Plate Washer, Bio-Plex Pro II Enspire Reader @BRDG Used .

BioPlex 200 Array Reader w HTFBy Bio-Rad @BRDGPlate Washer, Bio-Plex Pro II@BRDG used withBioPlex Array readerEnspire ReaderPerkin-ElmerPlate ReadersSOPMicroplate ReaderBio-RadSTLCC-CPLS;Morrison 7/30/2014Detect on Instrument Picture to Jump to that SectionPrepared by: Bob MorrisonSTLCC- Instrumentation SpecialistOriginal May 08 , Latest Rev Nov 2012Page 1

Array Reader: Bio-Plex 200, w HTF (high-throughput fluidics) Bio-RadBioPlex 200 @BRDG, R126BThe Bio-Plex suspension array system offers protein and nucleic acid researchers a reliable multiplexassay 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 differentcolor-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 monthlyvalidation 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 OfficeProfessional 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 applicationsPage 2STLCC-CPLS;Morrison 7/30/2014

Array Reader:Bio-Plex, TechnologyAPD: Avalanche photodiode detectorDD: Doublet discriminator channel, discriminates singlebeads from aggregated beadsCL1: Classify channel, allows multiplexing, detects dyeinside beadsRP1: Reporter channel, quantitates assay in this channelBio-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 bearinga 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 beadsinto 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 thePE 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 bylight scatter Optics — The Bio-Plex array reader contains an optics bench with four detection channels. Details are shown below. The duallasers are precisely focused to excite an individual bead within the quartz flow cell. Upon excitation, the fluorescent signal emittedfrom 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 theclassify CL1 and CL2 and doublet discrimination channelsSTLCC-CPLS;Morrison 7/30/2014Page 3

Array Reader: Bio-Rad Pro IIWash StationBio-Plex Pro wash stations eliminate manual wash steps from Bio-Plex assays, making Bio-Plex assaysas 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 interchangeablevacuum manifold for hands-free wash steps on any xMAP assay (magnetic andnonmagnetic bead-based assays)Both options include preset wash programs that have been optimized for Bio-Plex assays. Using BioPlex Pro wash stations in an assay workflow can reduce manual intervention and help decreasevariability between independent experimentsHotlink to STLCC Liquid Handling/Plate Washer SOPsSTLCC-CPLS;Morrison 7/30/2014Page 4

Array Reader: Bio-Plex Manuals and TrainingHotlink to Training OverviewHotlink to Best PracticesWindow Login: None required for normal use/functionalityAdmin account established by SS 12/4/12Morrison/15Biorad also setup by SS 12/4/12Hotlink to Bio-Plex 200 Array Reader Instruction Manual. .pdf (.6mb)Hotlink to Bio-Plex 200 Maintenance Guide. .pdf (.6mb)STLCC-CPLS;Morrison 7/30/2014Page 5

Array Reader: Bio-Plex, Front Panel, MaintenanceSee pg 8 for infoSTLCC-CPLS;Morrison 7/30/2014Page 6

Array Reader: BioPlex, Rear Panel, MaintenanceSee pg 8,9 for infoSTLCC-CPLS;Morrison 7/30/2014Page 7

Array Reader: BioPlex, Ventilation,Sheath Filters, Maint.STLCC-CPLS;Morrison 7/30/2014Page 8

Array Reader: BioPlex, Internal FluidicsSTLCC-CPLS;Morrison 7/30/2014Page 9

Array Reader: BioPlexMaintenancepg 1 of 5STLCC-CPLS;Morrison 7/30/2014Page 10

Array Reader:BioPlexMaintenancepg 2 of 5STLCC-CPLS;Morrison 7/30/2014Page 11

Array Reader:BioPlexMaintenancepg 3 of 5STLCC-CPLS;Morrison 7/30/2014Page 12

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Array Reader: Bio-Plex 200, Cytokine Assay KitHot link to Bio-Plex Pro Cytokine, Chemokine, and Growth Factor Manual pdfSTLCC-CPLS;Morrison 7/30/2014Page 15

Array Reader: Bioplex, Software Licensing to LaptopsFromSubject ReceivedCategoriesJim Rau Re: BioPlex Software Licenses at STLCC BRDGCC: Scott SchneiderThu 9/16/10Hello 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, JimJim Rau Instrumentation Systems SpecialistMO, NE, and Southern IllinoisBio-Rad LaboratoriesVM: 800-876-3425 x8302Jim Rau@Bio-Rad.comTechnical Support/Orders 800-4BIORAD CPLS;Morrison 7/30/2014Page 16

Plate Reader: EnSpire 2300, PerkinElmerEnSpire Alpha Multilabel Plate ReaderProduct # 2300-001A16H x 20D x 19 W (inches)SN: 23000302,Software Ver. 3.0 8/25/11 RGM(Setting/Options menu)Contract Specialist July 2014Jessica.Killmon@PERKINELMER.COM Fluorescence Intensity detection capability with monochromator Photometric UV/VIS technology for ELISA assays andEnspire2300 @BRDG, R126BDNA/protein quantitation with monochromator ELISA reference wavelength correctionThe EnSpire Multilabel Plate Reader is a compact, Up to 384-well platesconfigurable multimode reader with quad-monochromator for Touch screen: Easy to use; saves desktop space, can be usedfluorescence and absorbance, ultra-sensitive luminescencewith glovesand Alpha-technology. The instrument has been designed Pre-coded assay protocolsfor laboratories with diverse user needs, and includes an Data export in Excel /text formats to the network or into a USBintuitive and easy to use touch screen to reduce assay startmemory stick Easy access to the filter wheel with eight barcode identified filter up times and simplify protocols for all users.positions Integrated data-analysis software: curve fit (lin-reg, spline,Skip to Maintenance/Service Info4PL/5PL), background subtraction, ratio calculation, IC50calculation, Average, CV %, and Z’-value 21 CFR Part 11 supportHot link to DNA/RNA quantification brochure Windows Vista Operating SystemSTLCC-CPLS;Morrison 7/30/2014Hot link to Enspire User Manual . Pdf 188 pgsPage 17

Plate Reader: Enspire; Startup, Do Steps 1-6 in Order5. Logon ifrequired asEnspire User ( nopassword) usingtouchscreen pen6. Start EnspireManager Softwareby tapping iconwith pen1. Make sure screenis “on” push button2. Push long flatsilver colored buttonon rear to boot CPU,blue LED goes tosolid3. Wait for WindowsVista to start4. Push togglenear power plugon rear of deviceto turn on deviceSTLCC-CPLS;Morrison 7/30/2014Page 18

Plate Reader: Enspire Manager, Main ScreenSTLCC-CPLS;Morrison 7/30/2014Page 19

Plate Reader: Enspire; Settings, Restore Floating Menu Bar1. Select “Settings” to examineand configure items such aslocations for files andoptions to interface withWindows OS.2. Use Settings also to accessor restore (normalize) thefloating menu bar. Exit orMinimize windows and locatefloating bar in bottom tray.3. Use Settings-Windows pathalso to transfer Print-Screenimage dumps to applicationssuch as MS Word.STLCC-CPLS;Morrison 7/30/2014Page 20

Plate Readers: Enspire, Settings, General, Floating Menu Bar1. Select General Tab (above)and then2. “Show Floating Menu Bar” toadd navigation bar to allfuture Enspire menus.STLCC-CPLS;Morrison 7/30/2014Page 21

Plate Reader: Enspire; Database TabSTLCC-CPLS;Morrison 7/30/2014Page 22

Plate Reader: Enspire; Protocol TemplatesSTLCC-CPLS;Morrison 7/30/2014Page 23

Plate Reader: Enspire; Copy Protocol to EditSTLCC-CPLS;Morrison 7/30/2014Page 24

Plate Reader: Enspire; Plate Map OptionsSTLCC-CPLS;Morrison 7/30/2014Page 25

Plate Reader: Enspire; Plate Map OperationsSTLCC-CPLS;Morrison 7/30/2014Page 26

Plate Reader: Enspire; Plate Map, Unknown DesignationSTLCC-CPLS;Morrison 7/30/2014Page 27

Plate Reader: Enspire; Loading PlateA1 PositionSTLCC-CPLS;Morrison 7/30/2014Page 28

Plate Reader: Enspire; Run Protocol, Counts ScreenSTLCC-CPLS;Morrison 7/30/2014Page 29

Plate Reader: Enspire, BioTech Lab , Absorbance ELISA ExampleThese parameters were setusing the “Create Protocol”menu.STLCC-CPLS;Morrison 7/30/2014Page 30

Plate Readers: Enspire, Biotech Lab Ex, Run, then Show Results1.2.3.4.STLCC-CPLS;Morrison 7/30/2014Load PlateSelect Run ProtocolSelect Show ResultsSelecdt Counts/CalPage 31

Plate Readers: Enspire; Loss ofThese parameters were setusing the “Create Protocol”menu.STLCC-CPLS;Morrison 7/30/2014Page 32

Plate Readers: Enspire, Biotech Lab Ex, Show, Export to Flashdrive1. Select Show Results2. Select Export3. Select File Settings and enterpath and name to deviceSTLCC-CPLS;Morrison 7/30/2014Page 33

Plate Reader: Enspire, Maintenance, Service AgreementSTLCC-CPLS;Morrison 7/30/2014Page 34

Plate Reader: Enspire, Suggested Calibration MethodBB 4/2013STLCC-CPLS;Morrison 7/30/2014Page 35

On/Off lower rearMenu WindowKey Pad MembranecontrolsBio –Rad 680Microplate ReaderMicroplate ReaderStandard Operating ProceduresLink to BioRad Model 680 Users Manual (pdf)Prepared by: Bob MorrisonSTLCC-FV, Instrumentation SpecialistMay 2008, Revised Nov 08, Feb 09STLCC-CPLS;Morrison 7/30/2014Page 36

Microplate Reader: External Features and PortsSTLCC-CPLS;Morrison 7/30/2014Page 37

Microplate Reader: Membrane Keyboard MapSTLCC-CPLS;Morrison 7/30/2014Page 38