BD FACSCalibur Flow Cytometer - File.qums.ac.ir
BD FACSCalibur Flow CytometerThe Automated, Multicolor Flow Cytometry SystemA Complete System forToday’s LaboratoryBD BiosciencesClontechDiscovery LabwareImmunocytometry SystemsPharmingen
The BD FACSCalibur System:The Gold Standard thatContinues to EvolveThe Gold Standard That Continues to EvolveSince 1974, when the firstFluorescent Activated CellSorterBDintroduced,Biosciences has beentheofwasleadingproviderinnovativetechnologyand advanced solutions ldautomatedbenchtop sorting and duallaserBDtechnology,theFACSCalibur continuestosystembetheinstrument of choice vestmentinaBD Biosciences instrumentpays off over and over againbecausetheyprovidemodular, flexible solutionsthat can be adapted andevolvedasyourneedschange. You have the optionto choose a basic system,orcomplete,automatedsolutions, or a variety ofchoices in between. Mostimportant, you’ll always getthe industry’s most usablesoftware and the service andsupport that come only fromBD Biosciences.1www.bdbiosciences.comThe BD FACSCalibur flow cytometer is another BD Biosciences first:a multicolor benchtop flow cytometry system that is capable of bothanalyzing and sorting. Combined with your choice of sample loader,a wide variety of reagents, and powerful software, the BD FACSCalibursystem offers the high throughput necessary to meet productivityrequirements without compromising the flexibility and performanceessential for today’s busy, cost-conscious laboratory.
Compact and easy to use, the BD FACSCalibur system can quickly perform routinetasks including immunophenotyping, absolute counting, residual white blood cellenumeration, stem cell analysis, DNA analysis, and isolation by sorting. And withits superior multicolor analysis and sorting capabilities, the BD FACSCalibur systemis a highly sensitive tool, providing laboratories with the technological capabilitiescrucial for sophisticated cellular and molecular work in areas such as genetics,proteomics, multicolor analysis, and immune function.The BD FACSCalibur system.The modular instrument of choicefor research and clinical laboratoriesworldwide. The gold standard thatcontinues to evolve.
FACSCalibur8/22/025:46 PMPage 4B D FA C S L o a d e r * :Cell Sorting †:Routine Automation EnhancesInnovative Technology Makes CellP ro d u c t i v i t ySorting RoutineWith the BD FA C S Loader option,the BD FA C S Calibur system provideswalkaway automation for routineapplications. Featuring removable40-tube carousels, on-board mixing,and BD Wo r k l i s t M a n a g e r* software,the BD FA C S Loader option off e r sunsurpassed flexibility to eff i c i e n t l yaddress the workflow demands oft o d a y ’s laboratories.The BD FA C S Calibur system is thefirst benchtop flow cytometry systemavailable that is capable of sortingcells of interest for further study.With the BD FA C S Calibur system,sorting is routine—just gate thepopulation of interest and clickA C Q U I R E—the BD FA C S Calibursystem does the rest. Sorting occursin a completely enclosed, aerosol-freeenvironment for enhanced safety whenprocessing biohazardous samples. Cellssorted by the BD FA C S Calibur systemare collected into test tubes, or ifdesired, can be deposited directlyonto filters or cell culture inserts withthe Cell Concentrator Module option.BD Multiwell AutoSampler † :Flexible Easy-to-Use AutomationBD FA C S Calibur Flow Cytometer:A Complete System For To d a y ’sLaboratoryBD FA C S C alibur Cytometer*:A Modular Benchtop SystemThe BD FA C S Calibur flow cytometeris the first automated multicolorflow cytometry system that performsboth analysis and sorting in onebenchtop system. Designed specificallyto support a wide range of applications,the BD FA C S Calibur flow cytometer is afully integrated multiparameter systemthat ensures objective, reproducibleresults so vital in today’s clinical andresearch laboratories.The BD FA C S Calibur systems’ usercentered design approach goes beyondmere ergonomics. Software instrumentcontrol, auto–sample loading, andpushbutton fluidic control maximizeease-of-use and productivity. Simpleyet powerful software allows youto customize applications whileautomating redundant tasks. Finally,the BD FA C S Calibur system lets youleverage your investment througha modular design that is flexible andupgradeable to meet your future needs.3www.bdbiosciences.comFor those who prefer to work withmultiwell plates, BD Biosciences offersthe first walkaway sample introductiondevice for 96- and 384-well plates, bothstandard and deep-well. BD Multiwell Plate Manager software providesan easy-to-use interface for you toinput sample information and createcustomized protocols, making experiment runs as simple as click-and-go.Dual Lasers:Optimized for Multicolor AnalysisThe BD FA C S Calibur system offersthe industry standard in multicoloranalysis capability—two lasers spatiallyseparated for high sensitivity, minimalcompensation, and maximum flexibilityin fluorochrome selection. With multicolor analysis, you maximize theinformation gained from small samples,as well as increase the cost-effectivenessof each assay.BD FA C S tationData Management System:Powerful and Easy-to-UseThe ease-of-use of the BD FACSCalibursystem is enhanced by its user-friendlydata management system. TheBD FACStation workstation takesadvantage of the intuitive Macintosh environment by using simple pull-downmenus and icons. It performs all thecomputing tasks required for fast,accurate results, including instrumentsetup as well as data collection,analysis, and management.The BD FACStation workstationoffers automated software andhigh-performance acquisition andanalysis tools for plotting, gating,statistical analysis, and reporting.
To CollectionTo WasteCatcher TubeMoves In andOut of StreamSample InjectionFigure 1Unique Sorting TechnologyThe BD FACSCalibur system features a unique catcher tube mechanism that moves in and out ofthe sample core stream 300 times per second to capture designated cells after they pass throughthe laser. The cells are then directed to collection tubes or the Cell Concentrator Module (optional)for further processing.ReagentsBD Biosciences offers a broad range of reagent products designed to meet multipleapplication requirements — from reagent and software systems for routine clinicalanalyses to cutting-edge research reagents and custom conjugates to enhance yourresearch capabilities. BD Biosciences also offers a complete line of quality assuranceproducts and services for instrument validation, method control, reliable gating,absolute counting, and within-panel consistency checks.www.bdbiosciences.com4
Routine Applications:Productivity and SimplicityWhen it CountsLaboratory technologists will appreciate the BD FACSCalibur system forits user-friendly design, high samplethroughput, improved workflowmanagement through automation,and simple yet sophisticated softwareapplications that enhance routineanalyses.Whether you are performingimmunophenotyping of lymphocytesubsets, leukemia or lymphomastudies, transplantation research,residual white blood cell enumeration,or a wide range of other applications,the BD FACSCalibur system provides thetools you need to obtain results quickly,easily, and accurately.ImmunophenotypingCharacterizing cells at differentstages of development through theuse of fluorescent-labeled monoclonalantibodies against surface markers isone of the most common applicationsof flow cytometry. The BD FACSCalibursystem opens the window to this worldof information by providing multicolorcapability in an easy-to-use system.Instrument setup is as easy as a pushof a button and a click of the mouse.Efficient collection optics, coupledto the flow cell with an optical gel,permit the use of a low-powered,air-cooled argon laser and a reddiode laser to ensure the highsensitivity required for analysisof low-density antigens.BD Biosciences also provides softwareand reagent systems to maximize yourlaboratory’s productivity. BD MultiSET software, in combination withBD MultiTEST reagents andBD Multi-Check controls, provides acomplete system for automating dataacquisition and analysis.5www.bdbiosciences.comOur timesaving lyse/no-washmethodology‡ using BD MultiTESTreagents reduces sample handlingand provides more answers pertube, leaving you with more time toperform other valuable tests. By usingBD Biosciences exclusive Expert Gate,BD MultiSET software ensures that thecell population of interest is analyzed,increasing the accuracy and reliabilityof your results.Absolute CountsBD TruCOUNT Tubes make singleplatform absolute counting easy—nosyringes, additional pipetting steps, orneed to integrate data from anotheranalyzer. Designed for use with clinicalproducts such as BD MultiTEST andBD TriTEST lymphocyte subsetreagents, BD LeucoCOUNT rWBCenumeration kit, and BD ProCOUNT progenitor cell enumeration kit,BD TruCOUNT Tubes bring precisionto your counts and improvedproductivity to your lab. For researchapplications, BD Liquid Counting beads†are also available for quick estimationsof cell culture or bacterial concentration.Figure 2Clinical Lab ReportsThe BD MultiSET software Lab Report containslymphocyte subset information, includingoptional absolute counts, reference-rangechecks, T-helper/suppressor ratio, andtube-to-tube consistency checks.
Residual White Blood CellEnumerationThe BD LeucoCOUNT system is thefirst automated, IVD-cleared productfor monitoring leucoreduced bloodproducts. Together with BD TruCOUNTTubes and BD LeucoCOUNT processcontrols, the BD LeucoCOUNT systemprovides excellent accuracy, precision,and linearity. For high throughputwalkaway capability, the BD FACSLoader can also be added toyour system.Mean Observed LeucoCOUNT WBC/µL 2.607 0.871 Expected WBC/µL; R2 0.955Mean Observed LeucoCOUNT WBC/µL DNA Analysis †Quality AssuranceThe DNA Analysis System componentswork together, from sample preparationto analysis, to eliminate many of thevariables associated with ploidy andcell-cycle analysis. The BD FACSCalibursystem’s optical and fluidic designensures that high-resolution DNAploidy and S-phase determinationsare easily achievable. Electronic doubletdiscrimination, utilizing the area andwidth of the fluorescence pulse, allowsfor the removal of artifacts—doubletsand aggregates—from the analysis.High-resolution electronics and linearityensure accurate results, and DNA QCparticles verify optimal performance.BD Biosciences offers a complete familyof products designed to ensure thehighest quality clinical results fromthe BD FACSCalibur system.BD FACSComp software, inconjunction with BD CaliBRITE beads,provides a comprehensive system to setup the instrument, assess sensitivity, andset compensation for immunofluorescenceapplications. For clinical applications,BD Biosciences offers a wide variety ofcellular controls to verify performancefor immunophenotyping, reticulocyteanalysis, stem cell enumeration, andrWBC applications.2.420 0.829 Expected WBC/µL; R2 0.931Mean Observed LeucoCOUNT WBC/µL100010010Decision PointArea1 5 106 1 1060.10.11101001000Adjusted Expected WBC/µLFigure 4BD LeucoCOUNT KitDNA AnalysisThe BD LeucoCOUNT system provides excellentaccuracy, precision, and linearity at clinicallysignificant levels of detection for RBC (red)and platelet (blue) samples.Pulse processing is a powerful tool used in the discrimination of doublets for optimal DNA analysis.104104103103CD3 PerCPCD5 PEFigure 3102104101103100Peripheral T-cellLymphoma102101100100101102103104100101CD7 FITC102103104103104CD45RO APC102101100101102103FSC-Heightred: aberrant (neoplastic) T cellsblue: normal T cellsgreen: normal NK cells104104103103Two four-color tubesare shown: 1) CD7 FITCvs CD5 PE and CD45ROAPC vs CD3 PerCP, and2) CD2 PE vs CD3 PerCPand CD8 FITC vs CD4 APC.Aberrant T cells are colored red, normal T cellsare colored blue, andnormal NK cells arecolored green.104CD4 APC100CD3 PerCPSSC-HeightFigure 5102101102101100100100101102CD2 PE103104100101102CD8 FITCwww.bdbiosciences.com6
Advanced Applications:Multicolor AnalysisSorting †Performance and FlexibilityWhen it CountsFlow cytometry has rapidly evolvedtoward the use of multiple fluorochromesas an effective means of identifyingsubset populations within a singlesample—maximizing the informationgained from small samples, as well asincreasing the cost-effectiveness of eachassay. BD Biosciences pioneered thetechnology used in multicolor analysisand sets the industry standard with itsuse of two spatially separated laserbeams. This dual-laser technologyensures high sensitivity, minimalcompensation, and maximum flexibilityin fluorochrome selection. And theBD FACSCalibur system is the first toinclude diode laser technology andinterbeam compensation—all in analignment-free benchtop system.Multiparameter flow cytometrycombined with cell sorting adds anadditional dimension to the informationyou gain from your sample. Purificationof subpopulations through sorting isuseful, eliminating contaminants thatcan interfere with sensitive assays,increasing the number of target cellsin the sample, or confirming flowcytometric findings. Cell sorting isideal for verifying cell morphologyand performing subsequent moleculartechniques such as FISH or PCR, orfunctional assays. Cross-correlation ofthe flow cytometric information withresults of the post-sort assay is apowerful tool in cellular analysis.Researchers and pathologists willappreciate the BD FACSCalibur systemfor its flexibility, which allows them toexpand their menu of applications astheir needs change. From genes toproteins to cells, this high-performancesystem is a valuable tool for assaydevelopment, verification, and physicalseparation of cells of interest for furthertesting or analysis.High PerformanceSide-Scatter andFluorescencePhotomultiplierArgon LaserBeam-Shaping OpticsRed Diode Laser7www.bdbiosciences.comLow-Angle,Light-Scatter Detector
FACSCalibur8/23/0212:32 PMPage 9Lymphocyte Function Assessment †102CD69 PE1016.49%10092.75%100101102103104102101CD69 PE103104Vβ6 FITC95.28%4.61%100Figure 6AThe BD FastImmune assay system isan innovative flow cytometric activationassay using whole blood to directlyassess individual T-lymphocytesubset responses to a wide variety ofphysiological and nonphysiologicalstimuli. And unlike radioisotopemethods that are time and laborintensive, the BD FastImmune assay iscompleted in just hours. The system’sunique approach makes it the idealresearch tool for answering basicbiological questions regarding immuneregulation, T-cell subset responsesto viral and bacterial antigens,and immune status assessmentfor chemotherapy, transplantation,and parasitology.103104REST100101102103104Vβ8 FITCFigure 6B104SEB103Figures 6A&B15.45%1.35%101PBMC and Daudi fractions were sorted for puritybased on immunofluorescence and scatter.Female PBMC and male Daudi were mixed andhybridized with chromosome Y (green) probesand counterstained with DAPI (red). Thehybridization of the sorted fractions show purefemale PMBC (Figure 6A, two red dots) andmale Daudi cells (Figure 6B, a red and a greendot), as expected.(Courtesy of Nga Bui, Tom Frey, PhD, SharylIwata, and Kathy Yokobata, BD Biosciences.)102CD69 PESorting Results4.35%10078.82%R5Injured100101102Vβ6 FITC103104102CD69 PE18.90%101R4Live103104R6DeadAdditional Applications †A thorough understanding of thebiology of the system under study,combined with the power and flexibilityof the BD FACSCalibur system, reagents,and data analysis tools, ensures thesuccessful application of flow cytometryin broad areas of biological andmedical research. Fluorescencequantitation with BD QuantiBRITE reagents, platelet analysis, stem cellresearch, multiparameter DNA analysis,BD Cytometric Bead Array, fluorescentprotein research, and microbial analysisare just a few of the forward-focusedapplication possibilities using theBD FACSCalibur system.Figure 7100101102103104Vβ8 FITCBacterial ApplicationsViability and concentration of bacteria frompharmaceutical, food, or environmental samplescan be demonstrated using propidium iodide (PI),thiazole orange‡ (TO), and BD Liquid countingbeads.† This sample of P. aeruginosa wasprepared in staining buffer at 5 x 105 cells/uL.6.23%10074.81%Figure 8Superantigen Activation of T CellsThe BD FastImmune Assay system†can be employed to examine specificantigen responses in human wholeblood. Staphylococcal enterotoxinB (SEB) activates VB6 T-cell subsetsbut does not activate VB8 subsets,as determined by CD69 expression.‡www.bdbiosciences.com8
BD FACSCalibur SystemSpecificationsThe BD FACSCalibur System:Service and Support When it CountsLaser488-nm, air-cooledArgon-ion laser635-nm, air-cooled reddiode laser (optional)Fluorescence SensitivityEstimated detection limit is200 FITC equivalent moleculesper particle.§Fluorescence ResolutionCoefficient of variation of 3%, full peak, for propidiumiodide–stained chickenerythrocyte nucleiForward and Side ScatterSensitivitySensitivity enables theseparation of fixed plateletsfrom noise.Forward and Side ScatterResolutionScatter performance isoptimized for resolvinglymphocytes, monocytes,and granulocytes.Sorting SpecificationSorting Purity: 95%Sorting Rate: Up to300 particles per secondSort Modes: Single Cell,Exclusion, and RecoveryPowerUS: 120 VAC /- 10%50/60 Hz /-2 HzCurrent: 15 amps maximumOutside US: Externaltransformer provided for100 VAC /-10%; 50/60 Hzand 220/240 VAC /-10%;50/60 Hz /-2 HzSizeSensor Module:Width 91.4 cm (36 in)Depth 61.5 cm (24.2 in)Height 67.3 cm (26.5 in)Cover Open: 124.5 cm (49 in)Computer: 48 H x 41 D x 54 W cm(19 x 16 x 11 in)WeightSensor Module: 109.1 kg(270 lbs)Computer:20 kg (44 lbs)10www.bdbiosciences.comBecause we understand how critical it is to have your system up and running,BD Biosciences backs its instruments, software, and reagents with BD FACService ,the most highly rated service and support program in the industry. BD FACServiceensures maximum instrument up-time and optimal performance with guaranteedresponse times, preventative maintenance programs, and highly trained service andapplication specialists.In addition, we provide a variety of training packages and customizededucational programs for your laboratory personnel that can be tailored tomeet the requirements of users at any skill level. This ensures a shorter learningcurve and increased productivity for your laboratory.The BD FACSCalibur System:The Gold Standard That Continues to EvolveWith its innovative reagents, powerful application software, and advancedcytometry hardware, the BD FACSCalibur system is the answer to your laboratoryneeds. Its built-in versatility meets the current demands of both clinical andresearch environments; its modularity takes you into the future. Best of all, theBD FACSCalibur system is backed by the leader in flow cytometry, BD Biosciences.To find out more about how the BD FACSCalibur system can contribute to your lab’sproductivity and performance, contact your local BD Biosciences sales representative.
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The BD FACSCalibur System: The Gold Standard That Continues to Evolve The BD FACSCalibur flow cytometer is another BD Biosciences first: a multicolor benchtop flow cytometry system that is capable of both analyzing and sorting. Combined with your choice of sample loader, a wide variety of reagents, and powerful software, the BD FACSCalibur
The BD FACSCalibur flow cytometer is another BD Biosciences first: a multicolor benchtop flow cytometry system that is capable of both . the BD FA C S C alibur flow cytometer is a fully integrated multiparameter system that ensures objective, reproducible results so vital in today's clinical and
Becton-Dickinson FACSCalibur Flow Cytometer Instructions Copyright 2013 University Instrumentation Center UNH W123 Parsons Hall 23 Academic Way, Durham, NH 03824 -). ).
1.2 Flow cytometer and Sorter (FACS) Analysis Summary The Becton-Dickinson (BD) FACSVantage SE flow cytometer sorter can sort/isolate or measure subpopulations of cells from a variety of sources including cells with specific reporters, stem cells, or primary tissues (blood or other human or mouse tissues or tumors). Only RG 1 and 2
1 Start the BD FACSCalibur instrument and then turn on the computer. 2 Set up the cytometer using BD Calibrite beads and BD FACSComp software with 3-color or 4-color lyse/no-wash (LNW) settings. Verify that all parameters pass. NOTE You can use either 3-color or 4-color LNW settings to set up the cytometer.
The BD Accuri C6 flow cytometer offers an entirely new vision for the role of flow cytometry in life science research. Designed from the ground up to be compact and easy to use, the BD Accuri C6 flow cytometer fits in any lab and is readily accessible to all researchers—experts and novices alike. Figure 1-1. The BD Accuri C6 Flow Cytometer .
conventional flow cytometer and on the Attune Acoustic Focusing Cytometer using a 488 nm blue laser (top row) and a 405 nm violet laser excitation (bottom row). The conventional flow cytometer was run at its highest sensitivity setting, at a sample input rate of 15 μL/min (left column).
10 Aurora User's Guide Cytometer Overview The Aurora spectral flow cytometer is an air-cooled , multi-laser, compact benchtop flow cytometer. It is equipped with three lasers and up to 48 detection channels for fluorescence and up to two detection channels for scatter (FSC [forward scatter] and violet laser SSC [side scatter]). Solid-state
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