXMSRSJXLIQIXLSHMRGPYHMRKER]TVIVYR IVVSVQIWWEKIWWIIRun:Prerun Method =SYGERWEZIEVIWYPXJMPIEXER]XMQIHYVMRKEVYR[MXLSYX MRXIVVYTXMRKXLIVYR8LIVIWYPXWEVIWEZIHEXXLITSMRX[LIR]SY GLSSWIXLIGSQQERH-JAutosaveMWEGXMZIYRHIVTools:Preferences VIWYPXW[MPPFIWEZIHEYXSQEXMGEPP]EXTVIWIXMRXIVZEPW 6IWYPXJMPIWEVIWEZIHMR&-%'36)JSVQEX9WI File:ExportMRXLI WIRWSVKVEQ[MRHS[XSI\TSVXWIRWSVKVEQHEXEXSEXI\XJMPIVIEHEFPI F]SXLIVTVSKVEQW *SVEJMPI[LMGLLEWEPVIEH]FIIRWEZIHXLISaveGSQQERHWEZIWXLI HEXE[MXLXLISVMKMREPJMPIREQISZIV[VMXMRKXLISVMKMREPGST]9WI Save AsMJ]SY[ERXXSWEZIXLIHEXE[MXLEHMJJIVIRXJMPIREQIIKMJ ]SYLEZIQEHIGLERKIWXSXLIRSXIFSSOGSRXIRXWERH[ERXXSOIITXLI SVMKMREPVIWYPXJMPI

167 BIACORE Control Software reference Save As 7EZIWXLIGSRXIRXWSJXLIGYVVIRXP]EGXMZI[MRHS[MRERI[JMPI*SVE JMPI[LMGLLEWRSXTVIZMSYWP]FIIRWEZIHSave AsMWIUYMZEPIRXXS Save Export =SYGERI\TSVXIMXLIVWIRWSVKVEQHEXESVVITSVXTSMRXXEFPIXSXEF WITEVEXIHXI\XJMPIWJSVXVERWJIVXSSXLIVETTPMGEXMSRW'LSSWI[LMGL X]TISJHEXEXSI\TSVXJVSQXLIWYFQIRY 7IRWSVKVEQHEXEEVII\TSVXIHEWXEFWITEVEXIHXEFPISJXMQIERH VIWTSRWIZEPYIW-RQYPXMGLERRIPQSHISRISVQSVIWIRWSVKVEQWEVI I\TSVXIHEGGSVHMRKXSXLIGYVVIRXHMWTPE]QSHIWIIView:OverlayERH View:Plot Curve Classes8LI[LSPIXMQIVERKISJXLIWIRWSVKVEQ HEXEMWI\TSVXIHVIKEVHPIWWSJXLIVERKIGYVVIRXP]HMWTPE]IHMRXLI WIRWSVKVEQ[MRHS[ 6ITSVXTSMRXHEXEEVII\TSVXIHEWEXEFWITEVEXIHXEFPI6ITSVXTSMRXW JSVG]GPIWERHEPPWIRWSVKVEQWEVII\TSVXIHVIKEVHPIWWSJXLIGYVVIRX HMWTPE]QSHI )\TSVXIHHEXEQE]FIMQTSVXIHHMVIGXP]MRXSQSWXWTVIEHWLIIX TVSKVEQWIK1MGVSWSJX)\GIP SVGYVZIJMXXMRKTVSKVEQWIK 3VMKMRF]GLSSWMRKXEFWITEVEXIHXI\XJSVQEXJSVXLIMQTSVX STIVEXMSR2SXIXLEX&-%IZEPYEXMSRWSJX[EVIGERSTIR&06JMPIW HMVIGXP]ERHHSIWRSXVIUYMVIHEXEXSFII\TSVXIH 8LIIZIRXPSKGERRSXFII\TSVXIH2SXIFSSOGSRXIRXWGERRSXFI I\TSVXIHFYXGERFIGSTMIHXSSXLIVETTPMGEXMSRWYWMRKXLI;MRHS[W GPMTFSEVH File:ExportMWSRP]EZEMPEFPI[LIREWIRWSVKVEQ[MRHS[MWEGXMZI=SY GERI\TSVXWIRWSVKVEQHEXEEXER]XMQIHYVMRKEVYR

168 BIACORE Control Software reference Show Info (MWTPE]WMRJSVQEXMSREFSYXXLIGYVVIRXWIRWSVKVEQ8LMWGSQQERHMW SRP]EZEMPEFPI[LIREWIRWSVKVEQ[MRHS[MWEGXMZI 8LIMRJSVQEXMSRHMWTPE]IHMRGPYHIWXLIHEXIERHXMQI[LIRXLI WIRWSVKVEQ[EWVYRXLI&-%'36)'SRXVSP7SJX[EVIZIVWMSR MRWXVYQIRX-HERHGSRJMKYVEXMSRERHJMPIHIXEMPWJSVXLIVIWYPXJMPI [LIVIETTPMGEFPIJMPIHIXEMPWEVIVITPEGIHF]XLIXI\X#N/AJSV WIRWSVKVEQWXLEXLEZIRSXFIIRWEZIH 'PMGOSRPrintMRXLIHMEPSKYIFS\XSTVMRXXLIWIRWSVKVEQ MRJSVQEXMSR Print (ctrl-p) 4VMRXWXLIGSRXIRXWSJXLIEGXMZI[MRHS[ 8LIJSPPS[MRKSTXMSRWEVIEZEMPEFPIJSVXLIWIRWSVKVEQ[MRHS[

169 BIACORE Control Software reference System Information 'LIGOXLMWFS\XSTVMRXXLIWIRWSVKVEQMRJSVQEXMSRWII File:Show Info Graph 'LIGOXLMWFS\XSTVMRXXLIGSRXIRXWSJXLIWIRWSVKVEQ[MRHS[ 7IRWSVKVEQWEVITVMRXIHEWGYVVIRXP]HMWTPE]IH[MXLVIWTIGXXS SZIVPE]QSHIQEVOIVWVIJIVIRGIPMRIFEWIPMRIERH^SSQ WIXXMRKW-JAll CyclesMWGLIGOIHHEXEJSVEPPG]GPIWMRXLIVYR [MPPFITVMRXIH3XLIV[MWIHEXEJSVXLIGYVVIRXG]GPISRP]MW TVMRXIH Table 'LIGOXLMWFS\XSTVMRXXLIVITSVXTSMRXXEFPI8LIOI][SVHWMR XLITVMRXIHVITSVXTSMRXXEFPIEVIXLSWIHMWTPE]IHSRXLIWGVIIR WIIView:Keywords Event Log 'LIGOXLMWFS\XSTVMRXXLIIZIRXPSK8LIRSVQEPSVI\XIRHIH IZIRXPSKMWTVMRXIHEGGSVHMRKXSXLIGYVVIRXIZIRXPSKQSHI All Cycles 'LIGOXLMWFS\XSTVMRXHEXEJVSQEPPG]GPIWMREQYPXMG]GPIVYR -JXLMWFS\MWRSXGLIGOIHSRP]XLIGYVVIRXP]HMWTPE]IHG]GPI[MPP FITVMRXIH Wizard results 'LIGOXLMWFS\XSTVMRXXLI[M^EVHVIWYPXW3RP]EZEMPEFPISJXLI WIRWSVKVEQ[EWSFXEMRIHF]VYRRMRKERETTPMGEXMSR[M^EVH Fit To Page 'LIGOXLMWFS\XSTVMRXIEGLGSQTSRIRXSREWITEVEXITEKI Draft 2SXI 'LIGOMRKXLMWFS\VIHYGIWXLIRYQFIVSJTSMRXWTVMRXIHSRXLI WIRWSVKVEQ9WIXLMWJIEXYVIMJQIQSV]WLSVXEKIMR]SYVTVMRXIV GEYWIWHMJJMGYPX]MRTVMRXMRKWIRWSVKVEQW[MXLPEVKIRYQFIVWSJ HEXETSMRXWSVMJTVMRXMRKXEOIWERYRHYP]PSRKXMQI 4VMRXMRKMRHVEJXQSHIGERSGGEWMSREPP]LMHIWMRKPITSMRX WTMOIWMRXLIWIRWSVKVEQF]WOMTTMRKXLISYXPMIVTSMRX(SRSX YWIHVEJXQSHIYRPIWW]SYLEZITVSFPIQWTVMRXMRKMRRSVQEP QSHI 'PMGOSRSetupXSWIXXLIGYVVIRXTVMRXIVTVSTIVXMIWTVMRXIVHVMZIV TEKISVMIRXEXMSRTETIVXVE]IXG 'PMGOSRXLIMore>>FYXXSRXSWIXXLIQEVKMRWERHXSWIPIGXLIEHIVERH JSSXIVGSQTSRIRXWJSVXLITVMRXSYX

170 BIACORE Control Software reference 'SRXVSPPMRKWIRWSVKVEQTVMRXWM^I 7IRWSVKVEQWEVITVMRXIHEXXLIGYVVIRX[MHXLSJXLITEKIFIX[IIRXLI QEVKMRW8STVMRXEXEWQEPPIV[MHXLMRGVIEWIXLIQEVKMRWIXXMRKW -JFit to PageMWRSXGLIGOIHWIRWSVKVEQWEVITVMRXIH[MXLXLIWEQI TVSTSVXMSRWEWXLIGYVVIRXWIRWSVKVEQ[MRHS[SRXLIWGVIIR8LYWMJ XLI[MRHS[MWXEPPERHREVVS[XLITVMRXSYX[MPPEPWSFIXEPPERH REVVS[(VEKEGSVRIVSJXLIWIRWSVKVEQ[MRHS[XSEHNYWXXLI [MRHS[TVSTSVXMSRW -JFit to PageMWGLIGOIHWIRWSVKVEQW[MPPFIWGEPIHXSJMX[MXLMRXLI QEVKMRWSJXLITVMRXIHTEKIVIKEVHPIWWSJXLITVSTSVXMSRWSJXLI WIRWSVKVEQ[MRHS[SRXLIWGVIIR8SGLERKIXLITVMRXIHTVSTSVXMSRW MRXLMWQSHIGLERKIXLIQEVKMRWIXXMRKEGGSVHMRKP] Exit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oolsQIRY

171 BIACORE Control Software reference 9.5 Edit menu 8LMWQIRYMWSRP]HMWTPE]IH[LIREWIRWSVKVEQSVQIXLSHIHMXSV [MRHS[MWEGXMZI8LIQIRYSTXMSRWVIPEXIXSXLIX]TISJEGXMZI [MRHS[ Sensorgram window 8LISTXMSRWXLEXJSPPS[VIPEXIXSXLIWIRWSVKVEQ[MRHS[ Copy Graph (ctrl-h) 'STMIWXLIGYVVIRXWIRWSVKVEQHMWTPE]XSXLI;MRHS[WGPMTFSEVH8LI GSTMIHHMWTPE]GERXLIRFITEWXIHMRXSERSXLIV;MRHS[WETTPMGEXMSR IK;SVHSV;VMXI 8LMWSTXMSRHSIWRSXGST]XLIVITSVXTSMRXXEFPISVXLIIZIRXPSK Add Report Point 3TIRWEHMEPSKYIFS\JSVEHHMRKERI[VITSVXTSMRXXSXLIGYVVIRX WIRWSVKVEQ[MRHS[ )RXIVZEPYIWJSVTimeERHWindowERHEVITSVXTSMRXMHIRXMJMIVJSVId -JXLIVIJIVIRGIPMRIMWEGXMZIXLIXMQIEXXLIVIJIVIRGIPMRIMWWYKKIWXIH EWXLIXMQIJSVXLIVITSVXTSMRX6IWTSRWIZEPYIWVIGSVHIHMRXLIVITSVX TSMRXXEFPI[MPPFIXLIEZIVEKIVIWTSRWISZIVXLIXMQI[MRHS[ GIRXIVIHSRXLIVITSVXTSMRXXMQI 'LIGOBaselineMJXLMWVITSVXTSMRXMWXSFIYWIHEWEFEWIPMRIJSV WYFWIUYIRXTSMRXW'LIGOApply to all curves in windowmj]sy[erxxs VIGSVHEVITSVXTSMRXJSVEPPWIRWSVKVEQWMRXLI[MRHS[MRQYPXM GLERRIPQSHI-JXLMWFS\MWRSXGLIGOIHXLIVITSVXTSMRX[MPPFI VIGSVHIHJSVXLIEGXMZIWIRWSVKVEQSRP]EWMRHMGEXIHF]XLICurve PMWX FS\MRXLIXSSPFEV

172 BIACORE Control Software reference 8LIMRJSVQEXMSRHMWTPE]IHMRXLIVITSVXTSMRXXEFPIMWHIXIVQMRIHF] XLIWIXXMRKWYRHIVView:Keywords8LIWIWIXXMRKWGERFIGLERKIHEX ER]XMQI 2SXI =SYGEREPWSEHHVITSVXTSMRXWXSEHHMXMSREPGYVZIWGVIEXIH [MXLXLICURVEGSQQERHMREQIXLSHEPPS[MRKVITSVXTSMRXW XSVIGSVHIKXIQTIVEXYVISVFYJJIVJPS[EXWTIGMJMIHXMQIW HYVMRKXLIEREP]WMW 8SIHMXSVHIPIXIEVITSVXTSMRXHSYFPIGPMGOSRXLIGSVVIWTSRHMRKPMRI MRXLIVITSVXTSMRXXEFPI 'PMGOSRDeleteXSVIQSZIXLIVITSVXTSMRXSVGLERKIXLIWIXXMRKWERH GPMGOSROKXSIHMXXLIVITSVXTSMRXHIJMRMXMSR Method editor Undo (ctrl-z) 6IZIVWIWXLIPEWXIHMXMRKEGXMSR Cut (ctrl-x) 6IQSZIWWIPIGXIHXI\XERHWXSVIWMXMRXLI;MRHS[W'PMTFSEVHJVSQ [LIVIMXGERFITEWXIHXSERI[PSGEXMSRMRXLIQIXLSHSVXSERSXLIV QIXLSHHIJMRMXMSR8LIVIQSZIHXI\XVITPEGIWER]TVIZMSYWGSRXIRXWSJ XLI'PMTFSEVH 8SWIPIGXXI\XHVEK[MXLXLIPIJXQSYWIFYXXSRSVLSPHHS[R7LMJXERH QSZIXLIGYVWSV[MXLXLIEVVS[OI]W Copy (ctrl-c) 'STMIWWIPIGXIHXI\XERHWXSVIWMXMRXLI;MRHS[W'PMTFSEVHJVSQ [LIVIMXGERFITEWXIHXSERI[PSGEXMSRMRXLIQIXLSHSVXSERSXLIV QIXLSHHIJMRMXMSR8LIGSTMIHXI\XVITPEGIWER]TVIZMSYWGSRXIRXWSJ XLI'PMTFSEVH 8SWIPIGXXI\XHVEK[MXLXLIPIJXQSYWIFYXXSRSVLSPHHS[R7LMJXERH QSZIXLIGYVWSV[MXLXLIEVVS[OI]W

173 BIACORE Control Software reference Paste (ctrl-v) 'STMIWXLIGSRXIRXWSJXLI;MRHS[W'PMTFSEVHMRXSXLIGYVVIRX QIXLSHJMPIEXXLIPSGEXMSRSJXLIGYVWSV%GST]SJXLITEWXIHXI\X VIQEMRWMRXLI'PMTFSEVHYRXMPERI[WIPIGXMSRMWGYXSVGSTMIHSVXLI QIXLSHJMPIMWGPSWIH Delete (IPIXIWQEVOIHXI\X 8SWIPIGXXI\XHVEK[MXLXLIPIJXQSYWIFYXXSRSVLSPHHS[R7LMJXERH QSZIXLIGYVWSV[MXLXLIEVVS[OI]W Adjust Method %HNYWXWXLIQIXLSHHIJMRMXMSRXSEGLSWIRXMQIFEWIERHE WXERHEVHM^IHXI\XPE]SYX7IPIGXXLIXMQIFEWIWIGSRHWSVQMRYXIW JVSQXLIAdjust Method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

174 BIACORE Control Software reference &IJSVIAdjust Method %JXIVAdjust Method:Minutes

175 BIACORE Control Software reference Find (Ctrl-F) 7IEVGLJSVEWTIGMJMIHXI\X8LIWIEVGLGERFIWTIGMJMIHXSQEXGL[LSPI [SVHWSRP]ERHSVQEXGLGEWI Replace (Ctrl-R) 6ITPEGIEWTIGMJMIHXI\XWXVMRK[MXLIRXIVIHXI\X8LIWIEVGLGERFI WTIGMJMIHXSQEXGL[LSPI[SVHWSRP]ERHSVQEXGLGEWI Select All 7IPIGXWEPPXLIXI\XMRXLIJMPIJSVCopyCutERHDeleteSTIVEXMSRW

176 BIACORE Control Software reference 9.6 View menu 'SQQERHWMRXLMWQIRYVIPEXITVMQEVMP]XSWIRWSVKVEQ[MRHS[ HMWTPE] Scale 7IXWXLIWGEPISJXLIWIRWSVKVEQHMWTPE]=SYGEREPWSIRXIVEXMXPIJSV XLIWIRWSVKVEQ8LIXMXPIMWHMWTPE]IHMRXLIWIRWSVKVEQ[MRHS[ERH MWMRHITIRHIRXSJJMPIREQI 7GEPMRKSTXMSRWQE]FIWIXMRHITIRHIRXP]JSVXLI\E\MWERHXLIPIJX ERHVMKLX]E\IWSTXMSRWJSVXLIVMKLX]E\MWEVIHMWTPE]IHSRP]JSV QIXLSHGSRXVSPPIHVYRW[LMGLYWIXLICURVE -ogsqqerhwii %TTIRHM\* Auto Method Manual 7GEPIWXLIHMWTPE]EYXSQEXMGEPP]*SVEWIRWSVKVEQ [MRHS[HMWTPE]MRKGSQTPIXIHVIWYPXWMI[LIRXLI WIRWSVKVEQMWRSXVYRRMRKEYXSWGEPMRKEHNYWXWXLIWGEPI XSEGGSQQSHEXIXLIIRXMVIWIRWSVKVEQ(YVMRKEVYR EYXSQEXMGWGEPMRKETTPMIWSRP]XSHEXEGSPPIGXIHEJXIVXLI WIXXMRKMWGLERKIHXSAuto8SEGGSQQSHEXIXLIIRXMVI WIRWSVKVEQHYVMRKEVYRYWIView:Adjust Scale 7GEPIWXLIHMWTPE]EGGSVHMRKXSXLISCALEGSQQERHMR XLIQIXLSHAPROGF][LMGLXLIWIRWSVKVEQ[EW KIRIVEXIHRSXEZEMPEFPIMJXLIAPROGHSIWRSXMRGPYHIE SCALEGSQQERH 7GEPIWXLIHMWTPE]EGGSVHMRKXSQE\MQYQERHQMRMQYQ ZEPYIWIRXIVIHMRXLIHMEPSKYIFS\ Logarithmic 7IXWEPSKEVMXLQMGMRWXIEHSJEPMRIEVWGEPI 2SXIW ;MXLPSKEVMXLQMGWGEPMRKXLIQE\MQYQERH QMRMQYQZEPYIWIRXIVIHJSVManualVIJIVXSXLI EFWSPYXIVIWTSRWIMR69RSXXSXLIPSKEVMXLQWSJ XLIWIZEPYIW >IVSSVRIKEXMZIZEPYIWGERRSXFITPSXXIH GSVVIGXP]SREPSKEVMXLQMGWGEPI

177 BIACORE Control Software reference 8LIHIJEYPXWIXXMRKMWAutoI\GITXJSVQIXLSHGSRXVSPPIHVYRW[LIVI XLISCALEGSQQERHMWYWIH'LERKMRKXLIWGEPISJXLIWIRWSVKVEQ HMWTPE]HSIWRSXEJJIGXXLIRYQFIVSJHEXETSMRXWGSPPIGXIHJSVXLI WIRWSVKVEQW 8LIScaleHMEPSKYIFS\GEREPWSFIHMWTPE]IHF]HSYFPIGPMGOMRKSR ER]SJXLIWIRWSVKVEQE\IW Adjust Scale %HNYWXWXLIWGEPISJXLIWIRWSVKVEQ[MRHS[MRFSXL\ERH] HMVIGXMSRWXSHMWTPE]XLIIRXMVIWIRWSVKVEQ 'LSSWMRKAdjust ScaleMWIUYMZEPIRXXSWIXXMRKScaleXSAutoJSVFSXL E\IW[LIREWIRWSVKVEQMWRSXVYRRMRK'LSSWMRKAdjust Scale[LMPIE WIRWSVKVEQMWVYRRMRKJMXWXLIIRXMVIGSPPIGXIHHEXEMRXLIHMWTPE] GSQTEVIEYXSQEXMGWGEPMRK[LMGLEHNYWXWXLIWGEPIEWRIGIWWEV]JSV MRGSQMRKHEXE Unzoom (ctrl-u) 6IWXSVIWE^SSQIHWIRWSVKVEQHMWTPE]XSXLITVIZMSYW^SSQJEGXSV (SYFPIGPMGOMRK[MXLXLIPIJXQSYWIFYXXSRMWEWLSVXGYXXSUnzoom 8LIHMWTPE]MWl^SSQIHzF]HVEKKMRKXLIGYVWSVSZIVXLIEVIEXSFI IRPEVKIHUnzoom[MPPVIWXSVIYTXSTVIZMSYW^SSQIHWM^IW;LIR XLISVMKMREPWM^ILEWFIIRVIWXSVIHXLIUnzoomGSQQERHMWKVI] Reference Line (F8) 8SKKPIWHMWTPE]SJXLIVIJIVIRGIPMRIMRXLIWIRWSVKVEQ[MRHS[8LI VIJIVIRGIPMRIEPPS[WI\EGXGSSVHMREXIW69ERHIPETWIHXMQIXSFI VIEHJVSQXLIWIRWSVKVEQ8LIGSSVHMREXIWEVIWLS[RMRXLIWXEXYW [MRHS[[LIRXLIVIJIVIRGIPMRIMWHMWTPE]IH8LIVIWTSRWIMWWLS[REW EFWSPYXI69MJXLIFEWIPMRIMWRSXHMWTPE]IHERHEW69VIPEXMZIXSXLI FEWIPMRIMJXLIFEWIPMRIMWHMWTPE]IHWIIView:Baseline

178 BIACORE Control Software reference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dit:Add Report Point8LITSWMXMSRSJXLIVIJIVIRGIPMRI[MPPFIWYKKIWXIHEWXLIXMQI JSVXLIVITSVXTSMRX

179 BIACORE Control Software reference Baseline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dit:Add Report Point Plot Single (MWTPE]WSRP]XLIGYVVIRXP]EGXMZIGYVZIMRXLIWIRWSVKVEQ[MRHS[ 9WIXLICurvePMWXXSGLERKIXLIGYVVIRXGYVZI Plot Curve Classes 4PSXWIMXLIVSVMKMREPWIRWSVKVEQWJVSQMRHMZMHYEPJPS[GIPPWSV VIJIVIRGIWYFXVEGXIHGYVZIWEGGSVHMRKXSXLIX]TISJGYVZIXLEXMW GYVVIRXP]EGXMZI7IPIGXMRKEHMJJIVIRXX]TISJGYVZIJVSQXLI CurvePMWX [MPPGLERKIXLIGYVZIGPEWWXLEXMWTPSXXIH 2SXI %HHMXMSREPGYVZIWGVIEXIHF]XLICURVEGSQQERHMREQIXLSH JSVQEHHMXMSREPGPEWWIWJSVPlot Curve Classes

180 BIACORE Control Software reference Plot Overlay 8SKKPIWFIX[IIRHMWTPE]SJSRIERHWIZIVEPGYVZIWMRXLIWIRWSVKVEQ [MRHS[8LIVITSVXTSMRXXEFPIPMWXWVITSVXTSMRXWJSVXLIGYVZIWXLEX EVIHMWTPE]IH -RWMRKPIWIRWSVKVEQHMWTPE]QSHIGLSSWIXLIWIRWSVKVEQXSHMWTPE] MRXLICurvePMWXSRXLIXSSPFEV-JXLIOI]FSEVHJSGYWMWSRXLIGYVZI PMWX]SYGERW[MXGLFIX[IIRGYVZIWYWMRKXLIEVVS[OI]W Method Trace (MWTPE]WEQIXLSHXVEGI[MRHS[[LIVIXLITVSKVIWWSJXLIQIXLSH GERFIJSPPS[IH7II7IGXMSRJSVHIXEMPW Method (MWTPE]WXLIQIXLSHEWWSGMEXIH[MXLXLIGYVVIRXWIRWSVKVEQ[MRHS[ 8LMWGSQQERHMWSRP]EZEMPEFPIMJXLIEREP]WMW[EWVYRJVSQEQIXLSH Wizard results (MWTPE]WXLIETTPMGEXMSR[M^EVHVIWYPXW8LMWGSQQERHMWSRP] EZEMPEFPIMJXLIEREP]WMW[EWVYRJVSQERETTPMGEXMSR[M^EVH Wizard template 4VSZMHIWSTXMSRWJSVVYRRMRKERHSVTVMRXMRKXLI[M^EVHWIXXMRKWYWIH XSGVIEXIXLIVIWYPXJMPI8LMWGSQQERHMWSRP]EZEMPEFPIMJXLIEREP]WMW [EWVYRJVSQERETTPMGEXMSR[M^EVH7II7IGXMSRJSVQSVIHIXEMPW

181 BIACORE Control Software reference 2SXI -XMWRSXRIGIWWEV]XSWEZIXLI[M^EVHWIXXMRKWI\TPMGMXP]EWE XIQTPEXIMRSVHIVXSYWIXLMWGSQQERH8LIWIXXMRKWEVIWEZIH MRXLIVIWYPXJMPIERHQE]FIEGGIWWIH[MXLView:Wizard TemplateIZIRMJRSWITEVEXIXIQTPEXI[EWWEZIH Notebook 3TIRWXLIRSXIFSSOJSVXLIGYVVIRXWIRWSVKVEQ -JXLIRSXIFSSOLEWRSXTVIZMSYWP]FIIRSTIRIHJSVXLMWWIRWSVKVEQ ]SYQE]GLSSWIXSSTIRXLIRSXIFSSOXIQTPEXIWIIFIPS[SVSTIRE FPERORSXIFSSO-RXLIRSXIFSSO]SYGERQSZIUYMGOP]XSYWIV HIJMRIHlJMIPHWzYWMRKXLIXEFJSV[EVHWSV7LMJXXEFFEGO[EVHW OI]W%JMIPHMWHIJMRIHF]XLIJMVWXSGGYVVIRGISJEGSPSRSREPMRI 8LIRSXIFSSO[MRHS[LEWEQIRYFEV[MXLFileERHEditQIRYW8LI FileQIRYGSRXEMRWSTXMSRWJSVWEZMRKXLIGYVVIRXRSXIFSSOEWE XIQTPEXIERHTVMRXMRKXLIRSXIFSSO8LIEditQIRYGSRXEMRWWXERHEVH

182 BIACORE Control Software reference ;MRHS[WXI\XIHMXMRKGSQQERHWUndoCutCopyPasteDelete Select All 8LIWEQIRSXIFSSOXIQTPEXIMWEZEMPEFPIJSVEPPVYRW -J]SY[ERXXSGLERKIXLIRSXIFSSOXIQTPEXI 7XEVXEQERYEPWIRWSVKVEQ 3TIRXLIRSXIFSSOERHQEOIXLIVIUYMVIHGLERKIWXSXLI XIQTPEXI 'LSSWIFile:Save As TemplateJVSQXLIQIRYMRXLIRSXIFSSO HMEPSKYIFS\ 7XSTXLIQIXLSHERHHMWGEVHXLIWIRWSVKVEQ 'LERKIW]SYQEOIXSXLIXIQTPEXIHSRSXEJJIGXXLIRSXIFSSOJSVVYRW XLEXLEZITVIZMSYWP]FIIRWEZIH Markers 8SKKPIWXLIHMWTPE]SJVITSVXTSMRXWERHIZIRXQEVOIVWMRXLI WIRWSVKVEQ[MRHS[ %VITSVXTSMRXMWHMWTPE]IHEWERxSRXLIWIRWSVKVEQ[MXLER MRHMGEXMSRSJXLIVITSVXTSMRX[MRHS[*SVJYVXLIVMRJSVQEXMSRSR VITSVXTSMRXWVIJIVXS7IGXMSR %RIZIRXQEVOIVMWHMWTPE]IHEWEFPEGOXVMERKPISRXLI\E\MW)ZIRX QEVOIVWEVIEHHIHXSXLIHMWTPE]EXJYRGXMSREPFVIEOTSMRXWWYGLEW MRNIGXMSRWSVGLERKIWMRXLIJPS[VEXI%HIWGVMTXMSRSJXLIIZIRXMW VIGSVHIHMRXLIIZIRXPSK Extended Event Log 8SKKPIWXLIIZIRXPSKHMWTPE]FIX[IIRWXERHEVHERHI\XIRHIH8LI I\XIRHIHIZIRXPSKMRGPYHIWEHHMXMSREPIZIRXW[LMGLQE]FIYWIJYPMR XVSYFPIWLSSXMRKERHWIVZMGIGSRXI\XW

183 BIACORE Control Software reference Keywords 7IPIGXWXLIOI][SVHWXSMRGPYHIEWLIEHIVWMRXLIVITSVXTSMRXXEFPI ERHXLIRYQFIVSJHIGMQEPWHMWTPE]IHJSVVITSVXTSMRXZEPYIW 'LSSWIXSWLS[EPPRSRISVWIPIGXIHOI][SVHW-J]SYGLSSWINone XLIVITSVXTSMRXXEFPIMWLMHHIR 8LIOI][SVHWEVIEWJSPPS[W SYSTEM defined Time Window AbsResp SD FcJPS[GIPPJSVEPPVITSVXTSMRXWERHAPROG REQISJXLIAPROGJSVVITSVXTSMRXWKIRIVEXIHF] XLIRPOINTGSQQERHMREQIXLSH 8MQIJSVXLIVITSVXTSMRXQIEWYVIHJVSQXLIWXEVX SJXLIWIRWSVKVEQ 8LIXMQI[MRHS[JSVGEPGYPEXMSRSJVIWTSRWI ZEPYIW8LI[MRHS[I\XIRHWIUYEPP]SRIMXLIVWMHI SJXLIVITSVXTSMRXXMQI7IXXMRKXLI[MRHS[XS WIPIGXWEWMRKPIHEXETSMRX %FWSPYXIVIWTSRWIMR69GEPGYPEXIHEWXLIQIER ZEPYISZIVXLIXMQI[MRHS[ 7XERHEVHHIZMEXMSRSJHEXETSMRXWMRXLIXMQI [MRHS[GEPGYPEXIHEW 7( = (] M ]) ( R ) [LIVIR!RYQFIVSJTSMRXW ERH]!VIWTSRWIMR69

184 BIACORE Control Software reference Slope LRSD Quality Baseline RelResp Id APROG keywords 7PSTIHYVMRKXMQI[MRHS[MR69uW GEPGYPEXIHEW WPSTI = ((] M ]) (\ M \)) (\ \) M %PMKRQIRXSJWPSTIXSEWXVEMKLXPMRIVIKVIWWMSR GSIJJMGMIRXGEPGYPEXIHEW LRSD = [LIVI Q 0 ( n 2) ( ) Q = y y 0 i 2 ( ( yi y)( xi x) ) 2 ( x x) OKJSVRSVQEPWMKREPUYEPMX]BadJSVFEHHEXE =)7JSVVITSVXTSMRXWHIJMRIHEWFEWIPMRI 3XLIV[MWI23 6IPEXMZIVIWTSRWIHMJJIVIRGIFIX[IIR%FW6IWTERH FEWIPMRIMR692%MJRSFEWIPMRILEWFIIRWIX 6ITSVXTSMRXMHXI\XWXVMRK *SVQIXLSHGSRXVSPPIHVYRWEHHMXMSREPOI][SVHW QE]FIHIJMRIHMRXLIQIXLSH i 2 Status 8SKKPIWHMWTPE]SJXLIWXEXYW[MRHS[FIX[IIRXLIJYPP[MRHS[ERHE WMRKPIWXEXYWPMRIWLS[MRKXMQIVIWTSRWIERHWXEXYWQIWWEKIW

185 BIACORE Control Software reference 9.7 Command menu Inject -RNIGXWWEQTPISZIVXLIWIRWSVGLMTWYVJEGI 9WIXLIJMIPHWMRXLMWHMEPSKYIFS\EWJSPPS[W Inject command 'LSSWIXLIETTVSTVMEXIMRNIGXGSQQERHJVSQXLIPMWXFS\%VERKISJ MRNIGXGSQQERHWMWTVSZMHIHJSVHMJJIVIRXETTPMGEXMSRTYVTSWIWEW WYQQEVM^IHMRXLIXEFPIFIPS[%PPMRNIGXGSQQERHWI\GITX1%29%0-2.)'8TVITEVIERHTIVJSVQXLIMRNIGXMSREYXSQEXMGEPP]ERH[EWLXLI -*'EXXLIIRHSJXLIMRNIGXMSR7II7IGXMSRJSVEXEFPISJ TVITEVEXMSRERHJMRMWLMRKXMQIWJSVXLIHMJJIVIRXMRNIGXXMQIW

186 BIACORE Control Software reference Command Comments Injected volume Sample consumption Max flow rate INJECT General purpose inject µl Injected vol +30 µl 100 µl/min KINJECT QUICKINJECT Low dispersion inject for kinetics and for samples sensitive to dilution with running buffer. Controlled dissociation time after sample injection. Low sample consumption. Less preparation time than INJECT but more sample dispersion µl Injected vol +40 µl 100 µl/min µl Injected vol +10 µl 100 µl/min COINJECT Injection of second sample immediately following the first (e.g. for dissociation in the presence of ligand). Variable contact times for first sample only. Recovery of either first or second sample, but not both. Sample 1: µl Sample 2: µl Sample 1: injected vol +40 µl Sample 2: injected vol +40 µl 100 µl/min BIGINJECT Large volume inject. Direct inject up to 325 µl, sample loop between 326 and 425 µl, direct between 426 and 750 µl µl Injected vol +52 µl 50 µl/min* MANUAL INJECT Manually controlled inject. See Section µl / loading Loaded vol +30 µl *BIGINJECT cannot be performed at higher flow rates because the autosampler tubing must be refilled during injection from the sample loop for samples larger than 425 µl. Variable Contact Times 'LIGOXLMWFS\JSVZEVMEFPIGSRXEGXXMQIMRNIGXMSRWMRQYPXMGLERRIP QSHI8LIFS\MWSRP]EZEMPEFPIMJEPPJSYVJPS[GIPPWEVIMRGPYHIHMR XLIHIXIGXMSR:SPYQIJMIPHWETTIEVJSVXLIJPS[GIPPWMRXLIGYVVIRX JPS[TEXLIKMJ]SYLEZIWIXJPS[TEXLXLIVI[MPPFIJSYV ZSPYQIJMIPHWJSVER-2.)'8GSQQERH)EGLZSPYQIQYWXFIPIWW XLERSVIUYEPXSXLIZSPYQIMRXLITVIGIHMRKJPS[GIPP*SVI\EQTPI [MXLJSYVJPS[GIPPW 100 µl/min 4IVQMXXIH 2SXTIVQMXXIH

187 BIACORE Control Software reference Position )RXIVXLIVEGOTSWMXMSRJSVXLIWEQTPIXSFIMRNIGXIHSVWIPIGXXLI TSWMXMSRJVSQXLITYPPHS[RPMWX Volume )RXIVXLIZSPYQISJWEQTPIXSFIMRNIGXIH8LIHMEPSKYIFS\EPWS HMWTPE]WGSRXEGXXMQISJXLIWEQTPI[MXLXLIWIRWSVWYVJEGIGSRXEGX XMQI!MRNIGXIHZSPYQIJPS[VEXIERHXLIWEQTPIGSRWYQTXMSRJSVXLI MRNIGXMSRWIIXEFPIEFSZI 8LIJMIPHWMRXLIHMEPSKYIFS\EVIEHNYWXIHXSGSVVIWTSRHXSXLI GLEVEGXIVMWXMGWSJXLIGYVVIRXP]WIPIGXIHMRNIGXGSQQERHIK '3-2.)'8VIUYMVIWX[SWEQTPITSWMXMSRW[MXLZEVMEFPIGSRXEGXXMQIW TIVQMXXIHSRP]JSVXLIJMVWXMRNIGXMSR 8LIMore>>FYXXSRSTIRWEHHMXMSREPSTXMSRWJSVXLIMRNIGXMSR 'LIGOPredip NeedleERHIRXIVEVEGOTSWMXMSRXSHMTXLIRIIHPIMRXLI WTIGMJMIHTSWMXMSRFIJSVIEWTMVEXMRKWEQTPI8LMWSTIVEXMSREPWS EWTMVEXIW PJVSQXLITVIHMTTSWMXMSRMRXSXLIEYXSWEQTPIVXYFMRKXS TVIZIRXXLIWEQTPIJVSQGSQMRKMRXSGSRXEGX[MXLVYRRMRKFYJJIV9WI XLMWJIEXYVIXSQMRMQM^IGEVV]SZIVFIX[IIRWEQTPIZMEPWIK[LIR WSPYXMSRWSJ[MHIP]HMJJIVIRXGSQTSWMXMSREVIYWIHMRWYGGIWWMSRERH EPWS[LIRMXMWMQTSVXERXXLEXXLIWEQTPIMRXLIZMEPHSIWRSXFIGSQI GSRXEQMREXIH[MXLVYRRMRKFYJJIV9WISJPredip NeedleMRERI\XVE ZMEPSJWEQTPIGEREPWSLIPTXSQMRMQM^IWEQTPIHMWTIVWMSRMRXLI MRNIGXMSRF]MQTVSZMRKXLIWITEVEXMSRFIX[IIRMRNIGXIHWEQTPIERH VYRRMRKFYJJIV 'LIGOExtra CleanupXSTIVJSVQEREHHMXMSREP[EWLSJXLIRIIHPIERH -*'EJXIVXLIMRNIGXMSR9WIXLMWJIEXYVIXSQMRMQM^IGEVV]SZIV[LIR XLIRI\XMRNIGXMSRYWIWWEQTPISJ[MHIP]HMJJIVIRXGSQTSWMXMSR8LI -*'MWVSYXMRIP][EWLIHEWTEVXSJXLIWXERHEVHMRNIGXMSRWIUYIRGI Extra CleanupWLSYPHFIYWIHMJTEVXMGYPEVGEVIMWRIGIWWEV]IKEJXIV MRNIGXMSRSJVIKIRIVEXMSRWSPYXMSR 'LIGORecoverySVFractionsERHIRXIVZEPYIWMRXLIETTVSTVMEXIJMIPHW XSEGXMZEXIWEQTPIVIGSZIV]SVJVEGXMSRGSPPIGXMSRJSVXLIMRNIGXMSR

188 BIACORE Control Software reference %28 &IJSVIWXEVXMRKERMRNIGXMSR[MXLVIGSZIV]SVJVEGXMSRGSPPIGXMSR TIVJSVQEWash[MXLRecoverySTXMSRXS[EWLXLIVIGSZIV]GYTERH -*'VIGSZIV]GLERRIPERHIQTX]XLIVIGSZIV]GLERRIP-J]SYHSRSX HSXLMWXLIVIGSZIVIHWEQTPIQE]FIHMPYXIH[MXLYTXS P FYJJIVJVSQXLIVIGSZIV]GLERRIP Recovery Fractions To PositionMWXLIVEGOTSWMXMSRXS[LMGLXLIVIGSZIVIH WEQTPI[MPPFIHMWTIRWIHVolumeMWXLIZSPYQISJPMUYMH XLEX[MPPFIVIGSZIVIHJVSQXLIVIGSZIV]GYTVERKI PVIGSQQIRHIH" PJSVFIWXTVIGMWMSR Start PositionMWXLIVEGOTSWMXMSRJSVXLIJMVWXJVEGXMSR VolumeMWXLIZSPYQISJIEGLJVEGXMSRVERKI P VIGSQQIRHIH" PJSVFIWXTVIGMWMSRNo of fractionsmw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njectGSQQERHMWSRP]EZEMPEFPI[LMPIEQERYEPP]GSRXVSPPIH WIRWSVKVEQMWVYRRMRK Stop Inject 7XSTWXLIGYVVIRXMRNIGXMSRMQQIHMEXIP]=SYGEREPPS[XLII\GIWW WEQTPI[LMGLLEWRSX]IXFIIRMRNIGXIHXSKSXS[EWXISVVIGSZIVMXMR EWIPIGXIHVEGOTSWMXMSR

189 BIACORE Control Software reference 8LIStop InjectGSQQERHMWRSXEZEMPEFPIHYVMRK1%29%0-2.)'8 8SIRHEQERYEPMRNIGXMSRGPMGOSRXLIExitFYXXSRMRXLI1%29%0-2.)'8HMEPSKYIFS\WII7IGXMSR Transfer 8VERWJIVWWEQTPIJVSQSRIVEGOTSWMXMSRXSERSXLIV )RXIVVEGOTSWMXMSRWERHZSPYQIXSFIXVERWJIVVIHMRXLIETTVSTVMEXI JMIPHW 'LIGOPredip NeedleERHIRXIVEVEGOTSWMXMSRXSHMTXLIRIIHPIMRXLI WTIGMJMIHTSWMXMSRFIJSVIXLIXVERWJIVSTIVEXMSR8LMWJYRGXMSREPWS EWTMVEXIW PJVSQXLITVIHMTTSWMXMSRMRXSXLIEYXSWEQTPIVXYFMRKXS TVIZIRXXLIWEQTPIJVSQGSQMRKMRXSGSRXEGX[MXLVYRRMRKFYJJIV9WI XLMWJIEXYVIXSQMRMQM^IGEVV]SZIVFIX[IIRWEQTPIZMEPWIK[LIR WSPYXMSRWSJ[MHIP]HMJJIVIRXGSQTSWMXMSREVIYWIHMRWYGGIWWMSRERH EPWS[LIRMXMWMQTSVXERXXLEXXLIWEQTPIMRXLIZMEPHSIWRSXFIGSQI GSRXEQMREXIH[MXLVYRRMRKFYJJIV Mix 1M\IWWEQTPIMRSRITSWMXMSR9WIXLMWGSQQERHJSVI\EQTPIEJXIV XVERWJIVVMRKX[SHMJJIVIRXWSPYXMSRWXSEGSQQSRZMEPXSTVITEVIE LSQSKIRISYWQM\XYVI

190 BIACORE Control Software reference )RXIVXLIVIUYMVIHVEGOTSWMXMSRERHETTVSTVMEXIZSPYQIXSFIQM\IH QE\MQYQ PXLIRGPMGOSROK%WEKIRIVEPKYMHIPMRIQM\ YWMRKSJXLIXSXEPZSPYQIMRXLIZMEP8LIQM\MRKTVSGIHYVI EWTMVEXIWXLIWTIGMJMIHZSPYQISJWEQTPIERHXLIRHMWTIRWIWMXXSXLI WEQIZMEP8LMWMWTIVJSVQIHXLVIIXMQIW 'LIGOPredip NeedleERHIRXIVEVEGOTSWMXMSRXSHMTXLIRIIHPIMRXLI WTIGMJMIHTSWMXMSRFIJSVIXLIQM\STIVEXMSR8LMWJYRGXMSREPWS EWTMVEXIW PJVSQXLITVIHMTTSWMXMSRMRXSXLIEYXSWEQTPIVXYFMRKXS TVIZIRXXLIWEQTPIJVSQGSQMRKMRXSGSRXEGX[MXLVYRRMRKFYJJIV9WI XLMWJIEXYVIXSQMRMQM^IGEVV]SZIVFIX[IIRWEQTPIZMEPWIK[LIR WSPYXMSRWSJ[MHIP]HMJJIVIRXGSQTSWMXMSREVIYWIHMRWYGGIWWMSRERH EPWS[LIRMXMWMQTSVXERXXLEXXLIWEQTPIMRXLIZMEPHSIWRSXFIGSQI GSRXEQMREXIH[MXLVYRRMRKFYJJIV Dilute 8LMWGSQQERHTIVJSVQWXLIIUYMZEPIRXSJTransferJSPPS[IHF]Mix [MXLEJM\IHJMREPZSPYQISJ PERHQE]FIYWIHJSVHMPYXMRK WEQTPIW[MXLFYJJIVSVJSVTVITEVMRKEHIJMRIHQM\XYVISJX[SWEQTPIW

191 BIACORE Control Software reference Sample 1 Position Sample 2 Position Mix Position Total volume Dilute Sample 1 to 6EGOTSWMXMSRJSVXLIWEQTPIXSFIHMPYXIH 6EGOTSWMXMSRJSVXLIWSPYXMSRXSHMPYXI[MXL 6EGOTSWMXMSRMR[LMGLXLIHMPYXIHWEQTPI[MPPFI TVITEVIH 8LIJMREPXSXEPZSPYQISJHMPYXIHWEQTPIMWJM\IH EX P *MREPGSRGIRXVEXMSRSJWEQTPII\TVIWWIHEWE TIVGIRXEKI 8LII\EQTPIMRXLIMPPYWXVEXMSREFSZI[MPPTVITEVIEQM\XYVIGSRWMWXMRK SJ PWEQTPIJVSQTSWMXMSRVE PWSPYXMSRJVSQTSWMXMSR VE8LIQM\XYVI[MPPFITVITEVIHMRTSWMXMSRVE 'LIGOPredip NeedleERHIRXIVEVEGOTSWMXMSRXSHMTXLIRIIHPIMRXLI WTIGMJMIHTSWMXMSRFIJSVIXLIHMPYXISTIVEXMSR8LMWJYRGXMSREPWS EWTMVEXIW PJVSQXLITVIHMTTSWMXMSRMRXSXLIEYXSWEQTPIVXYFMRKXS TVIZIRXXLIWEQTPIJVSQGSQMRKMRXSGSRXEGX[MXLVYRRMRKFYJJIV9WI XLMWJIEXYVIXSQMRMQM^IGEVV]SZIVFIX[IIRWEQTPIZMEPWIK[LIR WSPYXMSRWSJ[MHIP]HMJJIVIRXGSQTSWMXMSREVIYWIHMRWYGGIWWMSRERH EPWS[LIRMXMWMQTSVXERXXLEXXLIWEQTPIMRXLIZMEPHSIWRSXFIGSQI GSRXEQMREXIH[MXLVYRRMRKFYJJIV Wash )\XVE[EWLSJEYXSWEQTPIVRIIHPI-*'SVVIGSZIV]GYT *SVRIIHPI[EWLVYRRMRKFYJJIVMWTYQTIHXLVSYKLXLIRIIHPIXS [EWXI *SVVIGSZIV][EWLXLIVIGSZIV]GLERRIPMRXLI-*'ERHXLIVIGSZIV] GYTEVIVMRWIH[MXLPMUYMHJVSQXLIWTIGMJMIHTSWMXMSR'LSSWIXLI TSWMXMSRBUFFERXSVMRWI[MXLVYRRMRKFYJJIV 2SXI 8LIPMUYMHLERHPMRKW]WXIQMW[EWLIHEYXSQEXMGEPP]EW VIUYMVIHJSVRSVQEPSTIVEXMSR9WIXLIWashGSQQERH[LIR EHHMXMSREPWIGYVMX]EKEMRWXGVSWWGSRXEQMREXMSRMWVIUYMVIH

192 BIACORE Control Software reference Flow 7IXWXLIJPS[VEXIJSVVYRRMRKFYJJIV )RXIVXLIVIUYMVIHJPS[VEXIMR PQMRTIVQMXXIHVERKI PQMRMR WXITWSJ P8SWXSTXLIJPS[IMXLIVGLSSWICommand:Stop FlowSV WIXXLIJPS[XS PQMR[MXLCommand:Flow -JXLIRefillFS\MWGLIGOIHXLITYQT[MPPFIVIJMPPIHFIJSVIXLIRI[ JPS[VEXIMWWIX8LIVIJMPPSTIVEXMSRXEOIWEFSYXWIGSRHW 8LMWGSQQERHMWRSXEZEMPEFPIYRPIWWEWIRWSVKVEQMWVYRRMRK Stop Flow 7XSTWXLIJPS[SJVYRRMRKFYJJIV8LMWGSQQERHMWIUYMZEPIRXXS WIXXMRKXLIJPS[XS PQMR 8LMWGSQQERHMWSRP]EZEMPEFPI[LIRXLIJPS[MWWIXXSERSR^IVS ZEPYI8LIJPS[MWWXSTTIHEYXSQEXMGEPP]EXXLIIRHSJEQERYEP WIRWSVKVEQSVQIXLSHGSRXVSPPIHVYR8LIJPS[GERRSXFIWXSTTIH HYVMRKERMRNIGXMSRI\GITX1%29%0-2.)'8 Flow path 'LERKIWXLIJPS[TEXLHYVMRKEWIRWSVKVEQ

193 BIACORE Control Software reference 'LSSWIXLIVIUYMVIHJPS[TEXLEWMRHMGEXIHF]XLIMGSRW=SYGERSRP] GLERKIXLIJPS[TEXLXSEWYFWIXSJXLISVMKMREPP]WIPIGXIHJPS[GIPPW IKMJ]SYWXEVX[MXL*G]SYGERGLERKIXS*GSV*GFYXXSER] SJXLISXLIVSTXMSRW'LERKMRKXLIJPS[TEXLHSIWRSXGLERKIXLI HIXIGXMSRQSHI 8LMWGSQQERHMWSRP]EZEMPEFPI[LIREWIRWSVKVEQMWVYRRMRK Set Temperature 7IXWXLIVIUYMVIHXIQTIVEXYVIEXXLIJPS[GIPP 8LIXIQTIVEXYVIHMWTPE]MRXLIWXEXYW[MRHS[XSKKPIWFIX[IIRXLIWIX XIQTIVEXYVIERHXLIEGXYEPXIQTIVEXYVIYRXMPXLIQIEWYVIH XIQTIVEXYVIMWWXEFPI%XIQTIVEXYVIGLERKISJ 'XEOIWEFSYX QMRYXIW ;EMXYRXMPXLIWIRWSVGLMTWYVJEGIXIQTIVEXYVIMWWXEFPIFIJSVI TIVJSVQMRKEREREP]WMWEWWLS[RF]XLIXIQTIVEXYVI0)(SRXLI TVSGIWWMRKYRMXERHXLIXIQTIVEXYVIHMWTPE]MRXLI&-%'36)'SRXVSP 7SJX[EVIWXEXYW[MRHS[ Dock (SGOWXLIWIRWSVGLMT 8LIHSGOMRKSTIVEXMSRXEOIWEJI[WIGSRHWXSGSQTPIXI%JXIV HSGOMRKXLIWorking ToolsHMEPSKYIFS\ETTIEVWEYXSQEXMGEPP][MXL PrimeLMKLPMKLXIHEWEVIQMRHIVXLEX]SYWLSYPHVYRPrimeEJXIV HSGOMRKERI[GLMT*SVFIWXVIWYPXW]SYWLSYPHEPWSVYR Working Tools:NormalizeEJXIVHSGOMRKERI[GLMT

194 BIACORE Control Software reference Undock 9RHSGOWXLIWIRWSVGLMT -JXLIEmpty flow cellsfs\mwgligoihxlijps[gippw[mppfiiqtxmih SJPMUYMHFIJSVIXLIWIRWSVGLMTMWYRHSGOIH8LMWSTXMSRMWGLIGOIHF] HIJEYPXERHWLSYPHEP[E]WFIYWIHYRPIWW]SYVWIRWSVGLMTWYVJEGIMW TEVXMGYPEVP]WIRWMXMZIXSI\TSWYVIXSEMVIK]SYVPMKERHMWVIEHMP] S\MHM^IHMREMV ;LIRXLIYRHSGOMRKSTIVEXMSRMWGSQTPIXIXLIDockHMEPSKYIFS\MW HMWTPE]IHEYXSQEXMGEPP]MRTVITEVEXMSRJSVHSGOMRKERI[WIRWSVGLMT -J]SYHSRSX[ERXXSHSGOERI[GLMTIK]SYMRXIRHXSWLYXHS[R XLIMRWXVYQIRXWMQTP]GLSSWICancelJVSQXLMWHMEPSKYIFS\ -QTSVXERX (SRSXEXXIQTXXSVIQSZIXLIWIRWSVGLMTJVSQXLIMRWXVYQIRXYRXMP XLIYRHSGOMRKSTIVEXMSRMWGSQTPIXI Rack Base (IJMRIWXLIX]TIERHTPEGMRKSJVEGOWSRXLIEYXSWEQTPIVVEGOFEWI 8LIHMEPSKYIFS\WLS[WXLIGYVVIRXWIXXMRKWJSVVEGOFEWIRYQFIVW ERHPIJXERHVMKLXSRXLIEYXSWEQTPIVVIWTIGXMZIP]'LSSWIXLI WIXXMRKWXLEXGSVVIWTSRHXSXLIVEGOW]SYMRXIRHXSYWI 8LIWIXXMRKWMRXLMWHMEPSKYIFS\HIXIVQMRIXLITSWMXMSRWEZEMPEFPIJSV QERYEPGSQQERHWXLEXYWIXLIEYXSWEQTPIVIKInjectDilute TransferMixERHEPWSXLIZEPMHTSWMXMSRWJSVXLIGSVVIWTSRHMRK

195 BIACORE Control Software reference GSQQERHWMREQIXLSHXLEXHSIWRSXMRGPYHIERACKGSQQERH 4SWMXMSRWMREQIXLSHEVIGLIGOIHHYVMRKTVIVYR7II7IGXMSRJSV HIXEMPWSJZEPMHTSWMXMSRWMREQIXLSH 2SXI 8LIVIMWRSEYXSQEXMGWIRWMRKSJVEGOX]TIMRXLIWSJX[EVI-J ]SYWIXXLI[VSRKVEGOX]TIXLIEYXSWEQTPIVRIIHPIQE]FI HMVIGXIHXSXLI[VSRKSVRSRI\MWXIRXTSWMXMSRW6IQIQFIVXS VIHIJMRIXLIVEGOFEWIYWMRKXLMWGSQQERHMJ]SYGLERKIXLI X]TISJVEGOMRXLIEYXSWEQTPIVHYVMRKQERYEPSTIVEXMSR Command Queue 7XEVXWSVWXSTWXLIGSQQERHUYIYIEGGSVHMRKXSXLIGYVVIRXWXEXYWSJ XLIUYIYI8LIGSQQERHMWGLIGOIHMJXLIUYIYIMWWXEVXIH8LMW GSQQERHGERSRP]FIYWIH[LMPIEWIRWSVKVEQMWVYRRMRK 9.8 Run menu Run Sensorgram 7XEVXWEQERYEPP]GSRXVSPPIHWIRWSVKVEQ8LIDetectionERHFlow HMEPSKYIFS\IWEVIEYXSQEXMGEPP]HMWTPE]IHFIJSVIXLIWIRWSVKVEQ WXEVXWXSIRWYVIXLEXXLIWIXXMRKWEVIGSVVIGXJSVXLIMRXIRHIHVYR&] HIJEYPXXLIWIHMEPSKYIFS\IWHMWTPE]XLIQSWXVIGIRXWIXXMRKW %PPQIXLSHIHMXSV[MRHS[W[MPPFIGPSWIHFIJSVIXLIWIRWSVKVEQ WXEVXW-JER]QIXLSHIHMXSV[MRHS[GSRXEMRWYRWEZIHGLERKIW]SY [MPPFITVSQTXIHXSWEZIXLIQIXLSH'LERKIW[LMGLEVIRSXWEZIHEX XLMWWXEKIEVIPSWX 8LMWGSQQERHGLERKIWXSStop Sensorgram[LMPIEWIRWSVKVEQMW VYRRMRK 2SXI (YVMRKWIRWSVKVEQWXEVXXLIWXEXYW[MRHS[XVERWMIRXP] HMWTPE]WZEVMSYWWIXXMRKWJSVJPS[VEXIERHJPS[TEXL VIKEVHPIWWSJXLIWIXXMRKWVIUYIWXIHJSVXLIVYR8LMWMWE RSVQEPTEVXSJXLIWIRWSVKVEQWXEVXYTTVSGIHYVI8LI VIUYIWXIHWIXXMRKW[MPPFIIWXEFPMWLIHFIJSVIHEXEGSPPIGXMSR WXEVXW Stop Sensorgram 7XSTWXLIGYVVIRXQERYEPP]GSRXVSPPIHWIRWSVKVEQ3RGI]SYLEZI WXSTTIHEWIRWSVKVEQ]SYGERRSXVIWYQIVIGSVHMRKI\GITXEWERI[ WIRWSVKVEQ 8LIPMUYMHLERHPMRKW]WXIQMWJPYWLIH[MXLFYJJIVERHXLIJPS[VEXIMW WIXXSEXXLIIRHSJXLIWIRWSVKVEQ

196 BIACORE Control Software reference 8LIGSQQERHGLERKIWXSRun Sensorgram[LIREWIRWSVKVEQMW WXSTTIH 2SXIW (YVMRKWIRWSVKVEQIRHXLIWXEXYW[MRHS[XVERWMIRXP]HMWTPE]W ZEVMSYWWIXXMRKWJSVJPS[VEXIERHJPS[TEXLVIKEVHPIWWSJXLI WIXXMRKWYWIHHYVMRKXLIVYR8LMWMWERSVQEPTEVXSJXLI WIRWSVKVEQXIVQMREXMSRTVSGIHYVI 8SWXSTEQIXLSHGLSSWICommand:Stop Method Start new cycle 7XEVXWERI[G]GPIMREQERYEPP]GSRXVSPPIHVYR8LMWGSQQERHMWSRP] EZEMPEFPI[LMPIEQERYEPWIRWSVKVEQMWVYRRMRK Run Application Wizard 4VIWIRXWEPMWXSJETTPMGEXMSR[M^EVHWXLEXQE]FIVYR7II7IGXMSR JSVQSVIHIXEMPW Run Method 6YRWEQIXLSHJVSQXLIGYVVIRXP]EGXMZIQIXLSHIHMXSV[MRHS[SV JVSQEJMPISRHMWO -JEQIXLSHIHMXSV[MRHS[MWGYVVIRXP]EGXMZIXLIRun Method GSQQERH[MPPWXEVXXLIQIXLSHMRXLEXIHMXSV[MRHS[3XLIV[MWIXLI GSQQERH[MPPTVSQTXJSVEQIXLSHJMPI 4VIVYRMWTIVJSVQIHEYXSQEXMGEPP][LIRIZIVEQIXLSHVYRMW VIUYIWXIHXSTVIZIRXW]RXEGXMGEPP]MRGSVVIGXQIXLSHWJVSQWXEVXMRK

197 BIACORE Control Software reference &IJSVIXLIQIXLSHWXEVXWERYQFIVSJHMEPSKYIFS\IWQE]ETTIEV HITIRHMRKSRXLIGYVVIRXMRWXVYQIRXWXEXYWERHXLIQIXLSHGSRXIRXW -JAutosaveMWWIXJSVQIXLSHVYRWWIIView:Preferences]SY[MPP FIEWOIHXSWTIGMJ]EVIWYPXJMPIREQIJSVXLIEYXSWEZIJIEXYVI -JXLIQIXLSHHSIWRSXGSRXEMREDETECTIONGSQQERH]SY[MPPFI EWOIHXSWIPIGXXLIHIXIGXMSRQSHIERHJPS[TEXL1EOIWYVIXLEX ]SYWIPIGXEHIXIGXMSRQSHIXLEXMWGSQTEXMFPI[MXLXLIQIXLSH TYVTSWI-JMRHSYFXGLSSWIFC WMRGIXLMW[MPPIRWYVIXLEX HEXEMWVIGSVHIHJVSQEPPJSYVGLERRIPW -JXLIHEXEGSPPIGXMSRVEXIMWGYVVIRXP]WIXXSLowSVHigh]SY[MPPFI KMZIRERSTTSVXYRMX]XSVIWIXXLIVEXIXSMedium =SY[MPPFI[EVRIHMJXLIVIMWPMQMXIHWTEGIEZEMPEFPISRXLI GSQTYXIVLEVHHMWO=SYWLSYPHQEOIQSVIWTEGIEZEMPEFPIFIJSVI WXEVXMRKXLIVYRTEVXMGYPEVP]MJAutosave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top Method 7XSTWEVYRRMRKQIXLSH8LIQIXLSH[MPPWXSTEXXLIIRHSJXLI GYVVIRXG]GPIMIXLIIRHSJXLIGYVVIRXAPROGMRXLIQIXLSH2SXI XLEXXLIQIXLSH[MPPVYRXSGSQTPIXMSRMJStop MethodMWVIUYIWXIH HYVMRKXLIPEWXG]GPIMRXLIQIXLSHStop MethodEYXSQEXMGEPP][EWLIW XLIW]WXIQEWJSVERSVQEPQIXLSHGSQTPIXMSR 8SWXSTEQIXLSHMQQIHMEXIP]YWIXLIIQIVKIRG]WXST'XVP&VIEOWII 7IGXMSR=SY[MPPFIEWOIHMJ]SY[ERXXS[EWLXLIPMUYMH LERHPMRKW]WXIQ

198 BIACORE Control Software reference Data Collection Rate (IXIVQMRIWXLIRYQFIVSJTSMRXWGSPPIGXIHTIVYRMXXMQIHYVMRKXLI WIRWSVKVEQ8LIEZEMPEFPIWIXXMRKWEVILowMediumERHHigh GSVVIWTSRHMRKXSEGXYEPHEXEGSPPIGXMSRVEXIWEWWYQQEVM^IHFIPS[ (IXIGXMSRQSHI (EXEGSPPIGXMSRVEXI 7MRKPI 1YPXMGLERRIP 0S[ TSMRXW TSMRXW 1IHMYQ TSMRXW TSMRXW,MKL TSMRXWW TSMRXWW 2SXI :EPYIWMRQYPXMGLERRIPQSHIVIJIVXSXLIGSPPIGXMSRVEXITIV GLERRIP 8LIHIJEYPXWIXXMRKSJMedium MWEHIUYEXIJSVQSWXTYVTSWIW7IXXLI VEXIXSHighJSVETTPMGEXMSRWVIUYMVMRKZIV]LMKLVIWSPYXMSR7IXXLI VEXIXSLowXSVIHYGIXLIWM^ISJXLIWIRWSVKVEQJMPIJSVPSRKVYRW% VIQMRHIVMWMWWYIHMJ]SYWXEVXEVYR[MXLData Collection RateWIXXS HighSVLow 7IRWSVKVEQHEXEMWVIGSVHIHMRXIVREPP]MRWIKQIRXWSJQE\MQYQ TSMRXW8LYWEXEGSPPIGXMSRVEXISJQIEWYVIQIRXWWSRI WIKQIRX[MPPXEOIW!QMRYXIW;LIREWIKQIRXMWJYPP EYXSQEXMGHEXEVIHYGXMSRSGGYVW[LIVIF]TEMVWSJEHNEGIRXTSMRXWEVI EZIVEKIHXSSRITSMRXERHHEXEGSPPIGXMSRGSRXMRYIWEXLEPJXLI TVIZMSYWVEXI%RI[WIKQIRXMWWXEVXIH[LIRIZIVXLIData Collection RateMWWIXIZIRMJXLIGYVVIRXWIXXMRKMWRSXGLERKIH8SEZSMHHEXE VIHYGXMSRXLIVIJSVIGLSSWIData Collection RateWLSVXP]FIJSVIXLI IRHSJEWIKQIRX (EXEVIHYGXMSRMWVIGSVHIHEWERIZIRXMRXLIIZIRXPSK Prerun Method (ctrl-p) 'LIGOWXLIW]RXE\SJXLIQIXLSHMRXLIEGXMZIQIXLSHIHMXSV[MRHS[ -JW]RXE\IVVSVWEVIHIXIGXIH]SY[MPPFI[EVRIHERHERI\TPEREXSV] GSQQIRX[MPPFIMRGPYHIHMR]SYVQIXLSHXSLIPT]SYJMRHXLIIVVSV 8LIMPPYWXVEXMSRFIPS[WLS[WEW]RXE\IVVSVVIWYPXMRKJVSQER MRGSVVIGXP]WTIPPIHOI][SVH7II7IGXMSR)JSVJYVXLIVHIXEMPW

199 BIACORE Control Software reference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

200 BIACORE Control Software reference 9.9 Tools menu BIAsimulation 4VSZMHIWEGGIWWXSXLI&-%WMQYPEXMSRWSJX[EVI[LMGLGYVVIRXP] TVSZMHIWJEGMPMXMIWJSVWMQYPEXMRK OMRIXMGEREP]WMWSJEWWSGMEXMSRERHHMWWSGMEXMSRTLEWIW[MXLSTXMSREP WMKREPRSMWIERHFEWIPMRIHVMJXJSVVIEPMWXMGWMQYPEXMSR OMRIXMGEREP]WIW[MXLLIXIVSKIRISYWPMKERHSVEREP]XI QYPXMTPIFMRHMRKWXYHMIW ITMXSTIQETTMRKQEXVMGIW GSRGIRXVEXMSRHIXIVQMREXMSR &-%WMQYPEXMSRWSJX[EVIMWHIWGVMFIHMRJYPPMRXLI&-%WMQYPEXMSR 7SJX[EVI,ERHFSSO 8LMWGSQQERHMWRSXEZEMPEFPIMJ&-%WMQYPEXMSRWSJX[EVILEWRSXFIIR MRWXEPPIH Evaluation (ctrl-e) 4VSZMHIWEGGIWWXS&-%IZEPYEXMSRWSJX[EVI[LMGLGYVVIRXP]TVSZMHIW JEGMPMXMIWJSV SZIVPE]TPSXWJSVGSQTEVMWSRSJWIRWSVKVEQW IZEPYEXMSRSJFMRHMRKVEXIW HIVMZEXMSRSJOMRIXMGGSRWXERXW IZEPYEXMSRSJVITSVXTSMRXXEFPIWJSVGSRGIRXVEXMSRQIEWYVIQIRX &-%IZEPYEXMSRWSJX[EVIMWHIWGVMFIHMRXLI&-%IZEPYEXMSR7SJX[EVI,ERHFSSO 8LMWGSQQERHMWRSXEZEMPEFPIMJ&-%IZEPYEXMSRWSJX[EVILEWRSXFIIR MRWXEPPIH Working Tools 8LI[SVOMRKXSSPWEVIEWIXSJVSYXMRIWJSVWXEVXMRKYTERHWLYXXMRK HS[RXLIW]WXIQERHJSVWTIGMEPGPIERMRKTVSGIHYVIW

201 BIACORE Control Software reference 8SYWIE[SVOMRKXSSPGLSSWIXLIXSSPJVSQXLIPMWXERHGPMGOSRStart -RWXVYGXMSRWJSVIEGLWXITMRXLITVSGIHYVIEVITVSZMHIHSRXLIWGVIIR 8LIPEWXXMQIXLILMKLPMKLXIHXSSP[EWYWIHMWHMWTPE]IHEXXLIFSXXSQ SJXLIHMEPSKYIFS\ 7II7IGXMSRJSVEHIWGVMTXMSRSJXLI[SVOMRKXSSPW Test Tools 8LIXIWXXSSPWEVIEWIXSJVSYXMRIWJSVXIWXMRKXLIW]WXIQTIVJSVQERGI 8SYWIEXIWXXSSPGLSSWIXLIXSSPJVSQXLIPMWXERHGPMGOSR Start -RWXVYGXMSRWJSVIEGLWXITMRXLITVSGIHYVIEVITVSZMHIHSRXLIWGVIIR 8LIPEWXXMQIXLILMKLPMKLXIHXSSP[EWYWIHMWHMWTPE]IHEXXLIFSXXSQ SJXLIHMEPSKYIFS\ 7II7IGXMSRJSVEHIWGVMTXMSRSJXLIXIWXXSSPW

202 BIACORE Control Software reference Service Tools 8LIWIVZMGIXSSPWEVIEWIXSJVSYXMRIWJSVW]WXIQQEMRXIRERGI 8SYWIEWIVZMGIXSSPGLSSWIXLIXSSPJVSQXLIPMWXERHGPMGOSRStart -RWXVYGXMSRWJSVIEGLWXITMRXLITVSGIHYVIEVITVSZMHIHSRXLIWGVIIR 8LIPEWXXMQIXLILMKLPMKLXIHXSSP[EWYWIHMWHMWTPE]IHEXXLIFSXXSQ SJXLIHMEPSKYIFS\ 7II7IGXMSRJSVEHIWGVMTXMSRSJXLIWIVZMGIXSSPW Preferences 7IXWTVIJIVIRGIWJSVGSRXVSPWSJX[EVISTIVEXMSRPreferencesWIXXMRKW ETTP]JSVEPPVYRWYRXMPXLIWIXXMRKWEVIGLERKIH 8LIAutosaveJYRGXMSR[MPPWEZIVIWYPXWEXXLIWTIGMJMIHMRXIVZEPHYVMRK EQERYEPVYRSVEJXIVIZIV]G]GPIMREQIXLSHGSRXVSPPIHVYR TVSZMHMRKEWEJIKYEVHEKEMRWXPSWWSJHEXEMRXLIIZIRXSJETS[IV JEMPYVISVGSQTYXIVQEPJYRGXMSR'LIGOXLIGSVVIWTSRHMRKFS\ERH IRXIVEXMQIMRXIVZEPXSXYVRXLIAutosaveJYRGXMSRSRJSVQERYEP ERHSVQIXLSHGSRXVSPPIHVYRW7IXXMRKAutosaveXSSRMW VIGSQQIRHIHTEVXMGYPEVP]JSVYREXXIRHIHQIXLSHGSRXVSPPIHVYRW

203 BIACORE Control Software reference 2SXI -JAutosaveMWXYVRIHSR]SY[MPPFIEWOIHJSVEJMPIREQIMR [LMGLXSWEZIHEXEFIJSVIXLIVYRMWWXEVXIH 9.10 Window menu New window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ascade, Tile Horizontal, Tile vertical 8LIWIGSQQERHWEVIWXERHEVH;MRHS[WGSQQERHWXLEXGSRXVSPXLI [E]WITEVEXI[MRHS[WEVIEVVERKIH6IJIVXSXLI;MRHS[WSRPMRI LIPTJSVEHIWGVMTXMSRSJXLIWIJYRGXMSRW Arrange Icons %VVERKIWMGSRWJSVQMRMQM^IH[MRHS[WMRXLI&-%'36) ETTPMGEXMSR[MRHS[8LMWMWEWXERHEVH;MRHS[WGSQQERHERHLEW RSIJJIGXMJ[MRHS[WEVIRSXQMRMQM^IH <Open window list> 3TIRSVQMRMQM^IH[MRHS[WEVIPMWXIHEXXLIFSXXSQSJXLIWindow QIRY8LIEGXMZI[MRHS[MWGLIGOQEVOIH7IPIGXER]MREGXMZI[MRHS[ XSEGXMZEXIMX

204 BIACORE Control Software reference 9.11 Help menu Help on Application Wizards 3TIRWXLISRPMRILIPTJSVETTPMGEXMSR[M^EVHW About (MWTPE]WMRJSVQEXMSREFSYXXLIZIVWMSRSJ&-%'36)'SRXVSP7SJX[EVI GYVVIRXP]FIMRKYWIHEW[IPPEWXLIMRWXVYQIRX-(ERHGSRJMKYVEXMSR

205 BIACORE Control Software reference

206 System specifications A. System specifications A.1 System performance Measuring range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

207 System specifications A.2 Technical specifications Processing Unit (MQIRWMSRWP [ L 2IX[IMKLX 1EMRWWYTTP]X]TI 1EMRWWYTTP]X]TI 1IQSV]TVSXIGXMSR %QFMIRXXIQTIVEXYVIVERKI %QFMIRXLYQMHMX] 7EJIX]WXERHEVH QQ OK :b:bw[mxglih,^:%qe\ *YWIW Τ6.25Α / 2 Τ3.15Α :b%yxsverki,^:%qe\ *YWIW Τ5.0Α Η PMXLMYQFEGOYTFEXXIV] ' 6, IPIGXVSRMGWHIWMKRIHXS-)' VIUYMVIQIRXW %GSYWXMGRSMWIPIZIP0)5 H&%QIEWYVIHYWMRKWXERHEVH TVSGIHYVIW Detection Unit 0MKLXWSYVGI 0)(0MKLX)QMXXMRK(MSHI ;EZIPIRKXL RQ &ERH[MHXL RQ 6IJVEGXMZIMRHI\VERKI 69 &EWIPMRIHVMJX X]TMGEPP] ±69QMRYXI (EXESYXTYX SRIJPS[GIPP SVQIEWYVIQIRXWW,^ X[SJPS[GIPPW SVQIEWYVIQIRXWW,^ JSYVJPS[GIPPW SVQIEWYVIQIRXWW,^ 2SMWI X]TMGEPP] 69617,^ 69617,^ 8IQTIVEXYVIVERKI 'RSXPS[IVXLER ' FIPS[EQFMIRXXIQTIVEXYVI 8IQTIVEXYVIWXEFMPMX] &IXXIVXLER± 'QMR 8IQTIVEXYVIEGGYVEG] ± 'IRXMVIXIQTVERKI ± 'EX ' -RWXVYQIRXWEVIJMXXIH[MXLSRISJX[STS[IVWYTTP]X]TIW6IJIVXSXLI QEMRWMRTYXTERIPXSHIXIVQMRIXLIWTIGMJMGEXMSRWJSV]SYVMRWXVYQIRX %

208 System specifications 8IQTIVEXYVIIUYMPMFVEXMSRXMQI EJXIV 'WXIT EJXIV 'WXIT X]TMGEPP] QMRYXIW X]TMGEPP] QMRYXIW Pumps *PS[VEXIVERKI -RGVIQIRXW PQMR PQMRYWIVGSRXVSPPIH 7XITWSJ PQMR *PS[VEXIEGGYVEG] ± PQMRPMUYMHEX ' *PS[VEXITVIGMWMSR PQMRPMUYMHEX ' 4VIWWYVIVERKI 14E 8YFMRK -RPIXFSXLTYQTW 4))/MHQQ 3YXPIXGSRXMRYSYWJPS[ 4))/MHQQ 3YXPIXEYXSWEQTPIV 8IJ^IPMHQQ Autosampler (MWTIRWMRKZSPYQIVERKI PMRGPYHMRKEMVWIKQIRX -RGVIQIRXW 7XITWSJ P %GGYVEG] HIZMEXMSRJSVZSPYQIW P 4VIGMWMSR P ': P ': 'EVV]SZIVFIX[IIRZMEPW X]TMGEPP] P[MXLSYXWITXYQ X]TMGEPP] P[MXLWITXYQ 7EQTPIGETEGMX]TIVVEGO 1MGVSXMXVITPEXI 6EGOX]TI% 6EGOX]TI& 6EGOX]TI' 1E\MQYQZMEPLIMKLX 3TXMSREPXLIVQSWXEXXMRK QP[IPP ZMEPWHMEQQQQP ZMEPWHMEQQQQP ZMEPWHMEQQQQP ZMEPWHMEQQQQP ZMEPWHMEQQQ )TTIRHSVJJX]TIQP QQ šgsrrigxmsrw QE\MQYQMRPIXTVIWWYVI14E 2SXI 3RP]WSQIFVERHWSJQMGVSXMXVITPEXIWEVIGSQTEXMFPI[MXLXLI EYXSWEQTPIV'EPP]SYV&MEGSVIXIGLRMGEPWYTTSVX VITVIWIRXEXMZIJSVQSVIMRJSVQEXMSR %

209 System specifications Integrated -Fluidic Cartridge (IFC) *PS[GIPPW RYQFIV :EPZIW 4RIYQEXMGQIQFVERIZEPZIW -RNIGXMSRZSPYQIXSJPS[GIPP PWXITWSJ P 'EVV]SZIVJVSQ TVIZMSYWMRNIGXMSR 7EQTPIPSSTGETEGMX] P QE\MQYQMRNIGXMSRZSPYQI P 'LERRIPERHZEPZIGSRJMKYVEXMSREPPS[WEGGIWWXSJPS[GIPP SVJVSQPSSTSVMRNIGXMSRTSVX *SVMRNIGXMSRZSPYQIWERHWEQTPIGSRWYQTXMSRWII7IGXMSR Sample recovery :SPYQI P" PJSVFIWXTVIGMWMSR (MPYXMSRSJVIGSZIVIHWEQTPI WII8IGLRMGEPHIWGVMTXMSRFIPS[ 'EVV]SZIV JVSQTVIZMSYWVIGSZIVIHWEQTPI Fractions :SPYQI 1E\MQYQRYQFIV P" PJSVFIWXTVIGMWMSR YTXSPMQMXIHF]RYQFIVSJZMEPW MRSRIEYXSWEQTPIVVEGO %

210 System specifications A.3 Technical description 8LMWWIGXMSRHIWGVMFIWXIGLRMGEPEWTIGXWSJ&-%'36)8LI MRJSVQEXMSRLIVIMWRSXVIUYMVIHJSVRSVQEPMRWXVYQIRXYWI Fuses )\XIVREPJYWIWEVIPSGEXIHMRXLIQEMRWGSRRIGXSVSRXLIPIJXWMHISJXLI MRWXVYQIRXERHQE]FIVITPEGIHF]XLIYWIV*SVVIUYMVIHJYWIVEXMRKW WII7TIGMJMGEXMSRWEFSZI -RXIVREPJYWIWEVIPSGEXIHMRWMHIXLITVSGIWWMRKYRMX8LIWIJYWIWQE] SRP]FIVITPEGIHF]ETTVSZIH&MEGSVIWIVZMGITIVWSRRIP LED display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etection unit 8LISTXMGEPHIXIGXMSRYRMXYWIWJSYVWITEVEXIPMRIEVEVVE]WSJPMKLX WIRWMXMZIHMSHIWSRIJSVIEGLJPS[GIPP8LIWTEGMRKFIX[IIRHMSHIW GSVVIWTSRHWXSEHMJJIVIRGISJETTVS\MQEXIP] MRMRGMHIRXPMKLX ERKPI'SQTYXIVEPKSVMXLQWEYXSQEXMGEPP]GEPGYPEXIXLIERKPIEX[LMGL QMRMQYQVIJPIGXMSRSGGYVWXLI746ERKPI[MXLEVIWSPYXMSRSJEFSYX %

211 System specifications 'SRWXERXXIQTIVEXYVIEXXLIHIXIGXSVMWQEMRXEMRIHF]GSRXMRYSYW GMVGYPEXMSRSJEMVMREXLIVQEPP]MRWYPEXIHFS\8LIXIQTIVEXYVIMW VIKYPEXIHF]4IPXMIVIPIQIRXW[LMGLEVIGSRXVSPPIHF]XIQTIVEXYVI WIRWSVWMRXLISTXMGEPYRMXGPSWIXSXLIJPS[GIPPWERHMRXLIGMVGYPEXIH EMV Flow cells 8LIZSPYQISJIEGLJPS[GIPPMWEFSYX P8LIXSXEPWIRWSVGLMT EVIEMRIEGLJPS[GIPPMWQQ 8LIJSGYWIHMRGMHIRXPMKLXMPPYQMREXIWEREVIEQIEWYVMRKEFSYX QQ EPSRKXLIJPS[GIPPMRXLIHMVIGXMSRSJJPS[8LIHIXIGXSVEVVE]VIEHW VIJPIGXIHPMKLXJVSQE[MHXLSJQQEGVSWWXLIJPS[GIPP 8LIHIEHZSPYQIFIX[IIRXLIMRPIXZEPZIERHXLIJPS[GIPPMW P -RWIVMEPMRNIGXMSRQSHIXLIHIEHZSPYQIMW PXSJPS[GIPPERH EREHHMXMSREP PTIVWYFWIUYIRXJPS[GIPP8LMWKMZIWEXMQIHIPE] MRWIRWSVKVEQWFIX[IIRJPS[GIPPWGSVVIWTSRHMRKXSEFSYXWEX PQMRWIIEPWS7IGXMSR Pumps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utosampler 8LIXYFMRKFIX[IIRRIIHPIERHTYQT-(!QQLEWEZSPYQISJ PERHEGXWEWEGSRXEMRIVJSVEWTMVEXIHPMUYMH8LIQE\MQYQ EWTMVEXMRKZSPYQIMW PMRGPYHMRKEMVWIKQIRXW8LI VIGSQQIRHIHVERKIMW P8LI&-%'36)GSRXVSPWSJX[EVI GSQQERHWIKDILUTETRANSFERMIXERHXLIZEVMSYWINJECTWEVI HIWMKRIHXSQEMRXEMREMVWIKQIRXWSJEFSYX P %

212 System specifications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hermo-racks 8LVIIX]TIWSJXLIVQSVEGOWEVIEZEMPEFPI[MXLZMEPTSWMXMSRWEW MPPYWXVEXIHFIPS[2SXIXLEXVS[JSRXLIVQSVEGO%LEWSRP] TSWMXMSRW 7YTTSVXIHQMGVSXMXVITPEXIWLEZIVS[WELSJTSWMXMSRWTIVVS[ IFC 0MUYMHJPS[MWHMVIGXIHXLVSYKLXLI-*'GLERRIPWF]TRIYQEXMGQMGVS TMRGLZEPZIWSTIVEXIHF]EMVTVIWWYVIJVSQERMRXIVREPGSQTVIWWSV %

213 System specifications 'LERRIPWJVSQXLIEMVFYWXSXLI-*'ZEPZIWEVIJMPPIH[MXLKP]GIVSPXS TVIZIRXHMJJYWMSRSJEMVJVSQXLIEMVFYWMRXSXLI-*'GLERRIPW 8LI-*'GLERRIPWEVIQSYPHIHMRWMPMGSRITPEWXMG8LIWQEPPIWXGLERRIP GVSWWWIGXMSREPEVIEMWQQ WSXLEXEZSPYQIJPS[VEXISJ PQMRGSVVIWTSRHWXSEPMRIEVJPS[VEXISJ GQW Recovery and fractions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utomatic preparation and finishing routines %YXSQEXMGTVITEVEXMSRERHJMRMWLMRKVSYXMRIW[EWLXLIPMUYMHLERHPMRK W]WXIQEXXLIFIKMRRMRKERHIRHSJQIXLSHWQIXLSHG]GPIWQERYEPP] GSRXVSPPIHWIRWSVKVEQWERHMRNIGXMSRW8LIWYQQEV]FIPS[HSIWRSX MRGPYHIGSQQERHWTIGMJMGSTIVEXMSRWWYGLEWEWTMVEXMRKWEQTPIFIJSVI ERMRNIGXMSRERHEXTRACLEANDIPNEEDLEFRACTIONSERHRECOVER STIVEXMSRW %

214 System specifications % 1IXLSHGSRXVSP 7XEVXQIXLSH MAIN *VIWLFYJJIVMWHIPMZIVIHXS-*'ERH[EWL WXEXMSR8LIJPS[MWWIXXS PQMRERHEPP-*' ZEPZIWEVISTIRIH 7XEVXQIXLSHG]GPI APROG 8LIGSRXMRYSYWJPS[TYQTMWJMPPIHERHXLI WEQTPIPMRIMRXLI-*'MWTVMQIH[MXLJVIWL FYJJIV*PS[VEXIERHJPS[TEXLEVIWIX 7XEVXMRNIGXMSR 4YQTWEVIVIJMPPIHMJVIUYMVIHEWIWXMQEXIH JVSQFYJJIVGSRWYQTXMSRHYVMRKXLIMRNIGXMSR )RHMRNIGXMSR 8LIRIIHPIERH-*'WEQTPIPMRIEVI[EWLIH )RHQIXLSHG]GPI APROG 8LI-*'WEQTPIPMRIERHJPS[GIPPWEVI[EWLIH [MXLFYJJIV %R]JPS[TEXLGLERKIWVIUYIWXIH[MXL FLOWPATHMRXLIAPROGEVIVIWXSVIH &YJJIVJPS[MWWIXXS PQMR )RHQIXLSH MAIN 8LI-*'WEQTPIPMRIERHJPS[GIPPWEVI[EWLIH [MXLFYJJIV*PS[MWWIXXS^IVSERHEPP-*' ZEPZIWEVISTIRIH 1ERYEPGSRXVSP 7XEVXWIRWSVKVEQ %WLSVXTYPWISJFYJJIVMWTYQTIHXLVSYKLEPP JPS[GIPPW 8LIRIIHPIMW[EWLIH 7XEVXMRNIGXMSR 4YQTWEVIVIJMPPIHMJVIUYMVIHEWIWXMQEXIH JVSQFYJJIVGSRWYQTXMSRHYVMRKXLIMRNIGXMSR )RHMRNIGXMSR 8LIRIIHPIERH-*'WEQTPIPMRIEVI[EWLIH )RHWIRWSVKVEQ %PP-*'ZEPZIWEVISTIRIH *PS[MWWIXXS^IVSERHTYQTWEVIVIWIXXS WXEVXMRKTSWMXMSR -*'GLERRIPWEVI[EWLIHMJERMRNIGXMSRLEW FIIRTIVJSVQIH -QTSVXERXRSXIW %YXSQEXMGVSYXMRIWEVIOITXXSEQMRMQYQYRHIVQERYEPGSRXVSPXS VIHYGIXLIXMQIWXEOIRJSVWIRWSVKVEQWXEVXIRHERHMRNIGXMSRW-XMWXLI YWIVWVIWTSRWMFMPMX]XSIRWYVIXLEXETTVSTVMEXI[EWLGSQQERHWEVI I\IGYXIHFIJSVIWEQTPIMRNIGXMSRYRHIVQERYEPGSRXVSP6IQIQFIVMR TEVXMGYPEVXSTIVJSVQEWash[MXLRecovery STXMSRFIJSVIMRNIGXMSRW [LMGLYWIFractionsSVRecover-JXLMWMWRSXHSRIXLIVIGSZIV]PMRI [MPPRSXFIIQTXMIHERHXLIWEQTPIHIPMZIVIVHXSXLIVIGSZIV]GYT[MPP FIHMPYXIH[MXLYTXS PFYJJIV

215 System specifications %

216 Application guidelines B. Application guidelines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eneral considerations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

217 Application guidelines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ypes of application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

218 Application guidelines B.3 Manual versus automated operation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pecial features of BIACORE LIEVXMJEGXWHMWGYWWIHFVMIJP]EFSZIGERFIGMVGYQZIRXIHSV GSQTIRWEXIHF]WYMXEFPIGSRXVSPI\TIVMQIRXWMREPP&-%MRWXVYQIRXW,S[IZIVQYPXMGLERRIPEREP]WMWMR&-%'36)SJJIVWWTIGMEP JEGMPMXMIWJSVMRPMRIGSRXVSPI\TIVMQIRXW B.4.1 Gradient surfaces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

219 Application guidelines XLIJSVQIVZEVMIW[MXLXLIPMKERHHIRWMX][LMPIXLIPEXXIVQE]FI XEOIREWGSRWXERXWMRGIMXMWMRXLIVERKISJEJI[LYRHVIH69 'SQTEVMWSRWQEHIJVSQWMRKPIGLERRIPQIEWYVIQIRXWEVISJXIR HMJJMGYPXFIGEYWISJQMRSVZEVMEXMSRWMRXLIVIWTSRWIFIX[IIR WITEVEXIWEQTPIMRNIGXMSRW8LIYWISJEKVEHMIRXWYVJEGIXSMQTVSZI XLIHIXIGXMSRPMQMXMWHIWGVMFIHMR%TTPMGEXMSR2SXIJVSQ &MEGSVI%& B.4.2 In-line reference cell 8LIWYVJEGIMRSRIJPS[GIPPGERTVSZMHIEVIJIVIRGIGIPPMR QYPXMGLERRIPEREP]WIW8LMWVIJIVIRGIQE]FIEFPEROGIPPJSV GSVVIGXMSRSJFYPOVIJVEGXMZIMRHI\IJJIGXWXIQTIVEXYVIHVMJXRSR WTIGMJMGFMRHMRKIXGSVElJYRGXMSREPFPEROzEMQIHXSIPMQMREXIE TEVXMGYPEVJIEXYVISJXLIFMSQSPIGYPEVMRXIVEGXMSRFIMRKWXYHMIH3R PMRIVIJIVIRGIWYFXVEGXMSRTVSZMHIWHMVIGXHMWTPE]SJXLIFPERO GSVVIGXIHVIWYPXW7II7IGXMSR&JSVTVEGXMGEPGSRWMHIVEXMSRW GSRGIVRMRKVIJIVIRGIGIPPW B.5 Experimental considerations B.5.1 Sample preparation -RTVITEVMRKXLIEREP]XIWEQTPIXV]XSVIHYGIXLIHMJJIVIRGIFIX[IIR WEQTPIERHVYRRMRKFYJJIVGSQTSWMXMSREWQYGLEWTSWWMFPI (MJJIVIRGIWMRVIJVEGXMZIMRHI\KMZIFYPOIJJIGXW[LMGLGERSFWGYVI VETMHFMRHMRKIZIRXW(MJJIVIRGIWMRT,ERHMSRMGWXVIRKXLGERKMZI QEXVM\IJJIGXW[LMGLQE]XEOIJVSQWIGSRHWXSLSYVWXSWXEFMPM^I8LI I\XIRXSJQEXVM\IJJIGXWMJER][MPPZEV][MXLXLIREXYVIERHEQSYRX SJXLIMQQSFMPM^IHPMKERH -JXLIWEQTPIGSRXEMRWEHHMXMZIWWYGLEWHIXIVKIRXWMRORS[R GSRGIRXVEXMSREHHXLIWEQIGSRGIRXVEXMSRXSXLIVYRRMRKFYJJIV -JHMJJIVIRGIWMRGSQTSWMXMSRFIX[IIRWEQTPIERHVYRRMRKFYJJIV GERRSXFIEZSMHIHERHQEXVM\IJJIGXWEVIWYWTIGXIHWIIREWEVIPEXMZIP] PSRKXIVQHVMJXMRXLIWMKREPMRGPYHIEWAITGSQQERHWMRXLIAPROG XSEPPS[WMKREPWXEFMPM^EXMSR=SYQE]EPWSYWIEVIJIVIRGIGIPPXS GSQTIRWEXIJSVXLIQEXVM\IJJIGXW-HIEPP]XLIVIJIVIRGIGIPPWLSYPH GSRXEMREGSRXVSPPMKERHMQQSFMPM^IHEXXLIWEQIPIZIPEWXLIXVYI PMKERH-XMWSXLIV[MWIHMJJMGYPXXSGSQTIRWEXIJYPP]JSVQEXVM\IJJIGXW YWMRKEVIJIVIRGIGIPPWMRGIXLII\XIRXERHVEXISJQEXVM\IJJIGXWEVI WIRWMXMZIXSXLIEQSYRXSJMQQSFMPM^IHQEXIVMEP *SVWEQTPIWXLEX[MPPFIOITXJSVVIPEXMZIP]PSRKTIVMSHW"LMRXLI EYXSWEQTPIVYWIGETTIHZMEPWXSIPMQMREXIFSXLWEQTPIIZETSVEXMSR ERHGSRHIRWEXMSRMRWMHIXLIZMEPWTEVXMGYPEVP]MJXLIEYXSWEQTPIVVEGOW EVIGSSPIH &

220 Application guidelines B.5.2 Ligand immobilization %QSYRXSJMQQSFMPM^IHPMKERH 8LISTXMQEPPIZIPSJMQQSFMPM^IHPMKERHHITIRHWSRXLITYVTSWISJXLI EREP]WMW-RKIRIVEPYWIXLIPS[IWXVIEWSREFPIPIZIPJSVOMRIXMGEREP]WIW XSQMRMQM^IXLIIJJIGXSJQEWWXVERWJIVPMQMXEXMSRW*SVGSRGIRXVEXMSR QIEWYVIQIRXXLISTXMQEPPMKERHPIZIPMWSJXIRVIPEXIHXSXLIVIUYMVIH H]REQMGVERKISJXLIEWWE]7II&-%ETTPMGEXMSRW,ERHFSSO'LETXIVW ERHJSVJYVXLIVHIXEMPW 4VITEVMRKKVEHMIRXWYVJEGIW +VEHMIRXWYVJEGIWJSVQYPXMGLERRIPETTPMGEXMSRWGERFIGSRZIRMIRXP] TVITEVIHF]YWMRKZEVMEFPIGSRXEGXXMQIWJSVIMXLIVXLIWYVJEGI EGXMZEXMSRWXITSVXLIPMKERHFMRHMRKWXIT8LIGLSMGISJ[LMGLWXITXS ZEV][MPPHITIRHSRXLIGLEVEGXIVMWXMGWSJXLIMQQSFMPM^EXMSR TVSGIHYVI[MXLWXERHEVHEQMRIGSYTPMRKZEV]MRKXLIWYVJEGI EGXMZEXMSRXMQIMWKIRIVEPP]VIGSQQIRHIHJSVGSRXVSPPMRKXLIEQSYRX SJMQQSFMPM^IHPMKERH %XPS[JPS[VEXIWWEQTPIHITPIXMSRFIX[IIRJPS[GIPPWGERFIGSQI WMKRMJMGERX*PS[VEXIWSJ PQMRSVLMKLIVHYVMRKPMKERH MQQSFMPM^EXMSREVIVIGSQQIRHIHXSQMRMQM^IWEQTPIHITPIXMSR %KVEHMIRXWYVJEGIGVIEXIHYWMRKZEVMEFPIGSRXEGXXMQIWMRXLI MQQSFMPM^EXMSRTVSGIHYVILEWXLILMKLIWXPMKERHGSRGIRXVEXMSRMRXLI JMVWXJPS[GIPP-XQE]WIIQXLEXXLMW[SYPHEKKVEZEXIXLITVSFPIQSJ WEQTPIHITPIXMSRJSVOMRIXMGQIEWYVIQIRXW[LIVIEGGYVEXIORS[PIHKI SJXLIEREP]XIGSRGIRXVEXMSRMWVIUYMVIH,S[IZIVOMRIXMG QIEWYVIQIRXWYWILMKLJPS[VEXIWERHPS[PMKERHPIZIPWWSXLEXWEQTPI HITPIXMSRMWVEVIP]ETVSFPIQ-J]SYHS[ERXXSGVIEXIEKVEHMIRX WYVJEGI[MXLXLIPS[IWXMQQSFMPM^EXMSRPIZIPMRXLIJMVWXGIPPTIVJSVQ XLIMQQSFMPM^EXMSRMRIEGLJPS[GIPPWITEVEXIP] -QQSFMPM^MRKHMJJIVIRXPMKERHWJSVQYPXMGLERRIPETTPMGEXMSRW 1YPXMGLERRIPETTPMGEXMSRWQE]FIHIWMKRIH[MXLHMJJIVIRXPMKERHW MQQSFMPM^IHMRHMJJIVIRXJPS[GIPPWJSVQSRMXSVMRKQYPXMTPIEREP]XIWMR GSQTPI\WEQTPIW-RTVITEVMRKWYVJEGIWSJXLMWOMRHMXMWIWWIRXMEPXS EZSMHGSRXEQMREXMSRSJXLIWYVJEGISJSRIJPS[GIPP[MXLPMKERHJVSQ ERSXLIVJPS[GIPP8LIWYVJEGIWWLSYPHFITVITEVIHMRWITEVEXIG]GPIW IMXLIVQERYEPVYRWSVWITEVEXIAPROGWMREQIXLSHXLIEYXSQEXMG GPIERMRKTVSGIHYVIWFIX[IIRG]GPIW[MPPIPMQMREXIGEVV]SZIV8LI ETTEVIRXP]GSRZIRMIRXXIGLRMUYISJEGXMZEXMRKEPPWYVJEGIW[MXLE WMRKPIQYPXMGLERRIPMRNIGXMSRERHXLIRMRNIGXMRKXLIHMJJIVIRXPMKERHW WITEVEXIP]MWRSXVIGSQQIRHIHWMRGIXLIVIMWWMKRMJMGERXVMWOJSV GEVV]SZIVSJPMKERHWFIX[IIRJPS[GIPPW-REHHMXMSRXLIHIPE] FIX[IIREGXMZEXMSRERHGSYTPMRKWLSYPHFIOITXEWWLSVXEWTSWWMFPI WMRGIXLI2,7IWXIVWSREWYVJEGIEGXMZEXIH[MXL2,7)('FVIEO HS[RSZIVXMQI &

221 Application guidelines B.5.3 Flow rate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

222 Application guidelines Response (RU) Rise time Fall time time (s) Figure 5-1. Illustration of sample dispersion. The values indicate the extent of dispersion that can be expected for an injection of sucrose at different flow rates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

223 Application guidelines 7EQTPIHITPIXMSRMWVEVIP]ETVSFPIQMRWMRKPIGLERRIPETTPMGEXMSRW FYXQE]FIWMKRMJMGERXMRQYPXMGLERRIPEREP]WIW[LIVIXLIWEQTPI TEWWIWSZIVWIZIVEPJPS[GIPPWMRWIVMIW[MXLXLIWEQIPMKERH MQQSFMPM^IH-RKIRIVEPYWIJPS[VEXIWEFSZI PQMRJSV QYPXMGLERRIPETTPMGEXMSRWXSQMRMQM^IXLMWIJJIGX7EQTIHITPIXMSRMW TVSTSVXMSREPXS ( flow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rimeSVFlushXSGPIEVEMVFYFFPIWJVSQXLIW]WXIQ EJXIVTVSPSRKIHSTIVEXMSREXPS[JPS[VEXIW8LITVSFPIQMWPIWW QEVOIHEXLMKLIVJPS[VEXIW7II7IGXMSRJSVJYVXLIVHIXEMPWSJ LS[XSQMRMQM^ITVSFPIQW[MXLEMVFYFFPIW B.5.4 Injection techniques &-%'36)SJJIVWEVERKISJETTPMGEXMSRSVMIRXIHMRNIGXMSR GSQQERHWEWHIWGVMFIHMR7IGXMSR8LIWIGSQQERHWHMJJIVMR GETEGMX]BIGINJECTJYRGXMSREPMX]COINJECTERHXLIFEPERGISJ TVITEVEXMSRXMQIERHMRNIGXMSRTIVJSVQERGIQUICKINJECTKINJECT 9WIXLIGSQQERHXLEXFIWXWYMXWXS]SYVETTPMGEXMSR &

224 Application guidelines 8MQIGSRXVSPPIHMRNIGXMSRW (YVMRKEQIXLSHMRNIGXMSRWEVITVITEVIHEWWSSREWXLIEYXSWEQTPIVMW EZEMPEFPIIZIRMJXLIMRNIGXMSRMWXMQIHXSWXEVXEXEPEXIVTSMRX*SV I\EQTPIMRXLI%463+FIPS[XLIMRNIGXMSR[MPPFITVITEVIHEWWSSR EWXLI%463+WXEVXWFYX[MPPFIWXEVXIHEXXMQIQMRYXIW DEFINE APROG analyse FLOW 30 10:00 INJECT r1a1 30 END *SVETTPMGEXMSRWVIUYMVMRKLMKLMRNIGXMSRTIVJSVQERGIPS[WEQTPI HMWTIVWMSRXLMWGERFIEHMWEHZERXEKIWMRGIXLIWEQTPI[EMXWMRXLI EYXSWEQTPIVXYFMRKJSVEFSYXQMRYXIWERHWEQTPIHMWTIVWMSR[MPP MRGVIEWI8SEZSMHXLMWTVSKVEQEXMQIGSRXVSPPIHEYXSWEQTPIVIZIRX IK[EWLXLI-*'GLERRIPWNYWXFIJSVIXLIMRNIGXMSR DEFINE APROG analyse FLOW 30 10:00 WASH s INJECT r1a1 30 END ;MXLXLMWGSRWXVYGXMSRXLIMRNIGXMSR[MPPRSXFITVITEVIHYRXMPXLI [EWLMWGSQTPIXI8MQIGSRXVSPSJXLIEGXYEPMRNIGXMSRWXEVXMWPIWW TVIGMWIXLIMRNIGXMSR[MPPWXEVXEXQMRYXIWXMQIJSV[EWLXMQI JSVTVITEVMRKXLIMRNIGXFYXXLIVIWMHIRGIXMQISJXLIWEQTPIMRXLI EYXSWEQTPIVXYFMRKMWOITXEWWLSVXEWTSWWMFPI9WIXLIIZIRXPSKXS HIXIVQMRIXLIEGXYEPXMQISJMRNIGXMSRWXEVX 2SXI %XMQIHRPOINTGSQQERHGERRSXFIYWIHJSVXLMWTYVTSWI WMRGIRPOINTHSIWRSXSGGYT]XLIEYXSWEQTPIVERHXLI MRNIGXMSR[MPPFITVITEVIHEXXLIFIKMRRMRKSJXLIAPROG B.5.5 Using a reference cell 1YPXMGLERRIPHIXIGXMSRMR&-%'36)EPPS[WSRIGIPPXSFIYWIH EWERMRPMRIVIJIVIRGIXSGSQTIRWEXIJSVFYPOVIJVEGXMZIMRHI\IJJIGXW RSRWTIGMJMGFMRHMRKIXG-RHIWMKRMRKETTPMGEXMSRW[MXLEVIJIVIRGIGIPP XSGSQTIRWEXIJSVEVXMJEGXWFIEVXLIJSPPS[MRKGSRWMHIVEXMSRWMRQMRH 9WIEGIPPEHNEGIRXXSXLIQIEWYVMRKGIPPEWEVIJIVIRGI%HNYWXXLI XMQIE\MWSJXLIVIJIVIRGIWIRWSVKVEQXSGSMRGMHI[MXLXLIWEQTPI GIPPMR&-%IZEPYEXMSR8LIHIPE]FIX[IIREHNEGIRXGIPPWMRXLIVE[ HEXEGSVVIWTSRHWXS PPMUYMHWIGSRHWEX PQMR -JTSWWMFPIMQQSFMPM^IElRIYXVEPzQSPIGYPIMRXLIVIJIVIRGIGIPPEX XLIWEQIPIZIPEWMRXLIWEQTPIGIPP'LSSWIEQSPIGYPI[MXLWMQMPEV &

225 Application guidelines QSPIGYPEV[IMKLXERHGLEVKIGLEVEGXIVMWXMGWXSXLIPMKERH8LMW[MPP LIPTMRYWMRKXLIVIJIVIRGIGIPPXSGSQTIRWEXIJSVQEXVM\IJJIGXW B.5.6 Reducing carry-over 8LIXIVQlGEVV]SZIVzMRGPYHIWERYQFIVSVTSXIRXMEPEVXMJEGXW XVERWJIVSJWEQTPIFIX[IIRWYGGIWWMZIMRNIGXMSRW XVERWJIVSJWEQTPISVFYJJIVFIX[IIRHMJJIVIRXZMEPW XVERWJIVSJWEQTPIFIX[IIRJPS[GIPPW MRGSQTPIXIVIKIRIVEXMSRSJXLIWYVJEGI PIEOEKISJGSRXEQMRERXWJVSQWYVJEGIWMRXLIPMUYMHLERHPMRKW]WXIQ -RKIRIVEPXLIJMVWXJI[EREP]WMWG]GPIW[MXLERI[WYVJEGIEVIQSVI WIRWMXMZIXSGEVV]SZIVERHlQIQSV]IJJIGXWzXLERPEXIVG]GPIW[LIR XLIWYVJEGILEWWXEFMPM^IH-XMWSJXIREHZERXEKISYWXSVYRSRISVQSVI FPEROEREP]WMWG]GPIWGSQTPIXIG]GPIWMRGPYHMRKVIKIRIVEXMSRr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

226 Application guidelines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igsqqerherhxlirywiewaitgsqqerhxsepps[fyjjivjps[ YRXMPXLIFEWIPMRIMWWXEFPIEJXIVI\TSWYVIXSXLIGSQTSYRHMRUYIWXMSR 6YRDesorbEXPIEWXIZIV][IIOMJFMSQSPIGYPIWJVSQWEQTPIWEHWSVF XSXLI-*'[EPPWERHMRXIVJIVI[MXLEREP]WIW -RGPIEVMRKGEVV]SZIVJVSQXLI-*'GLERRIPWXMQIMWSJXIRQSVI MQTSVXERXXLERXLI[EWLMRKZSPYQIWMRGIHMWWSPZIHQEXIVMEPWPIEGL SYXEXEGLEVEGXIVMWXMGVEXI%YWIJYPXIWXSJGEVV]SZIVMWEWSGEPPIH ^IVSMRNIGXMSRMIEREYXSWEQTPIVXVERWJIVERHMRNIGXMSRYWMRK FYJJIVMRWXIEHSJWEQTPI DEFINE APROG zero_inject FLOW 30 WASH n!wash needle TRANSFER B r1a1 450! 'B' is buffer INJECT r1a1 30 END &

227 Application guidelines 9WIERI[KPEWWZMEP[MXLEWITXYQJSVXLIXVERWJIVERHMRNIGXTSWMXMSR ;MXLTL]WMSPSKMGEPWEPXGSRGIRXVEXMSRMRXLIFYJJIVEVIWTSRWISJ EFSYX69JVSQE^IVSMRNIGXMSRMWRSXYRGSQQSRERHGERFI XVEGIHXSWEPXHITSWMXWSRXLIRIIHPI B.5.7 Temperature changes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analyse FLOW 25 10:00 WASH s! 10 minutes stabilization... END &

228 Application guidelines DEFINE LOOP temp_series LPARAM %degrees TIMES END MAIN! Start method at 5 C or below LOOP temp_series STEP TEMP %degrees PRIME NORMALIZE r2f1! glycerol position APROG analyse ENDLOOP END B.5.8 Regeneration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

229 Application guidelines B.5.9 Washing the liquid handling system (YVMRKRSVQEPSTIVEXMSRXLIPMUYMHLERHPMRKW]WXIQMW[EWLIH EYXSQEXMGEPP]EXXLIIRHSJIEGLG]GPI[LIVIXLIEYXSWEQTPIVMWYWIH *SVLMKLTIVJSVQERGI[SVOXLIVIEVIERYQFIVSJEHHMXMSREP[EWLMRK JEGMPMXMIW WASH n[ewliwxlieyxsweqtpivriihpi WASH s[ewliwxlimrnigxmsrglerripmrxli-*' WASH l[ewliwxliweqtpipsstsrp]vipizerx[mxlcoinject WASH f[ewliwxlijps[gippeviemrxli-*' WASH i[ewliwxlimrnigxmsrglerripweqtpipssterhjps[gippevie MRXLI-*'8LITVSGIWWSJ[EWLMRKXLIJPS[GIPPEVIEQE]GEYWI FEWIPMRIHMWXYVFERGIW FLUSHERHPRIME[EWLXLITYQTXYFMRKERH-*'GLERRIPWEXELMKL JPS[VEXI8LMWGERLIPTXSVIQSZIEMVFYFFPIWXVETTIHMRXLIW]WXIQ EXTRACLEANTIVJSVQWEREHHMXMSREP[EWLSJXLI-*'GLERRIPW MQQIHMEXIP]EJXIVERMRNIGXMSRERHQE]FIYWIHXSQMRMQM^IGEVV] SZIVFIX[IIRMRNIGXMSRW 8LIWASHERHWASHRECGSQQERHWGERFIYWIHIMXLIVMRXLIMAIN FPSGOSVMRERAPROGMREQIXLSHFLUSHERHPRIMEQE]SRP]FIYWIH MRMAINEXTRACLEANQYWXFITPEGIHMQQIHMEXIP]EJXIVERMRNIGXMSR ERHQE]SRP]FIYWIHMRERAPROG &

230 Chemical resistance C. Chemical resistance and recommended solutions 8LMWETTIRHM\GSRXEMRWGLIQMGEPVIWMWXERGIMRJSVQEXMSRJSV&-%'36) GSQTSRIRXWXSKIXLIV[MXLVIGSQQIRHIHFYJJIVWERHWSPYXMSRW JSVETTPMGEXMSRW 7EJIX]QEXIVMEPGLEVXWJSVXLIGLIQMGEPWMRXLI%QMRI'SYTPMRK/MXEVI MRGPYHIH 'LIQMGEPEFFVIZMEXMSRW[LMGLEVIRSXI\TPEMRIHMRXLMWETTIRHM\QE] FIJSYRHMRXLI&MSGLIQMGEPW3VKERMG'SQTSYRHWJSV6IWIEVGLERH (MEKRSWXMG6IEKIRXWGEXEPSKYIJVSQ7MKQE'LIQMGEP'SQTER]7X 0SYMW1S97% Terminology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

231 Chemical resistance C.1 General chemical resistance 8LIXEFPIFIPS[PMWXWWSQISJXLIQSWXGSQQSRWSPYXMSRWYWIHJSV &-%'36)ETTPMGEXMSRWEVIJSYRH8LIXEFPIQE]FIYWIHEWE KYMHIJSVYWEKISJSXLIVWYFWXERGIWWMQMPEVXSXLISRIWPMWXIH 6IWMWXERGIXSGLIQMGEPWEXLMKLIVGSRGIRXVEXMSRWXLERXLSWIWTIGMJMIH MRXLIXEFPIMWRSXKYEVERXIIH 7TEVMRKYWISJSVKERMGEGMHWERHEPGSLSPWMWKIRIVEPP]VIGSQQIRHIH,S[IZIVJSVQMGEGMHEXGSRGIRXVEXMSRWYTXSERHIXLERSPEX GSRGIRXVEXMSRYTXSGERFIYWIHMRWLSVXTYPWIW 7SPYXMSR 'SRGIRXVEXMSR T, 6IWMWXERGI %GIXMGEGMH HSRSXYWI %GIXSRMXVMPI TYPWI %QQSRMYQWYPTLEXI 1 GSRXMRYSYW &IR^EQMHMRI Q1 GSRXMRYSYW &SVEXIFYJJIV Q1 GSRXMRYSYW 'LPSVLI\MHMRI TYPWI 'LPSVSFYXERSP TYPWI 'MXVEXIFYJJIV Q1 GSRXMRYSYW (MXLMSIV]XLVMXSPMR WSHMYQFMGEVFSREXI Q1Q1 TYPWI (1* TYPWI (173 HSRSXYWI (173 TYPWI (173 GSRXMRYSYW )XLERSP )XLERSP TYPWI GSRXMRYSYW )XLERSPEQMRI L]HVSGLPSVMHI WSHMYQL]HVS\MHI 1 GSRXMRYSYW '

232 Chemical resistance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

233 Chemical resistance -WSTVSTERSP HSRSXYWI 0MXLMYQPEYV]P WYPTLEXI GSRXMRYSYW 1)7 Q1 GSRXMRYSYW 1IXLERSP 1IXLERSP HSRSXYWI TYPWI 4LSWTLSVMGEGMH Q1 TYPWI 7SHMYQEGIXEXI FYJJIV Q1 GSRXMRYSYW 7SHMYQGEVFSREXI Q1 GSRXMRYSYW 7SHMYQGLPSVMHI 1 GSRXMRYSYW 7SHMYQL]HVSKIR GEVFSREXI Q1 GSRXMRYSYW 7SHMYQL]HVS\MHI Q1 TYPWI 7SHMYQL]HVS\MHI EGIXSRMXVMPI Q1 TYPWI 7SHMYQL]HVS\MHI 1 TYPWI 7SHMYQL]TSGLPSVMXI TYPWI 7(7MR[EXIV TYPWI 7SHMYQTIVGLPSVEXI 1 TYPWI 9VIE 1 TYPWI '

234 Chemical resistance C.2 Buffer solutions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

235 Chemical resistance 4LSWTLEXI Q1 GSRXMRYSYW 4-4)7 Q1 GSRXMRYSYW Q1 GSRXMRYSYW 8%47 Q1 GSRXMRYSYW 8%473 Q1 GSRXMRYSYW 8)( Q1 GSRXMRYSYW 8)7 Q1 GSRXMRYSYW 86-'-2) Q1 GSRXMRYSYW 86-7,'P%274 Q1Q1 GSRXMRYSYW 86->1%&%7) Q1 GSRXMRYSYW '

236 Chemical resistance C.3 Regeneration solutions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

237 Chemical resistance C.4 Disinfectants 7ERMXM^EXMSRWLSYPHFITIVJSVQIHJSVHIGSRXEQMREXMSREJXIVI\TSWYVI XSMRJIGXERXWERHFIJSVIWXSVMRKXLIW]WXIQXSMRLMFMXFEGXIVMEPKVS[XL HYVMRKWXSVEKI'LSSWIEHMWMRJIGXERX[LMGLLEWXLIFIWXIJJIGXSRXLI X]TISJGSRXEQMREXMRKQEXIVMEPEXLERH&IE[EVIXLEXXLIWERMXM^EXMSR TVSGIHYVIQE]HIWXVS]XLIWIRWSVGLMT 7SPYXMSR 'SRGIRXVEXMSR T, 6IWMWXERGI )XLERSP TYPWI 'LPSVLI\MHMRI TYPWI 'LPSVSFYXERSP TYPWI +IVQEPP-- TYPWI +P]GMRIPMXLMYQ WYPTLEXI 11 TYPWI 7SHMYQL]HVS\MHI 1 TYPWI 7SHMYQL]TSGLPSVMXI TYPWI '

238 Status and event log messages D. Status and event log messages D.1 Status window messages 8LMWWIGXMSRPMWXWQIWWEKIW[LMGLQE]ETTIEVMRXLIWXEXYW[MRHS[ ERH[LMGLVIUYMVII\TPEREXMSRSVYWIVEGXMSR7IPJIZMHIRXQIWWEKIW PMOIMethod runningevirsxpmwxih Not Connected to Instrument 8LMWMRWXERGISJXLIWSJX[EVIMWRSXGSRRIGXIHXSXLIMRWXVYQIRX 8LMWMWRSVQEPJSVWIGSRHSVWYFWIUYIRXMRWXERGIWSJXLIWSJX[EVI ERHJSVMRWXERGIWWXEVXIHJVSQXLI;MRHS[W*MPI1EREKIVWII 7IGXMSR Instrument Offline 8LIWSJX[EVILEWEXXIQTXIHXSGSRRIGXXSXLIMRWXVYQIRXFYXLEW JEMPIH8LMWX]TMGEPP]SGGYVWMJXLIMRWXVYQIRXMWXYVRIHSJJSVXLI GSQQYRMGEXMSRGEFPIMWRSXTVSTIVP]GSRRIGXIH'SRRIGXMSR[MPPFI IWXEFPMWLIHEYXSQEXMGEPP]MJXLIGSQQYRMGEXMSRIVVSVMWVIGXMJMIH Instrument Disconnected 8LIWSJX[EVILEWFIIRPSKMGEPP]HMWGSRRIGXIHJVSQXLIMRWXVYQIRX ERH[MPPRSXXV]XSIWXEFPMWLERI[GSRRIGXMSR8LMWMWRSVQEPP] GEYWIHF]TVIWWMRK'XVP&VIEO%FSVXHYVMRKWIPJHMEKRSWXMGWSVF] ERMRZEPMHLEVH[EVIGSRJMKYVEXMSRMR4'SVMRWXVYQIRX8LMWQIWWEKI MWRSVQEPP]TVIGIHIHF]ERI\TPEREXSV]QIWWEKIMREQIWWEKIFS\ Performing Self Diagnostics 8LMWQIWWEKIETTIEVW[LIRXLIGSRXVSPWSJX[EVIMWWXEVXIHERH IWXEFPMWLIWGSRRIGXMSR[MXLXLIMRWXVYQIRX Instrument Self Diagnostics Failed :EPMHEXMSRSJMRWXVYQIRXWSJX[EVIERHGSRJMKYVEXMSRSVXLIVIWYPXMRK EYXSQEXMGTVSKVEQPSEHMRKJEMPIH8LMWQE]FIGEYWIHF]MRWXVYQIRX WXEVXYTJEMPYVIERHSVMRZEPMHMRWXVYQIRXWSJX[EVI8LIWSJX[EVILEW RSGSRRIGXMSRXSXLIMRWXVYQIRX Initiating System ;LIRGSRRIGXMSRLEWFIIRIWXEFPMWLIHERHMRWXVYQIRXWSJX[EVI ZEPMHEXIHXLIWSJX[EVIMWMRMXMEPMWIHEGGSVHMRKXSXLIGYVVIRXP] GSRRIGXIHMRWXVYQIRX8LMWMRMXMEPMWEXMSRMRGPYHIWZIVMJ]MRKXLEXXLI LEVH[EVIGSRJMKYVEXMSRMRXLI4'GSVVIWTSRHW[MXLMRWXVYQIRX GSRJMKYVEXMSR Initialization failed 8LIGSRXVSPWSJX[EVIMRMXMEPM^EXMSRJEMPIH8LIWSJX[EVI[MPPFI GSRRIGXIHXSXLIMRWXVYQIRXFYXW]WXIQGSRXVSPQE]FIYRVIPMEFPI 8LITVSFPIQWLSYPHFIMHIRXMJMIHERHGSVVIGXIHFIJSVIXLI MRWXVYQIRXMWYWIH8LIQIWWEKI[MPPVIQEMRHMWTPE]IHYRXMPXLI WSJX[EVIMWGPSWIHSVEGSRRIGXMSRMWWYGGIWWJYPP]MRMXMEPM^IH (

239 Status and event log messages D.2 Event log messages 8LMWWIGXMSRI\TPEMRWXLIQIWWEKIW[LMGLQE]FIIRXIVIHMRXLIIZIRX PSK1IWWEKIWEVIPMWXIHMREPTLEFIXMGEPSVHIV Connection lost -RHMGEXIWXLEXXLIVYR[EWXIVQMREXIHFIGEYWIGSRRIGXMSRFIX[IIR XLI4'ERHXLIMRWXVYQIRX[EWPSWX Data collection 6IGSVHWXLIHEXEGSPPIGXMSRVEXIMR,^EXXLIFIKMRRMRKSJXLI QIXLSHG]GPISVQERYEPWIRWSVKVEQERH[LIREGLERKIMRHEXE GSPPIGXMSRVEXIMWVIUYIWXIH Data reduction in sensorgram -RHMGEXIWXLEXHEXEVIHYGXMSRLEWSGGYVVIHMRXLIWIRWSVKVEQJVSQ XLIWTIGMJMIHJPS[GIPP7II7IGXMSRJSVEHIWGVMTXMSRSJHEXE VIHYGXMSR Detection mode -HIRXMJMIWWMRKPISVQYPXMGLERRIPHIXIGXMSRQSHI2SXIXLEXXLMWMW MRHITIRHIRXSJXLIJPS[TEXL Dilute: End -RHMGEXIWGSQTPIXMSRSJEHMPYXISTIVEXMSR Dilute: Start -RHMGEXIWXLIWXEVXSJEHMPYXISTIVEXMSR8LIJMREPGSRGIRXVEXMSRERH WSYVGIERHHIWXMREXMSRTSWMXMSRWEVIVIGSVHIHIKDilute: Start 30% RlA1+R1A2->R1A3MRHMGEXIWXLEXTSWMXMSRVE[MPPGSRXEMRE QM\XYVISJ PJVSQVEERH PJVSQVE Dissociation: Ready -RHMGEXIWXLIIRHSJXLIHMWWSGMEXMSRXMQIWTIGMJMIHMREKINJECT GSQQERH End OPERATION -RHMGEXIWGSQTPIXMSRSJERMRNIGXMSRVIUYIWXIHJVSQXLIQERYEP GSRXVSPMRXIVJEGIMRGPYHMRKXLEYXSQEXMGGPIERYTVSYXMRIWJSPPS[MRK XLIMRNIGXMSR2SXIXLEXEnd INJECTHSIWRSXMRHMGEXIXLIIRHSJXLI EGXYEPWEQTPIMRNIGXMSRWIIInject: Ready <Error messages> 7SJX[EVIIVVSVWEVIVIGSVHIHMRXLIIZIRXPSK[MXLXLIETTVSTVMEXI MRJSVQEXMSR)VVSVWEVMWMRKMRXLIMRWXVYQIRXQMGVSTVSGIWWSVEVI HSGYQIRXIHF]I\MXGSHIW)VVSVWEVMWMRKMRXLIGSRXVSPWSJX[EVIEVI HIWGVMFIHF]EXI\XQIWWEKI Flow -RHMGEXIWXLIGYVVIRXJPS[VEXIJSVGSRXMRYSYWJPS[ (

240 Status and event log messages Flow path -RHMGEXIWXLIGYVVIRXJPS[TEXL2SXIXLEXXLMWMWMRHITIRHIRXSJXLI HIXIGXMSRQSHI Flowfill:Ready -RHMGEXIWXLIIRHSJETYQTVIJMPPSTIVEXMSR2SVQEPP]WIGSRHW EJXIVFlowfill:Start Flowfill:Start -RHMGEXIWXLIWXEVXSJETYQTVIJMPPSTIVEXMSR Fraction: Ready -RHMGEXIWXLIIRHSJEGSPPIGXIHJVEGXMSR8LIEYXSWEQTPIVTSWMXMSR JSVXLIJVEGXMSRMWVIGSVHIH Inject: Accumulated Volume 6IGSVHWXLIEGGYQYPEXIHMRNIGXIHZSPYQIJSVEQERYEPMRNIGXMSR 8LMWQIWWEKIMWKIRIVEXIH[LIRIZIVXLIPSSTMWVIPSEHIHERHEXXLI IRHSJXLIMRNIGXMSR Inject: Ready -RHMGEXIWXLIIRHSJWEQTPIMRNIGXMSRSZIVXLIWIRWSVWYVJEGI8LI ZSPYQISJWEQTPIMRNIGXIHMWVIGSVHIH8LIX]TISJMRNIGXMSR GSQQERHMWVIGSVHIHJSVMRNIGXMSRWTIVJSVQIHYRHIVQERYEP GSRXVSP Inject: Start -RHMGEXIWXLIWXEVXSJWEQTPIMRNIGXMSRSZIVXLIWIRWSVWYVJEGI8LI JPS[GIPPJSV[LMGLXLIMRNIGXMSRETTPMIWERHEYXSWEQTPIVTSWMXMSR JSVXLIWEQTPIEVIVIGSVHIH*SVEZEVMEFPIGSRXEGXXMQIMRNIGXMSRMR QYPXMGLERRIPQSHIERInject: StartQIWWEKIMWVIGSVHIHEWIEGL WYGGGIWWMZIJPS[GIPPMWW[MXGLIHMRXSXLIJPS[TEXL8LIX]TISJ MRNIGXMSRGSQQERHMWVIGSVHIHJSVMRNIGXMSRWTIVJSVQIHYRHIV QERYEPGSRXVSP Inject: Stop -RHMGEXIWXLEXERMRNIGXMSR[EWWXSTTIHF]XLIYWIVMRQERYEP GSRXVSP8LIZSPYQISJWEQTPIMRNIGXIHMWVIGSVHIH8LIX]TISJ MRNIGXMSRGSQQERHMWVIGSVHIHJSVMRNIGXMSRWTIVJSVQIHYRHIV QERYEPGSRXVSP Load Loop: End -RHMGEXIWGSQTPIXMSRSJEPSSTPSEHMRKSTIVEXMSR Load Loop: Start -RHMGEXIWXLIWXEVXSJEPSSTPSEHMRKSTIVEXMSRQERYEPMRNIGXMSR SRP]8LIPSEHMRKZSPYQIERHEYXSWEQTPIVTSWMXMSREVIVIGSVHIH IKLoad loop: Start 80 µl R1A1 (

241 Status and event log messages Low data quality -RHMGEXIWPS[HEXEUYEPMX]MRXLIWIRWSVKVEQJVSQXLIWTIGMJMIHJPS[ GIPP8LMWQIWWEKIMWSRP]HMWTPE]IHMRXLII\XIRHIHIZIRXPSK Mix: End -RHMGEXIWGSQTPIXMSRSJEQM\STIVEXMSR Mix: Start -RHMGEXIWXLIWXEVXSJEQM\STIVEXMSR8LIQM\MRKZSPYQIERH EYXSWEQTPIVTSWMXMSREVIVIGSVHIHIKMix: Start 120 µl R1A1 Normal data quality -RHMGEXIWXLEXHEXEUYEPMX]LEWVIXYVRIHJVSQPS[XSRSVQEPMRXLI WIRWSVKVEQJVSQXLIWTIGMJMIHJPS[GIPP8LMWQIWWEKIMWSRP] HMWTPE]IHMRXLII\XIRHIHIZIRXPSK Run aborted -RHMGEXIWXLEXXLIYWIVEFSVXIHXLIVYRF]TVIWWMRK'XVP&VIEOWII 7IGXMSR Set temperature 6IGSVHWXLIVIUYIWXIHQIEWYVMRKXIQTIVEXYVIEXXLIXMQIXLIVIUYIWX MWQEHI2SVIGSVHMWIRXIVIHMJXLIVIUYIWXIHXIQTIVEXYVIMWXLI WEQIEWXLIEGXYEPXIQTIVEXYVI Start <injection> -RHMGEXIWXLITSMRXEX[LMGLXLIYWIVVIUYIWXIHERMRNIGXGSQQERHMR XLIQERYEPGSRXVSPMRXIVJEGI8LIX]TISJMRNIGXGSQQERHMW VIGSVHIHMRXLIQIWWEKI2SXIXLEXStart INJECTHSIWRSXMRHMGEXI XLIWXEVXSJXLIEGXYEPWEQTPIMRNIGXMSRWIIInject: Start Temperature 6IGSVHWXLIEGXYEPQIEWYVMRKXIQTIVEXYVIEXXLIFIKMRRMRKERHIRH SJXLIQIXLSHG]GPISVQERYEPWIRWSVKVEQ Temperature stable -RHMGEXIWXLEXXLIQIEWYVMRKXIQTIVEXYVILEWFIGSQIWXEFPI Temperature unstable -RHMGEXIWXLEXXLIQIEWYVMRKXIQTIVEXYVILEWFIGSQIYRWXEFPI Terminate run -RHMGEXIWXLEXXLIVYR[EWXIVQMREXIHEWEVIWYPXSJEGSRXVSP WSJX[EVIIVVSV Timestamp 6IGSVHWXLIHEXIERHXMQIEXXLIFIKMRRMRKSJXLIQIXLSHG]GPISV QERYEPWIRWSVKVEQ Transfer: End -RHMGEXIWGSQTPIXMSRSJEXVERWJIVSTIVEXMSR (

242 Status and event log messages Transfer: Start -RHMGEXIWXLIWXEVXSJEXVERWJIVSTIVEXMSR8LIZSPYQIXVERWJIVVIH ERHXLIWSYVGIERHHIWXMREXMSRTSWMXMSRWEVIVIGSVHIHIKTransfer: Start 80 µl R1A1->R1A2 Wash: <item> -RHMGEXIWXLIWXEVXSJE[EWLSTIVEXMSR8LIMXIQXSFI[EWLIHMW VIGSVHIHIKWash: Recovery R1A1MRHMGEXIWXLEXXLIVIGSZIV]GYT MW[EWLIH[MXLWSPYXMSRJVSQTSWMXSMRVE Wash:Ready -RHMGEXIWXLIIRHSJE[EWLSTIVEXMSR (

243 Status and event log messages (

244 Writing BIACORE 2000 methods E. Writing BIACORE 2000 methods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nstrument Handbook E-1

245 Writing BIACORE 2000 methods E.1 Basic method programming 8LMWWIGXMSRGSZIVWXLIFEWMGVIUYMVIQIRXWJSV[VMXMRKQIXLSHWMR 1(0-J]SYEVIEPVIEH]EGUYEMRXIH[MXLTVSKVEQQMRKXIGLRMUYIW XLIWITVMRGMTPIW[MPPFIPEVKIP]JEQMPMEVXS]SY-J]SYEVIRI[XS TVSKVEQQMRKSRXLISXLIVLERHVIEHXLMWWIGXMSRGEVIJYPP]rMX[MPP LIPT]SYXSEZSMHQER]IVVSVWMR]SYVQIXLSHW E.1.1 Language syntax 'SQQERHWMR1(0GSRWMWXSJOI][SVHWTEVEQIXIVWERHSTXMSRW /I][SVHWEVI[SVHW[LMGLEVIWXVMGXP]HIJMRIHMRXLIPERKYEKIWYGLEW FLOWXLI]QYWXEP[E]WFI[VMXXIRSYXMRJYPPI\EGXP]EWWTIGMJMIHMR %TTIRHM\*/I][SVHW[LMGLEVIMRGSVVIGXP]WTIPPIHGEYWIER l9rvigskrm^ihgsqqerhzivvsv[lireqixlshmwtvivyr/i][svhw EVI[VMXXIRMR%TTIRHM\*MRYTTIVGEWIPIXXIVWXSMHIRXMJ]XLIQEWWYGL FYXXLIGEWIYWIHMRQIXLSHWHSIWRSXQEXXIV 4EVEQIXIVWEVIZEPYIW[LMGLUYEPMJ]XLIEGXMSRSJEGSQQERHWYGLEW MRXLIGSQQERH FLOW 304EVEQIXIVWEP[E]WJSPPS[XLI OI][SVHJSVXLIGSQQERHXS[LMGLXLI]ETTP] 7SQIGSQQERHWQE]EPWSFIUYEPMJMIHF]STXMSRWWYGLEWJMRXLI GSQQERHFLOW 30 -fxlmwstxmsrvijmppwxlifyjjivtyqt3txmsrw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nstrument Handbook

246 Writing BIACORE 2000 methods E.1.2 Constants and variables 4EVEQIXIVWEVIZEPYIWWYTTPMIHXSGSQQERHWEWI\TPEMRIHEFSZI-XMW QSWXREXYVEPXSTVSZMHIXLIWIZEPYIWHMVIGXP]EWRYQFIVWSVGLEVEGXIV WIUYIRGIWWYGLEWFLOW 30SVINJECT r1a1 108LIWIZEPYIWEVI GEPPIHGSRWXERXW 1IXLSHWEVIQSVIJPI\MFPIMJZEVMEFPIWEVIYWIHMRWXIEHSJGSRWXERXW% ZEVMEFPIMWETPEGILSPHIVJSVEZEPYI[LMGLMWEWWMKRIHEREGXYEPZEPYI MRXLIGYVVIRXGSRXI\XSJXLIQIXLSH*SVMRWXERGIXLIGSQQERH INJECT %sample %vol[livi%sampleerh%volevizevmefpiw[mpp MRNIGXEZSPYQIHIJMRIHF]XLIZEPYIGYVVIRXP]EWWMKRIHXS%volJVSQ XLITSWMXMSRMHIRXMJMIHF]XLIZEPYIGYVVIRXP]EWWMKRIHXS %sample :EVMEFPIREQIW =SYGLSSWI]SYVS[RREQIWJSVZEVMEFPIW[MXLXLIJSPPS[MRK VIWXVMGXMSRWREQIWQE]FIYTXSGLEVEGXIVWPSRKERHGSRXEMRER] SJXLI)RKPMWLPIXXIVWE^ERH%>XLIHMKMXWERHXLIWTIGMEP GLEVEGXIVWCERH 2EQIWQE]RSXFIKMR[MXLEHMKMXSV YRHIVWGSVIGLEVEGXIV2SXIXLEXREQIWQE]RSXGSRXEMRWTEGIW9WI YRHIVWGSVIGLEVEGXIVWMRWXIEHSJWTEGIWMRREQIWPMOI %mixing_positionsv%flow_rate 'SQQERHOI][SVHWPMOIFLOWERHINJECTWLSYPHRSXFIYWIHEW REQIW %YWIJYPGSRZIRXMSRJSPPS[IHMRXLMWQERYEPMWXSWXEVXEPPZEVMEFPI REQIW[MXLEWTIGMEPGLEVEGXIVWYGLEW8LMWWIVZIWX[STYVTSWIW XLIVIMWRSVMWOXLEX]SY[MPPGSRJYWIOI][SVHWERHZEVMEFPIREQIWRS OI][SVHWFIKMR[MXLERHMXMWIEW]XSWIIMREQIXLSHJMPI[LMGL [SVHWEVIZEVMEFPIREQIW E.1.3 Comments =SYGEREHHGSQQIRXWXS]SYVQIXLSHXSLIPT]SYYRHIVWXERH[LEX MXMWQIERXXSHS'SQQIRXWEVIMRXVSHYGIHF]ERI\GPEQEXMSRQEVO ERHGSRWMWXSJEPPGLEVEGXIVWJVSQXLII\GPEQEXMSRQEVOXSXLIIRH SJXLIPMRI'SQQIRXWQE]FI[VMXXIRSRXLIWEQIPMRIEWGSQQERHW SVSRWITEVEXIPMRIW =SYGEREPWSYWIERI\GPEQEXMSRQEVOXSVIQSZIGSQQERHW XIQTSVEVMP]JVSQEQIXLSH'SQTEVIXLIX[SQIXLSHJVEKQIRXW FIPS[VIJIVXS%TTIRHM\*MJ]SY[ERXXSYRHIVWXERHXLIHIXEMPWSJXLI GSQQERHJSVQYPEXMSRW &-%'36) 2 Instrument Handbook E-3

247 Writing BIACORE 2000 methods... TRANSFER r2a1 r2b1 10 MIX r2b1 200 INJECT r2b TRANSFER r2a1 r2b1 10! MIX r2b1 200 INJECT r2b VERWJIV PJVSQTSWMXMSRVEXS VFQM\MRVFF]EWTMVEXMRKERH HMWTIRWMRK PXLIRMRNIGX P 8VERWJIV PJVSQTSWMXMSRVEXS VFXLIRMRNIGX PMQQIHMEXIP] 8LI1-<GSQQERHMWRS[EGSQQIRX ERHHSIWRSXLMRK E.2 Method blocks &-%'36)'SRXVSP7SJX[EVIQIXLSHWEVIHMZMHIHMRXSERYQFIVSJ WITEVEXIWIGXMSRWSVFPSGOW8LIVIEVIFPSGOX]TIWIEGL[MXLE HIJMRIHJYRGXMSRMRXLIQIXLSH &PSGOOI][SVH DEFINE ALERT DEFINE APROG DEFINE LOOP DEFINE MESSAGE MAIN *YRGXMSR +MZIWXLIYWIVEGLERGIXSXIVQMREXISV GSRXMRYIXLIQIXLSH %REP]WMWTVSKVEQ(IJMRIWXLIWIUYIRGISJ STIVEXMSRWMRSRIVYRSVWIVMIWSJVYRW% WIRWSVKVEQMWGSPPIGXIHERHEREP]^IH (IJMRIWXLITEVEQIXIVWXSFIYWIHJSVXLI VITIEXIHSTIVEXMSRWMRE0334FPSGO (MWTPE]WEQIWWEKISRXLIWGVIIR (IJMRIWXLIWIUYIRGISJFPSGOI\IGYXMSRMRE QIXLSH 8LIFIKMRRMRKSJEFPSGOMWHIJMRIHF]EPMRIGSRXEMRMRKXLIFPSGO OI][SVHERHER]TEVEQIXIVWXLEXQE]FIVIUYMVIH8LIIRHSJIEGL FPSGOMWMHIRXMJMIHF]EPMRIGSRXEMRMRKXLIWMRKPIOI][SVH END%PP SXLIVGSQQERHWQYWXFI[VMXXIR[MXLMRFPSGOWMRSXLIV[SVHWXLIVI QE]SRP]FIGSQQIRXWSVIQTX]PMRIWFIX[IIRXLIENDXIVQMREXMRK SRIFPSGOERHXLIFPSGOOI][SVHWXEVXMRKXLIRI\X 8LIMAINFPSGOMWVIUYMVIHMRIZIV]QIXLSHERHQE]SRP]SGGYVSRGI %PPSXLIVFPSGOWEVISTXMSREPERHQE]SGGYVEWQER]XMQIWEW VIUYMVIH(MJJIVIRXFPSGOWSJXLIWEQIX]TIEVIMHIRXMJMIHF]REQIW 8LISVHIVMR[LMGLFPSGOWEVI[VMXXIRMREQIXLSHHSIWRSXQEXXIV E-4 &-%'36) 2 Instrument Handbook

248 Writing BIACORE 2000 methods E.3 Constructing methods 8LMWWIGXMSRWLS[W]SYLS[XSGSRWXVYGXJEMVP]WMQTPIQIXLSHWYWMRK 1(0XSKMZI]SYEJIIPJSVLS[XS[VMXIQIXLSHW8LII\EQTPIWMR %TTIRHM\&MPPYWXVEXIGSQTPIXIQIXLSHWYWMRKEVERKISJ1(0 GSQQERHW E.3.1 The MAIN block 8LIMAINFPSGOHIJMRIWXLIWIUYIRGISJSTIVEXMSRWMRXLI[LSPI QIXLSH8LMWMWXLISRP]VIUYMVIHFPSGOMREQIXLSHERHMWXLISRP] FPSGO[LMGLQE]RSXFIYWIHQSVIXLERSRGI 8LI MAINOI][SVHMWRSXJSPPS[IHF]ER]TEVEQIXIVW8LIIRHSJXLI MAINFPSGOMWMHIRXMJMIHF]XLIENDOI][SVH 1SWXGSQQSRP]XLIMAINFPSGOGEPPWEWIVMIWSJAPROGWWII7IGXMSR )[LMGLGSRWXMXYXIXLIlFSH]zSJXLIQIXLSH8LIMAINFPSGOGER EPWSWIXXLIHIXIGXMSRQSHIERHXLIHIJEYPXJPS[TEXLJSVXLIAPROGW [MXLXLIDETECTIONGSQQERHWII7IGXMSR)JSVQSVIHIXEMPW MAIN DETECTION 2 APROG immob END E.3.2 The analysis program The DEFINE APROG block 7MQTPIEREP]WMWTVSKVEQW 8LIWIUYIRGISJSTIVEXMSRWJSVSRIG]GPIMW[VMXXIRMREDEFINE APROGEREP]WMWTVSKVEQFPSGO%QIXLSHQE]GSRXEMRER]RYQFIV SJDEFINE APROGFPSGOW)EGLSRIMWMHIRXMJMIHF]EYRMUYIREQI %ZIV]WMQTPII\EQTPIWIXWXLIJPS[VEXIXS PQMRERHMRNIGXW P WEQTPIJVSQTSWMXMSRVE DEFINE APROG injection FLOW 20 INJECT r2a1 20 END %RAPROGMWRSXEGSQTPIXIQIXLSHMXMWI\IGYXIHJVSQXLIMAIN FPSGOF][VMXMRKXLIGSQQERHAPROGJSPPS[IHF]XLIMHIRXMJ]MRK REQIWII7IGXMSR) &-%'36) 2 Instrument Handbook E-5

249 Writing BIACORE 2000 methods MAIN DETECTION 1 APROG injection END 8LIGSQTPIXIQIXLSHMWXLIWIX[SFPSGOWXSKIXLIV DEFINE APROG injection FLOW 20 INJECT r2a1 20 END MAIN END DETECTION 1 APROG injection 2SXIXLEXXLIDEFINE APROGFPSGOWXEVXW[MXLEFLOWGSQQERHXS WIXXLIJPS[VEXI)ZIV]APROGQYWXWIXXLIJPS[VEXIEXXLIFIKMRRMRK SJXLIAPROG )ZIV]APROGWXEVXW[MXLEREYXSQEXMGMRMXMEXMSRWIUYIRGIXLEXVIJMPPW XLIFYJJIVTYQTMJRIGIWWEV]ERHWXEVXWWIRWSVKVEQVIGSVHMRKERHIRHW [MXLEXIVQMREXMSRWIUYIRGIXLEX[EWLIWXLIEYXSWEQTPIVRIIHPIERH -*'GLERRIPWERHVIWIXWEPP-*'ZEPZIW %MAINFPSGOGERGEPPQSVIXLERSRIAPROGXSVYRQYPXMTPIEREP]WMW G]GPIW)EGLAPROGVITVIWIRXWSRIG]GPIMRXLIQIXLSHERHKIRIVEXIW SRISVQSVIWIRWSVKVEQWHITIRHMRKSRXLIHIXIGXMSRQSHI8LI I\EQTPIFIPS[MQQSFMPM^IWEPMKERHXSXLIWIRWSVGLMTWYVJEGIYWMRK EQMRIGLIQMWXV]XLIRMRNIGXWEREP]XIWEQTPISZIVXLIPMKERHWYVJEGI E-6 &-%'36) 2 Instrument Handbook

250 Writing BIACORE 2000 methods DEFINE APROG immob FLOW 10 DILUTE r1a1 r1a2 r1a3 50!Mix EDC/NHS * INJECT r1a3 70!Inject EDC/NHS -0:30 RPOINT Baseline -b DIPNEEDLE r1b1!ligand buffer INJECT r1b2 70!Ligand * INJECT r1c1 70!Deactivation EXTRACLEAN 8:00 RPOINT immob!immobilized level END DEFINE APROG injection FLOW 20 INJECT r2a1 20 END MAIN DETECTION 1!Sensorgram in flow cell 1 APROG immob APROG injection END 9WMRKZEVMEFPIW 8SQEOIXLIAPROGQSVIJPI\MFPI]SYGERYWIZEVMEFPIWMRWXIEHSJ GSRWXERXZEPYIWJSVIKWEQTPITSWMXMSRWERHZSPYQIW8LIZEVMEFPIW EVIVITPEGIHF]ZEPYIW[LIRXLIAPROGMWGEPPIHJVSQEMAINFPSGO WIIFIPS[ 8SYWIZEVMEFPIWMRXLIDEFINE APROGFPSGO]SYVITPEGIXLIGSRWXERX TEVEQIXIVZEPYIW[MXLZEVMEFPIREQIW%XXLIWEQIXMQI]SYQYWXEHH SRISVQSVIPARAMGSQQERHWMQQIHMEXIP]EJXIVXLIDEFINE APROG PMRIXSXIPPXLIWSJX[EVI[LMGLZEVMEFPIREQIW]SY[MPPYWIMRXLI EREP]WMWTVSKVEQ*SVI\EQTPI DEFINE APROG injection PARAM %position %volume FLOW 20 INJECT %position %volume END 2SXIXLEXEGSRWXERXMWYWIHJSVXLIJPS[VEXIMRXLMWI\EQTPI[LMPI ZEVMEFPIWEVIYWIHJSVWEQTPITSWMXMSRERHZSPYQI 8SYWIEREREP]WMWTVSKVEQ[MXLZEVMEFPIWEHHXLIZEPYIWXLEX]SY [ERXXSKMZIXLIZEVMEFPIWXSXLIAPROGGSQQERHPMRIEJXIVXLIREQI SJXLITVSKVEQ*SVMRWXERGIYWMRKXLIJSPPS[MRKQIXLSH[MPPMRNIGX JMVWX PJVSQTSWMXMSRVEXLIR PJVSQTSWMXMSRVE &-%'36) 2 Instrument Handbook E-7

251 Writing BIACORE 2000 methods DEFINE APROG injection PARAM %position %volume FLOW 20 INJECT %position %volume END MAIN DETECTION 1 APROG injection r1a1 20 APROG injection r2a5 35 END 8LISVHIVMR[LMGL]SYKMZIXLITEVEQIXIVZEPYIWMRXLIAPROGQYWX FIXLIWEQIEWXLISVHIVMR[LMGLXLI]EVIWTIGMJMIHMRXLIPARAM GSQQERHW[MXLMRDEFINE APROG8LIJMVWXZEPYIMWEWWMKRIHXSXLI JMVWXZEVMEFPIREQIXLIWIGSRHZEPYIXSXLIWIGSRHZEVMEFPIREQIERH WSSR =SYGERYWIXLIWEQIZEVMEFPIREQIWMRWIZIVEPHMJJIVIRXAPROGW [MXLMRXLIWEQIQIXLSH8LIREQIWEVIPSGEPXSXLIAPROG=SY GERRSXSRXLISXLIVLERHYWIXLIWEQIREQIJSVX[SHMJJIVIRX ZEVMEFPIW[MXLMRXLIWEQIAPROG 7IXXMRKXLIWIRWSVKVEQGETXMSR 8LICAPTIONGSQQERHEPPS[W]SYXSWIXEGETXMSRXI\XJSV]SYV WIRWSVKVEQ=SYGERYWIEZEVMEFPIREQIEWTEVXSJXLIGETXMSRSVEW XLI[LSPIGETXMSRMJETTVSTVMEXI8LII\EQTPIFIPS[WIXWXLIXI\X l-rnigxmsrsj PzEWXLIWIRWSVKVEQGETXMSR[MXLXLIVIWTIGXMZI WEQTPIZSPYQIJMPPIHMRJSVIEGLWIRWSVKVEQ DEFINE APROG injection PARAM %position %volume CAPTION Injection of %volume µl FLOW 20 INJECT %position %volume END MAIN DETECTION 1 APROG injection r1a1 20 APROG injection r2a5 35 END E-8 &-%'36) 2 Instrument Handbook

252 Writing BIACORE 2000 methods 8MQIHIZIRXWMREREP]WMWTVSKVEQW 7SJEVXLIAPROGI\EQTPIWLEZII\IGYXIHGSQQERHWMRE WXVEMKLXJSV[EVHWIUYIRGI3RIGSQQERHWXEVXWEWWSSREWXLITVIZMSYW GSQQERHMWGSQTPIXIH =SYGERWTIGMJ]XLIXMQI[LIREKMZIRGSQQERH[MPPWXEVXVIPEXMZI IMXLIVXSXLIFIKMRRMRKSJXLIEREP]WMWTVSKVEQSVXSWSQIHIJMRIH TSMRX[MXLMRXLITVSKVEQ=SYHSXLMWF]TYXXMRKEXMQIWTIGMJMGEXMSR FIJSVIXLIGSQQERH DEFINE APROG injection FLOW 20! time 0 0:50 INJECT r1a1 20! start at 50 s 6:00 INJECT r1a2 30! start at 6 min END -RXLMWAPROGXLIX[SMRNIGXMSRW[MPPFITIVJSVQIHWIGSRHWERH QMRYXIWVIWTIGXMZIP]EJXIVXLIWXEVXSJXLIAPROG %GSQQERH[MXLEXMQIWTIGMJMGEXMSRMWI\IGYXIHEXXLIXMQIWTIGMJMIH SVEWWSSREWTSWWMFPIXLIVIEJXIV-RXLIAPROGEFSZIJSVI\EQTPIXLI JMVWXMRNIGXMSRSGGYTMIWXLIEYXSWEQTPIVJSVEFSYXQMRYXIW-JXLI WIGSRHMRNIGXMSR[IVIXMQIHXSWXEVXEXQMRYXIWMQTSWWMFPIWMRGIXLI EYXSWEQTPIVMWSGGYTMIHXLIWIGSRHMRNIGXMSR[SYPHFITIVJSVQIHEW WSSREWXLIJMVWX[EWGSQTPIXIH8MQIWTIGMJMGEXMSRWMRAPROGWEVIXLYW QMRMQYQXMQIWERHHSRSXRIGIWWEVMP]VITVIWIRXXLIXMQISJI\IGYXMSR SJXLIGSQQERH8LIEGXYEPXMQI[LIRXLIGSQQERHMWI\IGYXIHMW VIGSVHIHMRXLIIZIRXPSK 8SWIXERMRNIGXMSRXVERWJIVSVQM\MRKXMQIEWERI[^IVSXMQITSMRX [MXLMRETVSKVEQTYXEREWXIVMWOFIJSVIXLIXMQIWTIGMJMGEXMSR(S RSXX]TIEWTEGIFIX[IIRXLIEWXIVMWOERHXLIXMQIWTIGMJMGEXMSR DEFINE APROG injection FLOW 20! time 0 *0:30 INJECT r1a1 20! start at 30 s 5:00 INJECT r1a2 30! start 5 min after! first inject END,IVIXLIWIGSRHMRNIGXMSRMWXMQIHXSWXEVXQMRYXIWEJXIVXLI FIKMRRMRKSJXLIJMVWXMRNIGXMSR =SYGERRSXYWIZEVMEFPIWJSVXMQIWTIGMJMGEXMSRW &-%'36) 2 Instrument Handbook E-9

253 Writing BIACORE 2000 methods 'VIEXMRKVITSVXXEFPIW 1(0KMZIW]SYJYPPJVIIHSQMRGVIEXMRKVITSVXTSMRXXEFPIWPMWXWSJ 746WMKREPPIZIPWEXWTIGMJMIHXMQIWrWII7IGXMSRJVSQ WIRWSVKVEQW=SYWTIGMJ]XLIGSRXIRXWSJ]SYVVITSVXXEFPIXLVSYKL GSQQERHWMRXLIEREP]WMWTVSKVEQ 6ITSVXTSMRXWEVIWIX[MXLXLIRPOINTGSQQERH[LMGLWLSYPHEP[E]W FITVIGIHIHF]EXMQIWTIGMJMGEXMSRSXLIV[MWIXLIWSJX[EVIHSIWRSX ORS[[LIRXSWIXXLITSMRX8LIGSQQERHOI][SVHMWJSPPS[IHF] ERMHIRXMJMGEXMSR[LMGLETTIEVWMRXLIl-HzGSPYQRMRXLIVITSVXTSMRX XEFPI%GSQQSRP]YWIHSTXMSR[MXLRPOINTMW-b[LMGLWIXWERI[ FEWIPMRIEXXLIVITSVXTSMRX%WMQTPII\EQTPIJSPPS[W DEFINE APROG report FLOW 20 * INJECT r1a1 10! set this as time 0-0:15 RPOINT baseline -b! set baseline 0:30 RPOINT plateau! record plateau level END 2SXIXLIRIKEXMZIXMQIWTIGMJMGEXMSRJSVXLIJMVWXRPOINTWIXXMRKXLI VITSVXTSMRXXSWFIJSVIXLIJMVWXMRNIGXMSR8LMWMWSJXIRE GSRZIRMIRX[E]XSWIXVITSVXTSMRXWVIPEXMZIXSERIZIRX %VITSVXTSMRXXEFPI[MPPEP[E]WGSRXEMREQMRMQYQWXERHEVHWIXSJ GSPYQRW=SYGERWTIGMJ]EHHMXMSREPGSPYQRW[MXLXLIKEYWORD GSQQERH[LMGLVIUYMVIWELIEHIVJSVXLIGSPYQRERHEZEPYIXSFI TPEGIHMRXLIGSPYQR8LIAPROGFIPS[[MPPKIRIVEXIEVITSVXTSMRX XEFPI[MXLEREHHMXMSREPGSPYQRLIEHIHl%RXMFSH]zGSRXEMRMRKXLI ZEPYIl1%Fz-REHHMXMSRXLIOI][SVHl%RXMFSH]z[MPPFI MRGPYHIHEWEWIEVGLOI]MRXLIVIWYPXJMPI DEFINE APROG report KEYWORD Antibody MAb-451 FLOW 20 * INJECT r1a1 10! set this as time 0-15 RPOINT baseline -b! set baseline 30 RPOINT plateau! record plateau level END 8LIRPOINTERHKEYWORDGSQQERHWGERYWIZEVMEFPIWXSLIPTMR HSGYQIRXEXMSRSJXLIVITSVXTSMRXXEFPI E-10 &-%'36) 2 Instrument Handbook

254 Writing BIACORE 2000 methods E.3.3 Repetitive analyses The DEFINE LOOP block &EWMGPSSTGSRWXVYGXMSRW 8LILOOPGSQQERHERHGSVVIWTSRHMRKDEFINE LOOPFPSGOEPPS[]SY XSGSRWXVYGXQIXLSHWJSVWIVMIWSJVITIXMXMZIEREP]WIW 8LIJSPPS[MRKQIXLSHEREP]WIWJSYVHMJJIVIRXWEQTPIWMRHYTPMGEXIrE XSXEPSJVYRW;IEWWYQIXLEXPMKERHLEWTVIZMSYWP]FIIRGSYTPIHXS XLIWIRWSVGLMTWYVJEGI DEFINE LOOP cycle LPARAM %sample %position %volume TIMES 2 Sample1 r1a1 10 Sample2 r1a2 15 Sample3 r1a3 10 Sample4 r1a4 20 END DEFINE APROG analyse PARAM %sample %position %volume KEYWORD sample %sample FLOW 20 * INJECT %position %volume -0:15 RPOINT baseline -b 2:00 RPOINT plateau 5:00 INJECT r2a1 10! regenerate END MAIN DETECTION 1 LOOP cycle STEP APROG analyse %sample %position %volume ENDLOOP END 8LIIWWIRXMEPJIEXYVIWSJXLMWGSRWXVYGXMSREVIEWJSPPS[W :EVMEFPIWEVIHIJMRIH[MXLXLILPARAMGSQQERHMRXLIDEFINE LOOPFPSGO8LMWFPSGOIWXEFPMWLIWWIXWSJZEVMEFPIZEPYIWWIXWSJ ZEVMEFPIWIEGLMRXLMWI\EQTPI8LITIMES 2GSQQERHMRDEFINE LOOPWTIGMJMIWXLEXXLIPSST[MPPKSXLVSYKLXLIZEVMEFPIWIXWX[MGI 8LIWEQIZEVMEFPIREQIWEVIWIXYTEWTEVEQIXIVWJSVAPROG analyseywmrkxliparamgsqqerhmrdefine APROG -RXLIMAINFPSGOXLILOOP cycle...endloopgsrwxvygxmsr I\IGYXIWXLIGSQQERHWFIX[IIRLOOPERHENDLOOPSRGIJSVIZIV] ZEVMEFPIWIXMRDEFINE LOOPG]GPI-RXLMWI\EQTPIXLIGSQQERHW &-%'36) 2 Instrument Handbook E-11

255 Writing BIACORE 2000 methods [MPPFIVITIEXIHXMQIWZEVMEFPIWIXW[MXLETIMES 2VITIEX :EVMEFPIWHIJMRIHMRXLIDEFINE LOOPFPSGOGERFIYWIHEW TEVEQIXIVWJSVER]GSQQERHMRXLILOOP...ENDLOOPGSRWXVYGXMSR MRXLMWGEWIXLI]EVITEWWIHXSAPROG analyse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cycle LPARAM %sample %pos %vol TIMES 2! do the following rows twice Sample1 r1a1 10 Sample2 r1a2 15 Sample3 r1a3 10 TIMES 1! do the last row only once Sample4 r1a4 20 END 6ITIEXMRKQYPXMTPIGSQQERHW -RXLII\EQTPIWEFSZIXLILOOPGSQQERHEGXMZEXIWNYWXSRIEREP]WMW TVSKVEQ=SYGERLS[IZIVMRGPYHIER]GSQQERH[LMGLMWTIVQMXXIH MRMAIN[MXLMREPSST8LI[LSPIGSQQERHWIUYIRGIFIX[IIRLOOP ERHENDLOOP[MPPFIVITIEXIHJSVXLIVS[WMRDEFINE LOOP8LI I\EQTPIFIPS[YWIWEPSSTGSRWXVYGXMSRXSVYREWEQTPIXLVSYKL HMJJIVIRXJPS[GIPPWMRJSYVWITEVEXIMRNIGXMSRW[MXLEHMJJIVIRXMRNIGXMSR ZSPYQIMRIEGLJPS[GIPP E-12 &-%'36) 2 Instrument Handbook

256 Writing BIACORE 2000 methods DEFINE APROG analyse PARAM %volume FLOW 20 INJECT r1a1 %volume... END DEFINE LOOP flowcells LPARAM %cell %volume TIMES END MAIN LOOP flowcells STEP DETECTION %cell APROG analyse %volume ENDLOOP END %HZERGIH0334GSRWXVYGXMSRW 0SSTGSRWXVYGXMSRWGERFIRIWXIHXLEXMWEPSSTGERFIGEPPIHJVSQ [MXLMRERSXLIVPSST8LMWXIGLRMUYIMWTEVXMGYPEVP]YWIJYPJSV ETTPMGEXMSRWXLEX[SVO[MXLEQEXVM\SJWEQTPIWXIWXMRKEPPTEMV[MWI GSQFMREXMSRWIKITMXSTIQETTMRK8LII\EQTPIFIPS[VYRWE QEXVM\[MXLXLIWEQTPIW%&ERH'MRTSWMXMSRWVEVE &-%'36) 2 Instrument Handbook E-13

257 Writing BIACORE 2000 methods DEFINE LOOP cycle1 LPARAM %s1 %p1 A r1a1 B r1a2 C r1a3 END DEFINE LOOP cycle2 LPARAM %s2 %p2 A r1a1 B r1a2 C r1a3 END MAIN DETECTION 1 FLOW 20 LOOP cycle1 STEP LOOP cycle2 STEP APROG analyse %s1 %s2 %p1 %p2 APROG regenerate ENDLOOP ENDLOOP END 8LMW[SVOWEWJSPPS[W %XLOOPcycle1STEPZEPYIWJSV%s1ERH%p1EVIXEOIRJVSQXLI JMVWXVS[MRDEFINE LOOP cycle1l%zerhlvez VIWTIGXMZIP] 8LIQIXLSHXLIRVYRWXLVSYKLEPPXLVIIVS[WMRDEFINE LOOP cycle2ywmrkxlizepyiwl%zerhlvezjsv%s1erh%p18lmw VYRWXLIGSQFMREXMSRW%%%&ERH%' ;LIRLOOP cycle2mwgsqtpixihxliri\xwxitsjloop cycle1 MWI\IGYXIHXEOMRKl&zERHlVEzJSV%s1ERH%p1LOOP cycle2rs[vyrwekemrkirivexmrkxligsqfmrexmsrw&%&& ERH&' -RXLIWEQI[E]XLIPEWXWXITSJLOOP cycle1kirivexiwxli GSQFMREXMSRW'%'&ERH'' 8LMWI\EQTPIMWSRP]E QEXVM\FYXXLIIGSRSQ]SJRIWXIHPSST GSRWXVYGXMSRWMWVIEHMP]ETTEVIRXMJ]SYGSRWMHIVJSVI\EQTPIE QEXVM\[MXLGSQFMREXMSRW E-14 &-%'36) 2 Instrument Handbook

258 Writing BIACORE 2000 methods E.3.4 Communicating with the user The DEFINE ALERT and DEFINE MESSAGE blocks 8LIX[SGSQQERHWALERTERHMESSAGE[MXLXLIMVGSVVIWTSRHMRK DEFINE ALERTERHDEFINE MESSAGEFPSGOWEVIYWIHXSMRJSVQXLI YWIVSJXLITVSKVIWWSJEQIXLSHALERTEGXMZEXIWEHMEPSKYI[MRHS[ [LMGLEPPS[WXLIYWIVXSEFSVXSVGSRXMRYIXLIQIXLSHMESSAGE WMQTP]HMWTPE]WEXI\X[MRHS[8LIGSQQERHBEEPWSYRHWEFIITXS GEPPXLISTIVEXSVk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nstrument Handbook E-15

259 Writing BIACORE 2000 methods DEFINE ALERT insert_sample TITLE Unstable sample BEEP LTEXT LTEXT Continuation of this method requires LTEXT sample in position r2a1. Make sure that LTEXT the sample is completely thawed before LTEXT placing it in the autosampler rack. LTEXT LTEXT Close the door and click on Continue LTEXT when you are ready. GO Continue STOP Cancel END MAIN RACK 1 thermo_a RACK 2 thermo_b ALERT insert_sample APROG cycle1... END E.3.5 Additional data curves The CURVE command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nstrument Handbook

260 Writing BIACORE 2000 methods DEFINE APROG monitor CURVE TEMP -o CURVE bufflow FLOW END! use right-hand axis! use default axis =SYGERWIXVITSVXTSMRXWSRWXEXYWGYVZIWYWMRKXLI-cSTXMSRJSVXLI RPOINTGSQQERH E.3.6 Closing down The APPEND command 8LI)2(GSQQERHEXXLIIRHSJIZIV]APROGMRGPYHIWEYXSQEXMG JYRGXMSRWJSV[EWLMRKXLI-*'ERHJPS[GIPPW[MXLFYJJIV=SYGER ETTIRHXLI[SVOMRKXSSPStandbyXSEQIXLSHMJXLIMRWXVYQIRXMWRSX KSMRKXSFIYWIHJSVYTXSHE]WEJXIVGSQTPIXMSRSJXLIQIXLSH8LMW [MPPJPYWLXLI-*'ERHQEMRXEMREPS[FYJJIVJPS[JSVYTXSHE]WWII 7IGXMSR*SVI\EQTPI DEFINE APROG analyse... END DEFINE APROG regenerate... END MAIN DETECTION 1 APROG analyse APROG regenerate APPEND standby END 2SXI *SVGSQTEXMFMPMX][MXLIEVPMIVZIVWMSRWSJ1(0XLIGSQQERH APPEND continuemwiuymzepirxxsappend standby &-%'36) 2 Instrument Handbook E-17

261 Writing BIACORE 2000 methods E.4 Method techniques 8LMWWIGXMSRGSRWMHIVWWSQIJIEXYVIWSJQIXLSHTVSKVEQQMRKJVSQXLI ZMI[TSMRXSJETTPMGEXMSRVIUYMVIQIRXW E.4.1 Detection mode and flow path (IXIGXMSRQSHI &-%'36)STIVEXIWMRWMRKPISVQYPXMGLERRIPHIXIGXMSRQSHI [MXLHEXEGSPPIGXMSRJVSQSRIX[SSVJSYVJPS[GIPPW =SYWIXXLIHIXIGXMSRQSHIERHXLIMRMXMEPJPS[TEXL[MXLE DETECTIONGSQQERHMRXLIMAINFPSGOSJXLIQIXLSH=SYGER GLERKIXLIHIXIGXMSRQSHIFIX[IIRAPROGWMRXLIMAINFPSGO MAIN DETECTION 1 APROG single DETECTION 1,2,3,4 APROG multi END! single channel mode! multichannel mode -JXLIVIMWRSDETECTIONGSQQERHFIJSVIXLIJMVWXAPROGMRXLIMAIN FPSGOXLIWSJX[EVI[MPPTVSQTXJSVHIXIGXMSRQSHI[LIRXLIQIXLSH MWVYR 8LIWIPIGXMSRMRXLMWHMEPSKYIFS\GSVVIWTSRHWHMVIGXP]XSXLI DETECTIONGSQQERHMRXLIMAINFPSGO8LIWIRWSVKVEQ[MPPRSXWXEVX YRXMP]SYGPMGOSROKMRXLMWHMEPSKYIFS\ 1IXLSHW[VMXXIR[MXLSYXEDETECTIONGSQQERHEVIJPI\MFPIMRXLEX ]SYGERYWIXLIWEQIQIXLSHJSVEREP]WMWMRHMJJIVIRXJPS[GIPPW 8LIDETECTIONGSQQERHQE]SRP]FIYWIHMRXLI MAINFPSGO E-18 &-%'36) 2 Instrument Handbook

262 Writing BIACORE 2000 methods *PS[TEXL 8LIJPS[TEXLWIXF]XLIDETECTIONGSQQERHVIQEMRWSTIRYRXMPSRI SJXLIJSPPS[MRKSGGYVW ERI[DETECTIONGSQQERHMWMWWYIHMRXLIMAINFPSGO EFLOWPATHGSQQERHMWMWWYIHMRERAPROGWIIFIPS[ ERINJECTGSQQERHMWMWWYIHMRERAPROG[MXLEJPS[GIPP VIUYMVIQIRXHMJJIVIRXJVSQXLIGYVVIRXJPS[TEXL =SYGERYWIXLIFLOWPATHGSQQERHMRERAPROGXSGLERKIXLI JPS[TEXLHYVMRKXLIG]GPI=SYQE]JSVMRWXERGI[MWLXSFMRHEREP]XI XSXLIPMKERHMREPPJSYVJPS[GIPPWERHXLIRXIWXXLIFMRHMRKSJJSYV HMJJIVIRXWIGSRHEV]PMKERHWSRIMRIEGLJPS[GIPP MAIN DETECTION 1,2,3,4 APROG epitope END DEFINE APROG epitope PARAM %pos1 %pos2 %pos3 %pos5 %pos5 FLOW 5 FLOWPATH 1,2,3,4 INJECT %pos1 5! first Mab INJECT r1c1 5! analyte FLOWPATH 1 INJECT %pos2 5! second Mab 1... FLOWPATH 2 INJECT %pos3 5! second Mab 2... FLOWPATH 3 INJECT %pos4 5! second Mab 3... FLOWPATH 4 INJECT %pos5 5! second Mab 4... FLOWPATH 1,2,3,4 INJECT r1d3 15! regenerate... END 8LIFLOWPATHGSQQERHQE]SRP]FIYWIHMRERAPROGFPSGO &-%'36) 2 Instrument Handbook E-19

263 Writing BIACORE 2000 methods %28 8LIDETECTION GSQQERHWIXWFSXLXLIHIXIGXMSRQSHIERHXLIMRMXMEP JPS[TEXL,S[IZIVXLIFLOWPATHGSQQERHHSIWRSXEJJIGXXLI HIXIGXMSRQSHI8EOIGEVIRSXXSWIXXLIJPS[TEXLXSMRGPYHIGIPPW [LIVIHEXEMWRSXFIMRKGSPPIGXIHIK MAIN 4 END DETECTION 4 APROG missing_data! single channel detection in Fc DEFINE APROG missing_data FLOW 20 FLOWPATH 1,2,3! sample flow through Fc 1-2-3! detection only in Fc 4... END 8LIWEQIQMWXEOIGERIEWMP]FIQEHI[MXLEZEVMEFPIGSRXEGXXMQI MRNIGXMSR[LMGLWIXWXLIJPS[TEXLMQTPMGMXP]WII7IGXMSR) 8LIHIWMKRSJXLIDetectionERHInjectHMEPSKYIFS\IWTVIZIRXWXLMW QMWXEOIYRHIVQERYEPGSRXVSPFYXXLIVIMWRSIUYMZEPIRXWEJIKYEVHMR QIXLSHW8SEZSMHQMWWMRKHEXEMRQIXLSHVYRW 9WIFLOWPATH[MXLGEVI*I[ETTPMGEXMSRWVIUYMVIGLERKIWMRJPS[ TEXLHYVMRKERAPROG-XMWSJXIRTVIJIVEFPIXSYWIWITEVEXIAPROGW WIXXMRKXLIJPS[TEXLJSVIEGLSRI[MXLEDETECTIONGSQQERHMR MAIN 7IXDETECTION 1,2,3,4EXXLIFIKMRRMRKSJXLIMAINFPSGOMJ]SY EVIYRWYVISJXLIVIUYMVIQIRXWSJ]SYVAPROGW8LMW[MPPWIXQYPXM GLERRIPHIXIGXMSRQSHIERHRSHEXE[MPPFIPSWX 3RPMRIVIJIVIRGIWYFXVEGXMSR 8SYWISRPMRIVIJIVIRGIWYFXVEGXMSRYWISRISJXLIJSPPS[MRK TEVEQIXIVW[MXLXLIDETECTIONGSQQERH 4EVEQIXIV (IXIGXMSRQSHI *PS[TEXL 6IJIVIRGI WYFXVEGXMSR E-20 &-%'36) 2 Instrument Handbook

264 Writing BIACORE 2000 methods %RI\EQTPIMWKMZIRFIPS[ MAIN DETECTION 2-1,4-3! single channel mode APROG subtract_reference END E.4.2 Sample injection types &-%'36)SJJIVWEVERKISJWEQTPIMRNIGXMSRX]TIWJSVHMJJIVIRX ETTPMGEXMSRTYVTSWIW8LIWIEVIEZEMPEFPIJVSQXLIQERYEPGSRXVSP MRXIVJEGIWII7IGXMSRERHEPPI\GITXQERYEPMRNIGXMSRGERFI TVSKVEQQIHMRQIXLSHW 8SQEOIWYVIXLEXERMRNIGXMSR[MPPRSXFIHMWXYVFIHF]ETYQTVIJMPP STIVEXMSRMWWYIEFLOWGSQQERH[MXLXLI-fSTXMSRNYWXFIJSVIXLI INJECTGSQQERHIK DEFINE APROG smooth_inject FLOW 35 10:00 WASH n FLOW 35 -f! refill pump KINJECT r1a END %R]INJECTGSQQERHMWWYIH[LIRXLIJPS[VEXIMW^IVSGEYWIWEVYR XMQIIVVSV8LMWWMXYEXMSRMWRSXHIXIGXIHF]QIXLSHTVIVYR &-%'36) 2 Instrument Handbook E-21

265 Writing BIACORE 2000 methods Command Comments Injected volume Sample consumption Max flow rate INJECT General purpose inject µl Injected vol +30 µl 100 µl/min KINJECT QUICKINJECT Low dispersion inject for kinetics and for samples sensitive to dilution with running buffer. Controlled dissociation time after sample injection. Low sample consumption. Less preparation time than INJECT but more sample dispersion µl Injected vol +40 µl 100 µl/min µl Injected vol +10 µl 100 µl/min COINJECT Injection of second sample immediately following the first (e.g. for dissociation in the presence of ligand). Variable contact times for first sample only. Recovery of either first or second sample, but not both. Sample 1: µl Sample 2: µl Sample 1: injected vol +40 µl Sample 2: injected vol +40 µl 100 µl/min BIGINJECT Large volume inject. Direct inject up to 325 µl, sample loop between 326 and 425 µl, direct between 426 and 750 µl µl Injected vol +52 µl 50 µl/min* *BIGINJECT cannot be performed at higher flow rates because the autosampler tubing must be refilled during injection from the sample loop for samples larger than 425 µl. E.4.3 Variable contact time injections 8LIWEQTPIZSPYQITEVEQIXIVJSVINJECTGSQQERHWGERFIIMXLIVE WMRKPIZSPYQISVEPMWXSJSVZSPYQIWWITEVEXIHF]GSQQEW 7MRKPIZSPYQI :SPYQIPMWX -RWMRKPIGLERRIPQSHIXLMWMWXLIWEQTPIZSPYQI MRNIGXIHXLVSYKLXLIGYVVIRXP]WIPIGXIHJPS[GIPP-R QYPXMGLERRIPQSHIXLMWZSPYQI[MPPFIMRNIGXIHSZIV EPPJPS[GIPPWMRXLIGYVVIRXP]WIPIGXIHJPS[TEXL %PMWXSJZSPYQIWWTIGMJMIWZEVMEFPIGSRXEGXXMQIWMR QYPXMGLERRIPQSHI8LIJMVWXWIGSRHXLMVHERHJSYVXL ZSPYQIWETTP]XSJPS[GIPPWERHVIWTIGXMZIP] )EGLZSPYQIMRXLIZSPYQIPMWXQYWXFIPIWWXLERSVXLIWEQIEWXLI TVIGIHMRKZSPYQI8LYWINJECT 35,25,20,10ERHINJECT 35,25,25,10EVIZEPMHFYXINJECT 35,20,25,10MWRSX E-22 &-%'36) 2 Instrument Handbook

266 Writing BIACORE 2000 methods %RINJECTGSQQERH[MXLEZSPYQIPMWXWIXWXLIJPS[TEXLEWVIUYMVIH JSVXLIHYVEXMSRSJXLIMRNIGXMSR;LIRXLIMRNIGXMSRMWGSQTPIXIXLI JPS[TEXLMWVIWXSVIHXSXLITVIZMSYWWIXXMRK MAIN DETECTION 1,2,3,4 path APROG immob END! All four flow cells in flow DEFINE APROG immob FLOW 5... INJECT r1a1 35,25! Flow cells 1 and 2 for INJECT...! Back to all four flow cells END 6IQIQFIVXSYWIDETECTION 1,2,3,4MRXLIMAINFPSGOXSWIXQYPXM GLERRIPHIXIGXMSRMJ]SYYWIZEVMEFPIGSRXEGXXMQIWMRINJECT GSQQERHW-J]SYWIXWMRKPIGLERRIPHIXIGXMSRQSHI]SY[MPPMRNIGX WEQTPISZIVQYPXMTPIJPS[GIPPW[MXLZEVMEFPIGSRXEGXXMQIWFYX]SY [MPPVIGSVHXLIWIRWSVKVEQJVSQSRIGIPPSRP]WIIRSXIEFSZI 2SXI %RINJECTGSQQERH[MXLEWMRKPIZSPYQITEVEQIXIVHSIW RSXEJJIGXXLIJPS[TEXL E.4.4 Sample recovery and fraction collection 8STVSKVEQWEQTPIVIGSZIV]SVJVEGXMSRGSPPIGXMSRTPEGIXLIRECOVER SVFRACTIONSGSQQERHHMVIGXP]EJXIVXLIINJECTGSQQERHXS[LMGL MXETTPMIW=SYGERLS[IZIVIRXIVEGSQQIRXSVEREXTRACLEAN GSQQERHFIX[IIRXLIINJECTERHXLIRECOVERSVFRACTIONS GSQQERH E.4.5 The EXTRACLEAN command %YXSQEXMGVSYXMRIW[EWLXLIPMUYMHLERHPMRKW]WXIQEJXIVIEGLINJECT GSQQERHERHEJXIVIEGLAPROG-JQSVIXLSVSYKLGPIERMRKMWVIUYMVIH YWIXLIEXRACLEANGSQQERHGSVVIWTSRHMRKXSExtra CleanupMRXLI InjectHMEPSKYIFS\MRXLIQERYEPGSRXVSPMRXIVJEGI7II%TTIRHM\& JSVVIGSQQIRHEXMSRWGSRGIVRMRK[LIRXSYWIXLII\XVEGPIERYT VSYXMRIW 8LIEXTRACLEANGSQQERHQYWXFITPEGIHHMVIGXP]EJXIVXLIINJECT GSQQERHXS[LMGLMXETTPMIW=SYGERLS[IZIVIRXIVEGSQQIRXSVE FRACTIONSSVRECOVERGSQQERHFIX[IIRXLIINJECTERHXLI EXTRACLEANGSQQERH &-%'36) 2 Instrument Handbook E-23

267 Writing BIACORE 2000 methods E.4.6 User tools in methods 8LI[SVOMRKXSSPWFlush, Normalize, PrimeERH RinseERHXLIWIVZMGI XSSPUncloggingGERFIYWIHFIX[IIRG]GPIWMREQIXLSH)RXIVXLI GSVVIWTSRHMRKGSQQERHMRXLIMAINFPSGOWII%TTIRHM\*JSV GSQQERHHIXEMPW 8LIYWIVXSSPStandbyQE]FIEHHIHEXXLIIRHSJEQIXLSHYWMRKXLI APPENDGSQQERHMRXLIMAINFPSGO8LMWTYXWXLIMRWXVYQIRXMR WXERHF]QSHI[LIRXLIQIXLSHMWGSQTPIXIH E.5 Common errors and error handling 8LMWWIGXMSRHIWGVMFIWWSQISJXLIQSWXGSQQSRQMWXEOIW]SY[MPP QEOI[LIR[VMXMRKQIXLSHWERHLS[&-%'36)'SRXVSP7SJX[EVI LERHPIWMRGSVVIGXQIXLSHW E.5.1 General error handling %PPQIXLSHWEVIXIWXIHMRXLI&-%'36)'SRXVSP7SJX[EVITVIVYR JEGMPMX]FIJSVIXLI]EVIVYR=SYQE]GLSSWIXSHSXLMWI\TPMGMXP]EX ER]WXEKIEPXIVREXMZIP]]SYQE]XV]XSVYRXLIQIXLSHHMVIGXP]MR [LMGLGEWIXLITVIVYRMWHSRIEYXSQEXMGEPP]FIJSVIXLIQIXLSHMW EGXYEPP]EGXMZEXIH 8LITVIVYRJEGMPMX]GLIGOWXLIW]RXE\ERHGSRWMWXIRG]SJ]SYVQIXLSH -JXLIVIEVIER]IVVSVWIVVSVQIWWEKIWEVI[VMXXIRMRXLIETTVSTVMEXI TPEGIWMRXLIEGXYEPQIXLSHJMPI%RIVVSVQIWWEKIMWMHIRXMJMIHF]XLI X[SGLEVEGXIVWl""zEXXLIFIKMRRMRKSJXLIPMRI%QIWWEKIEFSZIXLI IVVSVMHIRXMJMIWXLIREXYVISJXLIIVVSVERHEPMRIFIPS[XLIIVVSVLEWE GEVIXBTSMRXMRKXSXLIMRGSVVIGX[SVH... >> Invalid vial position * INJECT r4a1 20 >> ^! r4 is not a valid rack number... -J]SYGSVVIGXIVVSVWMREQIXLSHFYXPIEZIXLIIVVSVQIWWEKIWMRTPEGI EPPPMRIWFIKMRRMRK[MXLl""zEVIWXVMTTIHSYXSJXLIQIXLSHJMPIEXXLI RI\XTVIVYR=SYHSRSXRIIHXSVIQSZIXLIIVVSVQIWWEKIPMRIW QERYEPP]=SYGEREPWSYWIEdit:Adjust MethodXSVIQSZIIVVSV QIWWEKIWJVSQXLIQIXLSH E-24 &-%'36) 2 Instrument Handbook

268 Writing BIACORE 2000 methods E.5.2 Syntax errors %QSRKXLIQSWXGSQQSRIVVSVWEVIW]RXE\IVVSVWFVIEOMRKXLIVYPIW SJ1(0*SVMRWXERGIJSVKIXXMRKXLIENDEXXLIIRHSJEDEFINE APROGMWEW]RXE\IVVSV =SYQE]QEOIW]RXE\IVVSVWXLVSYKLMRGSVVIGXYWEKIJSVMRWXERGI XV]MRKXSTYXERACKGSQQERHMREDEFINE APROGFPSGO)SVXLVSYKL X]TMRKQMWXEOIWJSVMRWXERGI[VMXMRKAPORG MRWXIEHSJAPROG 2SVQEPP]&-%'36)'SRXVSP7SJX[EVIGERRSXHMWXMRKYMWLFIX[IIR XLIWIWMXYEXMSRWrFSXLKMZIERl9RVIGSKRM^IHGSQQERHz DEFINE APROG test FLOW 20 >> Unrecognized command RACK 1 Thermo_a >> ^! RACK cannot be used in APROG MAIN >> Unrecognized command APORG test >> ^! APORG should be APROG 2SXIXLEXWSQIIVVSVWQE]EVMWIEWEHMVIGXGSRWIUYIRGISJTVIZMSYW QMWXEOIWIZIRXLSYKLXLIW]RXE\MRMXWIPJMWGSVVIGX-RXLII\EQTPI FIPS[XLIENDGSQQERHMWQMWWMRKJVSQXLIDEFINE APROG[MXLXLI VIWYPXXLEXXLIW]RXE\GLIGOIVLEWHMJJMGYPX]MRXIVTVIXMRKXLIGSVVIGXP] [VMXXIRMAINFPSGO DEFINE APROG test FLOW 20 INJECT r1a1 10! END missing >> Expecting keyword END MAIN >>^ DETECTION 2 >> Characters outside DEFINE-END or MAIN-END blocks APROG test >> ^ END &-%'36) 2 Instrument Handbook E-25

269 Writing BIACORE 2000 methods 8SQEOIMRXIVTVIXEXMSRSJGSRWIUYIRXMEPIVVSVWIEWMIV[LIR]SYLEZI WIZIVEPIVVSVQIWWEKIWMRSRIQIXLSHGSVVIGXXLIIVVSVWSRIEXEXMQI WXEVXMRKEXXLIFIKMRRMRKSJXLIQIXLSHERHVITIEXPrerunEJXIVIEGL GSVVIGXMSR E.5.3 Incorrect variable names 1MWWTIPPMRKEZEVMEFPIREQIGEYWIWERIVVSVMRX[SGSRXI\XW ;MXLMREDEFINEFPSGOHIJMRMRKALERTAPROGSVMESSAGEXLI ZEVMEFPIREQIWHIGPEVIHMRXLIPARAMGSQQERHQYWXFIXLIWEQIEW XLSWIYWIHMRGSQQERHW[MXLMRXLIFPSGO 8LIZEVMEFPIREQIWHIGPEVIHMRXLILPARAMGSQQERHMREDEFINE LOOPQYWXFIXLIWEQIEWXLSWIYWIHMRGSQQERHW[MXLMRXLILOOP GSRWXVYGXMSR 1MWXEOIWMRXLIWIWMXYEXMSRWKIRIVEXIXLIJSPPS[MRKIVVSVQIWWEKIW DEFINE APROG test PARAM %flow %pos %vol >> Expecting an integer value, min=0 max=100 FLOW %folw >> ^! variable name should be %flow END INJECT %pos %vol DEFINE LOOP cycle LPARAM %flow %pos %vol 5 r1a r1a1 10 END MAIN DETECTION LOOP cycle STEP >> Expecting an integer value, min=5 max=325 APROG test %flow %pos %volume >> ^! variable name should be %vol ENDLOOP END E-26 &-%'36) 2 Instrument Handbook

270 Writing BIACORE 2000 methods E.5.4 Incorrect parameter values 1ER]TEVEQIXIVWLEZIEVIWXVMGXIHVERKISJZEPYIW=SYGERRSXJSV MRWXERGIWIPIGXJPS[GIPPRYQFIVXLIVIEVISRP]JPS[GIPPWSVQSZI XLIEYXSWEQTPIVRIIHPIXSTSWMXMSRVEXLITSWMXMSRHSIWRSXI\MWX 4IVQMXXIHVERKIWEVIKMZIRMRXLIHIXEMPIHHIWGVMTXMSRWJSVIEGL GSQQERH%TTIRHM\*&-%'36)'SRXVSP7SJX[EVI[EVRW]SYJSV MRGSVVIGXTEVEQIXIVZEPYIW... >> Invalid vial position * INJECT r4a1 20 >> ^! r4 is not a valid rack number... l-rzepmhzmeptswmxmsrzmwewtigmepgewimrxlexxlihmjjivirxtswmxmsrw EVIZEPMHJSVHMJJIVIRXVEGOW-JXLIQIXLSHHSIWRSXGSRXEMRERACK GSQQERHXLIW]RXE\[MPPFIGLIGOIHMRXLIGSRXI\XSJXLIGYVVIRX VEGOWIXXMRK E.5.5 Incorrect parameter ordering 4EVEQIXIVWZEPYIWSVZEVMEFPIREQIWQYWXFIKMZIRMRXLIVMKLXSVHIV [LIR]SYGEPPERAPROGALERTSVMESSAGEJVSQXLIMAINFPSGO8LI SVHIVQYWXFIXLEXKMZIRMRXLIPARAMHIGPEVEXMSR -J]SYQEOIEQMWXEOIMRXLISVHIVMRKXLIGLERGIWEVIXLEX]SY[MPP KIRIVEXIEW]RXE\IVVSV8LIJSPPS[MRKMRGSVVIGXQIXLSHJSVMRWXERGI XVMIWXSYWIXLIZEPYIJSVEYXSWEQTPIVTSWMXMSRERHVEJSVZSPYQI DEFINE APROG test PARAM %vol %pos FLOW 20 INJECT %pos %vol END MAIN DETECTION 1 >> Expecting a value value, min=5 max=325 APROG test r1a1 10 >> ^! APROG test is defined with volume before position END &-%'36) 2 Instrument Handbook E-27

271 Writing BIACORE 2000 methods -RWSQIGEWIWMRGSVVIGXTEVEQIXIVSVHIVMRKQMKLXKSYRHIXIGXIHF] &-%'36)'SRXVSP7SJX[EVI8LIJSPPS[MRKQIXLSHMWW]RXEGXMGEPP] ZEPMH DEFINE APROG test PARAM %flow %pos %vol FLOW %flow INJECT %pos %vol END DEFINE LOOP cycle LPARAM %vol %pos %flow 10 r1a r1a1 10 END MAIN DETECTION 1 LOOP APROG test %vol %pos %flow ENDLOOP END 8LITEVEQIXIVSVHIVMWVIZIVWIHFIX[IIRXLIAPROGGEPPERHXLIPARAM HIGPEVEXMSRFYXXLIQIXLSHTEWWIWTIVJIGXP]ZEPMHTEVEQIXIVZEPYIWXS XLIAPROG[MXLXLIVIWYPXXLEX PMWMRNIGXIHEX PQMRMRWXIEHSJ PEX PQMR)VVSVWSJXLMWOMRHGERFIHIXIGXIHF]MRWTIGXMSRSJ XLIIZIRXPSKJSVXLIVYR E.5.6 Incorrect number of variables =SYQYWXHIGPEVIEPPZEVMEFPIWYWIHMRETVSKVEQ%RIVVSVVIWYPXW SXLIV[MWI DEFINE APROG test PARAM %pos %vol >> Expecting an integer value, min=0 max=100 FLOW %flow >> ^! variable %flow not declared END INJECT %pos %vol =SYQE]HIGPEVIQSVIZEVMEFPIWXLER]SYYWIrJSVMRWXERGIHYVMRK HIZIPSTQIRX[SVO]SYQE]lGSQQIRXSYXzGSQQERHW[LMGLYWI ZEVMEFPIWFYX]SYHSRSXRIIHXSVIQSZIXLIZEVMEFPIREQIWJVSQXLI HIGPEVEXMSR E-28 &-%'36) 2 Instrument Handbook

272 Writing BIACORE 2000 methods DEFINE APROG test PARAM %sample %mix %vol FLOW 20! TRANSFER %sample %mix %vol! MIX %mix 100 INJECT %mix 10 END! parameters %sample and %volume not used -RXLIWEQI[E]TEVEQIXIVZEPYIWQYWXFIWYTTPMIHJSVEPPZEVMEFPIW YWIHMRETVSKVEQ-J]SYHSRSXWYTTP]IRSYKLZEPYIWERIVVSV VIWYPXW DEFINE APROG test PARAM %flow %pos %vol FLOW %flow INJECT %pos %vol END MAIN DETECTION 1 >> Missing parameter APROG test 25 r1a1 >> ^! APROG test requires 3 parameters.! No value given for %vol END E.5.7 Prerun error messages 8LMWWIGXMSRPMWXWXLIIVVSVQIWWEKIWXLEXGERFIMRWIVXIHMRXSXLI QIXLSHJMPI[LIRTVIVYRIRGSYRXIVWEW]RXE\IVVSV GSQQERHWQYWXTVIGIHIEPPSXLIVMRWXVYQIRXGSQQERHX]TIW :EVMEFPIHIGPEVEXMSRWPARAMMRAPROGLPARAMMRLOOPQYWXFI XLIJMVWXGSQQERHWMRXLIFPSGOKEYWORDMODECURVEERHFLOW QYWXTVIGIHISXLIVGSQQERHWMRAPROG LEWEPVIEH]FIIRYWIH =SYLEZIVITIEXIHSRISJRECOVERFRACTIONSSVEXTRACLEAN EJXIVERMRNIGXMSRGSQQERH &-%'36) 2 Instrument Handbook E-29

273 Writing BIACORE 2000 methods QYWXFIXLIJMVWXGSQQERHMR RACKQYWXFIXLIJMVWXGSQQERHMRMAIN[MXLXLII\GITXMSRSJ ALERT[LMGLGERTVIGIHIRACK RSXEPPS[IHEJXIV%44)2( 2SGSQQERHWQE]FI[VMXXIREJXIVAPPENDMRXLIMAINFPSGO RSXEPPS[IH[MXL =SYLEZIYWIHEGSQFMREXMSRSJGSQQERHW[LMGLMWRSX TIVQMXXIH RECOVERGERRSXFIYWIH[MXLFRACTIONS RECOVERSVFRACTIONSGERRSXFIYWIH[MXLBIGINJECT FRACTIONSGERRSXFIYWIH[MXLCOINJECT &YJJIVSZIVJPS[ 8LMWIVVSVGEREVMWIMJEPMRIMRXLIQIXLSHHIJMRMXMSRI\GIIHW GLEVEGXIVW 'LEVEGXIVWSYXWMHI()*-2))2(SV1%-2)2(FPSGOW %PPGLEVEGXIVWMREQIXLSHJMPIQYWXFI[MXLMRQIXLSHHIJMRMXMSR FPSGOW%GSQQSRWSYVGISJXLMWIVVSVMWSQMWWMSRSJDEFINEEX XLIFIKMRRMRKSJEFPSGO'SRWIUYIRXMEPIVVSVQIWWEKIWGERIEWMP] EVMWIEWEVIWYPX 'SRJPMGXMRK04%6%1 2IWXIHPSSTWYWITEVEQIXIVREQIWMRGSQQSR8LIIVVSVQIWWEKI TSMRXWXSXLIMRRIVLOOPGSQQERH )2(0334[MXLSYXQEXGLMRK0334 8LILOOPGSQQERHQYWXFIFEPERGIH[MXLERENDLOOPEJXIVXLI GSQQERHWXSFIVITIEXIHF]XLIPSST3QMXXMRKXLILOOPKMZIW XLMWIVVSVQIWWEKI )\TIGXMRK (ITIRHMRKSRXLIGSRXI\XSJXLIIVVSVXLIW]RXE\GLIGOIVQE]FI EFPIXSWYKKIWXERI\TIGXIHOI][SVHZEPYISVGLEVEGXIV )\TIGXMRKEVIEPRYQFIVQMR!QE\! 8LIGSQQERHVIUYMVIWEVIEPRYQFIVMIMRXIKIVSVHIGMQEP ZEPYIEWETEVEQIXIV8LMWQIWWEKIGEREVMWIMJEZEVMEFPIREQIMW MRGSVVIGXWSXLEXXLITEVEQIXIVZEPYIWYTTPMIHXSXLIGSQQERHMW RSXEVIEPRYQFIVMRXLIGSVVIGXVERKI )\TIGXMRKEXMQIZEPYIQMR!QE\! %XMQITEVEQIXIVLEWFIIRKMZIREZEPYI[LMGLMWSYXWMHIXLI VERKITIVQMXXIHMRXLIGSRXI\XJSVI\EQTPIEXMQI[LMGLMW PSRKIVXLERW )\TIGXMRKERMRXIKIVZEPYIQMR!QE\! 8LIGSQQERHVIUYMVIWERMRXIKIVZEPYIEWETEVEQIXIV8LMW QIWWEKIGEREVMWIMJEZEVMEFPIREQIMWMRGSVVIGXWSXLEXXLI E-30 &-%'36) 2 Instrument Handbook

274 Writing BIACORE 2000 methods TEVEQIXIVZEPYIWYTTPMIHXSXLIGSQQERHMWRSXERMRXIKIVMRXLI GSVVIGXVERKI *VEGXMSREPXMQIMWRSXEPPS[IH 8LMWQIWWEKIETTIEVW[LIREHIGMQEPXMQIMWWTIGMJMIH[LIVISRP] MRXIKIVXMQIWTIGMJMGEXMSRWEVIEPPS[IHIKWAIT -RGSQTEXMFPITEVEQIXIVW =SYLEZIYWIHEZEVMEFPIMRHMJJIVIRXGSRXI\XW[LMGLEVIRSX GSQTEXMFPI[MXLIEGLSXLIV8LMWQIWWEKIEPWSETTIEVWMJ]SYYWI XLIWEQIZEVMEFPIJSVXLIZSPYQI[MXLBIGINJECTERHERSXLIV MRNIGXMSRGSQQERH -RZEPMHFPSGOREQI =SYLEZIYWIHEREQI[LMGLMWRSXZEPMHEWXLIREQISJE QIXLSHHIJMRMXMSRFPSGO2EQIWQE]GSRXEMRPIXXIVWHMKMXWERH XLIWTIGMEPGLEVEGXIVW CERHFYXQE]RSXFIKMR[MXLEHMKMX SVYRHIVWGSVIGLEVEGXIV -RZEPMHVEGOX]TI 8LITEVEQIXIV]SYLEZITVSZMHIHJSVXLIRACKGSQQERHMWRSXE ZEPMHVEGOX]TI:EPMHX]TIWEVITHERMO_ATHERMO_BTHERMO_C ERHMICRO -RZEPMHXMQI =SYLEZIYWIHERMRZEPMHXMQIWTIGMJMGEXMSR+MZIXMQIWMR WIGSRHWWWWWQMRYXIWERHWIGSRHWQQQWWSVLSYVWQMRYXIW ERHWIGSRHWLLQQWW8LIWTIGMJMGEXMSRQMRYXIERH WIGSRHWMWMRZEPMH -RZEPMHZEVMEFPIREQI =SYLEZIYWIHEREQI[LMGLMWRSXZEPMHEWXLIREQISJE ZEVMEFPI2EQIWQE]GSRXEMRPIXXIVWHMKMXWERHXLIWTIGMEP GLEVEGXIVW CERHFYXQE]RSXFIKMR[MXLEHMKMXSV YRHIVWGSVIGLEVEGXIV -RZEPMHZMEPTSWMXMSR =SYLEZIWTIGMJMIHEVEGOTSWMXMSRXLEXMWRSXZEPMHJSVXLIGYVVIRX VEGOX]TI4SWMXMSRWEVIlZEPMHzMRXLIGSRXI\XSJXLIGYVVIRXVEGO HIJMRMXMSR'LIGOXLITSWMXMSRWTIGMJMGEXMSRERHXLIVEGOX]TI GYVVIRXP]HIJMRIHMRCommand:Define Rack Base 1IXLSHRSXJSYRH =SYLEZIYWIHAPPEND[MXLERMRGSVVIGXQIXLSHREQI3RP]XLI MRXIVREPQIXLSHSTANDBYQE]FIGEPPIH[MXLAPPEND 1MWWMRK 5YSXEXMSRQEVOWEVSYRHXI\XIKMRLTEXTCTEXTRTEXT CAPTIONGSQQERHWQYWXFIQEXGLIH &-%'36) 2 Instrument Handbook E-31

275 Writing BIACORE 2000 methods 1MWWMRKTEVEQIXIV =SYLEZIGEPPIHEFPSGO[MXLJI[IVTEVEQIXIVWXLEREVIHIGPEVIH MRXLIGSVVIWTSRHMRKFPSGOHIJMRMXMSR 1MWWMRKVIJIVIRGIXMQI =SYLEZIWTIGMJMIHERIKEXMZIXMQIJSVERRPOINTGSQQERHMR APROG[MXLRSTVIGIHMRKQEVOIHGSQQERHXSWIXXLIVIJIVIRGI XMQI 2EQIHYTPMGEXIH =SYLEZIYWIHXLIWEQIREQIX[MGIMRXLIWEQIGSRXI\X8[S FPSGOWSJXLIWEQIX]TIQE]RSXLEZIXLIWEQIREQIERH ZEVMEFPIWHIGPEVIH[MXLPARAMQYWXFIYRMUYI[MXLMRSRIFPSGO 9WIXLIEdit:FindGSQQERHMRXLIQIXLSHIHMXSVXSJMRHHMJJIVIRX SGGYVVIRGIWSJXLIWEQIREQI 3RP]SRIHIGMQEPEPPS[IH 8LMWQIWWEKIETTIEVW[LIRQSVIXLERSRIHIGMQEPMWWTIGMJMIH JSVERYQFIVXLEXSRP]WYTTSVXWSRIHIGMQEP 3RP]SRI1%-2EPPS[IH %QIXLSHQE]RSXGSRXEMRQSVIXLERSRIMAINFPSGOHIJMRMXMSR 3TXMSR[MXLSYXZEPYI =SYLEZIYWIHEGSQQERHSTXMSR[MXLSYXWTIGMJ]MRKEZEPYI [LIVISRIMWVIUYMVIHJSVI\EQTPICURVEXIQTT 7XVMRKXSSPSRK =SYLEZIYWIHEWXVMRKMRJSVI\EQTPIETITLECTEXTLTEXTSV RTEXTGSQQERHXLEXMWPSRKIVXLERXLIQE\MQYQTIVQMXXIH PIRKXL 7]RXE\IVVSV 8LIVIMWERIVVSVMRXLIQIXLSHHIJMRMXMSRXLEXXLIW]RXE\GLIGOIV GERRSXMHIRXMJ] 8LMWGSQQERHGERRSXFIYWIHEWEVIJIVIRGITSMRX =SYGERSRP]WIXEVIJIVIRGITSMRXJSVXMQIHIZIRXWMRER APROG MIJSVXLIGSQQERHWBIGINJECTCOINJECTDILUTEINJECT KINJECTQUICKINJECTMIXERHTRANSFER 8LMWGSQQERHQYWXJSPPS[MQQIHMEXIP]EJXIV =SYLEZIYWIHEGSQQERHMRXLI[VSRKGSQQERHSVHIV8LI GSQQERHMRUYIWXMSRQYWXJSPPS[HMVIGXP]EJXIVERMRNIGX GSQQERH8LMWIVVSVQE]SGGYV[MXLXLIGSQQERHW RECOVER FRACTIONSERHEXTRACLEAN 8MQIQE]RSXI\GIIHW 8LIQE\MQYQXMQIEPPS[IHJSVG]GPIMW WEFSYX HE]W=SYLEZITVSFEFP]QEHIEQMWXEOIMREXMQIWTIGMJMGEXMSR E-32 &-%'36) 2 Instrument Handbook

276 Writing BIACORE 2000 methods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nstrument Handbook E-33

277 Writing BIACORE 2000 methods E-34 &-%'36) 2 Instrument Handbook

278 Method language reference F. Method language reference 8LMWGLETXIVMWEGSQTPIXIVIJIVIRGIXSXLIQIXLSHIHMXSVGSQQERHW /I][SVHWEVI[VMXXIRMRYTTIVGEWI 4EVEQIXIVMHIRXMJMIVWEVI[VMXXIRMRPS[IVGEWIMXEPMGW 3TXMSREPGSQTSRIRXWEVIIRGPSWIHMRWUYEVIFVEGOIXW?A %RIPPMTWMWMWYWIHXSMRHMGEXIXLEXEPMWXSJGSQTSRIRXWQE]FI I\XIRHIHMRHIJMRMXIP] 3TXMSRWEVIYWIHSRP]JSVWTIGMEPTYVTSWIWWYGLEWQEMRXIRERGIERH XIWXQIXLSHW%PPGSQQERHWLEZIHIJEYPXSTXMSRWIXXMRKW[LMGLEVI WYMXEFPIJSVRSVQEPETTPMGEXMSRW l(ijeypxzepyiwzevizepyiwxlex[mppfiywihjsvteveqixivwerh STXMSRWMJRSSXLIVZEPYIMWWTIGMJMIH8LIHIJEYPXZEPYIWETTP]IZIR MJXLISTXMSRMWSQMXXIHJVSQXLIGSQQERH Command summary 8LIXEFPIWFIPS[PMWXXLIGSQQERHW[LMGLGERFIYWIHMRIEGLQIXLSH FPSGOXSKIXLIV[MXLEFVMIJMRHMGEXMSRSJXLIGSQQERHTYVTSWI8LI VIQEMRHIVSJXLMWGLETXIVHIWGVMFIWXLIGSQQERHWMRHIXEMPPMWXIHMR EPTLEFIXMGEPSVHIV ()*-2)%0)68FPSGO BEEP 7SYRHWEFIITXSEPIVXXLISTIVEXSV CTEXT ;VMXIWXI\XGIRXIVIHLSVM^SRXEPP]MRFS\ GO (MWTPE]WEFYXXSRJSVlGSRXMRYIzJYRGXMSR LTEXT ;VMXIWXI\XPIJXEPMKRIHMRFS\ PARAM (IGPEVIWZEVMEFPIWYWIHMRALERT RTEXT ;VMXIWXI\XVMKLXEPMKRIHMRFS\ STOP (MWTPE]WEFYXXSRJSVlEFSVXzJYRGXMSR TITLE 7IXWXLIXMXPIFEVXI\XJSVXLIALERTFS\ *

279 Method language reference * ()*-2)%463+FPSGO BIGINJECT 0EVKIZSPYQIMRNIGXMSR PXS-*' CAPTION 7IXWGETXMSRJSVWIRWSVKVEQSVGYVZI COINJECT (YEPMRNIGXMSRXS-*' CURVE 4PSXWZEPYIWJSVEWXEXYWMXIQ DILUTE (MPYXIWSRIWEQTPI[MXLERSXLIV DIPNEEDLE 4VIHMTWRIIHPIMRZMEP EXTRACLEAN )\XVE[EWLTVSGIHYVIEJXIVMRNIGXMSR FLOW 7IXWJPS[VEXI FLOWPATH (IJMRIWJPS[TEXL FRACTIONS 6IGSZIVWHMWWSGMEXMRKWEQTPIHYVMRKFYJJIVJPS[ INJECT -RNIGXWWEQTPIXS-*' KEYWORD (IJMRIWGSPYQRMRVITSVXTSMRXXEFPI KINJECT 0S[HMWTIVWMSRMRNIGXMSR MIX 1M\IWWEQTPI MODE 7IXWHEXEGSPPIGXMSRTEVEQIXIVW PARAM (IGPEVIWZEVMEFPIWYWIHMRAPROG QUICKINJECT 5YMGOMRNIGXMSRXS-*' RECOVER 6IGSZIVWMRNIGXIHWEQTPISVVIEKIRX RPOINT 7IXWVITSVXTSMRX SCALE 7IXWHIJEYPXWGEPIJSVWIRWSVKVEQERHWXEXYWGYVZIW TRANSFER 8VERWJIVWWEQTPIFIX[IIREYXSWEQTPIVZMEPW WAIT ;EMXWEWTIGMJMIHXMQI WASH ;EWLIW-*'GLERRIPWERHSVEYXSWEQTPIVRIIHPI WASHREC ;EWLIWVIGSZIV]GYT ()*-2)0334FPSGO LPARAM (IGPEVIWZEVMEFPIWYWIHMRDEFINE LOOP TIMES 7IXWXLIVITIEXGSYRXJSVWIXWSJZEVMEFPIW ()*-2)1)77%+)FPSGO BEEP 7SYRHWEFIITXSEPIVXXLISTIVEXSV CLOCK (MWTPE]WGSYRXHS[RXMQIV CTEXT ;VMXIWXI\XGIRXIVIHLSVM^SRXEPP]MRFS\ LIFETIME 7LS[WXMQIJSV[LMGLQIWWEKILEWFIIRHMWTPE]IH LTEXT ;VMXIWXI\XPIJXEPMKRIHMRFS\ PARAM (IGPEVIWZEVMEFPIWYWIHMRMESSAGE RTEXT ;VMXIWXI\XVMKLXEPMKRIHMRFS\ TITLE 7IXWXLIXMXPIFEVXI\XJSVXLIMESSAGE FS\

280 Method language reference * 1%-2FPSGO ALERT (MWTPE]WERALERTFS\ APPEND 6YRWEXIVQMREXMSRVSYXMRIEJXIVXLIQIXLSH APROG 6YRWEREREP]WMWTVSKVEQAPROG BEEP 7SYRHWEFIITXSEPIVXXLISTIVEXSV DETECTION 7IPIGXWHIXIGXMSRQSHIERHMRMXMEPJPS[TEXL DILUTE (MPYXIWSRIWEQTPI[MXLERSXLIV ENDLOOP )RHWEPSSTHIJMRMXMSR FLOW 7IXWJPS[VEXI FLUSH 6YRWXLI[SVOMRKXSSPFlush LOOP 6ITIEXWSRISVQSVIGSQQERHW MESSAGE (MWTPE]WEMESSAGEFS\ MIX 1M\IWWEQTPI NORMALIZE 6YRWXLI[SVOMRKXSSPNormalize PRIME 6YRWXLI[SVOMRKXSSPPrime RACK 7IXWXLIVEGOFEWIX]TI RINSE 6YRWXLI[SVOMRKXSSPRinse TEMP 7IXWXLIHIXIGXMSRXIQTIVEXYVI TRANSFER 8VERWJIVWWEQTPIFIX[IIREYXSWEQTPIVZMEPW UNCLOG 6YRWXLI[SVOMRKXSSPUnclogging WAIT ;EMXWEWTIGMJMIHXMQI WASH ;EWLIWPMUYMHLERHPMRKW]WXIQGSQTSRIRXW WASHREC ;EWLIWVIGSZIV]GYT

281 Method language reference ALERT name [parameter...] (MWTPE]WERALERTFS\ 9WEKIMAINFPSGO 4EVEQIXIVW REQI TEVEQIXIV 2EQISJXLIALERTFS\EWWTIGMJMIHMR DEFINE ALERT 4EVEQIXIVWYTTPMIHXSXLIALERTFS\ 4EVEQIXIVWWYTTPMIHXSXLIALERTQYWXGSVVIWTSRHWIUYIRXMEPP]XSXLI ZEVMEFPIWHIGPEVIHMRXLIPARAMGSQQERHMRDEFINE ALERT ;MXLMRELOOPGSRWXVYGXMSRTEVEQIXIVWQE]FIWYTTPMIHEWIMXLIV GSRWXERXZEPYIWSVZEVMEFPIREQIW:EVMEFPIREQIWQYWXFIXLIWEQI EWXLSWIHIGPEVIHMRXLIGSVVIWTSRHMRKDEFINE LOOP3YXWMHIELOOP TEVEQIXIVWQYWXFIWYTTPMIHEWGSRWXERXZEPYIW )\EQTPI DEFINE ALERT check_solution PARAM %pos LTEXT Place test solution in position %pos GO OK END MAIN ALERT check_solution r1a1 END *

282 Method language reference APPEND standby 6YRWXLI[SVOMRKXSSPStandbyEJXIVXLIQIXLSH 9WEKIMAINFPSGO 4EVEQIXIVW WXERHF] 2EQISJXLIXSSPXSVYREJXIVXLIQIXLSH%X TVIWIRXSRP]XLIStandbyXSSPMWEGGITXIH 8LIAPPENDGSQQERHQYWXFIXLIPEWXGSQQERHMRXLIMAINFPSGO 2SXI )\EQTPI 8LI[SVOMRKXSSPContinueTVSZMHIHMRTVIZMSYWZIVWMSRW SJ&-%'36)GSRXVSPWSJX[EVILEWFIIRVIREQIHXS Standby*SVFEGO[EVHGSQTEXMFMPMX]XLIREQIContinue QE]WXMPPFIYWIH[MXLXLIAPPENDGSQQERH MAIN DETECTION 1 APROG immob APROG analyse... APROG regenerate APPEND standby END *

283 Method language reference APROG name [parameter...] 6YRWEREREP]WMWTVSKVEQAPROG 9WEKIMAINFPSGO 4EVEQIXIVW REQI TEVEQIXIV 2EQISJXLITVSKVEQEWWTIGMJMIHMRDEFINE APROG 4EVEQIXIVWYTTPMIHXSXLIAPROG 4EVEQIXIVWWYTTPMIHXSXLIAPROGQYWXGSVVIWTSRHWIUYIRXMEPP]XSXLI ZEVMEFPIWHIGPEVIHMRXLIPARAMGSQQERHMRDEFINE APROG ;MXLMRELOOPGSRWXVYGXMSRTEVEQIXIVWQE]FIWYTTPMIHEWIMXLIV GSRWXERXZEPYIWSVZEVMEFPIREQIW:EVMEFPIREQIWQYWXFIXLIWEQI EWXLSWIHIGPEVIHMRXLIGSVVIWTSRHMRKDEFINE LOOP3YXWMHIELOOP TEVEQIXIVWQYWXFIWYTTPMIHEWGSRWXERXZEPYIW )\EQTPI DEFINE APROG injection PARAM %position %volume FLOW 20 INJECT %position %volume END MAIN DETECTION 1 APROG injection r1a1 20 APROG injection r2a5 35 END *

284 Method language reference [*] [time] BIGINJECT position volume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large_vol FLOW 50 *0:30 BIGINJECT r1a :00 RPOINT response!rpoint 4 min!after injection start END *

285 Method language reference BEEP 7SYRHWEFIITJVSQXLIGSQTYXIVXSEPIVXXLISTIVEXSV 9WEKIDEFINE ALERTDEFINE MESSAGEERHMAINFPSGOW 4EVEQIXIVW 2SRI )\EQTPI MAIN APROG immob BEEP APROG analyse END DEFINE MESSAGE box BEEP BEEP LTEXT Don't miss this message! END CAPTION text 7IXWXLIGETXMSRSJXLIGYVVIRXKVETL 9WEKIDEFINE APROGFPSGO 4EVEQIXIVW XI\X 8I\XXSFIYWIHEWGETXMSR )\EQTPI DEFINE APROG temp_check CAPTION Temperature check CURVE temp FLOW 10 INJECT r1a1 10 END *

286 Method language reference CLOCK time [option] (MWTPE]WGSYRXHS[RXMQIV 9WEKIDEFINE MESSAGEFPSGO 4EVEQIXIVW XMQI 7XEVXMRKXMQIJSVXMQIV*SVQEX,, SV777 3TXMSR Q (MWTPE]XMQIMRQMRYXIWSRP] )\EQTPI DEFINE MESSAGE countdown RTEXT minutes remaining CLOCK 5:00 -m END *

287 Method language reference [*] [time] COINJECT position1 volume1 position2 volume2 0S[HMWTIVWMSRMRNIGX[MXLX[SHMVIGXJSPPS[MRKMRNIGXMSRWIKJSV GSQTIXMRKPMKERHHYVMRKHMWWSGMEXMSRTLEWI 9WEKIDEFINE APROGFPSGO 4EVEQIXIVW XMQI TSWMXMSR ZSPYQI TSWMXMSR ZSPYQI 8MQIWTIGMJMGEXMSRMRJSVQEX,, SV777%REWXIVMWOFIJSVIXLI XMQIWTIGMJMGEXMSRWIXWXLMWIZIRXXMQIEWE ^IVSXMQIVIJIVIRGIJSVWYFWIUYIRXXMQI WTIGMJMGEXMSRW 6EGOTSWMXMSRJSVWEQTPI :SPYQISJWEQTPISVEPMWXSJYTXSJSYV ZSPYQIWWITEVEXIHF]GSQQEWJSVZEVMEFPI GSRXEGXXMQIMRNIGXMSRWWII7IGXMSR) 6ERKI P 6EGOTSWMXMSRJSVWEQTPI :SPYQISJWEQTPIVERKI P 7EQTPIMWMRNIGXIHJVSQXLI-*'WEQTPIPSST7EQTPIMWMRNIGXIHMR HMVIGXMRNIGXQSHI:EVMEFPIGSRXEGXXMQIWGERSRP]FIWTIGMJMIHJSV WEQTPI FRACTIONSGERRSXFIYWIH[MXLCOINJECTRECOVERGERFIYWIHJSV IMXLIVWEQTPISVWEQTPIFYXRSXFSXL 7EQTPIGSRWYQTXMSRMWMRNIGXIHZSPYQI PJSVFSXLWEQTPIERH WEQTPI )\EQTPI DEFINE APROG double_inject FLOW 50 *0:30 COINJECT r1a1 50 r1a :50 RPOINT resp1!rpoint 1st inject 3:50 RPOINT resp2!rpoint 2nd inject END *

288 Method language reference CTEXT [text] ;VMXIWGIRXIVIHXI\XMRERALERTSVMESSAGEFS\ 9WEKIDEFINE ALERTERHDEFINE MESSAGEFPSGOW 4EVEQIXIVW XI\X 8I\XGSRXEMRMRKYTXSGLEVEGXIVW% FPEROPMRIMWMRWIVXIHMRXLIMESSAGEFS\MJ XI\XMWSQMXXIH )\EQTPI DEFINE MESSAGE demo TITLE DEMONSTRATION BOX LTEXT This text is left-adjusted CTEXT This text is centered RTEXT This text is right-adjusted END *

289 Method language reference CURVE item [option...] 4PSXWXLIZEPYISJEWXEXYWMXIQMRXLIWIRWSVKVEQ[MRHS[ 9WEKIDEFINE APROGFPSGO 4EVEQIXIVW MXIQ 2EQISJXLIWXEXYWMXIQXSTPSX XIQT HIXIGXMSRYRMXXIQTIVEXYVI FYJJPS[ FYJJIVJPS[VEXIGSRXMRYSYW JPS[TYQT FYJJZSP ZSPYQISJFYJJIVVIQEMRMRKMR XLIGSRXMRYSYWJPS[TYQT 3TXMSRW S 3TXMSREPTPSX6IPEXIZEPYIWXSXLIVMKLX LERHE\MW TR HR 4IRGSPSYVMHIRXMJMIHF]R FPEGO KVI] FPYI PMKLXFPYI KVIIR PMKLXKVIIR G]ER PMKLXG]ER VIH PMKLXVIH QEKIRXE PMKLXQEKIRXE ]IPPS[ PMKLX]IPPS[ [LMXI FVMKLX[LMXI 6IEHMRKVEXIMRXIVZEP:EPYIWJSVRMR WIGSRHW6ERKIW(IJEYPXZEPYIWEVI JSVXIQTERHJSVFYJJPS[ERHFYJJZSP )\EQTPI DEFINE APROG temp_check CURVE temp -o FLOW 10 INJECT r1a1 10 END *

290 Method language reference DEFINE ALERT (IJMRIWEHMEPSKYIFS\TVSQTXMRK[MXLlGSRXMRYIzERHlEFSVXz FYXXSRW 9WEKI&PSGOHIJMRMXMSR 7]RXE\ DEFINE ALERT name [PARAM variable...] other commands END 8LIGOERHSTOPGSQQERHWVIWTIGXMZIP]EVIYWIHMRXLI DEFINE ALERTFPSGOXSWTIGMJ]XI\XSRXLIlGSRXMRYIzERHlEFSVXzFYXXSRWMR XLIALERTFS\ -JXLIFPSGOGSRXEMRWRIMXLIV STOPRSVGOGSQQERHWXLIFS\[MPP MRGPYHIElGSRXMRYIzFYXXSR[MXLXLIXI\XContinueERHERlEFSVXz FYXXSR[MXLXLIXI\XAbort-JXLIFPSGOGSRXEMRWIMXLIV STOPSVGOFYX RSXFSXLXLIFS\[MPPMRGPYHISRIFYXXSREWETTVSTVMEXIFYXRSXXLI SXLIV2SXIXLEXEDEFINE ALERTFPSGO[LMGLGSRXEMRWSRP]ESTOP GSQQERH[MPPHMWTPE]EHMEPSKYIFS\[LIVIXLISRP]STXMSRMWXS EFSVXXLIQIXLSH 8LIMRWXVYQIRXHSSVMWYRPSGOIHERHXLIQIXLSHMWTEYWIH[LMPIXLI EPIVXFS\MWHMWTPE]IH'PMGOMRKSRXLIlGSRXMRYIzFYXXSRVIWYQIW QIXLSHSTIVEXMSR'PMGOMRKSRXLIlEFSVXzFYXXSRWXSTWXLIQIXLSH MQQIHMEXIP] *

291 Method language reference DEFINE APROG (IJMRIWEWIVMIWSJSTIVEXMSRWEWEREREP]WMWTVSKVEQ 9WEKI&PSGOHIJMRMXMSR 7]RXE\ DEFINE APROG name FLOW flowrate other commands END )EGLAPROGQYWXGSRXEMREFLOWGSQQERH8LIFLOWGSQQERHQYWX FIXLIJMVWXSTIVEXMSREPGSQQERHMRXLIAPROG'SQQERHWXLEXEJJIGX WIXXMRKWWYGLEWCAPTIONMODECURVEEVITPEGIHFIJSVIXLIFLOW GSQQERH DEFINE LOOP 7IXWYTEXEFPISJZEPYIWJSVVITIXMXMZIEREP]WIW 9WEKI&PSGOHIJMRMXMSR 7]RXE\ DEFINE LOOP name LPARAM variable... [TIMES count] value END 7II7IGXMSR)JSVHIXEMPWSJPSSTHIJMRMXMSR *

292 Method language reference DEFINE MESSAGE (IJMRIWEQIWWEKIFS\ 9WEKI&PSGOHIJMRMXMSR 7]RXE\ DEFINE MESSAGE name [PARAM variable...] other commands END %QIWWEKIFS\HMWTPE]WMRJSVQEXMSRJSVXLISTIVEXSV8LIQIWWEKI VIQEMRWHMWTPE]IHYRXMPERI[MESSAGEGSQQERHMWMWWYIHERALERT FS\MWHMWTPE]IHSVXLIQIXLSHIRHW (MWTPE]MRKEQIWWEKIFS\HSIWRSXMRXIVVYTXXLIQIXLSHSVYRPSGOXLI MRWXVYQIRXHSSVGSQTEVIDEFINE ALERT %QIWWEKIFS\GERRSXFIGPSWIHMRXIVEGXMZIP]HYVMRKQIXLSH STIVEXMSR-J]SY[ERXXSHIJMRIEQIWWEKIXSFIEGORS[PIHKIHF]XLI STIVEXSVYWIEDEFINE ALERTFPSGOERHMRGPYHIEGOGSQQERHMR XLIFPSGO *

293 Method language reference DETECTION fc 7IXWHIXIGXMSRQSHIERHMRMXMEPJPS[TEXL 9WEKIMAINFPSGO 4EVEQIXIVW JG 1SHIWIXXMRKW :EPYIWWIIXEFPIFIPS[ 8LIDETECTIONGSQQERHWIXWHIXIGXMSRQSHIJPS[TEXLERH VIJIVIRGIWYFXVEGXMSREGGSVHMRKXSXLIJSPPS[MRKXEFPI JGZEPYI (IXIGXMSR *PS[TEXL 6IJIVIRGI WYFXVEGXMSR RSXETTPMGEFPI RSXETTPMGEFPI RSXETTPMGEFPI RSXETTPMGEFPI RSRI RSRI 3XLIVGSQFMREXMSRWSJJPS[GIPPRYQFIVWMRXLIJGTEVEQIXIVEVI MRZEPMH 8LIJPS[TEXLFYXRSXXLIHIXIGXMSRQSHISVVIJIVIRGIWYFXVEGXMSR GERWYFWIUYIRXP]FIGLERKIH[MXLXLIFLOWPATHGSQQERHMRER APROG8LIINJECTGSQQERH[MPPEPWSXIQTSVEVMP]WIXXLIJPS[TEXL MJVIUYMVIH7II7IGXMSR)JSVQSVIHIXEMPW DETECTIONVITPEGIWXLIFLOWCELLGSQQERHYWIHMRIEVPMIVZIVWMSRW SJXLIWSJX[EVIFLOWCELLMWWXMPPEGGITXIHJSVGSQTEXMFMPMX][MXL QIXLSHW[VMXXIRMRIEVPMIVZIVWMSRW )\EQTPI MAIN DETECTION 1,2,3,4 APROG immob APROG analyse END *

294 Method language reference [*] [time] DILUTE position1 position2 mixpos percent 1M\IWX[SWEQTPIWMREXLMVHTSWMXMSR9WIHJSVHMPYXMRKERHQM\MRK 9WEKIDEFINE APROGERHMAINFPSGOW 4EVEQIXIVW XMQI TSWMXMSR TSWMXMSR QM\TSW TIVGIRX 8MQIWTIGMJMGEXMSRMRJSVQEX,, SV777%REWXIVMWOFIJSVIXLI XMQIWTIGMJMGEXMSRWIXWXLMWIZIRXXMQIEWE ^IVSXMQIVIJIVIRGIJSVWYFWIUYIRXXMQI WTIGMJMGEXMSRW 6EGOTSWMXMSRJSVWEQTPI 6EGOTSWMXMSRJSVWEQTPI 6EGOTSWMXMSRJSVXLIQM\IHTVSHYGX *MREPTIVGIRXEKIGSRGIRXVEXMSRSJWEQTPI 6ERKI 8LIJMREPZSPYQISJQM\IHTVSHYGXMWJM\IHEX P8LYWDILUTE r1a1 r1a2 r1a3 20XEOIW PJVSQVEERH PJVSQVEERH QM\IWXLIQMRTSWMXMSRVE %DILUTEGSQQERHMWIWWIRXMEPP]IUYMZEPIRXXSX[STRANSFER GSQQERHWJSPPS[IHF]EMIXGSQQERHDILUTEMWLS[IZIVSTXMQM^IH JSVFIWXTIVJSVQERGI[MXLXLIJM\IHJMREPZSPYQISJ P DILUTEMWWYIHMREMAINFPSGO[MPPRSXFIVIGSVHIHMRXLIIZIRXPSK 8LIHMPYXMSREGGYVEG]QE]FIVIHYGIHMJXLIX[SWSPYXMSRWFIMRKQM\IH LEZIWMKRMJMGERXP]HMJJIVIRXZMWGSWMXMIW )\EQTPI DEFINE APROG incubation FLOW 25 *0:30 DILUTE r1a1 r1a2 r1b1 50 5:00 INJECT r1b1 50! inject 5 min after! mixing sample END *

295 Method language reference DIPNEEDLE position 4VIHMTWRIIHPIMRSTIVEXSVHIJMRIHZMEPXSHIGVIEWIGEVV]SZIVIJJIGXW 9WEKIDEFINE APROGFPSGO 4EVEQIXIVW TSWMXMSR 4SWMXMSRMR[LMGLRIIHPIMWXSFIHMTTIH PWSPYXMSRMWEWTMVEXIHMRXSXLIRIIHPIJVSQXLITVIHMTTSWMXMSRXS VMRWIXLIRIIHPI9WIXLMWGSQQERHMRGSRRIGXMSR[MXLVITIEXIH WEQTPMRKJVSQSRITSWMXMSR[LIVIXLIWEQTPIWSPYXMSRHMJJIVWKVIEXP] JVSQFYJJIVMRVIJVEGXMZIMRHI\T,MSRMGWXVIRKXLIXG )\EQTPI DEFINE APROG predip FLOW 25 DIPNEEDLE r1a2 INJECT r1a1 50 END!sample buffer END (IJMRIWXLIIRHSJEFPSGO 9WEKI&PSGOHIJMRMXMSR 4EVEQIXIVW 2SRI *

296 Method language reference ENDLOOP (IJMRIWXLIIRHSJEPSSTGSRWXVYGXMSRMRXLIMAINFPSGO 9WEKIMAINFPSGO 4EVEQIXIVW 2SRI EXTRACLEAN 4IVJSVQWI\XVEGPIERMRKSJXLI-*' 9WEKIDEFINE APROGFPSGO 4EVEQIXIVW 2SRI ;LIRYWIHEXTRACLEANQYWXEP[E]WFITPEGIHHMVIGXP]EJXIVXLI MRNIGXMSRGSQQERHXS[LMGLMXVIJIVW=SYGERLS[IZIVIRXIVE GSQQIRXSVERECOVERSVFRACTIONS GSQQERHFIX[IIRXLIINJECT ERHXLIEXTRACLEANGSQQERH8LIGPIERMRKTVSGIHYVI[MPPFI I\IGYXIHHMVIGXP]EJXIVXLIMRNIGXMSR )\EQTPI DEFINE APROG injection FLOW 20 INJECT r1a1 30 EXTRACLEAN!extra clean-up after injection END *

297 Method language reference [*] [time] FLOW flowrate [option...] 7IXWJPS[VEXI 9WEKIDEFINE APROGERHMAINFPSGOW 4EVEQIXIVW XMQI JPS[VEXI 8MQIWTIGMJMGEXMSRMRJSVQEX,, SV777%REWXIVMWOFIJSVIXLI XMQIWTIGMJMGEXMSRWIXWXLMWIZIRXXMQIEWE ^IVSXMQIVIJIVIRGIJSVWYFWIUYIRXXMQI WTIGMJMGEXMSRW *PS[VEXIMR PQMR6ERKI PQMR 3TXMSRW J *MPPGSRXMRYSYWJPS[TYQTFIJSVIWXEVXMRK JPS[ 8LIFLOWGSQQERHMWSFPMKEXSV]MREDEFINE APROGFPSGOERHQE] SRP]FITVIGIHIHF]XLIGSQQERHWCAPTIONCURVEKEYWORDMODE, PARAMERHSCALE 8LIXMQIWTIGMJMGEXMSRMWRSXWYTTSVXIHF]FLOWYWIHMREMAINFPSGO SVF]XLIJMVWXFLOWGSQQERHMRERAPROG &IX[IIRAPROGWMRXLIMAINFPSGOXLIJPS[VEXIMWWIXXS PQMRFI HIJEYPX9WIEFLOWGSQQERHMRXLIMAINFPSGOMJEHMJJIVIRXJPS[VEXI MWVIUYMVIH )\EQTPI DEFINE APROG sample_inject FLOW 10 INJECT r1a1 10 END *

298 Method language reference [*] [time] FLOWPATH fc 7IXWXLIFYJJIVJPS[TEXL 9WEKIDEFINE APROGFPSGO 4EVEQIXIVW XMQI JG 8MQIWTIGMJMGEXMSRMRJSVQEX,, SV777%REWXIVMWOFIJSVIXLI XMQIWTIGMJMGEXMSRWIXWXLMWIZIRXXMQIEWE ^IVSXMQIVIJIVIRGIJSVWYFWIUYIRXXMQI WTIGMJMGEXMSRW *PS[GIPPWXSFIMRGPYHIHMRXLITEXL:EPYIW ERH FLOWPATHHMVIGXWFYJJIVJPS[XLVSYKLXLIWTIGMJMIHJPS[GIPPWFYXHSIW RSXEJJIGXXLIHIXIGXMSRQSHI9WIDETECTIONMRXLIMAINFPSGOXSWIX FSXLHIXIGXMSRQSHIERHJPS[TEXL %28 FLOWPATHHSIWRSXEPXIVXLIHIXIGXMSRQSHIWIXF]DETECTIONMR MAIN-RWMRKPIGLERRIPHIXIGXMSRQSHIMXMWTSWWMFPIXSWIXXLIJPS[ TEXLXSGIPPW[LIVIHEXEMWRSXGSPPIGXIH7II7IGXMSR)JSVQSVI HIXEMPW *PS[TEXLGLERKIWEVIPSGEPXSXLIGYVVIRXAPROG;LIRXLIAPROGMW GSQTPIXIHXLIJPS[TEXLMWVIWXSVIHXSXLIWIXXMRKXLEXETTPMIHEXXLI WXEVXSJXLIAPROG 8LIJPS[TEXLWIXXMRKGERFIXIQTSVEVMP]SZIVVMHHIRF]EZEVMEFPI GSRXEGXXMQIINJECTGSQQERH8LIJPS[TEXLMWWIXXSXLI VIUYMVIQIRXWSJXLIINJECTGSQQERHJSVXLIHYVEXMSRSJXLIWEQTPI MRNIGXMSRWII7IGXMSR) )\EQTPI DEFINE APROG analytes FLOW 25 FLOWPATH 1 INJECT r1a1 50!from r1a1 to Fc1 FLOWPATH 2 INJECT r1a2 50!from r1a2 to Fc2 FLOWPATH 3 INJECT r1a3 50!from r1a3 to Fc3 END MAIN DETECTION 1,2,3,4 APROG analytes END!multichannel mode *

299 Method language reference FLUSH 6YRWXLI[SVOMRKXSSPFlushWII7IGXMSR 9WEKIMAINFPSGO 4EVEQIXIVW 2SRI *

300 Method language reference FRACTIONS startpos volume no 6IGSZIVWHMWWSGMEXMRKWEQTPIHYVMRKFYJJIVJPS[*VEGXMSRWGERSRP]FI GSPPIGXIHHYVMRKHMWWSGMEXMSRTLEWI 9WEKIDEFINE APROGFPSGO 4EVEQIXIVW WXEVXTSW ZSPYQI RS 6EGOTSWMXMSRJSVXLIJMVWXJVEGXMSR *VEGXMSRZSPYQI6ERKI P 2YQFIVSJJVEGXMSRW6ERKI 8LMWGSQQERHQYWXEP[E]WJSPPS[HMVIGXP]EJXIVSRISJXLIMRNIGXMSR GSQQERHWINJECTKINJECTSVQUICKINJECT=SYGERLS[IZIV IRXIVEGSQQIRXSVEREXTRACLEANGSQQERHFIX[IIRXLIINJECT ERHXLIFRACTIONSGSQQERHFRACTIONSMWRSXWYTTSVXIHJSV BIGINJECTSVCOINJECT 8LIQMRMQYQXMQIXSGSPPIGXSRIJVEGXMSRMWWJVEGXMSRWQMR-J XLIVIUYIWXIHJVEGXMSRWM^IMWWQEPPIVXLERXLIZSPYQIHIPMZIVIHMRXLMW XMQIXLIEGXYEPJVEGXMSRWM^I[MPPFIXLIWQEPPIWXXLEXGERFIGSPPIGXIH EXXLIGYVVIRXJPS[VEXI*SVI\EQTPIVIUYIWXMRK PJVEGXMSRWEXE JPS[VEXISJ PQMR[SYPHVIUYMVIJVEGXMSRWQMRXLIEGXYEP JVEGXMSRWM^I[MPPFI P -JXLIVEGOTSWMXMSRJSVXLIJMVWXJVEGXMSRMWWTIGMJMIHWSXLEXXLIVIEVI RSXIRSYKLTSWMXMSRWJSVEPPJVEGXMSRWXLIJVEGXMSRGSPPIGXMSR[MPPWXST EXXLIPEWXTSWMXMSRMRXLIVEGOERHVIQEMRMRKIJJPYIRX[MPPFIPSWXXS [EWXI )\EQTPI DEFINE APROG collect FLOW 20 INJECT r1a1 50 FRACTIONS r2a1 20 5!collect 5 fractions!of 20 µl EXTRACLEAN END *

301 Method language reference GO name (MWTPE]WEFYXXSRJSVlGSRXMRYIzJYRGXMSRMRERALERTFS\ 9WEKIDEFINE ALERTFPSGO 4EVEQIXIVW REQI 8I\XSJYTXSGLEVEGXIVW[LMGL[MPPFI HMWTPE]IHEWXLIREQISJXLIFYXXSR )\EQTPI DEFINE ALERT demo LTEXT This text is left-adjusted CTEXT This text is centered RTEXT This text is right-adjusted GO OK STOP CANCEL END *

302 Method language reference [*] [time] INJECT position volume +IRIVEPTYVTSWIWEQTPIMRNIGXMSR 9WEKIDEFINE APROGFPSGO 4EVEQIXIVW XMQI TSWMXMSR ZSPYQI 8MQIWTIGMJMGEXMSRMRJSVQEX,, SV777%REWXIVMWOFIJSVIXLI XMQIWTIGMJMGEXMSRWIXWXLMWIZIRXXMQIEWE ^IVSXMQIVIJIVIRGIJSVWYFWIUYIRXXMQI WTIGMJMGEXMSRW 6EGOTSWMXMSRJSVWEQTPI :SPYQIXSFIMRNIGXIHSVEPMWXSJYTXSJSY V ZSPYQIWWITEVEXIHF]GSQQEWJSVZEVMEFPI GSRXEGXXMQIMRNIGXMSRWWII7IGXMSR) 6ERKI P 7EQTPIGSRWYQTXMSRMWMRNIGXIHZSPYQI P )\EQTPI DEFINE APROG immob FLOW 5 DILUTE r1a5 r1a6 r1b1 50!NHS/EDC INJECT r1b1 35!NHS/EDC * INJECT r2a1 35!ligand 5:00 INJECT r1c1 35!ethanolamine END *

303 Method language reference KEYWORD header value (IJMRIWGSPYQRERHWIEVGLOI][SVHMRVITSVXTSMRXXEFPI 9WEKIDEFINE APROGFPSGO 4EVEQIXIVW LIEHIV ZEPYI 8I\XJSVXLIGSPYQRLIEHMRKMRXLIVITSVX TSMRXXEFPI 'SPYQRGSRXIRXWTIGMJMIHEWEZEVMEFPISVE GSRWXERXXI\XZEPYI 8LILIEHIVTEVEQIXIVQE]RSXFIWYTTPMIHEWEZEVMEFPI 8LMWGSQQERHGERFIYWIHXSVIGSVHGSRGIRXVEXMSRWJSVIZEPYEXMSR )\EQTPI DEFINE APROG report PARAM %conc KEYWORD Antibody Mab-451 KEYWORD Conc %conc FLOW 10 * INJECT r1a RPOINT baseline -b 30 RPOINT plateau END *

304 Method language reference [*][time] KINJECT position volume disstime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kinetics 1 FLOW 10 * KINJECT r1a !4 min dissociation -20 RPOINT baseline INJECT r1a3 5!Regeneration END *

305 Method language reference LIFETIME (MWTPE]WXLIXMQIJSV[LMGLEQIWWEKIFS\LEWFIIREGXMZI 9WEKIDEFINE MESSAGEFPSGO 4EVEQIXIVW 2SRI 8LMWGSQQERHMWVIGSQQIRHIHJSVHIZIPSTQIRXTYVTSWIWSRP]-XQE] FIYWIHXSHIXIVQMRIWYMXEFPIWXEVXMRKZEPYIWJSVXLICLOCKGSQQERH )\EQTPI DEFINE MESSAGE develop LTEXT This process has been running for LIFETIME END *

306 Method language reference LOOP name order command [...] ENDLOOP 6ITIEXWEWIUYIRGISJSRISVQSVIGSQQERHWYWMRKTEVEQIXIVWEW WTIGMJMIHMRDEFINE LOOP 9WEKIMAINFPSGO 4EVEQIXIVW REQI SVHIV GSQQERH 2EQISJXLILOOPEWWTIGMJMIHMRDEFINE LOOP 3RISJXLIOI][SVHWSTEPORDERSV RANDOM8LMWTEVEQIXIVHIJMRIWXLISVHIVMR [LMGLVS[WMRDEFINE LOOP[MPPFILERHPIH WIIEPWS7IGXMSR) STEP ORDER RANDOM WXITXLVSYKLXLIZEVMEFPIWIXWERH XLIRVITIEXEGGSVHMRKXSXLITIMES GSQQERH VITIEXXLIJMVWXZEVMEFPIWIX EGGSVHMRKXSXLITIMESGSQQERH XLIRXLIWIGSRHWIXERHWSSR [SVOXLVSYKLXLIZEVMEFPIWIXWMR VERHSQSVHIV %R]GSQQERH[LMGLMWZEPMH[MXLMRXLI MAINFPSGO 'SQQERHW[MXLMRXLILOOPGSRWXVYGXMSRVIUYMVITEVEQIXIVWMRXLI JSVQSJZEVMEFPIREQIW8LIREQIWYWIHMRXLIMAINFPSGOQYWXFIXLI WEQIEWXLSWIHIGPEVIHMRXLIGSVVIWTSRHMRKDEFINE LOOP )\EQTPI DEFINE LOOP cycle LPARAM %sample %pos %vol TIMES 3 Sample1 r1a1 10 Sample2 r1a2 15 Sample3 r1a3 10 END MAIN DETECTION 1 LOOP cycle RANDOM APROG analyse %sample %pos %vol ENDLOOP END *

307 Method language reference LPARAM variable... (IGPEVIWZEVMEFPIWYWIHMRDEFINE LOOP 9WEKIDEFINE LOOPFPSGO 4EVEQIXIVW ZEVMEFPI 2EQISJXLIZEVMEFPIYWIHMRXLIDEFINE LOOPFPSGO7II7IGXMSR)JSVZEVMEFPI REQMRKVYPIW 8LILPARAMGSQQERHQYWXEP[E]WFIXLIJMVWXGSQQERHMREDEFINE LOOPFPSGOERHQE]RSXFIYWIHQSVIXLERSRGIMRSRIFPSGO )\EQTPI DEFINE LOOP cycle LPARAM %sample %pos %vol TIMES 3 Sample1 r1a1 10 Sample2 r1a2 15 Sample3 r1a3 10 END MAIN DETECTION 1 LOOP cycle RANDOM APROG analyse %sample %pos %vol ENDLOOP END LTEXT [text] ;VMXIWPIJXEPMKRIHXI\XMRERALERTSVMESSAGEFS\ 9WEKIDEFINE ALERTERHDEFINE MESSAGEFPSGOW 4EVEQIXIVW XI\X 8I\XGSRXEMRMRKYTXSGLEVEGXIVW% FPEROPMRIMWMRWIVXIHMRXLIMESSAGEFS\MJ XI\XMWSQMXXIH )\EQTPI DEFINE MESSAGE demo TITLE DEMONSTRATION BOX LTEXT This text is left-adjusted CTEXT This text is centered RTEXT This text is right-adjusted END *

308 Method language reference MAIN (IJMRIWXLISVHIVMRKSJAPROGWERHGSQQERHWMREQIXLSH 9WEKI&PSGOHIJMRMXMSR 7]RXE\ MAIN other commands END )EGLQIXLSHQYWXGSRXEMRSRIERHSRP]SRIMAINFPSGO 'SQQERHWMRXLIMAINFPSGOEVIRSXVIGSVHIHMRXLIWIRWSVKVEQSV XLIIZIRXPSK7IRWSVKVEQERHIZIRXPSKVIGSVHMRKEVIEGXMZISRP] HYVMRKI\IGYXMSRSJERAPROG[LMGLMWGEPPIHJVSQXLIMAINFPSGO *

309 Method language reference MESSAGE [name] [parameter...] (MWTPE]WEMESSAGEFS\ 9WEKIMAINFPSGO 4EVEQIXIVW REQI TEVEQIXIV 2EQISJXLIMESSAGEFS\EWWTIGMJMIHMR DEFINE MESSAGE :EPYIJSVHMWTPE]MRXLIMESSAGEFS\ 4EVEQIXIVWWYTTPMIHXSXLIMESSAGEQYWXGSVVIWTSRHWIUYIRXMEPP]XS XLIZEVMEFPIWHIGPEVIHMRXLIPARAMGSQQERHMRDEFINE MESSAGE ;MXLMRELOOPGSRWXVYGXMSRTEVEQIXIVWQE]FIWYTTPMIHEWIMXLIV GSRWXERXZEPYIWSVZEVMEFPIREQIW:EVMEFPIREQIWQYWXFIXLIWEQI EWXLSWIHIGPEVIHMRXLIGSVVIWTSRHMRKDEFINE LOOP3YXWMHIELOOP TEVEQIXIVWQYWXFIWYTTPMIHEWGSRWXERXZEPYIW 8LIQIWWEKIFS\VIQEMRWSRXLIWGVIIRYRXMPERI[QIWWEKIFS\SVER EPIVXFS\MWHMWTPE]IHSVXLIQIXLSHIRHW%MESSAGEGSQQERH[MXL RSTEVEQIXIVW[MPPIVEWIER]I\MWXMRKQIWWEKIFS\JVSQXLIWGVIIR )\EQTPI DEFINE MESSAGE no_break LTEXT Do not interrupt this cycle! END MAIN MESSAGE no_break APROG unbroken MESSAGE!clear message box... END *

310 Method language reference [*][time] MIX position volume 1M\IWWEQTPIMREYXSWEQTPIVZMEP 9WEKIDEFINE APROGERHMAINFPSGOW 4EVEQIXIVW XMQI TSWMXMSR ZSPYQI 8MQIWTIGMJMGEXMSRMRJSVQEX,, SV777%REWXIVMWOFIJSVIXLI XMQIWTIGMJMGEXMSRWIXWXLMWIZIRXXMQIEWE ^IVSXMQIVIJIVIRGIJSVWYFWIUYIRXXMQI WTIGMJMGEXMSRW 6EGOTSWMXMSRJSVQM\MRK :SPYQIMR PXSQM\6ERKI P MIXMWWYIHMREMAINFPSGO[MPPRSXFIVIGSVHIHMRXLIIZIRXPSK )\EQTPI DEFINE APROG sample_mixing FLOW 10 TRANSFER r1a1 r1a2 60 TRANSFER B r1a2 30 MIX r1a2 75 INJECT r1a2 40 END *

311 Method language reference MODE option [...] 7IXWXLIHEXEGSPPIGXMSRTEVEQIXIVWJSVERAPROG 9WEKIDEFINE APROGFPSGO 4EVEQIXIVW 2SRI 3TXMSRW HR 6IEHMRKVEXIMRXIVZEPJSVIEGLWIRWSVKVEQMR WIGSRHW6ERKIrW PR 1E\MQYQRYQFIVSJGSPPIGXIHHEXETSMRXW EXXLIGYVVIRXVIEHMRKVEXIMRXIVZEPWII 7IGXMSR6ERKIr(IJEYPX -JMODEMWWIXMRAPROGER]WIXXMRKWWTIGMJMIH[MXLXLIRun:Data collection rateqirygsqqerheviszivvmhhir-jmodemwsqmxxihxli WIXXMRKWWTIGMJMIH[MXLXLIQIRYGSQQERHETTP]9WMRKEMODE GSQQERHEXXLIFIKMRRMRKSJERAPROG[MPPIRWYVIXLEXXLIVIEHMRK VEXIMWGSVVIGXJSVXLIQIXLSH MODEQE]SRP]FITVIGIHIHF]XLIGSQQERHW CAPTIONPARAM SCALEERHKEYWORD )\EQTPI DEFINE APROG low_resolution MODE -d10! reading rate 1 point/10 s FLOW 10 INJECT r1a1 10 END *

312 Method language reference NORMALIZE position 6YRWXLI[SVOMRKXSSPNormalizeWII7IGXMSR 9WEKIMAINFPSGO 4EVEQIXIVW TSWMXMSR 6EGOTSWMXMSRJSVXLIKP]GIVSPWSPYXMSR &-%RSVQEPM^MRKWSPYXMSR -JRSVQEPM^EXMSRHSIWRSXWYGGIIHXLIQIXLSH[MPPFIXIVQMREXIH PARAM variable... (IGPEVIWZEVMEFPIWYWIHMRERALERTAPROGSVMESSAGEFPSGOW 9WEKIDEFINE ALERTDEFINE APROGERHDEFINE MESSAGEFPSGO 4EVEQIXIVW ZEVMEFPI 2EQISJXLIZEVMEFPI7II7IGXMSR)JSV ZEVMEFPIREQMRKVYPIW %FPSGOQE]GSRXEMREWQER]PARAMGSQQERHWEWVIUYMVIH%PP PARAMGSQQERHWWLSYPHTVIGIHIER]SXLIVGSQQERHWMRXLIFPSGO *SVZEVMEFPIWYWIHMREDEFINE LOOPFPSGOWIILPARAM )\EQTPI DEFINE APROG injection PARAM %position %volume FLOW 5 INJECT %position %volume END MAIN DETECTION 1 APROG injection r1a1 20 APROG injection r2a5 35 END *

313 Method language reference PRIME 6YRWXLI[SVOMRKXSSPPrimeWII7IGXMSR 9WEKIMAINFPSGO 4EVEQIXIVW 2SRI [*] [time] QUICKINJECT position volume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quickie FLOW 20 QUICKINJECT r1a1 500 END *

314 Method language reference RACK base type 7IXWXLIEYXSWEQTPIVVEGOX]TI 9WEKIMAINFPSGO 4EVEQIXIVW FEWI!PIJXTSWMXMSR!VMKLXTSWMXMSR X]TI 8,)613C%!XLIVQSVEGOX]TI% 8,)613C&!XLIVQSVEGOX]TI& 8,)613C'!XLIVQSVEGOX]TI' 1-'63!QMGVSXMXVITPEXI8LIRACKGSQQERHHIJMRIWXLIVEGO TSWMXMSRWXLEXEVIEGGITXIHMRQIXLSHTVIVYR-JRSRACKGSQQERHMW YWIHTSWMXMSRWEVIEGGITXIHEGGSVHMRKXSXLIGYVVIRXWIXXMRKWYRHIV Command:Define Rack BaseMRXLIQERYEPGSRXVSPMRXIVJEGIWII 7IGXMSR8LMWIRWYVIWXLEXXLIGYVVIRXWIXXMRKWEVIZEPMHJSVXLI QIXLSH 8LIRACKGSQQERHGERFIVITIEXIHXSHIJMRIEPXIVREXMZITIVQMXXIH VEGOW8LIVEGOTSWMXMSRWEGGITXIHMRQIXLSHTVIVYR[MPPLS[IZIV XLIRFIVIWXVMGXIHXSXLITSWMXMSRWXLEXEVIGSQQSRXSXLIWTIGMJMIH VEGOW*SVI\EQTPI[MXLXLIGSQQERHWRACK 1 THERMO_AERHRACK 1 MICROXLITSWMXMSRVJMWRSXZEPMHJI\MWXWMREQMGVSXMXVI TPEXIFYXRSXMRXLIVQSVEGO% -JERACKGSQQERHMWYWIHEPPVEGOWXLEXEVIVIJIVIRGIHMRXLIQIXLSH QYWXFIWTIGMJMIHIKMJXLIMAINFPSGOMRGPYHIWSRP]RACK 1 THERMO_ATSWMXMSRWMRVEGO[MPPRSXFIZEPMH -JXLIGYVVIRXVEGOWIXXMRKW[LIRXLIQIXLSHMWWXEVXIHHSRSX GSVVIWTSRHXSXLIWIXXMRKWVIUYIWXIHMRXLIRACKGSQQERHXLIQIXLSH [MPPTVSQTXJSVVEGOWIXXMRKWYWMRKXLIDefine Rack Base HMEPSKYIFS\ JVSQXLIQERYEPGSRXVSPMRXIVJEGIWII7IGXMSR-J]SYGLSSWI CancelJVSQXLMWHMEPSKYIFS\XLIQIXLSH[MPPFIEFSVXIH8LMW TVSQTXWIVZIWEWEVIQMRHIVXLEXXLIVEGOWQE]RIIHXSFIGLERKIH XLISRP]WIXXMRKW[LMGLGERFIIRXIVIHMRXLIHMEPSKYIFS\EVIXLSWI WTIGMJMIHMRXLIRACKGSQQERH 8LIRACKGSQQERHQE]SRP]FITVIGIHIHF]ERALERTGSQQERH [MXLMRXLIMAINFPSGO8LYW]SYGERRSXGLERKIVEGOWIXXMRKWFIX[IIR APROGWMREQIXLSH )\EQTPI MAIN RACK 1 thermo_a RACK 2 thermo_b APROG analysis... END *

315 Method language reference RECOVER position volume [option] 6IGSZIVWMRNIGXIHWEQTPISVVIEKIRX8LIGSQQERHMWTPEGIHHMVIGXP] EJXIVERMRNIGXGSQQERH 9WEKIDEFINE APROGFPSGO 4EVEQIXIVW TSWMXMSR ZSPYQI 6EGOTSWMXMSRXS[LMGLXLIWEQTPI[MPPFI VIGSZIVIH :SPYQIXSFIVIGSZIVIH6ERKI P EJXIVCOINJECT6IGSQQIRHIH " PJSVFIWXTVIGMWMSRMRVIGSZIVIH ZSPYQI 3TXMSRW WR 7EQTPIXSVIGSZIVCOINJECTSRP] 4IVQMXXIHZEPYIWJSVREVIJMVWXWEQTPI ERHWIGSRHWEQTPI(IJEYPXZEPYI RECOVERQE]SRP]FIYWIH[MXLCOINJECTINJECTKINJECTERH QUICKINJECT=SYGERLS[IZIVIRXIVEGSQQIRXSVEREXTRACLEAN GSQQERHFIX[IIRXLIINJECTERHXLIRECOVERGSQQERHRECOVER MWRSXWYTTSVXIHJSVBIGINJECT 8LIRECOVERGSQQERHQYWXFITPEGIHMQQIHMEXIP]EJXIVXLIINJECT GSQQERHXS[LMGLMXETTPMIW-J]SY[ERXXSFIWYVISJVIGSZIVMRKEPP XLIMRNIGXIHWEQTPIWTIGMJ]EFSYX PQSVIXLERXLIZSPYQISJ WEQTPIMRNIGXIH ;MXLCOINJECTXLISTXMSRWQE]FIYWIHXSWTIGMJ]VIGSZIV]SJXLI WIGSRHMRNIGXIHWEQTPI8LMWSTXMSRMWRSXZEPMH[MXLSXLIV INJECT GSQQERHW-JRSSTXMSRMWWTIGMJMIH[MXLCOINJECTXLIJMVWXWEQTPIMW VIGSZIVIH RECOVERERHFRACTIONSQE]RSXFIYWIH[MXLXLIWEQIINJECT GSQQERH 8LIZSPYQIWTIGMJMIHJSVRECOVERMWXLIZSPYQI[LMGL[MPPFIXEOIR JVSQXLIVIGSZIV]GYT8LMWMWRSXRIGIWWEVMP]XLIWEQIEWXLIMRNIGXIH ZSPYQIIKMJXLIVIGSZIV]GYTMWTVMQIH[MXLFYJJIVEWMRXLI I\EQTPIFIPS[XLIVIGSZIVIHZSPYQIWLSYPHFIXLIWYQSJXLI TVMQMRKZSPYQIERHXLIMRNIGXIHZSPYQI )\EQTPI DEFINE APROG recovery FLOW 10 TRANSFER r1b1 R 200!prime recovery cup INJECT r1a1 100 RECOVER r2a1 310 END *

316 Method language reference RINSE 6YRWXLI[SVOMRKXSSPRinseWII7IGXMSR 9WEKIMAINFPSGO 4EVEQIXIVW 2SRI *

317 Method language reference time RPOINT id [option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report KEYWORD Mab-451 FLOW 10 * INJECT r1a RPOINT baseline -b 30 RPOINT plateau END *

318 Method language reference RTEXT [text] ;VMXIWVMKLXEPMKRIHXI\XMRERALERTSVMESSAGEFS\ 9WEKIDEFINE ALERTERHDEFINE MESSAGEFPSGOW 4EVEQIXIVW XI\X 8I\XGSRXEMRMRKYTXSGLEVEGXIVW% FPEROPMRIMWMRWIVXIHMRXLIFS\MJXI\XMW SQMXXIH )\EQTPI DEFINE MESSAGE demo TITLE DEMONSTRATION BOX LTEXT This text is left-adjusted CTEXT This text is centered RTEXT This text is right-adjusted END *

319 Method language reference SCALE axis min max [option...] 7IXWXLIHIJEYPXWGEPIJSVEWIRWSVKVEQSVGYVZI 9WEKIDEFINE APROGFPSGO 4EVEQIXIVW E\MW QMR QE\ %\MWXSWGEPI4IVQMXXIHZEPYIW \ EFWGMWWE ] PIJXLERHSVHMREXI S VMKLXLERHSTXMSREPSVHMREXI 1MRMQYQZEPYISRE\MW 1E\MQYQZEPYISRE\MW 3TXMSRW PXI\X 9WIXI\XEWPEFIPSRE\MW XZEPYI 7IXXMGOWEXMRXIVZEPZEPYI E %YXSWGEPI *SVG]GPIW[LIVIXLISCALEGSQQERHMWYWIHXLIWIXXMRKWHIJMRIXLI HIJEYPXWGEPIFSXLHYVMRKXLIVYRERH[LIRXLIVIWYPXJMPIMWSTIRIH %JXIVGLERKMRKXLIWGEPIXLIQIXLSHWGEPIGERFIVIWXSVIHYWMRKXLI MethodSTXMSRMRXLIView:ScaleHMEPSKYIFS\ *SVG]GPIW[LIVISCALEMWRSXYWIHXLIHIJEYPXWGEPIWIXXMRKMWAuto. 8LIVMKLXLERHSTXMSREPSVHMREXIMWYWIHJSVGYVZIWEGXMZEXIH[MXLXLI SSTXMSRMRXLI'96:)GSQQERH )\EQTPI DEFINE APROG fix_scale SCALE y FLOW 10 INJECT r1a1 10 END *

320 Method language reference STOP name (MWTPE]WEFYXXSRJSVlEFSVXzJYRGXMSRMRERALERTFS\ 9WEKIDEFINE ALERTFPSGO 4EVEQIXIVW REQI 8I\XSJYTXSGLEVEGXIVW[LMGL[MPPFI HMWTPE]IHEWXLIREQISJXLIFYXXSR )\EQTPI DEFINE ALERT demo TITLE DEMONSTRATION BOX LTEXT This text is left-adjusted CTEXT This text is centered RTEXT This text is right-adjusted GO OK STOP Cancel END *

321 Method language reference TEMP degrees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temp_series LPARAM %degrees TIMES END DEFINE APROG analysis... END MAIN LOOP temp_series STEP TEMP %degrees NORMALIZE r2f2 APROG analysis ENDLOOP END *

322 Method language reference TIMES count 7IXWXLIVITIEXGSYRXJSVWIXWSJZEVMEFPIW 9WEKIDEFINE LOOPFPSGO 4EVEQIXIVW GSYRX 6ITIEXGSYRX:EPYIWHIJEYPX 8LIVITIEXGSYRXETTPMIWXSWYFWIUYIRXVS[WSVZEVMEFPIW[MXLMRXLI DEFINE LOOPYRXMPERI[VITIEXGSYRXMWWIX8LIVITIEXGSYRXQE]FI WIXEWQER]XMQIWEWVIUYMVIHMREDEFINE LOOP 7IXXMRKEVITIEXGSYRXSJQIERWXLEXWYFWIUYIRXVS[W[MPPFI TVSGIWWIHXLVIIXMQIW )\EQTPI DEFINE LOOP cycle LPARAM %sample %pos %vol TIMES 3 Sample1 r1a1 10 Sample2 r1a2 15 Sample3 r1a3 10 END MAIN DETECTION 1 LOOP cycle RANDOM APROG analyse %sample %pos %vol ENDLOOP END TITLE text 7IXWXLIXMXPIFEVXI\XJSVERALERTSVMESSAGE FS\ 9WEKIDEFINE ALERTERHDEFINE MESSAGEFPSGOW 4EVEQIXIVW XI\X 8I\XGSRXEMRMRKYTXSGLEVEGXIVW8LI XMXPIQE]RSXFIPIJXFPERO )\EQTPI DEFINE ALERT demo TITLE DEMONSTRATION BOX LTEXT This text is left-adjusted CTEXT This text is centered RTEXT This text is right-adjusted GO OK STOP CANCEL END *

323 Method language reference [*][time] TRANSFER from-pos to-pos volume 8VERWJIVWWEQTPIFIX[IIREYXSWEQTPIVZMEPW 9WEKIDEFINE APROGERHMAINFPSGOW 4EVEQIXIVW XMQI JVSQTSW XSTSW ZSPYQI 8MQIWTIGMJMGEXMSRMRJSVQEX,, SV777%REWXIVMWOFIJSVIXLI XMQIWTIGMJMGEXMSRWIXWXLMWIZIRXXMQIEWE ^IVSXMQIVIJIVIRGIJSVWYFWIUYIRXXMQI WTIGMJMGEXMSRW 6EGOTSWMXMSRXSXVERWJIVJVSQ 6EGOTSWMXMSRXSXVERWJIVXS :SPYQIMR PXSXVERWJIV6ERKI P 8LIWTIGMEPZEPYI&JSVXLIJVSQTSWTEVEQIXIVXVERWJIVWPMUYMHJVSQ XLIIPYIRXZIWWIP 8LIWTIGMEPZEPYI6JSVXLIXSTSWTEVEQIXIVXVERWJIVWPMUYMHXSXLI VIGSZIV]GYT TRANSFERMWWYIHMREMAINFPSGO[MPPRSXFIVIGSVHIHMRXLIIZIRXPSK )\EQTPI DEFINE APROG recovery FLOW 10 TRANSFER r1b1 R 200!prime recovery cup INJECT r1a1 100 RECOVER r2a1 310 END *

324 Method language reference UNCLOG 6YRWXLI[SVOMRKXSSPUncloggingWII7IGXMSR 9WEKIMAINFPSGO 4EVEQIXIVW 2SRI WAIT time ;EMXWJSVEWTIGMJMIHXMQI 9WEKIDEFINE APROGERHMAINFPSGOW 4EVEQIXIVW XMQI ;EMXXMQIMRXLIJSVQEX,, SV777 *PS[GSRXMRYIWHYVMRKE[EMX ;MXLMRELOOPGSRWXVYGXMSRTEVEQIXIVWQE]FIWYTTPMIHEWIMXLIV GSRWXERXZEPYIWSVZEVMEFPIREQIW:EVMEFPIREQIWQYWXFIXLIWEQI EWXLSWIHIGPEVIHMRXLIGSVVIWTSRHMRKDEFINE LOOP3YXWMHIELOOP TEVEQIXIVWQYWXFIWYTTPMIHEWGSRWXERXZEPYIW )\EQTPI DEFINE LOOP wait_time LPARAM %delay TIMES 2 5:00 10:00 MAIN DETECTION 1 WAIT 2:00!2 minute delay LOOP wait_time STEP WAIT %w!vary wait time APROG analyse %sample %pos %vol APROG regenerate ENDLOOP END *

325 Method language reference [time] WASH item ;EWLIW-*'SVEYXSWEQTPIVRIIHPI 9WEKIDEFINE APROGERHMAINFPSGOW 4EVEQIXIVW XMQI MXIQ 8MQIWTIGMJMGEXMSRMRJSVQEX,, SV777 -XIQXS[EWL J!JPS[GIPPEVIE M!EPP-*'GLERRIPW P!-*'WEQTPIPSST R!RIIHPI W!-*'WEQTPIPMRI WASH ferhwash iviuymviersr^ivsjps[vexi%rexxiqtxxs I\IGYXIXLIWIGSQQERHW[MXLXLIJPS[VEXIWIXXS^IVSVIWYPXWMREVYR XMQIIVVSV )\EQTPI DEFINE APROG immob FLOW 5 DILUTE r1a5 r1a6 r1b1 50 TRANSFER r1a6 r1b1 70 MIX r1b1 115 INJECT r1b1 35 WASH n! extra needle wash INJECT r2a1 35 INJECT r1c1 35 END *

326 Method language reference WASHREC [option] ;EWLIWVIGSZIV]GYT;MXLERSTXMSRXLISTIVEXSVGERYWI[EWLMRK PMUYMHJVSQEZMEPTSWMXMSR 9WEKIDEFINE APROGERHMAINFPSGOW 3TXMSRW TTSW 7TIGMJ]VEGOTSWMXMSRJSV[EWLMRKWSPYXMSR -JXLITSTXMSRMWRSXWTIGMJMIHXLIVIGSZIV]GYT[MPPFI[EWLIH[MXL IPYIRXFYJJIV Example: DEFINE APROG recovery FLOW 25 2:00 WASHREC -pr2a1 INJECT r1a1 50 RECOVER r2b1 55 END *

327 Method language reference *

328 Index M Index EGXMZIWIRWSVKVEQ Add Report PointQIRYGSQQERH EHHMRKGSQQERHWXSXLIUYIYI EHHMRKVITSVXTSMRXW Adjust MethodQIRYGSQQERH Adjust ScaleQIRYGSQQERH EHWSVTXMSRXS-*'GLERRIPW& EMQJSVMQQSFMPM^IHPIZIP VIWYPXW EMVFYFFPIW EXPS[JPS[VEXIW& GEYWMRKFEWIPMRIWLMJXW GEYWMRKHVMJX GEYWMRKWTMOIW MRXLIJPS[GIPP QMRMQM^MRKTVSFPIQW EMVGMVGYPEXMSR EMVWIKQIRXW MRHMJJIVIRXMRNIGXMSRGSQQERHW MRPMUYMHLERHPMRK ALERTFPSGO)) ALERTQIXLSHGSQQERH* HSSVPSGO) EREP]WMWTVSKVEQWIIAPROG EREP]XI WM^IVERKI ETTEVIRXOMRIXMGGSRWXERXW APPENDQIXLSHGSQQERH)* ETTPMGEXMSR KYMHIPMRIW& TVSFPIQW X]TIW& [M^EVHW ETTPMGEXMSR[M^EVHQIRY ETTPMGEXMSR[M^EVHXIQTPEXIW ETTPMGEXMSR[M^EVHW FMRHMRKEREP]WMW MQQSFMPM^EXMSRT,WGSYXMRK MRVIWYPXJMPIW RSXIFSSO SRPMRILIPT VEGOIHMXSV VIKIRIVEXMSRWGSYXMRK WEQTPITSWMXMSRW WXEVXMRK WYVJEGITIVJSVQERGIXIWX WYVJEGITVITEVEXMSR APROGFPSGO)

329 Index MM EYXSQEXMGMRMXMEXMSRERHXIVQMREXMSRWIUYIRGIW) APROGOI][SVHW APROGQIXLSHGSQQERH* EYXSQEXMG GSQQERHUYIYI HEXEVIHYGXMSR JMRMWLMRKVSYXMRIW% TVITEVEXMSRVSYXMRIW% WGEPMRK EYXSWEQTPIV HMPYXMRKWEQTPIW MVVITVSHYGMFPIEMVWIKQIRXW QEPJYRGXMSR QM\MRKWEQTPIW TSWMXMSRMRKRIIHPI TYQT VEGOFEWIWIXXMRKW VEGOTSWMXMSRW) VEGOW VITPEGMRKRIIHPI XIQTIVEXYVIGSRXVSP XVERWJIVVMRKWEQTPIW EYXSWEZI QERYEPWIRWSVKVEQW QIXLSHVYRW EZIVEKIVIWTSRWI FEGXIVMEPGSRXEQMREXMSR FEWIPMRI JSVVITSVXTSMRXW MRWIRWSVKVEQW WLMJXW BaselineQIRYGSQQERH FEWIPMRIWXEFMPMX] BEEPQIXLSHGSQQERH)* &-%'36) W]WXIQHIWGVMTXMSR &-%'36)'SRXVSP7SJX[EVIWIIGSRXVSPWSJX[EVI &-%IZEPYEXMSR &-%QEMRXIRERGIOMX &-%WMQYPEXMSR BIAsimulationQIRYGSQQERH Biginject) BIGINJECTQIXLSHGSQQERH* FMRHMRKEREP]WMW FMRHMRKTEVXRIVW FMRHMRKVEXIW FMRHMRKWXYHMIW WMQYPEXMSR FMSLE^EVHW FPEROWYFXVEGXMSR

330 Index MMM FPEROWYVJEGIW FPSGOW MRQIXLSHHIJMRMXMSRW) &06JMPIW FYJJIVVIGSQQIRHEXMSRW' FYPOIJJIGXW& GERGIPPMRKEPIVXERHQIWWEKIHMWTPE]W) CAPTIONQIXLSHGSQQERH)* GEVV]SZIV& FIX[IIREREP]WMWG]GPIW& FIX[IIRFYJJIVERHZMEPW& FIX[IIRJPS[GIPPW& FIX[IIRMRNIGXMSRW& GLERKMRKJPS[TEXL) GLIGOMRKXLIXIQTIVEXYVIHYVMRKEVYR GLIQMGEPVIWMWXERGI' GPIEVMRK-*'GLERRIPW CLOCKQIXLSHGSQQERH)* CloseQIRYGSQQERH Close[SVOMRKXSSP GPSWMRK EQIRY E[MRHS[ GSRXVSPWSJX[EVI GPSWMRKHS[R) GPSWMRKXLIGSQQERHUYIYI Coinject COINJECTQIXLSHGSQQERH)* GSPPIGXMRKHEXE GSPPIGXMRKJVEGXMSRWWIIJVEGXMSRGSPPIGXMSR GSPSYVGSHMRK MRGYWXSQM^IHGSQQERHWIUYIRGIW CommandQIRY Define Rack Base Dilute Dock Flow Inject Mix Set Temperature Stop Flow Stop Inject Transfer Undock Wash GSQQERHUYIYI EHHMRKGSQQERHW GPSWMRKXLIUYIYI GSQQERHWXEXYW HIPIXMRKGSQQERHW

331 Index MZ IHMXMRKGSQQERHW KIRIVEXMRKQIXLSHW MRWIVXMRKGSQQERHW QIXLSHGSQQERHW WXEVXMRKXLIUYIYI WXSTTMRKXLIUYIYI [MRHS[ GSQQERHUYIYI[MRHS[QIRY GSQQERHWMRGYWXSQM^IHETTPMGEXMSRW GSQQERHWRSXEZEMPEFPI GSQQIRX MRGYWXSQM^IHGSQQERHWIUYIRGIW MRQIXLSHHIJMRMXMSRW) GSQQIRXMRKSYXQIXLSHGSQQERHW) GSQQYRMGEXMSR[MXLXLIMRWXVYQIRX GSQTYXIVTSVXGSRRIGXMSRW GSRGIRXVEXMSRQIEWYVIQIRX WMQYPEXMSR GSRHMXMSREPGSQQERHW MRGYWXSQM^IHGSQQERHWIUYIRGIW GSRJSVQERGI[MXLWEJIX]WXERHEVHW GSRJSVQEXMSREPGLERKIW& GSRRIGXSVFPSGO PIEOEKI VITPEGMRK GSRWIUYIRXMEPIVVSVWMRQIXLSHHIJMRMXMSRW) GSRWXERXWMRQIXLSHGSQQERHW) GSRXEGXXMQI GSRXEQMREXMSR[MXLFMSLE^EVHW Continue[SVOMRKXSSPWIIStandby GSRXMRYSYWJPS[TYQT GSRXVSPI\TIVMQIRXWJSVOMRIXMGEREP]WMW GSRXVSPWSJX[EVI GPSWMRK MRWXEPPEXMSR QYPXMTPIMRWXERGIW WGVIIRVIKMSRW ZIVWMSRRYQFIV GSRXVSPWSJX[EVITVSFPIQW GSSPMRKJER MRERHSYXPIX GSSPMRKXLIEYXSWEQTPIVGSQTEVXQIRX Copy GraphQIRYGSQQERH CopyQIRYGSQQERH GSYRXHS[RXMQIV* GVIEXMRKERI[QIXLSH CTEXTQIXLSHGSQQERH)* 'XVP&VIEO GYVZI GYVZIGPEWWIW CurvePMWXMRXSSPFEV CURVEQIXLSHGSQQERH)*

332 Index Z GYWXSQM^IHETTPMGEXMSRW GSQQERHWIUYIRGIHMWTPE] GSQQERHW GSRHMXMSREPGSQQERHW HIJMRMRKGSQQERHWIUYIRGIW HIXIGXMSRQSHI IQTX]ZMEPWJSVQM\MRK I\EQTPIW JPS[GIPPREQIW JPS[TEXL If-ThenGSQQERHW OI][SVHW QM\ VITSVXTSMRXW [EMX ZEVMEFPIW Vary By Cycle GYWXSQM^IHGSQQERHWIUYIRGIW GSPSYVGSHMRK CutQIRYGSQQERH GYXXMRKQEVOIHXI\X G]GPI) CyclePMWXMRXSSPFEV HEXEGSPPIGXMSRVEXI Data Collection RateQIRYGSQQERH HEXEUYEPMX] HEXEVIHYGXMSR HIEHZSPYQI% DEFINEALERTQIXLSHGSQQERH* DEFINEAPROG QIXLSHGSQQERH* DEFINELOOPQIXLSHGSQQERH* DEFINEMESSAGEQIXLSHGSQQERH* DEFINERACKBASEQIRYGSQQERH HIJMRMRKGYWXSQM^IHGSQQERHWIUYIRGIW HIJMRMRKVEGOW HIPE]FIX[IIRWIRWSVKVEQWMRQYPXMGLERRIPQSHI DeleteQIRYGSQQERH HIPIXMRKGSQQERHWJVSQXLIUYIYI HIPIXMRKVITSVXTSMRXW Desorb[SVOMRKXSSP HIXIGXMSR TVMRGMTPI YRMX DETECTIONQIXLSHGSQQERH* HIXIGXMSRQSHI) EXQIXLSHWXEVX MRGYWXSQM^IHGSQQERHWIUYIRGIW WTIGMJ]MRKEXQIXLSHWXEVX) HIXIGXSVEVVE]% HIXIVKIRXW&

333 Index ZM HMEPSKYIFS\MGSRW HMEPSKYIFS\IW MRQIXLSHW) DiluteQIRYGSQQERH DILUTEQIXLSHGSQQERH* HMPYXMSRGEPGYPEXSV HMSHIHIXIGXSVW% DIPNEEDLEQIXLSHGSQQERH* HMVIGXMRNIGXMSR HMWMRJIGXERXW' HMWTIVWMSR&& HMWTPE]MRKIRXMVIWIRWSVKVEQ HMWTPE]MRKQIWWEKIW) DockQIRYGSQQERH HSGOMRKTVSFPIQW HSGOMRKXLIWIRWSVGLMT HSSVPSGO MRQIXLSHW) H]REQMGVERKI OMRIXMGGSRWXERXW% EditQIRY Add Report Point Adjust Method Copy Copy Graph Cut Delete Find Paste REPLACE Select All Undo IHMXMRKGSQQERHWMRXLIUYIYI IHMXMRKRSXIFSSOIRXVMIW IHMXMRKVITSVXTSMRXW IQIVKIRG]WXST IQTX]ZMEPWJSVQM\MRK IQTX]MRKJPS[GIPPW IQTX]MRKXLIVIGSZIV]GLERRIP ENDOI][SVH) ENDQIXLSHGSQQERH* ENDLOOPQIXLSHGSQQERH** IR^]QEXMGTVSGIWWIW ITMXSTIQETTMRK WMQYPEXMSR IVVSVLERHPMRKMRQIXLSHIHMXSV) )6636MRHMGEXSV IVVSVQIWWEKIW) )YVSTIERHMVIGXMZIW

334 Index IZEPYEXMRK WIRWSVKVEQHEXE W]WXIQGLIGO EvaluationQIRYGSQQERH IZIRXPSK HMWTPE] I\XIRHIHZIVWMSR JVEGXMSRGSPPIGXMSRIRXVMIW QEVOIVW QIWWEKIW( [MRHS[ IZIRXQEVOIVW JSVJVEGXMSRW I\IGYXMRKAPROGW) ExitQIRYGSQQERH ExportQIRYGSQQERH I\TSVXMRKHEXE I\TSVXMRKRSXIFSSOGSRXIRXW I\XIRHIHIZIRXPSK Extended Event LogQIRYGSQQERH Extra Cleanup EXTRACLEANQIXLSHGSQQERH)* JMIPHWMRRSXIFSSO JMPIEWWSGMEXMSRWMR;MRHS[W)\TPSVIV FileQIRY Close Exit Export New Method Open Print Save Save As Show Info JMPPMRKXLITYQTW) FindQIRYGSQQERH JMRHMRKVIKIRIVEXMSRGSRHMXMSRW Fit To Page JPS[ MRGYWXSQM^IHGSQQERHWIUYIRGIW JPS[GIPPREQIW MRGYWXSQM^IHGSQQERHWIUYIRGIW JPS[GIPPW GSPSYVWMRWIRWSVKVEQ HIEHZSPYQI% HMQIRWMSRW% FlowQIRYGSQQERH FLOWQIXLSHGSQQERH)* JPS[TEXL EXQIXLSHWXEVX ZMM

335 Index ZMMM MRGYWXSQM^IHGSQQERHWIUYIRGIW MRZEVMEFPIGSRXEGXXMQIMRNIGXMSRW) WIPIGXMRK JPS[VEXI IJJIGXSRWEQTPIHMWTIVWMSR& VIGSQQIRHEXMSRW& FLOWCELLQIXLSHGSQQERH JSVZEVMEFPIGSRXEGXXMQIMRNIGXMSRW) FlowcellQIRYGSQQERH FLOWPATHQIXLSHGSQQERH)* ERHHIXIGXMSRQSHI) FLUSHQIXLSHGSQQERH* Flush[SVOMRKXSSP JVEGXMSRGSPPIGXMSR) EYXSWEQTPIVTSWMXMSRW IZIRXQEVOW [MXLStop Inject Fractions FRACTIONSQIXLSHGSQQERH* JYWIW% KIRIVEXMRKQIXLSHWJVSQXLIGSQQERHUYIYI GOQIXLSHGSQQERH)* KVEHMIRXWYVJEGIW&& LEVH[EVIIVVSV LE^EVHW,&7FYJJIV HelpQIRY LIXIVSKIRISYWMRXIVEGXMSRW WMQYPEXMSR LMKLWIRWMXMZMX]& MGSRWMRHMEPSKYIFS\IW MHIRXMJMIVJSVVITSVXTSMRXW MHIRXMJ]MRKIZIRXWMREWIRWSVKVEQ -*' WXERHF]ZEPZITSWMXMSRW ZEPZIQEPJYRGXMSR -*'ZEPZIW% If-ThenGSQQERHW MRGYWXSQM^IHGSQQERHWIUYIRGIW MQQSFMPM^EXMSRT,WGSYXMRK MQQSFMPM^MRKPMKERH MQTPMGXJPS[TEXLWIXXMRK) MRGSVVIGX EYXSWEQTPIVVEGOTSWMXMSRW) RYQFIVSJZEVMEFPIW) TEVEQIXIVSVHIVMRK)

336 Index M\ TEVEQIXIVZEPYIW) ZEVMEFPIREQIW) MRMXMEXMRKPMUYMHHIPMZIV]W]WXIQ Inject MRGYWXSQM^IHGSQQERHWIUYIRGIW INJECTQIXLSHGSQQERH)* MRNIGXMRKWEQTPIWIIWEQTPIMRNIGXMSR MRNIGXMSRQSHIW MRNIGXMSRSJEMV MRNIGXMSRTSVX PIEOEKI Injection PortWIVZMGIXSSP MRNIGXMSRXIGLRMUYIW& MRPMRIVIJIVIRGIGIPPW&& MRWIVXMRKEWIRWSVGLMT MRWIVXMRKGSQQERHWMRXLIUYIYI MRWXEPPEXMSR MRWXEPPMRKGSRXVSPWSJX[EVI MRWXVYQIRXSJJPMRI MRWXVYQIRXTVSFPIQW MRXIKVEXIH JPYMHMGGEVXVMHKIWII-*' MRZEPMHVEGOTSWMXMSRW MSRMGWXVIRKXLHMJJIVIRGIW& KEYWORDQIXLSHGSQQERH)* OI][SVHW MRGYWXSQM^IHGSQQERHWIUYIRGIW MRQIXLSHHIJMRMXMSRW) MRVITSVXTSMRXXEFPIW) KeywordsQIRYGSQQERH OMRIXMGEREP]WMW WMQYPEXMSR OMRIXMGEREP]WMW[M^EVH FEWIPMRIWXEFMPMX] GSRXVSPI\TIVMQIRXW VIWYPXW OMRIXMGGSRWXERXW Kinject KINJECTQIXLSHGSQQERH)* PEVKIZSPYQIMRNIGXMSRW PE]SYX QIXLSHHIJMRMXMSR) PIEOEKIEXXLIGSRRIGXSVFPSGO PIEZMRKXLIMRWXVYQIRXSZIVRMKLX 0)(MRHMGEXSVW% LIFETIMEQIXLSHGSQQERH* PMKERH MQQSFMPM^EXMSR PMKLXWSYVGI

337 Index \ PMRIEVJPS[VEXI% PMROIHVIEGXMSRWGSRXVSPI\TIVMQIRX PMUYMHHIPMZIV]W]WXIQ WXEVXYT PSEHMRKXLIWEQTPIPSST PSGO MRQIXLSHW) PSKEVMXLQMGWGEPI LOOPFPSGO)) LOOPQIXLSHGSQQERH)* RIWXIHPSSTW) PSSTGSRWXVYGXMSRW) PSSTMRNIGXMSR PSSTSVHIV) Lost Data Found PS[HEXEUYEPMX] GEYWIW QEVOMRKSRWIRWSVKVEQ LPARAMQIXLSHGSQQERH)* LTEXTQIXLSHGSQQERH)* MAIN* MAINFPSGO)) MAINQIXLSHGSQQERH* QEMRWJYWIW% QEMRWZSPXEKIWIXXMRK QEMRXIRERGI VIKYPEV WEJIX]VIUYMVIQIRXW QEMRXIRERGIOMX QERYEPGSRXVSP& QERYEPGSRXVSPMRXIVJEGI Manual Inject QERYEPQSHI WXSTTMRKXLIMRNIGXMSR ZSPYQIQSHI QERYEPMRNIGXMSR QERYEPWGEPMRK QERYEPWIRWSVKEQ WXSTWIRWSVKVEQ QERYEPWIRWSVKVEQ TVITEVEXMSR WEZIVIWYPXW QEVOIVW VITSVXTSMRXWERHIZIRXPSK MarkersQIRYGSQQERH QEWWXVERWJIVGSRXVSPI\TIVMQIRXW QEWWXVERWTSVX&& QEXVM\IJJIGXW& QIEWYVIQIRXJVIUYIRG] QIQSV]IJJIGXW

338 Index \M QIRYFEV MESSAGEFPSGO)) MESSAGEQIXLSHGSQQERH* QIXLSH EHNYWXQIRX EWWSGMEXIH[MXLVYR JMPIJSVQEX) STIVEXMSR TVSKVEQQMRK) WGEPI WXEXYWFIX[IIRG]GPIW WXVYGXYVI) XMQIFEWI YWMRKYWIVXSSPW) QIXLSHFPSGOW) QIXLSHGSQQERHW ALERT)* APPEND)* APROG* BEEP)* BIGINJECT* CAPTION)* CLOCK)* COINJECT* GSQQIRXMRKSYX) GSRWXERXW) CTEXT)* CURVE)* DEFINEALERT* DEFINEAPROG* DEFINELOOP* DEFINEMESSAGE* DETECTION* DILUTE* DIPNEEDLE* END* ENDLOOP** EXTRACLEAN)* FLOW)* FLOWPATH)* FLUSH* FRACTIONS* GO)* MRGSQQERHUYIYI INJECT* KEYWORD)* KINJECT* LIFETIME* LOOP)* LPARAM)* LTEXT)* STXMSRW)*

339 Index \MM PARAM)* TEVEQIXIVW) PRIME* QUICKINJECT* RACK* RECOVER* RINSE* RPOINT)* RTEXT)* SCALE* STOP)* TEMP* TIMES)* TITLE* TRANSFER* UNCLOG* WAIT* ZEVMEFPIW) WASH* WASHREC* QIXLSHGSRXVSP& QIXLSHHIJMRMXMSR GSQQIRXW) GSQQSRIVVSVW) GSRWIUYIRXMEPIVVSVW) HMEPSKYIFS\IW) W]RXE\) XI\XFS\IW) XI\XPE]SYX) QIXLSHHIJMRMXMSRPERKYEKI) QIXLSHIHMXSV GLIGOMRKQIXLSHW]RXE\ GVIEXMRKERI[QIXLSH IVVSVLERHPMRK) XI\XPE]SYX YWMRKHYVMRKEVYR [MRHS[ MethodQIRYGSQQERH QIXLSHXVEGI QMGVSFMEPKVS[XL QMGVSXMXVITPEXIW QMRMQM^MRKEMVFYFFPITVSFPIQW QMRMQM^MRKWEQTPIHMWTIVWMSR QM\* MRGYWXSQM^IHGSQQERHWIUYIRGIW MixQIRYGSQQERH MIXQIXLSHGSQQERH* MODEQIXLSHGSQQERH* QSZMRKXLIMRWXVYQIRX QSZMRKXLIVIJIVIRGIPMRI QYPXMGLERRIPETTPMGEXMSRW& QYPXMGLERRIPQSHI

340 Index \MMM HIPE]FIX[IIRWIRWSVKVEQW QYPXMTPIFMRHMRK QYPXMTPIMRWXERGIWSJGSRXVSPWSJX[EVI QYPXMTPIPMKERHW& QYPXMTPIWIRWSVKVEQ[MRHS[W REQMRKZEVMEFPIW) RIIHPI VITPEGMRKERHTSWMXMSRMRK RIIHPITSWMXMSRMRK Needle Replacement/PositioningWIVZMGIXSSP RIWXIHPSSTW) New MethodQIRYGSQQERH New WindowQIRYGSQQERH RSMWIMRWIRWSVKVEQW RSRVITVSHYGMFPIVIWYPXW EJXIVDesorb NORMALIZEQIXLSHGSQQERH* Normalize[SVOMRKXSSP RSVQEPM^MRK STXMGEPW]WXIQ RSVQEPM^MRKMREQIXLSH RSXIFSSO ETTPMGEXMSR[M^EVHW JMIPHW XIQTPEXI NotebookQIRYGSQQERH RYGPIMGEGMHW SRPMRILIPT OpenQIRYGSQQERH STIRMRKEWEZIHWIRWSVKVEQ STIRMRKXLIQIXLSHIHMXSV STXMGEPW]WXIQ STXMQM^MRKPMKERHMQQSFMPM^EXMSR& STXMSRW) MRQIXLSHGSQQERHW* STXSMRXIVJEGI ORDERPSSTSVHIV) OverlayQIRYGSQQERH PARAMQIXLSHGSQQERH)* TEVEQIXIVSVHIVMRK) TEVEQIXIVW MRQIXLSHGSQQERHW) TEVEQIXIVWMRGYWXSQM^IHGSQQERHW PasteQIRYGSQQERH TIVQMXXIHGSQQERHW MRDEFINEALERT*

341 Index \MZ MRDEFINEAPROG* MRDEFINELOOP* MRDEFINEMESSAGE* MRMAIN* T,HMJJIVIRGIW& TPSXGYVZIGPEWWIW PositionJMIPHWMRHMEPSKYIFS\IW TSWMXMSRMRKXLIEYXSWEQTPIVRIIHPI TS[IVYT Predip Needle MRWEQTPIMRNIGXMSRW PreferencesQIRYGSQQERH TVITEVMRKFPEROWYVJEGIW TVITEVMRKKVEHMIRXWYVJEGIW& TVITEVMRKWEQTPIMRNIGXMSR TVITEVMRKWSPYXMSRW TVIVYR) IVVSVQIWWEKIW) Prerun MethodQIRYGSQQERH PRIMEQIXLSHGSQQERH* Prime[SVOMRKXSSP TVMQMRKXLIVIGSZIV]GYT TVMQMRKXLIW]WXIQ PrintQIRYGSQQERH TVMRXMRK LIEHIVERHJSSXIV MRHVEJXQSHI WIRWSVKVEQTVMRXWM^I TVMRXMRKETTPMGEXMSR[M^EVHXIQTPEXIW TVMRXMRKEYXSWEQTPIVVEGOTSWMXMSRW TVMRXMRKHEXE TVMRXMRKWIRWSVKVEQW TVSGIWWMRKYRMX TVSHYGXOI] TVSKVEQQMRKQIXLSHW) TYQTVIJMPP WIRWSVKVEQHMWXYVFERGIW TYQTXYFMRK% TYQTW EMVMRW]VMRKI QEPJYRGXMSR VIJMPP VITPEGMRKW]VMRKIXMT Quickinject QUICKINJECTQIXLSHGSQQERH)* VEGOFEWI HIJMRMRK VEGOIHMXSVMRETTPMGEXMSR[M^EVHW

342 Index \Z RACKQIXLSHGSQQERH* VEGOTSWMXMSRW VEGOX]TIW RANDOMPSSTSVHIV) randomzevmexmsrw VEXIQIEWYVIQIRX& VIEHMRKVIPEXMZIVIWTSRWI VIEKIRXWXLEXKIRIVEXIKEW VIEPXMQI&-% ETTPMGEXMSREVIEW VIGSQQIRHIHWSPYXMSRW' RECOVERQIXLSHGSQQERH* VIGSZIVMRKPSWXHEXE VIGSZIVMRKWEQTPIWIIWEQTPIVIGSZIV] Recovery VIGSZIV]GYT VIHYGMRKGEVV]SZIV& VIJIVIRGIGIPP&& GSVVIGXMRKJSVHVMJX GSVVIGXMRKJSVVITVSHYGMFPIHMWXYVFERGIW VIJIVIRGIPMRI MHIRXMJ]MRKIZIRXW VITSVXTSMRXERHIZIRXPSKQEVOIVW YWIHMREHHMRKVITSVXTSMRXW Reference LineQIRYGSQQERH VIJIVIRGIWYFXVEGXMSR VIJIVIRGIXMQI) VIJMPPMRKXLITYQTW) VIJVEGXMZIMRHI\SJFYPOWSPYXMSR& VIKIRIVEXMSR& VIKIRIVEXMSRWGSYXMRK VIKIRIVEXMSRWSPYXMSRW' VIKVIWWMSRGSIJJMGMIRX VIKYPEVQEMRXIRERGI VIPEXMZIVIWTSRWI& VIQMRHIVJSVSanitizeERHDesorb VIQSZMRKIVVSVQIWWEKIW) VITIEXMRKPSSTW) VITIXMXMZIEREP]WIW) ReplaceQIRYGSQQERH VITPEGMRK EYXSWEQTPIVRIIHPI GSRRIGXSVFPSGO MRNIGXMSRTSVX TYQTW]VMRKIXMT VITSVXTSMRX FEWIPMRI GEPGYPEXIHZEPYIW MHIRXMJMIV XEFPI)* [MRHS[ [MRHS[

343 Index VITSVXTSMRXQEVOIVW VITSVXTSMRXXEFPI EHHMRKVITSVXTSMRXW EHHMRKVITSVXTSMRXWMRXIVEGXMZIP] HIPIXMRKVITSVXTSMRXW I\TSVXMRK OI][SVHW VITSVXTSMRXW FEWIPMRI IHMXMRK -( MRGYWXSQM^IHGSQQERHWIUYIRGIW MRQIXLSHW QEVOIVW SRWXEXYWGYVZIW) [MRHS[ VITVSHYGMFPIHMWXYVFERGIW VIWIVZIH[SVHW) VIWXSVMRK^SSQIHHMWTPE] VIWYPXJMPIW VIYWMRKEWIRWSVGLMT RINSEQIXLSHGSQQERH* Rinse[SVOMRKXSSP RpointMRWXVYGXMSR RPOINTQIXLSHGSQQERH)* RTEXTQIXLSHGSQQERH)* RunQIRY Data Collection Rate Prerun Method Run Application Wizard Run Method Run Sensorgram Start New Cycle Stop Method Stop Sensorgram VYRRMRKEQIXLSH VYRRMRKFYJJIV WEJIX] MRJSVQEXMSR VIUYMVIQIRXW WEJIX]VIUYMVIQIRXWJSVQEMRXIRERGI WEPXVIWMHYIW WEQTPIGSRWYQTXMSR& WEQTPIGSRXEGXXMQI WEQTPIHITPIXMSR& HYVMRKPMKERHMQQSFMPM^EXMSR& WEQTPIHMWTIVWMSR&& WEQTPIMRNIGXMSR EMVFYFFPIW EMVWIKQIRXW \ZM

344 Index \ZMM GSMRNIGX Extra Cleanup JMRMWLMRKXMQIW PEVKIZSPYQIW QERYEPMRNIGXMSR RSVIWTSRWI Predip Needle TVITEVEXMSR WEQTPIMRNIGXMSRQSHIW WEQTPIPSST WEQTPITSWMXMSRW WEQTPITVITEVEXMSR& WEQTPIVIGSZIV]) TVMQMRKXLIVIGSZIV]GYT VERKI VIWXVMGXMSRW [MXLStop Inject Sanitize Sanitize[SVOMRKXSSP WEZI JMPIGLERKIWRSXWEZIH Save AsQIRYGSQQERH SaveQIRYGSQQERH WEZMRK EWIRWSVKVEQ VIWYPXWSJW]WXIQGLIGO WEZMRKVIWYPXW ScaleQIRYGSQQERH SCALEQIXLSHGSQQERH* WGEPMRKWIRWSVKVEQHMWTPE] WGEPMRKXLIHMWTPE] WGLIHYPIVJSVSanitizeERHDesorb Select AllQIRYGSQQERH WIPIGXMRKEQIRY WIPIGXMRKQIRYGSQQERHW WIRWSVGLMT HSGOMRK MRWIVXMRK RSVQEPM^MRKXLIWMKREP TVSFPIQW VIYWMRK YRHSGOMRK WIRWSVGLMTGEWWIXXI WIRWSVKVEQ EGXMZIGYVZI FEWIPMRI FEWIPMRIWLMJXW GETXMSR) GSPSYVW GST]KVETL HEXEVIHYGXMSR I\TSVXMRK

345 Index JMPIWM^I MRJSVQEXMSR RSVIWTSRWISRMRNIGXMSR TVMRXWM^I TVMRXMRK TVSFPIQW VERHSQZEVMEXMSRW VITVSHYGMFPIHMWXYVFERGIW VMKLXLERHE\MW) WEZMRK WGEPMRK WTMOIW WXEVXMRKMRQERYEPGSRXVSP WXSTTMRK W]WXIQEXMGZEVMEXMSRW [MRHS[ ^SSQ WIRWSVKVEQ[MRHS[ WIRWSVKVEQ[MRHS[ WIVMEPTSVXHIWMKREXMSRW WIVZMGIXSSPWWIIXSSPW Service ToolsQIRYGSQQERH Set TemperatureQIRYGSQQERH WIXXMRKJPS[VEXI WLSVXGYXOI]W Show InfoQIRYGSQQERH Shutdown[SVOMRKXSSP WLYXXMRKHS[R WMKREPRSMWI Signal Quality Check WMQYPEXMRKWIRWSVKVEQW WMRKPIGLERRIPQSHI WMXMRKXLIMRWXVYQIRX Skip InjectQIRYGSQQERH WPSTI WSJX[EVIMRWXEPPEXMSR Software Problem Report WTMOIW GEYWIW HMWTPE] RSXWLS[RSRTVMRXSYX WMKREP RSVQEPM^EXMSR WXERHEVHHIZMEXMSR WXERHEVHW GSRJSVQERGI[MXL Standby[SVOMRKXSSP) FYJJIVVIUYMVIQIRX WXEVXMRKEQERYEPWIRWSVKVEQ WXEVXMRKEQIXLSH WXEVXMRKETTPMGEXMSR[M^EVHW \ZMMM

346 Index WXEXMWXMGEPTEVEQIXIVW WXEXYWGYVZIW) WXEXYWMXIQW MRWXEXYWGYVZIW)* StatusQIRYGSQQERH WXEXYW[MRHS[ HMWTPE] QIWWEKIW( STEPPSSTSVHIV) Stop FlowQIRYGSQQERH Stop inject MRManual Inject [MXLGSMRNIGX Stop InjectQIRYGSQQERH STOPQIXLSHGSQQERH)* Stop MethodQIRYGSQQERH Stop SensorgramQIRYGSQQERH WXSTTMRK EQIXLSH EVYR EWIRWSVKVEQ EWIWRWSVKVEQ XLIGSQQERHUYIYI WYFXVEGXMRKVIJIVIRGIVIWTSRWI WYVJEGIGSRGIRXVEXMSR WYVJEGITIVJSVQERGIXIWX VIWYPXW WYVJEGITPEWQSRVIWSRERGIWII746 WYVJEGITVITEVEXMSR[M^EVH MRPMRIVIJIVIRGIGIPPW 7YVJEGXERX4 W]RXE\ HIWGVMTXMSR* IVVSVW) TVIVYRQIXLSH W]RXE\IVVSVW IVVSVQIWWEKIW) W]VMRKIXMT VITPEGMRK Syringe TipWIVZMGIXSSP System Check Evaluation[SVOMRKXSSP System CheckXIWXXSSP W]WXIQGSRXVSP W]WXIQOI][SVHW W]WXIQTIVJSVQERGI% W]WXIQEXMGZEVMEXMSRW XIGLRMGEPWTIGMJMGEXMSRW% TEMPQIXLSHGSQQERH* XIQTIVEXYVI MRGYWXSQM^IHGSQQERHWIUYIRGIW \M\

347 Index WIXXMRK XIQTIVEXYVIGLERKIW& IJJIGXSRVIWTSRWIPIZIP XIQTIVEXYVIGSRXVSP EYXSWEQTPIV EYXSWEQTPIVVEGOW% HIXIGXMSRXIQTIVEXYVI% MREYXSWEQTPIVGSQTEVXQIRX [EVRMRK XIQTIVEXYVIWXEFMPMX] XIWXXSSPWWIIXSSPW Test ToolsQIRY Signal Quality Check Test ToolsQIRYGSQQERH XI\XFS\IW MRQIXLSHW) XLIVQSVEGOW% XMQIFEWIJSVQIXLSHW XMQIVIWSPYXMSR XMQIWTIGMJMGEXMSRWMRQIXLSHW XMQIGSRXVSPPIHMRNIGXMSRW& XMQIHIZIRXW) RIKEXMZIXMQIWTIGMJMGEXMSR) VIJIVIRGIXMQI) TIMESQIXLSHGSQQERH)* TITLEQIXLSHGSQQERH* XSSPFEV XSSPFEVFYXXSRW XSSPW Close Continue Desorb Flush Injection Port Normalize Prime Rinse Sanitize WIVZMGIXSSPW Shutdown Software Problem Report Syringe Tip System Check System Check Evaluation XIWXXSSPW Unclogging YWIVWIVZMGI [SVOMRKXSSPW ToolsQIRY BIAsimulation Evaluation Preferences \\

348 Index Service Tools Test Tools Working Tools XVEGMRKQIXLSHSTIVEXMSR XVERWJIV MRGYWXSQM^IHGSQQERHWIUYIRGIW TransferQIRYGSQQERH TRANSFERQIXLSHGSQQERH* XVSYFPIWLSSXMRK UNCLOGQIXLSHGSQQERH* UncloggingWIVZMGIXSSP UndoQIRYGSQQERH UndockQIRYGSQQERH YRHSGOMRK TVSFPIQW YRHSGOMRKEWIRWSVGLMT YRPSGOMRKXLIHSSVMREQIXLSH) YRWXEFPIXIQTIVEXYVI UnzoomQIRYGSQQERH YWIVWIVZMGIXSSPW YWIVXSSPW MRQIXLSHW) YWMRKXLIQIXLSHIHMXSVHYVMRKEVYR ZEVMEFPIGSRXEGXXMQIW&) MQTPIQIRXEXMSR ZEVMEFPIREQIW) IVVSV) ZEVMEFPIW EWWMKRMRKZEPYIW) MRGETXMSRW) MRGYWXSQM^IHGSQQERHWIUYIRGIW MRKEYWORDGSQQERHW) MRQIXLSHGSQQERHW) MRRpointGSQQERHW) PSGEPWXEXYW) YWMRKMRAPROGGEPP) ZEV]F]G]GPI MRGYWXSQM^IHGSQQERHWIUYIRGIW ZMEPHMWPSHKIV ViewQIRY Adjust Scale Baseline Extended Event Log Keywords Markers Method Method Trace New Window \\M

349 Index Notebook Overlay Plot Curve Classes Reference Line Scale Status Unzoom ZMVYWTVSXIGXMSRTVSKVEQW Wait MRGYWXSQM^IHGSQQERHWIUYIRGIW WAITQIXLSHGSQQERH* [EWL EYXSQEXMGVSYXMRIW% EYXSWEQTPIVRIIHPI KIRIVEPVIGSQQIRHEXMSRW -*'GLERRIPW MRGYWXSQM^IHGSQQERHWIUYIRGIW QIXLSHGSQQERHSTXMSRW& RIIHPI VIGSZIV]GYT VIGSZIV]GYT] WashQIRYGSQQERH WASHQIXLSHGSQQERH* WASHRECQIXLSHGSQQERH* [IIOP]QEMRXIRERGI [LMXIWTEGIGLEVEGXIVW) [MRHS[JSVVITSVXTSMRXW ;MRHS[W GPMTFSEVH )\TPSVIV W]WXIQVIWSYVGIW ;MRHS[WXIGLRMUYIW [M^EVHWWIIETTPMGEXMSR[M^EVHW [SVOMRKXSSPWWIIXSSPW Working ToolsQIRYGSQQERH ^SSQ ^SSQWIRWSVKVEQ \\MM

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