Beginners Guide for Magellan Document Part No. I 117 519 November 2001 Document revision: 2.0 Software level: 3.1x
TECAN Affiliates and Service Centers Austria Tecan Austria GmbH Untersbergstrasse 1a A-5082 Grödig / Salzburg Austria Tel.: +43 62 46 89 33 Fax: +43 62 46 72 770 Switzerland Tecan Schweiz AG Seestrasse 103 CH-8708 Männedorf Switzerland Tel.: + 41 1 922 81 11 Fax. : +41 1 922 81 12 France Tecan France S.A. Parc d'activités de Pissaloup Batiment Hermes II Rue Edouard Branly F-78190 Trappes France Tel.: +33 1 30 68 81 50 Fax: +33 1 30 68 98 13 Japan Tecan Japan Co. Ltd Meiji Seimei Fuchu Building 10F 1-40 Miyamachi Fuchu City, Tokyo Japan Tel.: +81 42 334 88 55 Fax: +81 42 334 04 01 Spain Tecan Spain Sabino de Arana, 32 E-08028 Barcelona Spain Tel.: +34 93 490 01 74 Fax: +34 93 411 24 07 USA Tecan Boston 200 Boston Avenue Suite 3000 Medford, MA 02155 USA Tel.: +1 781 306 08 27 Fax: +1 781 306 0837 United Kingdom Tecan UK Theale Court 11-13 High Street Theale UK-Reading RG7 5AH United Kingdom Tel.: +44 11 89 300 300 Fax: +44 11 89 305 671 Asia Tecan Asia (Pte) Ltd. 80, Marine Parade #13-04 Singapore 449269 Singapore Tel.: +65 44 41 886 Fax: +65 44 41 836 Belgium Tecan Benelux B.V.B.A. Vaartdijk 55 B-2800 Mechelen Belgium Tel.: +32 15 42 13 19 Fax. +32 15 42 16 12 Italy Tecan Italia S.r.l. Via F.lli Cervi Palazzo Bernini Centro Direzionale Milano2 20090 Segrate (Mi) Italy Tel.: +39 02 215 21 28 Fax: +39 02 215 97 441 USA Tecan US P.O. Box 13953 Research Triangle Park, NC 27709 USA Tel.: +1 919 361 5200 Fax: +1 919 361 5201 Germany Tecan Deutschland GmbH Theodor-Strom-Straße 17 D-74564 Crailsheim Germany Tel.: +49 79 51 94 170 Fax: +49 79 51 50 38 Netherlands Tecan Benelux B.V.B.A. Industrieweg 30, NL-4283 Giessen, Netherlands; Tel.: +31 018 34 48 17 4 Fax: +31 018 34 48 06 7 2 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 November 2001
Notice WARNING CAREFULLY READ AND FOLLOW THE INSTRUCTIONS PROVIDED IN THIS MANUAL BEFORE OPERATING THE INSTRUMENT. Every effort has been made to avoid errors in text and diagrams, however, TECAN Austria Ges.m.b.H. assumes no responsibility for any errors which may appear in this publication. It is the policy of TECAN Austria Ges.m.b.H. to improve products as new techniques and components become available.tecan Austria Ges.m.b.H. therefore reserves the right to change specifications at any time. We would appreciate any comments on this publication. TECAN Austria Ges.m.b.H. Untersbergstraße 1A A-5082 Grödig/Salzburg AUSTRIA / EUROPE Telephone: +43 6246/8933 FAX: +43 6246/72770 E-mail: office.austria@tecan.com Copyright Information The contents of this manual are the property of TECAN Austria Ges.m.b.H and are not to be copied, reproduced or transferred to another person or persons without our prior written permission. Copyright TECAN Austria Ges.m.b.H All rights reserved. Printed in Austria. About this Manual Magellan is a general data reduction package used to analyze data generated from microplate assays for general research and in vitro diagnostic applications. It is designed for professional use only. This manual instructs how to: Install the software Operate the software November 2001 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 3
Warnings, Cautions and Notes There are three types of informational notices used in this manual. These notices highlight important information or warn the user of a potentially dangerous situation. The following notices are: Note: Gives helpful information. Caution Indicates a possibility of instrument damage or data loss if instructions are not followed. WARNING INDICATES THE POSSIBILITY OF SEVERE PERSONAL INJURY, LOSS OF LIFE OR EQUIPMENT DAMAGE IF THE INSTRUCTIONS ARE NOT FOLLOWED. 4 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 November 2001
Table of Contents 1. Basics... 1-1 1.1 Area of Application...1-1 1.2 System Requirements...1-2 1.2.1 Hardware Requirements...1-2 1.2.2 Software Requirements...1-2 1.3 Software Installation Procedure...1-2 1.4 Start working with Magellan...1-4 1.4.1 Working with an Instrument...1-4 1.4.2 Working in Demo mode...1-4 1.5 Working with the Magellan Wizards...1-5 1.5.1 How to start a wizard...1-6 2. Create/Edit a Method Wizard... 2-1 2.1 Introduction...2-1 2.2 Qualitative ELISA...2-1 2.2.1 Test kit Description and Data Handling...2-1 2.2.2 Creating the Qualitative Method in Magellan...2-2 2.3 Quantitative ELISA...2-11 2.3.1 Test kit Description and Data Handling...2-11 2.3.2 Creating the Quantitative Method in Magellan...2-12 3. Run a Method Wizard... 3-1 3.1 Introduction...3-1 3.2 Working with the Run a Method Wizard...3-1 4. Evaluate Results Wizard... 4-1 4.1 Introduction...4-1 4.2 Working with the Evaluate Results Wizard...4-1 4.2.1 HIV-ELISA (qualitative example)...4-1 4.2.2 IgM-ELISA (quantitative example)...4-4 5. Create/Edit a Sample-ID s List Wizard... 5-1 5.1 Introduction...5-1 5.2 Working with the Create/Edit a Sample-ID s List Wizard...5-1 6. Licensing... 6-1 Index 6.1 Introduction...6-1 6.2 Licensing...6-1 6.3 Serial number...6-2 6.4 License Number...6-2 6.5 Registration Form...6-3 6.6 License Agreement...6-3 6.7 Sending the Registration Form to TECAN...6-4 6.8 Receiving a License Number from TECAN...6-5 November 2001 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 5
1.Basics 1. Basics 1.1 Area of Application Magellan is a universal reader control and data reduction software for analysing data generated from microplate tests using TECAN measuring devices. Magellan can be used as an accessory together with a TECAN measuring device certified as suitable for in-vitro diagnosis (IVD) purposes. A user administration system must be installed for in-vitro diagnostic use. The following levels of user rights can be selected: Supervisor: Training as laboratory supervisor. The Supervisor can define measurement parameters and evaluation regulations and carry out measurements accordingly. The Supervisor must always be trained by TECAN. User: Training as laboratory technician. The user can change measurement parameters and evaluation regulations and carry out measurements accordingly. The user must always be trained by the supervisor. Guest: Training as laboratory assistant. The guest can carry out measurements according to the specifications given. The guest must be trained by the supervisor. When using Magellan as an accessory with an IVD device, all results must be traceable through the user administration and automatic saving of all measurement data. Magellan is designed for use with the specified operating systems and specified Microsoft Office versions. If additional programs are installed, functionality in accordance with IVD regulations can not be guaranteed. November 2001 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 1-1
1.2 System Requirements 1.2.1 Hardware Requirements The following requirements are needed for using Magellan: Hardware Minimum Recommend Memory 32 MB 64 MB CPU Pentium II Pentium III Resolution 800 x 600 1024 x 768 Serial ports 1 2 1.2.2 Software Requirements The following requirements are needed for using Magellan: Operating system Minimum Recommend Microsoft Windows Windows 98 SE Windows XP Professional, Windows 2000 or Windows NT 4.0/SP6 Additional supported software: Microsoft Office 2000 or Micorsoft Office XP Note: The IVD version of Magellan is released only for Windows NT4/SP6 and Windows 2000 in combination with Microsoft Office 2000. 1.3 Software Installation Procedure Magellan is installed by an automatic software setup program. All necessary components are installed: Start Setup.exe to begin with the installation procedure. When using the Reader PC Package CD press the Setup button to start the installation procedure. A welcome window informs the user about the beginning of the installation process. Click Next to continue. The next window displays the License Agreement. Click I agree continue. The next window is the Choose Destination Location window. The default installation path is displayed. In case the default path is not suitable, the Browse function allows the definition of another path. Click Next to continue. The next window asks for the name of the program group, where a shortcut to the Magellan program will be placed. The default group name is TECAN. Click Next to continue. Windows and Excel are registered trademarks of the Microsoft Corporation 1-2 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 November 2001
1.Basics The next window gives the choice to install Magellan3, Magellan3 for In-Vitro- Diagnostic or Magellan3 for In-Vitro-Diagnostic (upgrade from Non-In-Vitro- Diagnostic version). Select the installation option according to your order at TECAN. Click Next to continue. Now the installation of the required files is carried out and files are copied to the computer. At the end of a successful installation the user can quit the installation wizard by clicking the Finish button. The software can now be started via the Windows Start menu by selecting Magellan3 in the above defined program group. Note: It is very important that the person who installs the software has administrator rights on the PC with operating systems Win NT, Win 2000 and Win XP. November 2001 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 1-3
1.4 Start working with Magellan You can work with Magellan either with an instrument connected or without an instrument using the demo mode. 1.4.1 Working with an Instrument Connect your instrument with your computer as described in the Instrument operating manual and switch the instrument on. After starting Magellan a message box appears, which tells you on which port your instrument is connected to Magellan. 1.4.2 Working in Demo mode In order to use Magellan without an instrument you first have to switch on the demo mode allowed check mark in the reader server (RdrOle4) following this procedure: 1. Start the reader server by selecting Start Programs TECAN - RdrOle4. The following window appears: 2. In the Service menu click Demo Mode Allowed. Now, a tick should be displayed next to the Demo Mode Allowed option. 3. Close the reader server. 1-4 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 November 2001
1.Basics If you start Magellan now, the following dialog box appears: Select the instrument you are usually working with from the list box in the instrument field. In the port field select Demo mode and press OK. Now you are running Magellan in the demo mode. With Magellan in Demo mode you can: Evaluate results Create and edit methods Create and edit sample ID lists 1.5 Working with the Magellan Wizards The Magellan software is equipped with a number of wizards. These wizards represent workflow modules, which define a sequential procedure and guide you step-by-step to perform a specific complex activity. By using the wizards, all required options of the regular Magellan menu structure can be selected. Magellan consists of the following wizard types: 1. Obtain Raw Data enables you to generate raw data quickly and easily by setting the required measurement parameters and starting the measurement. By doing this the Obtain Raw Data wizard creates a new workspace in which the obtained values are presented and stored. 2. Create/edit a Method helps you to define a method, which includes all necessary parameters for measurement, evaluation and data handling according to the performed assay. 3. Run a Method enables you to perform measurements based on already defined methods. The wizard creates a new workspace, that contains the selected method (which consists of all measurement parameters and test definitions for evaluation) and enables you to insert a Sample ID list. After the measurement, the workspace is completed with the obtained raw data that will be evaluated. 4. Evaluate Results enables you to view the raw data and the evaluation of data. The evaluation parameters can also be viewed and data can be reevaluated. November 2001 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 1-5
1.5.1 How to start a wizard 5. Create/edit a Sample ID list enables you to create new sample lists. Up to three sample IDs per well can be entered or imported. 6. Create/edit a Test enables you to design the layout, define transformations, set a standard curve and define cutoffs, validations and kinetic parameters. This wizard is not accessible with MagellanCE. 7. Reader Control provides you with quick access to several reader functions like moving plate and filter. 8. Options gives you access to the Global Settings dialog box. 9. Delete files gives you full control when deleting files of all types. MagellanCE also enables copying of files, therefore the wizard is called Copy/Delete files. 10. XChange files enables you to transfer files from and to a SUNRISE Touchscreen. (This option is available with MagellanCE only.) 11. About Magellan provides you with quick access to information about Magellan including version number and licensing information. All Magellan wizards are displayed and started in the Wizard List dialog box. Using the default global settings the Wizard List dialog box appears automatically after starting Magellan. If the Wizard List dialog box doesn t appear automatically there are two ways to open the Wizard List dialog box: In the File menu, by clicking Wizard... On the toolbar, by clicking the Wizard button. 1-6 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 November 2001
2. Create/Edit a Method Wizard 2. Create/Edit a Method Wizard 2.1 Introduction 2.2 Qualitative ELISA This section provides you two step-by-step examples (qualitative and quantitative test) of how to create a method in Magellan. By following the instructions you learn to define the evaluations from a test kit description in Magellan. We created the methods connected with a Sunrise in Demo mode, but it is also possible to define the methods connected with another TECAN instrument, which is able to measure absorbance. 2.2.1 Test kit Description and Data Handling In the manufacturer s test kit description of a qualitative HIV-ELISA the following instructions are found: Plate Layout Reagent blank Negative controls Positive control HIV 1 Positive control HIV 2 Samples A1 B1-C1 D1 E1 F1-H12 Measurement and Evaluation Read plate at a wavelength of 450nm. Blank reader/plate on well A1. Perform all further calculations with blanked values. Calculate the Cutoff value by adding 0.2 to the mean of the negative controls. The mean absorbance of the negative controls must be less than 0.3. The absorbance of each positive control must be more than 0.8 above the mean absorbance of the negative controls. Samples giving an absorbance less than the Cutoff value are considered negative in the assay. Samples giving an absorbance equal or higher than the Cutoff value are considered reactive. Such samples should be retested. November 2001 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 2-1
2. Create/Edit a Method Wizard Data handling After the measurement the data file (workspace) should be stored automatically and a report containing the measurement parameter, plate layout, blanked values, cutoff definition, qualitative results of the samples and validations should be created. 2.2.2 Creating the Qualitative Method in Magellan In the Wizard List dialog box, select Create/edit a method and click OK. After clicking next on the Welcome page of the Create/edit a method wizard the Select a file dialog box is displayed. Click Create check box and type a file name and Remarks for the new method. After clicking Next the Measurement parameter dialog box of the Create/edit a method wizard is displayed. 2-2 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 November 2001
2. Create/Edit a Method Wizard Click Customize measurement parameters button and the following dialog box is displayed. In the General tab select Absorbance. In the Meas. Params tab select 450nm as Measurement wavelength. After clicking OK you are back at the Measurement parameter dialog box of the Create/edit a method wizard. Click Next to reach the Define the evaluation dialog box of the Create/edit a method wizard. November 2001 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 2-3
2. Create/Edit a Method Wizard Click Design layout and set transformations and the Plate Layout Definition window is displayed. Now you first have to define the Layout using the Well assignment dialog box. In the Identifier box, select BL (Blank). In the Experimental group box the number 1 remains. In the Replicates box Contiguous is selected. Click the A1 well, which is then marked with a red boarder. Click Fill selection and the well is labeled with the selected identifier type. You can fill one well also with double clicking the well. Now choose the following settings in the Well assignment dialog box: In the Identifier box, select NC (Negative Control). In the Experimental group box the number 1 remains. In the Replicates box Contiguous is selected. Click B1 well and drag the mouse to C1. The wells B1 to C1 are then marked with a red boarder. Click Fill selection and the wells are labeled with the selected identifier type. Then fill the wells D1 and E1 with the Identifiers PC (Positive Control HIV1) and HPC (Positive Control HIV2). 2-4 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 November 2001
2. Create/Edit a Method Wizard Now the following window should be displayed. Now use from the right mouse menu Select all Unused to select all empty wells of the plate. In the Well Assignment dialog box, select SM (Sample) under Identifiers. In the Experimental group box the number 1 remains. In the Replicates box you now can choose between Fix number and Contiguous: Fix number radio button: Only enabled for standards and samples where IDs can be used. If this radio button is active a number can be entered in the corresponding text field. This number defines how many replicates are intended for this method. In the selected wells, the entered number of replicates for every ID is created. Therefore the number of selected wells must be a multiple of the entered number of replicates. Contiguous radio button All selected wells are defined as replicates. If an existing ID number for the samples and standards is chosen, the selected wells are then added as replicates to the existing replicates. With all other identifier types the selected wells are added as replicates to the existing replicates. Two Arrow buttons define the direction of counting up the replicate number. In our example select Fix Number and 1 and click Fill selection. Now the selected wells are labeled as Samples and the definition of the plate layout is finished. The next step of the definition procedure is to define the blank reduction using the Transformations dialog box. This can be achieved by firstly making sure that the Transformation Number is 1 and that Raw data is selected in the Input data box. In the Output box the name Formula 1 is displayed as default. Type blank reduction instead. November 2001 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 2-5
2. Create/Edit a Method Wizard In the Formula box type x-bl1, where x refers to the current Input data value in a well and BL1 is the mean value of the blank wells of experimental group 1.You can enter the formula also by using the available list boxes (Variables, Operators and Functions). The following Transformations dialog box is displayed. Now select the whole plate by clicking the asterisk in the upper left corner of the micro plate and click Set in the Transformations dialog box. The following window is displayed. In each well the following information is displayed (Example well A5): SM1_4 Sample, experimental group number of 1, sample ID number of 4. 1/1 Number of replicate is 1, total number of Replicates is 1. x-bl1 or 1.000 Assigned transformation x-bl1 (Transformations dialog box is displayed) or Dilution Factor value of 1 (Transformations dialog box is closed). With clicking OK you finish the definition of Layout and transformations and you are back on the Define the evaluation dialog box of the Create/edit a method wizard. Click Define cutoffs to reach the Cutoff Definition dialog box, where you can define the limits for the qualitative evaluation. 2-6 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 November 2001
2. Create/Edit a Method Wizard In our example the cutoff value is calculated by adding 0.2 to the mean of the negative controls. This can be achieved by: In the Input box, select blank reduction. In the middle of the Cutoff Definition dialog box is a scale indicating the high and the low end for the Cutoff Limits and - Types. On the right side of the scale the Cutoff Limits are defined and on the left side of the scale, the Cutoff Types are indicated. In Cutoff Limits, type NC1+0.2 into the first row or enter the formula using the available Variables, Operators and Functions. In the Cutoff Types, type the test interpretation Reactive into the first and Negative into the second box. For adding a color, select a color from the color plate situated on the left and then assign the color to the particular types. Assign the following colors to the Cutoff - Types: Negative - Green Reactive - Red The following Cutoff Definition dialog box is displayed. With clicking Advanced you reach the Advanced settings dialog box, where you can select for which identifier types the cutoff results should be displayed. Click OK to come back to the Define the evaluation dialog box of the Create/edit a method wizard. Click Define validations to reach the Validation Definition dialog box. In this dialog box, the validation criteria for the test must be defined, so that the validity of the test results is guaranteed. November 2001 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 2-7
2. Create/Edit a Method Wizard In our example the following requirements must be fulfilled: The mean absorbance of the negative controls must be less than 0.3. The absorbance of each positive control must be more than 0.8 above the mean absorbance of the negative controls. In the Input box, select blank reduction. In the first row, type NC1<0.3, or enter the formula using the available Variables, Operators and Functions. In the second row, type PC1>NC1+0.8. In the third row, type HPC1>NC1+0.8. The Validation Definition dialog box is now displayed like this. Click OK to come back to the Define the evaluation dialog box of the Create/edit a method wizard. Click Next to reach the Report and Export dialog box of the Create/edit a method wizard. 2-8 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 November 2001
2. Create/Edit a Method Wizard Click Report setup to reach the following dialog box. Main is the first tab of the Report Setup dialog box. Select your favorite options and switch to the Data tab. On the Data tab you find in the Available data box all available data for the report. Using the buttons Attach, Paste and Merge you can transfer the data into the Selected data box. In the Print as box you can decide between printing the data as a matrix or as a list with a special orientation. November 2001 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 2-9
2. Create/Edit a Method Wizard Before creating your Report you have to remove the default Matrix-Raw Data from the Selected Data box. After this just the Measurement parameters remain in the Selected Data box. Than select Layout in the Available data box and attach it as a Matrix to the Report. Now merge blank reduction and Cutoff results into the matrix. For this first select blank reduction in the Available data box and Layout in the Selected Data box and click Merge. Do the same with the Cutoff results. With attaching the Cutoffs and the Validation criteria to the selected data you have defined the asked report. The Data tab of the Report setup dialog box is now displayed like this. On the Header and Footer tabs you can define the layout of the Header and the Footer of your Report. Click OK to come back to the Report and Export dialog box of the Create/edit a method wizard. Now click Automated Data handling. Select Automatic print and save to workspace default path in the appearing dialog box and click OK. When Magellan is used under IVD-conditions, automatic saving can t be deselected. Click Next to reach the Save dialog box of the Create/edit a method wizard. With clicking Finish you save the method and reach the Wizard List dialog box. 2-10 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 November 2001
2. Create/Edit a Method Wizard 2.3 Quantitative ELISA 2.3.1 Test kit Description and Data Handling In the manufacturer s test kit description of a quantitative IgM - Antibody detections - ELISA the following instructions are found: Plate Layout 1 2 3 4 5 6 7 8 9 10 11 12 BLK C3 S1 A NC C4 S2 B NC C4 S2 C C1 C5 S3 D C1 C5 S3 E C2 C6 F C2 C6 G C3 S1 H BLK = Blank, NC = Negative control, C1 C6 = Calibrators (Standards), S1 S = Samples Read plate at a wavelength of 492nm, reference at 620nm. Blank reader/plate on well A1. Concentrations of the Calibrators (Standards): Calibrator 1 5 UA/mL Calibrator 2 10 UA/mL Calibrator 3 20 UA/mL Calibrator 4 40 UA/mL Calibrator 5 80 UA/mL Calibrator 6 160 UA/mL After the blank correction the optical densities (OD492 OD620) are plotted versus the concentration. The regression line that goes through these points is the Standard Curve. November 2001 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 2-11
2. Create/Edit a Method Wizard Interpretation of the test results: IgM < 18 UA/mL Negative 18 UA/mL <= IgM < 22 UA/mL Intermediate IgM >= 22 UA/mL Positive The calculated IgM concentration of both negative controls must be under 8 UA/mL. Data Handling After the measurement the data file (workspace) should be stored automatically and a report containing the measurement parameter, plate layout, blanked values, standard curve, IgM-concentreations, cutoff definition, qualitative results of the samples and validations should be created. Additionally the Layout and the qualitative results should be stored as ascii file. 2.3.2 Creating the Quantitative Method in Magellan In the Wizard List dialog box, select Create/edit a method and click OK. After clicking next on the Welcome page of the Create/edit a method wizard the Select a file dialog box is displayed. Click Create check box and type a file name and Remarks for the new method. After clicking Next the Measurement parameter dialog box of the Create/edit a method wizard is displayed. 2-12 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 November 2001
2. Create/Edit a Method Wizard Click Customize measurement parameters button and the following dialog box is displayed. In the General tab select Absorbance. In the Meas. Params tab select 492nm as Measurement wavelength and 620nm as Reference wavelength. After clicking OK you are back at the Measurement parameter dialog box of the Create/edit a method wizard. Click Next to reach the Define the evaluation dialog box of the Create/edit a method wizard. November 2001 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 2-13
2. Create/Edit a Method Wizard Click Design layout and set transformations and the Plate Layout Definition window is displayed. Now you first have to define the Layout using the Well assignment dialog box. In the Identifier box, select BL (Blank). In the Experimental group box the number 1 remains. In the Replicates box Contiguous is selected. Click the A1 well, which is then marked with a red boarder. Click Fill selection and the well is labeled with the selected identifier type. You can fill one well also with double clicking the well. Now choose the following settings in the Well assignment dialog box: In the Identifier box, select NC (Negative Control). In the Experimental group box the number 1 remains. In the Replicates box Contiguous is selected. Click B1 well and drag the mouse to C1. The wells B1 to C1 are then marked with a red boarder. Click Fill selection and the wells are labeled with the selected identifier type. Next you have to assign the Calibrators (Standards) to the wells D1 to G2. Therefore select the following settings in the Well assignment dialog box: In the Identifier box, select ST (Standard). In the Experimental group box the number 1 remains. In the Replicates box you now can choose between Fix number and Contiguous: 2-14 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 November 2001
2. Create/Edit a Method Wizard Fix number radio button Only enabled for standards and samples where IDs can be used. If this radio button is active a number can be entered in the corresponding text field. This number defines how many replicates are intended for this method. In the selected wells, the entered number of replicates for every ID is created. Therefore the number of selected wells must be a multiple of the entered number of replicates. Contiguous radio button All selected wells are defined as replicates. If an existing ID number for the samples and standards is chosen, the selected wells are then added as replicates to the existing replicates. With all other identifier types the selected wells are added as replicates to the existing replicates. Two Arrow buttons define the direction of counting up the replicate - and ID number. In our example select Fix Number and 2. In the ID-Number box and in the Replicates box select the vertical Arrows. Then select the wells D1 to G2 and click Fill selection. Select the wells as follows: Click D1 well and drag the mouse over the required wells to H1. Then hold down the control (Ctrl) key and drag the mouse over the required wells from A2 to G2. Now the following window should be displayed. November 2001 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 2-15
2. Create/Edit a Method Wizard Now use from the right mouse menu Select all Unused to select all empty wells of the plate. Then hold down Ctrl and deselect well H12. In the Well Assignment dialog box select SM (Sample) under Identifiers. In the Experimental group box the number 1 remains. In the Replicates box choose Fix number and 2. In the ID-Number box and in the Replicates box select the vertical Arrows and click Fill selection. The next step of the definition procedure is to define the blank reduction using the Transformations dialog box. This can be achieved by firstly making sure that the Transformation Number is 1 and that Raw data is selected in the Input data box. In the Output box the name Formula 1 is displayed as default. Type blank reduction instead. In the Formula box type x-bl1, where x refers to the current Input data value in a well and BL1 is the mean value of the blank wells of experimental group 1.You can enter the formula also by using the available list boxes (Variables, Operators and Functions). The following Transformations dialog box is displayed. Now select the whole plate by clicking the asterisk in the upper left corner of the micro plate and click Set in the Transformations dialog box. The following window is displayed. 2-16 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 November 2001
2. Create/Edit a Method Wizard In each well the following information is displayed (Example well A5): SM1_9 Sample, experimental group number of 1, sample ID number of 9. 2/2 Number of replicate is 2, total number of Replicates is 2. x-bl1 or 1.000 Assigned transformation x-bl1 (when Transformations dialog box is displayed) or Dilution Factor value of 1 (when Transformations dialog box is closed). With clicking OK you finish the definition of Layout and transformations and you are back on the Define the evaluation dialog box of the Create/edit a method wizard. Click Define conc., dilutions and ref. Values to reach the Concentration/Dilution/ Reference Definition dialog box, where you have to define the concentrations of the Standards as described in the test kit: Calibrator 1 5 UA/mL Calibrator 2 10 UA/mL Calibrator 3 20 UA/mL Calibrator 4 40 UA/mL Calibrator 5 80 UA/mL Calibrator 6 160 UA/mL This can be achieved by firstly making sure that the ST is selected in the Select Identifier list box and that in the Exp. group box the number 1 remains. Now, in the Identifier list, a list of the standards from the Exp. Group 1 is displayed. In the corresponding Concentration box of ST1_1 type the number 5 and in the Unit box, type UA/mL. In the corresponding Concentration box of ST1_2 type the number 10. The unit only needs to be defined once and is valid for all standards. Type the values for the ST1_3 to ST1_6 in the same way. Now this Concentration / Dilution / Reference Definition dialog box is displayed: November 2001 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 2-17
2. Create/Edit a Method Wizard Click OK to come back to the Define the evaluation dialog box of the Create/edit a method wizard. Click Set standard curve to reach the Standard curve dialog box, where you can define the required standard curve. In the test kit description you find this description: After the blank correction the optical densities (OD492 OD620) are plotted versus the concentration. The regression line that goes through these points is the Standard Curve. First you have to select blank reduction as Input data in the Data tab of the Standard curve dialog box. In the Analysis Type tab, click Linear Regression. In the Axes tab, you can define the graph title, the labeling and the scaling of the axis like this: Click OK to come back to the Define the evaluation dialog box of the Create/edit a method wizard. Click Define cutoffs to reach the Cutoff Definition dialog box, where you can define the limits for the qualitative evaluation. In the test kit description you find the following instructions: Interpretation of the test results: IgM < 18 UA/mL Negative 18 UA/mL <= IgM < 22 UA/mL Intermediate IgM >= 22 UA/mL Positive 2-18 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 November 2001
2. Create/Edit a Method Wizard This can be achieved by: In the Input box, select Mean conc. (UA/mL). In the middle of the Cutoff Definition dialog box is a scale indicating the high and the low end for the Cutoff Limits and - Types. On the right side of the scale the Cutoff Limits are defined and on the left side of the scale, the Cutoff Types are indicated. In Cutoff Limits, type 22 as first (higher) Limit and 18 as second (lower) Limit. In the Cutoff Types, type the test interpretation (Positive, Intermediate and Negative) in the individual boxes. For adding a color, select a color from the color plate situated on the left and then assign the color to the particular types. Assign the following colors to the Cutoff - Types: Negative Green Intermediate - Blue Positive - Red The following Cutoff Definition dialog box is displayed. With clicking Advanced you reach the Advanced settings dialog box, where you can select for which identifier types the cutoff results should be displayed. Click OK to come back to the Define the evaluation dialog box of the Create/edit a method wizard. Click Define validations to reach the Validation Definition dialog box. In this dialog box, the validation criteria for the test must be defined, so that the validity of the test results is guaranteed. November 2001 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 2-19
2. Create/Edit a Method Wizard In our example the following requirement must be fulfilled: The calculated IgM concentration of both negative controls must be under 8 UA/mL. In the Input box, select Single conc. (UA/mL). In the first row, type NC1_1<8, or enter the formula using the available Variables, Operators and Functions. NC1_1 means Negative control of experimental group 1, replicate 1. In the second row, type NC1_2<8. The Validation Definition dialog box is now displayed like this. Click OK to come back to the Define the evaluation dialog box of the Create/edit a method wizard. 2-20 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 November 2001
2. Create/Edit a Method Wizard Click Next to reach the Report and Export dialog box of the Create/edit a method wizard. Click Report setup to reach the following dialog box. Main is the first tab of the Report Setup dialog box. Select your favorite options and switch to the Data tab. November 2001 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 2-21
2. Create/Edit a Method Wizard On the Data tab you find in the Available data box all available data for the report. Using the buttons Attach, Paste and Merge you can transfer the data into the Selected data box. In the Print as box you can decide between printing the data as a matrix or as a list with a special orientation. In this example a report containing the measurement parameter, plate layout, blanked values, standard curve, IgM-concentrations, cutoff definition, qualitative results of the samples and validations should be created. Before creating your Report you have to remove the default Matrix-Raw Data from the Selected Data box. After this just the Measurement parameters remain in the Selected Data box. Than select Layout in the Available data box and attach it as a Matrix to the Report. Now merge blank reduction, Mean conc. (UA/mL) and Cutoff results into the matrix. For this first select blank reduction in the Available data box and Layout in the Selected Data box and click Merge. Do the same with Mean conc. (UA/mL) and the Cutoff results. With attaching Graph (Standard curve) Cutoffs and the Validation criteria to the selected data you have defined the asked report. The Data tab of the Report setup dialog box is now displayed like this. On the Header and Footer tabs you can define the layout of the Header and the Footer of your Report. Click OK to come back to the Report and Export dialog box of the Create/edit a method wizard. Now click Export setup to reach the File Export dialog box, where you can define the datasets, which should be stored as ascci file. In our example the Layout and the qualitative results should be stored as ascii file. 2-22 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 November 2001
2. Create/Edit a Method Wizard Therefore select Layout in the Available data List and transfer it to the Selected Data List by clicking the arrow, which points to this list. Do the same with the Cutoff results. The File Export dialog box is now displayed like this. Click OK to come back to the Report and Export dialog box of the Create/edit a method wizard. Now click Automated Data handling. Select Automatic print, save to workspace default path and export to export default path in the appearing dialog box and click OK. When Magellan is used under IVD-conditions, automatic saving can t be deselected. Click Next to reach the Save dialog box of the Create/edit a method wizard. With clicking Finish you save the method and reach the Wizard List dialog box. November 2001 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 2-23
3. Run a Method Wizard 3. Run a Method Wizard 3.1 Introduction Run a Method enables you to perform measurements based on already defined methods. The wizard creates a new workspace, that contains the selected method (which consists of all measurement parameters and test definitions for evaluation) and enables you to insert a Sample ID list. After the measurement, the workspace is completed with the obtained raw data that will be evaluated. 3.2 Working with the Run a Method Wizard In the Wizard List dialog box, select Run a Method and click OK. After clicking Next on the Welcome page of the Run a Method wizard the Select a file dialog box is displayed. Select the required method from the previously defined methods and click Next to reach the following dialog box: Check the measurement parameters and the method name to see if they are correct. November 2001 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 3-1
3. Run a Method Wizard Click Finish and the Start measurement dialog box is displayed. Type the name of the workspace. Place the microplate on the plate support of the instrument and click Start to start the measurement immediately. If you want to load a Sample ID list, you have to click Load Sample-ID s to reach the Load Sample-ID s dialog box. Select the required Sample ID list and press OK. Now a preview of the sample ID s together with the plate layout is displayed. Click OK to confirm the selection and come back to the Start measurement dialog box. Click Start and the measurement begin. 3-2 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 November 2001
5.Create/Edit a Sample-ID s List Wizard 4. Evaluate Results Wizard 4.1 Introduction Evaluate Results enables you to view the raw data and the evaluation of data. The evaluation parameters can also be viewed and data can be re-evaluated. Corresponding to the created methods (HIV- and IgM-ELISA) this section guides you through the Evaluate results wizard using two example workspace files created by running these methods. 4.2 Working with the Evaluate Results Wizard In the Wizard List dialog box, select Evaluate results and click OK. After clicking Next on the Welcome page of the Evaluate results wizard the Select a file dialog box is displayed. Now choose between qualitative (4.2.1) and quantitative (4.2.2) example. 4.2.1 HIV-ELISA (qualitative example) Select the workspace HIV-ELISA.wsp from the file list and click Next to reach the following dialog box: Clicking Raw data, Transformations or Cutoff results brings you to a plate view window, where the indicated results are displayed. November 2001 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 4-1
5.Create/Edit a Sample-ID s List Wizard Using the drop down list box in the tool bar you can display for example statistic data (mean, standard deviation, variation coefficient) of replicates. In order to display the summery of all data of one well you have to select the well and click the button. In order to zoom to a part of the plate you have to select zoom in the toolbar and drag a frame over the part of the plate you want to enlarge. With clicking the Def. button on the Cutoff results window you can view the Cutoff definitions. Using the arrow buttons << and >> you can switch between the plate view windows with Raw data, Transformations and Cutoff results. Click OK to come back to the main page of the Evaluate results wizard. With clicking Validation you reach the Validation Results dialog box, where the defined validation criteria with the results true or false are displayed. 4-2 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 November 2001
5.Create/Edit a Sample-ID s List Wizard Click OK to exit the dialog. Then click Next on the main page of the Evaluate results wizard to reach the Save dialog box of the Evaluate Results wizard: Using the buttons on this dialog you can: Save the workspace-file. View the History of the file. Edit the Report setup. Print the report. Reevaluate the data using another method. With clicking Finish you reach the Wizard List dialog box. November 2001 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 4-3
5.Create/Edit a Sample-ID s List Wizard 4.2.2 IgM-ELISA (quantitative example) Select the workspace IgM-ELISA.wsp from the file list and click Next to reach the following dialog box: Clicking Raw data, Transformations, Concentrations or Cutoff results brings you to a plate view window, where the indicated results are displayed. Using the drop down list box in the tool bar you can display for example statistic data (mean, standard deviation, variation coefficient) of replicates. 4-4 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 November 2001
5.Create/Edit a Sample-ID s List Wizard In order to display the summery of all data of one well you have to select the well and click the button. In order to zoom to a part of the plate you have to select zoom in the toolbar and drag a frame over the part of the plate you want to enlarge. With clicking the Def. button on the Cutoff results window you can view the Cutoff definitions. By clicking the Curve button on the Concentrations window you can display the Standard curve. Using the arrow buttons << and >> you can switch between the plate view windows with Raw data, Transformations, Concentrations and Cutoff results. Click OK to come back to the main page of the Evaluate results wizard. With clicking Validation you reach the Validation Results dialog box, where the defined validation criteria with the results true or false are displayed. Click OK to exit the dialog. Then click Next on the main page of the Evaluate results wizard to reach the Save dialog box of the Evaluate Results wizard: Using the buttons on this dialog you can: Save the workspace-file. View the History of the file. Edit the Report setup. Print the report. Reevaluate the data using another method. With clicking Finish you reach the Wizard List dialog box. November 2001 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 4-5
5.Create/Edit a Sample-ID s List Wizard 5. Create/Edit a Sample-ID s List Wizard 5.1 Introduction Create/edit a sample-id s list enables you to create new and to edit existing sample lists. Up to three sample IDs can be entered or imported per well. 5.2 Working with the Create/Edit a Sample-ID s List Wizard In the Wizard List dialog box, select Create/edit a sample-id s list and click OK. After clicking Next on the Welcome page the Select a sample-id s list dialog box is displayed. Click Create check box and type a file name and Remarks for the new sample- ID s list. Click Next to reach the Import/edit a sample-id s list dialog box, where you have to decide, whether you want to enter the sample ID s list manually or you want to import the sample ID s from a file. November 2001 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 5-1
5.Create/Edit a Sample-ID s List Wizard Clicking Import sample-id s list gives you the possibility to browse for different types of sample ID s files. Click Manual enter sample-id s list to reach the Sample ID Settings dialog box. Select 3 sample ID s per well and 96well plate and click OK to reach a plate view window with the Edit Well... dialog box in the upper left corner. Now you can enter the sample ID s well by well. You can reach the next well by pressing enter after writing the third sample ID of the actual well or by clicking the Left, Right, Up and Down keys. Select Autoincrement for automatically numbering up the sample ID of the next well. Another possibility to fill the plate with sample ID s is the Auto fill function: After closing the Edit Well... dialog box, select the part of the plate you want to fill with sample ID s. Then click Auto fill in the toolbar or select Autofill selection from the right mouse menu to display the Autofill selection dialog box. Enter the Sample IDs and the Start index and choose if and in which direction the index should be counted up. Click OK to fill the selected part of the plate according to your settings. After you entered all Sample ID s press OK in the toolbar to come back to the Import/edit a sample-id s list dialog box. Click Next to reach the Save dialog box of the Create/edit a sample-id s list wizard. With clicking Finish you save the sample-id s list and reach the Wizard List dialog box. 5-2 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 November 2001
6.Licensing 6. Licensing 6.1 Introduction 6.2 Licensing Licensing Magellan is a very simple process. There is no additional hardware as dongle required. Two different typical scenarios are described: the customer has bougth a version of the software at his local dealer. the customer has obtained a version from a CD distributed by TECAN, via Internet or any other channel. The user can work with Magellan for 30 days without limits until registering it. Without registration after this period the save and print option is disabled. Licensing of Magellan is performed with a registration wizard. To start the wizard select Register Magellan in the Help menu. The registration wizard starts with the introduction page, which shows the contents of the wizard. November 2001 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 6-1
6.Licensing 6.3 Serial number The next dialog box shows two different choices. The first one is for users, who have purchased the Magellan software. The serial number, found in the Magellan package, must be typed in. Users, who have not yet purchased Magellan software, must select the second alternative. Please ask your local TECAN dealer. 6.4 License Number After the serial number has been entered, the next dialog box that appears is the first one where the license number is entered. Users installing the software for a first time will not yet have a license number. The next step is to request a license number. 6-2 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 November 2001
6.Licensing 6.5 Registration Form The wizard constructs a registration form. The user should fill out the form. The fields marked with * are mandatory. 6.6 License Agreement Read the following licensing information carefully. Click I agree to continue. November 2001 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 6-3
6.Licensing 6.7 Sending the Registration Form to TECAN Click on Next to finish the process. A dialog box appears where the registration information is displayed. Clicking on E-Mail Form starts the standard e-mail program on the computer. All the necessary data is automatically filled in the field. After emailing the registration form to TECAN, the user should receive a response within 24 hours. This also applies if the user prints the form directly Print From... or via word processing system View From... and sends it as a fax. 6-4 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 November 2001
6.Licensing 6.8 Receiving a License Number from TECAN After receiving the license number from TECAN, the user should repeat the steps above until the dialog box for entering the license number is reached (see below). Instead of requesting an Magellan license number the user enters the number received from TECAN. The registration wizard then confirms the licensing number and summarises the user information. November 2001 Magellan Beginners Guide No: I 117 519 Rev No: 2.0 6-5