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BD FACSCaliburThe flow cytometer for yourroutine cell analysis needs

A system with a rich application basis anda modular approach that continues to meetevolving needs of cell analysis worldwide.The BD FACSCalibur platform offers a unique modular approach to flowcytometry and allows users to perform both cell analysis and cell sorting in aninnovative single bench-top system. Designed with multicolor application supportin mind, the BD FACSCalibur flow cytometer provides flexibility to support a widevariety of research and clinical applications.Engineered to meet evolving clinical and research requirements, the user-friendlyBD FACSCalibur system can be easily upgraded to address emerging automationor sorting requirements in a laboratory. Walkway automation can be achieved bythe BD FACS Loader option that handles tube-based assays or the BD HighThroughput Sampler that handles assays in 96 or 384-well microtiter plates.* Inaddition, the BD FACSCalibur system can be upgraded to support cell sorting toallow for the isolation of a population of interest.*Built-in modularity and simple upgrades make the BD FACSCalibur an excellentlong-term investment for today’s cost conscious laboratories.The BD FACSCalibur system is compact and easy to use. It is complemented by abroad suite of intuitive software solutions to streamline analysis for a wide rangeof applications, including enumeration of lymphocyte subsets, stem cells, residualwhite blood cells, reticulocytes, DNA analysis,* immune function studies,* beadbased immunoassays,* and multiplexed analysis of signal transduction* andphosphorylation targets.*Enabling fast, easy, and accurate results forroutine applications with a versatile, modulardesign has made the BD FACSCalibur theautomated benchtop cytometer of choicefor cell analysis in research and clinicallaboratories worldwide.*Some options and applications are for Research Use Only.

REPRODUCIBLEPerformance and simplicitywhen it countsThe BD FACSCalibur is a fully integrated multiparametersystem that has the performance and sensitivity to ensureobjective and reproducible results vital to laboratoriesworldwide.Optical path configurationFlow cytometry has evolved rapidly to incorporate the useof multiple fluorochromes to effectively identify and isolatesubset populations in a single sample. This maximizes theinformation gained from limited samples and makes theassays more cost effective.FL1SSC 488/10The first system to provide standard multicolor analysiscapabilities with the use of dual-laser technology, interbeamcompensation, and an alignment free optical design, theBD FACSCalibur system ensures high sensitivity, minimalcompensation, and maximum flexibility in choice offluorochromes.530/30FL2 585/42FL4 661/16This high performance system is a valued tool worldwide forroutine cell analysis, assay development, verification, andidentification of cellular populations of interest.FL3 670LPBlue laser488 nmflowcellRed diode laser 635 nm488/10FSC diodeBD FACSCalibur optical path configurationon R1Gated Gatedon R1 andR2and R210 210 110 0aR410 310 2R3R310 1A10 310 210 10 1101010 110 210 210 310 310 4CD123CD123PEPE10 4bb10 010 010 010 4R4Anti–HLA-DR PerCP10 3Anti–HLA-DR PerCPA growing number of flow cytometry–based assays depend onrare-event detection. For example, in the characterization ofsubpopulations of stem cells, millions of cells are analyzed toachieve a statistically significant sampling of the subset of interest.Advanced applications like this require a flow cytometer capableof high-speed analysis. The BD FACSCalibur system features amultiparameter triggering system that allows a user to acceleratethe analysis by electronically eliminating events from onboardprocessing.Anti–HLA-DR PerCPRare-event analysisaon R1Gated Gatedon R1 andR2and R210 4Anti–HLA-DR PerCP10 410 410 3R510 210 110 010 00 11010B10 110 210 210 310 310 4CD11c CD11cAPC APCA Region R3 defines basophils and region R4 defines CD123 DCs(0.14% of total).B Region R5 defines CD11c DCs (0.21% of total).4R510 4

RESULTSDual-laser design for multicolor analysisThe BD FACSCalibur dual-laser design provides the flexibilityand sensitivity needed for multicolor analysis. Two lasers,an air-cooled argon laser and a red diode laser, are spatiallyseparated for high sensitivity, minimal compensation, andmaximum flexibility in fluorochrome selection. This maximizesthe information obtained from limited samples and increasesthe cost-effectiveness of each assay.A fully integrated multiparameter system, the BD FACSCalibursystem ensures objective and reproducible results, which arevital to today’s laboratories.BD FACSCalibur optical benchUnique easy-to-use sorting technology (optional)The BD FACSCalibur cell sorting option allows users to identify and isolate a population ofinterest. Sorting on the BD FACSCalibur system occurs in a completely enclosed, aerosolfree environment for enhanced safety, which is especially important when processingbiohazardous samples. The sorting process is designed to be straightforward. After thepopulation of interest is gated, simply click “Acquire” to begin the sort. After the sampleis acquired and cells pass through the laser, a unique catcher tube mechanism moves inand out of the sample core stream at a rate of roughly 300 times per second to capturedesignated cells and direct them to a collection tube or to an optional cell concentratormodule for further processing. (The BD FACSCalibur cell sorting and cell concentratoroptions are for Research Use Only)BD FACSCalibur sorting mechanism5

PRODUCTIVITYFlexibility for sample introductionand enhanced productivityBD FACS Loader optionThe BD FACS Loader option available for theBD FACSCalibur system provides walkaway automationfor routine applications. Featuring a removable 40-tubecarousel, onboard mixing, and BD Worklist Managersoftware, the BD FACS Loader option offers superiorflexibility to efficiently address the workflow demandsof the busy laboratory.BD High Throughput Sampler (HTS) optionThe BD High Throughput Sampler (HTS) option providesflexible, easy-to-use automation. For those who prefer towork with multiwell plates, BD Biosciences offers the firstwalkaway sample introduction device for 96- and 384well plates. The HTS option allows users to rapidly acquiresamples with the BD FACSCalibur flow cytometer in amicrotiter plate. As a modular option for new or existingBD FACSCalibur flow cytometers, the HTS option is designedto speed through a variety of microtiter plates in less than15 minutes* and support a wide range of applicationsincluding the BD Cytometric Bead Array assay. The HTSoption supports 96 U-, V-, and flat-bottom plates as well as384-well microtiter plates.BD Plate Manager software allows you to createcustomized delivery protocols to be used repeatedlywith user-defined mixing, wash, and analysis parameters.(The BD High Throughput Sampler option and BD PlateManager are for Research Use Only)* Based on a 2-second sample acquisition.The BD Leucocount system is the firstautomated, IVD-cleared product formonitoring leucoreduced blood products.Together with BD Trucount tubes andBD Leucocount process controls, the BDLeucocount system provides excellentaccuracy, precision, and linearity. For highthroughput walkaway capability, the BDFACS Loader can also be added to your baseBD FACSCalibur system.1000Mean Observed Leucocount WBC/µLEfficient residual white blood cellenumerationBD Leucocount kitThe BD Leucocount system provides excellentaccuracy, precision, and linearity at clinicallysignificant levels of detection for RBC (red) andplatelet (blue) samples.10010Decision PointArea1 5 106 1 1060.10.1110Expected WBC/µL61001000

104ImmunophenotypingCharacterizing cells at different stages of developmentthrough the use of fluorescent-labeled monoclonal antibodies against surface markers is one of the most commonapplications of flow cytometry. The BD FACSCalibur systemprovides multicolor capability in an easy-to-use system.Instrument setup is as easy as the push of a button and a clickof the mouse. Efficient collection optics, coupled to the flowcell with an optical gel, permit the use of a low-powered,air-cooled argon laser and a red diode laser to ensure thehigh sensitivity required for analysis of low-density antigens.CD4-PE102101100BD Trucount tubes make single-platform absolute counting easy—no syringes, additionalpipetting steps, or need to integrate data from another analyzer. Designed for use withclinical products such as BD Multitest and BD Tritest lymphocyte subsetting reagents,BD Leucocount rWBC assay kit, BD Trucount tubes bring precision to cell counts andimproved productivity. For research applications, BD liquid counting beads are also available within the BD Cell Viability kit for quick estimations of cell cultures or bacterial concentrations. (The BD Cell Viability kit is for Research Use Only)103Accurate absolute counts with BD Trucount technologyDisplay of analysis for a sample prepared withBD Multitest reagents in a BD Trucount tubeusing the lyse/no-wash protocol in BD Multiset software.100101102103104CD3-FITC7

DATA MANAGEMENTDelivering results with a powerful,easy-to-use data management systemThe BD FACSCalibur system is appreciated for its userfriendly design, high sample throughput, improvedworkflow management through automation, andsimple yet sophisticated software applications thatenhance routine analyses.While performing any of a wide range of supportedapplications, the BD FACSCalibur system provides the toolsto obtain results quickly, easily, and accurately.The ease-of-use of the BD FACSCalibur system is enhancedby the BD FACStation data management system thathas become one of the most popular, user-friendly datamanagement systems in flow cytometry. The BD FACStationsystem automates many software functions and delivershigh-performance acquisition and analysis tools for creatingplots, gating, statistical analysis, and reporting.The BD FACStation workstation takes advantage of theintuitive Mac environment by using simple pull-downmenus and icons. It performs all the computing tasks requiredfor fast, accurate results including instrument setup, as wellas data collection, analysis, and 2004006008001000FL2-AreaDNA analysisPulse processing is a powerful tool used in the discrimination of doublets foroptimal DNA analysis.8Counts800600400200FL2-AreaR10The BD FACSCalibur system works together with BD DNA analysisquality control products to eliminate many of the variables associated with ploidy and cell-cycle analysis. The instrument’s opticaland fluidic design ensures that high-resolution DNA ploidy andS-phase determinations are easy to achieve. Electronic doubletdiscrimination, using the area and width of the fluorescence pulse,allows for the removal of artifacts—doublets and aggregates—from the analysis. High-resolution electronics and linearity ensureaccurate results. To ensure optimal performance, BD DNA QCproducts, including fixed cells and fluorescent particles, can beused for setup, verification of the doublet discrimination module,and monitoring daily operation. (BD DNA Analysis products are forResearch Use Only)1000DNA Analysis300A comprehensive quality assurance systemBD Biosciences offers a complete family of productsdesigned to ensure the highest quality clinical results fromthe BD FACSCalibur system. BD FACSComp software,in conjunction with BD Calibrite beads, provides acomprehensive system to set up the instrument, assesssensitivity, and set compensation for immunofluorescenceapplications. For clinical applications, BD Biosciences offersa wide variety of cellular controls to verify performancefor immunophenotyping, reticulocyte analysis, stem cellenumeration, and rWBC applications.