Plate Washer, Bio-Plex Pro used with BioPlex Array reader. Plate Readers SOP. Detect on Instrument Picture to Jump to that Section

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1 BioPlex 200 Array Reader w HTF By Plate Washer, Bio-Plex Pro used with BioPlex Array reader Plate Readers SOP Enspire Reader Perkin-Elmer Microplate Reader Bio-Rad Detect on Instrument Picture to Jump to that Section Prepared by: Bob Morrison STLCC- Instrumentation Specialist Original May 08, Latest Rev Nov 2012 Page 1

2 Array Reader: Bio-Plex 200, w HTF (high-throughput fluidics) Bio-Rad BioPlex R126B The Bio-Plex suspension array system offers protein and nucleic acid researchers a reliable multiplex assay solution that permits analysis of up to 100 biomolecules in a single sample. System Components Array reader combines 2 lasers, fluidics, and real-time digital signal processing to distinguish up to 100 different color-coded bead sets, each representing a different assay Microplate platform automates the reading of 96-well plates, yielding up to 9,600 data points in about 35 minutes Calibration and validation kits system tools for daily calibration of signal output and for IQ/OQ and monthly validation of primary performance parameters of the reader Bio-Plex MCV plate IV maintenance, calibration, and validation (MCV) plate customized for hands-free startup, shutdown, and performance testing Optional high-throughput fluidics (HTF) delivers up to 20 L of sheath fluid (40 plates) without user intervention PC and monitor includes Pentium 4 processor, CD-RW, Windows XP operating system, Microsoft Office Professional and Adobe Acrobat Reader software, and a 17" flat-screen monitor Bio-Plex Manager software controls the instrument, data acquisition and analysis for superior curve fitting, statistics reporting, and charting features, with no requirements to export or import files Bio-Plex SNP Manager data analysis utility simple-to-use data analysis tool for nucleic acid applications Page 2

3 Array Reader:Bio-Plex, Technology APD: Avalanche photodiode detector DD: Doublet discriminator channel, discriminates single beads from aggregated beads CL1: Classify channel, allows multiplexing, detects dye inside beads RP1: Reporter channel, quantitates assay in this channel Bio-Plex assays are bead-based assays that can be performed in a mixed array (multiplexed). The reader combines two lasers, fluidics, and real-time digital signal processing to distinguish up to 100 different sets of color-coded polystyrene beads, each bearing a different assay. The Bio-Plex array reader is an essential tool that performs the key functions of this multiplex technology: Fluidics The Bio-Plex array reader detects individual beads by flow cytometry. The fluidics system of the reader aligns the beads into single file as they enter a stream of sheath fluid and then enter a flow cell. Once the beads are in single file within the flow cell, each bead is individually interrogated for bead color (analyte) and assay signal strength (PE fluorescence intensity) Lasers The Bio-Plex array reader is a dual laser system. The 532 nm Nd-Yag laser (green "reporter" laser) is used to excite the PE dye of the assay, for example, streptavidin-pe used for sandwich immunoassays. The 635 nm solid state laser (red "classify" laser) is used to excite the dyes inside the beads to determine their "color" or "region" and is also used for doublet discrimination by light scatter Optics The Bio-Plex array reader contains an optics bench with four detection channels. Details are shown below. The dual lasers are precisely focused to excite an individual bead within the quartz flow cell. Upon excitation, the fluorescent signal emitted from the bead travels though the optics paths to the individual detectors Detectors The Bio-Plex array reader has four detectors, one for each of the optical paths shown in the figure below. A highsensitivity photomultiplier tube (PMT) detector is used for the reporter channel. Photodiodes are used for the stronger signals of the classify CL1 and CL2 and doublet discrimination channels Page 3

4 Array Reader: Bio-Rad Pro II Wash Station Bio-Plex Pro wash stations eliminate manual wash steps from Bio-Plex assays, making Bio-Plex assays as easy as ELISAs. The wash stations are specifically designed to perform Bio-Plex assay wash steps, but are compatible with any standard xmap assay. Benefits of Bio-Plex Pro wash stations include: Improved lab productivity Reliable and reproducible results Optimized on-board protocols Bio-Plex Pro II wash station includes both a magnetic carrier and an interchangeable vacuum manifold for hands-free wash steps on any xmap assay (magnetic and nonmagnetic bead-based assays) Both options include preset wash programs that have been optimized for Bio-Plex assays. Using Bio- Plex Pro wash stations in an assay workflow can reduce manual intervention and help decrease variability between independent experiments Hotlink to STLCC Liquid Handling/Plate Washer SOPs Page 4

5 Array Reader: Bio-Plex Manuals and Training Hotlink to Training Overview Hotlink to Best Practices Window Login: None required for normal use/functionality Admin account established by SS 12/4/12 Morrison/15Biorad also setup by SS 12/4/12 Hotlink to Bio-Plex 200 Array Reader Instruction Manual....pdf (.6mb) Hotlink to Bio-Plex 200 Maintenance Guide....pdf (.6mb) Page 5

6 Array Reader: Bio-Plex, Front Panel, Maintenance See pg 8 for info Page 6

7 Array Reader: BioPlex, Rear Panel, Maintenance See pg 8,9 for info Page 7

8 Array Reader: BioPlex, Ventilation,Sheath Filters, Maint. Page 8

9 Array Reader: BioPlex, Internal Fluidics Page 9

10 Array Reader: BioPlex Maintenance pg 1 of 5 Page 10

11 Array Reader: BioPlex Maintenance pg 2 of 5 Page 11

12 Array Reader: BioPlex Maintenance pg 3 of 5 Page 12

13 Array Reader: BioPlex Maintenance pg 4 of 5 Page 13

14 Array Reader: BioPlex Maintenance pg 5 of 5 Page 14

15 Array Reader: Bio-Plex 200, Cytokine Assay Kit Hot link to Bio-Plex Pro Cytokine, Chemokine, and Growth Factor Manual pdf Page 15

16 Array Reader: Bioplex, Software Licensing to Laptops From Subject Received Categories Jim Rau Re: BioPlex Software Licenses at STLCC BRDG Thu 9/16/10 CC: Scott Schneider Hello Bob, Yes, you can install the software on the laptops. On each system with key inserted, you will be able to launch the software. Thanks, Jim Jim Rau Instrumentation Systems Specialist MO, NE, and Southern Illinois Bio-Rad Laboratories VM: x8302 Technical Support/Orders 800-4BIORAD ( ) Page 16

17 Plate Reader: EnSpire 2300, PerkinElmer EnSpire Alpha Multilabel Plate Reader Product # A 16H x 20D x 19 W (inches) SN: , Software Ver /25/11 RGM (Setting/Options menu) Contract Specialist July 2014 Jessica.Killmon@PERKINELMER.COM Fluorescence Intensity detection capability with monochromator Photometric UV/VIS technology for ELISA assays and DNA/protein quantitation with monochromator ELISA reference wavelength correction Up to 384-well plates Touch screen: Easy to use; saves desktop space, can be used with gloves Pre-coded assay protocols Data export in Excel /text formats to the network or into a USB memory stick Easy access to the filter wheel with eight barcode identified filter positions Integrated data-analysis software: curve fit (lin-reg, spline, 4PL/5PL), background subtraction, ratio calculation, IC50 calculation, Average, CV %, and Z -value 21 CFR Part 11 support Windows Vista Operating System R126B The EnSpire Multilabel Plate Reader is a compact, configurable multimode reader with quad-monochromator for fluorescence and absorbance, ultra-sensitive luminescence and Alpha-technology. The instrument has been designed for laboratories with diverse user needs, and includes an intuitive and easy to use touch screen to reduce assay startup times and simplify protocols for all users. Skip to Maintenance/Service Info Hot link to DNA/RNA quantification brochure Hot link to Enspire User Manual.. Pdf 188 pgs Page 17

18 Plate Reader: Enspire; Startup, Do Steps 1-6 in Order 5. Logon if required as Enspire User ( no password) using touchscreen pen 6. Start Enspire Manager Software by tapping icon with pen 1. Make sure screen is on push button 2. Push long flat silver colored button on rear to boot CPU, blue LED goes to solid 3. Wait for Windows Vista to start 4. Push toggle near power plug on rear of device to turn on device Page 18

19 Plate Reader: Enspire Manager, Main Screen Page 19

20 Plate Reader: Enspire; Settings, Restore Floating Menu Bar 1. Select Settings to examine and configure items such as locations for files and options to interface with Windows OS. 2. Use Settings also to access or restore (normalize) the floating menu bar. Exit or Minimize windows and locate floating bar in bottom tray. 3. Use Settings-Windows path also to transfer Print-Screen image dumps to applications such as MS Word. Page 20

21 Plate Readers: Enspire, Settings, General, Floating Menu Bar 1. Select General Tab (above) and then 2. Show Floating Menu Bar to add navigation bar to all future Enspire menus. Page 21

22 Plate Reader: Enspire; Database Tab Page 22

23 Plate Reader: Enspire; Protocol Templates Page 23

24 Plate Reader: Enspire; Copy Protocol to Edit Page 24

25 Plate Reader: Enspire; Plate Map Options Page 25

26 Plate Reader: Enspire; Plate Map Operations Page 26

27 Plate Reader: Enspire; Plate Map, Unknown Designation Page 27

28 Plate Reader: Enspire; Loading Plate A1 Position Page 28

29 Plate Reader: Enspire; Run Protocol, Counts Screen Page 29

30 Plate Reader: Enspire, BioTech Lab, Absorbance ELISA Example These parameters were set using the Create Protocol menu. Page 30

31 Plate Readers: Enspire, Biotech Lab Ex, Run, then Show Results 1. Load Plate 2. Select Run Protocol 3. Select Show Results 4. Selecdt Counts/Cal Page 31

32 Plate Readers: Enspire; Loss of These parameters were set using the Create Protocol menu. Page 32

33 Plate Readers: Enspire, Biotech Lab Ex, Show, Export to Flashdrive 1. Select Show Results 2. Select Export 3. Select File Settings and enter path and name to device Page 33

34 Plate Reader: Enspire, Maintenance, Service Agreement Page 34

35 Plate Reader: Enspire, Suggested Calibration Method BB 4/2013 Page 35

36 On/Off lower rear Menu Window Key Pad Membrane controls Bio Rad 680 Microplate Reader Microplate Reader Standard Operating Procedures Link to BioRad Model 680 Users Manual (pdf) Prepared by: Bob Morrison STLCC-FV, Instrumentation Specialist May 2008, Revised Nov 08, Feb 09 Page 36

37 Microplate Reader: External Features and Ports Page 37

38 Microplate Reader: Membrane Keyboard Map Page 38

39 Microplate Reader: Setup and Basic Operations 1. Flip the switch located in the back to the on position. The device will go through a brief self-test mode and then display a screen showing: Login Common User: ***** (if administrator is shown, toggle the up/down arrow to Common User) Enter Password: (Enter five zeros) using keypad, then hit Enter key 2. Settings from a previous run will be displayed. Select EDIT to change protocols or to modify the protocol settings for : Cutoff, Report, Limit, STDs, Mode, Mapping, Kit Name. Typical default values are: The current filter setting (previously set to 450nm) The shaking setting (usually 3 sec) The incubation setting (usually none) 3. To change the filter or other settings, use the membrane keyboard EDIT function. For example to define filter wavelengths for the 1-8 positions that can be used in the reading: Select EDIT, then Filters, then use the up/down arrow and key-in desired wavelengths for each of the 1-8 position, use 0 for no filter. Filters can be selected using the: EDIT-Protocol-Mode-Photomode-Filters and then toggle through the values assigned above. 4. Load the array to be tested and press the red START button on the top left side of the membrane keyboard to initiate the test. 5. When the test is completed, press PRINT on the membrane keyboard to print the raw data results. 6. Other reports are available, instructions are in the User manual. Page 39