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BD FACSCanto IIA proven platform for maximumreliability and the highest quality results
A proven platform for maximumreliability and the highest quality resultsBuilt on more than 25 years of BD experience and leadership in flow cytometryand multicolor analysis, the BD FACSCanto II system is an easy-to-use benchtopanalyzer that delivers proven performance, accuracy, and high quality results.The BD FACSCanto II provides the ultimate in flexibility and can be configuredwith two or three lasers to detect up to eight colors.The BD FACSCanto II features many innovations. At the heart of the cytometer,for example, the fluidics system features a fixed alignment flow cell to minimizestartup time and improve reproducibility. The optical system features apatented design that maximizes signal detection and increases sensitivity andresolution for each color in a multicolor assay. These capabilities and many otherinnovations make the BD FACSCanto II one of the most powerful and versatilebenchtop analyzers available for both clinical and research applications.The BD FACSCanto II is designed to address the needs of today’s busy clinical lab.It provides a high degree of automation and quality control to reduce handson time for technicians and improve reliability of results. The newly designedBD FACS Loader features single-tube instrument setup and testing as wellas walkaway sample introduction to help operators rapidly adopt routineclinical applications freeing technican time for more important activities. Tofurther simplify operation, BD FACSCanto clinical software automates setup,compensation analysis, and quality control for predefined clinical applications.For the research lab, the BD FACSCanto II system has the versatility to meetdemanding research requirements. Sample introduction can be accomplishedin a single tube or via the BD High Throughput Sampler (HTS),* a microtiterplate loader well suited for 96- and 384-well plates that can accommodate highthroughput research applications. BD FACSDiva software efficiently controlsthe setup, acquisition, and analysis of flow cytometry data.The BD FACSCanto II is supported by a broadportfolio of reagents. A full complement of highlyqualified BD technical and application supportpersonnel is available to help streamline workand maintain optimal instrument performance.* For Research Use Only.
FLUIDICSSaves time, improves reproducibilityHigh Performance, Innovative SystemThe BD FACSCanto II fluidics system is designedto streamline work, save time, and improveperformance.In the fluidics system, the sample travels up the sampleinjection tube, and hydrodynamic focusing within the flowcell forces particles into a single-file stream where laser lightintercepts the stream at the sample interrogation point. Theunique flow cell design permits particles to flow throughthe center of the flow cell. Increasing the sample pressureincreases the core diameter and the flow rate.A fluidics cart holds large fluid tanks necessary to operateand maintain the instrument. For sample acquisition,positive-pressure pumps in the fluidics cart send sheath fluidpast a 0.2-µm filter to a pressurized interior reservoir insidethe instrument called the plenum. The plenum maintainsa nearly constant fluid level and dampens pump pulsationusing a new dynamic feedback pressure control systemdesigned to regulate pressure. As a result, sheath flow ratedoes not vary with the level of fluid in the sheath cubitainer,and the reservoir automatically removes small air bubblesfrom the sheath supply.Daily routine procedures, such as startup, shutdown, andcleaning routine cycles, are automated as a result of fluidicintegration with BD FACS Canto clinical software orBD FACSDiva software. BD FACS Shutdown solutionprevents salt crystal buildup in fluidics lines and is supplemented with a preservative to prevent bacterial growth.During the instrument shutdown procedure, the BD FACSShutdown solution replaces sheath fluid in all sample andsheath fluid lines.Figure A-1 Hydrodynamic focusing of the sample core through the flow cellHydrodynamic focusingA high flow rate is generally used formeasurements such as immunophenotyping, forwhich data is less resolved but is acquired morequickly. A lower flow rate is generally used inapplications for which optimal resolution andsensitivity are critical.low samplepressure(12 µL/min)sheathfluid4laser beamsamplesheathfluidhigh samplepressure(60 µL/min)sheathfluidlaser beamsamplesheathfluid
Flow cellLED alertsLEDs are located in the front door tomonitor each acquisition parameter. EachLED blinks when the signal level reaches aprescribed threshold.5
OPTICSMaximize multicolor performanceInnovative Optical DesignDelivers Highest SensitivityThe innovative designs for both the excitationand collection optics reduce excitation losses,yielding more information from each sample.The optics of the BD FACSCanto II system consist of anexcitation source with up to three lasers, a blue (488-nm,air-cooled, 20-mW solid state), a red (633-nm, 17-mW HeNe)and a violet (405-nm, 30-mW solid state). Laser excitationoptics illuminate cells in the sample and collection opticsdirect light scatter and fluorescence signals through spectralfilters to the detectors.ExcitationThe excitation optics consist of multiple fixed wavelengthlasers, fiber optics up to the beam-shaping prisms, andachromatic focusing lenses that produce spatially separatedbeam spots in the flow cell. Each lens focuses the laser lightinto the gel-coupled cuvette flow cell. Since the opticalpathway and the sample core stream are fixed, alignment isfixed from day to day and from experiment to experimentwith no need for user intervention.EmissionThe emission signals are transmitted from the flow cell tothe detector arrays, an octagon for the blue and a trigoneach for the red and the violet laser signals. The octagoncontains five PMTs and detects light from the 488-nm bluelaser. A PMT in the octagon collects side scatter signals. Thetrigons contain two PMTs each and detect light from the633-nm red and the 405-nm violet lasers.-Cy7APC/60780PerCP-Cy5.5, PerCPAPC660670 LP/20735 LPPE585/655 LP42556LPLP300/353P0L61C502735 LPFITSSC488/10Collection OpticsThe octagon and trigon are BD patented detector arrays that use serial lightreflections to guide signals to their target detectors, resulting in highly efficientlight collection and providing maximum signal retention at the detector level.P685 L780/60PE-Cy7This BD serial reflective design further enhances instrument sensitivity bycollecting the dimmest emission signals first, moving from the longestwavelengths (typically PE-Cy7 ) to the shortest (FITC).6
Excitation optics focus the laser beamsEmission spectra of commonly used fluorochromesInstrumentLaserExcitation LaserLine (nm)FluorescenceChannelFluorochromes provided by BD BiosciencesThe BD FACSCanto II system is designed for thesefluorophore combinations.BD FACSCanto IIflow cytometry systemSolid State (L1)488GreenYellowOrangeRedInfraredRedFar RedInfraredGreenBlueFITCPEPE-Texas Red †PerCPPE-Cy7APCAlexa Fluor 700†APC-Cy7AmCyan†BD Horizon V450HeNe (L2)633Violet (L3)†405Alexa Fluor 488PerCP-Cy5.5Alexa Fluor 647BD APC-H7Pacific Blue †7
Productivity supportOptions and Related Tools to SupportLab RequirementsSample introduction productivity can begained with the optional BD FACS Loaderor the BD High Throughput Sampler (HTS).BD High Throughput Sampler (HTS)*Designed for research use, the optional BD HighThroughput Sampler (HTS) provides fully automated andrapid sample acquisition that can speed through a microtiter plate in less than 15 minutes, with less than 1% samplecarryover. In high-throughput mode, fast acquisition speedis achieved by synchronizing two high-precision pumpsfor sample mixing, sample injection, and probe washing.Standard-throughput mode can be selected for acquisitionof larger sample volumes. The HTS supports standard 96 and384-well plates. It is operated using BD FACSDiva software,allowing users to define customized delivery protocols,mixing, wash, and analysis parameters.* For Research Use Only.Barcode ReaderThe optional BD FACSCanto hand-held barcodereader reads the two-dimensional barcode labelsand it is used to read the compensation matrixof the BD 7-color setup beads reagent intoBD FACSCanto clinical software.8
OPTIONSBD FACS LoaderThe BD FACS Loader is an instrument option that allowswalkaway sample introduction to further improveproductivity. The BD FACS Loader carousel accommodatesup to 40 12x75-mm tubes and automatically loads them onthe BD FACSCanto II system without operator intervention.Mounted directly on the cytometer, the device includes adrive system, a tube lifter mechanism, and sensors on thesliding drawer. Two sliding doors safely enclose the drawerto protect technicians from moving parts during operation.The BD FACS Loader utilizes compressed air to allow amore reliable tube load as well as an intelligent tube guidemechanism that automatically sends an alert if a tube is notproperly positioned for loading.A unique ID and optically read label are printed on eachcarousel for easy carousel identification. The BD FACS Loaderis operated through both BD FACSCanto clinical andBD FACSDiva software. The BD FACS Loader is compatiblewith the BD FACS Sample Prep Assistant (SPA) and theBD FACS Lyse Wash (LWA) Assistant.Sample Preparation SupportBD Biosciences offers two BD FACSCantocomplementary instruments to help busylaboratories increase speed, reduce variability,and streamline sample preparation.The BD FACS Sample Preparation Assistant supportswalkaway lyse-no-wash sample automationprocessing. The BD FACS Lyse/Wash Assistant (LWA)lyses, mixes, washes, and resuspends samples inPBS. Pre-programmed as well as custom protocolsare available.9
CLINICALSample prep, setup, and analysisA Turnkey System for Clinical UseDesigned to address the needs of today’sbusy clinical lab, the BD FACSCanto II has aproven track record of reducing hands-on timeand improving reliability of results. A highdegree of automation and quality controlhelps save time, reduce cost, and improvereproducibility of results.Cytometer settings are tracked over time by Levey-Jenningsgraphs to monitor cytometer performance. Adjustmentsare made automatically to ensure consistent data integrityand reproducible results from day to day and experiment toexperiment. Integrated quality control features furthersupport data integrity by notifying operators if an assayfails predefined standards.BD FACSCanto clinical software includes specific applicationmodules optimized for use with specific IVD reagent kits.The modules feature automated gating, calculations, andreport generation to deliver a consistent, reproducible, andstandardized analysis.As a leading provider of tools for flow cytometry,BD Biosciences is committed to offering comprehensive,innovative software solutions, including third-partysolutions that support application specific customerrequirements.For example, optional Laboratory Information System(LIS) software easily connects the BD FACSCanto II withan existing customer Laboratory Information System toenable direct, bi-directional transfer of data. The softwaresimplifies laboratory workflow by customizing data reporting,securing validation, and transferring data to reduce manualtranscription. The solution automates the process fromrequest to reporting to help reduce errors, improve dataquality and laboratory productivity.CD3/CD16 56/CD45/CD4/CD19/CD8 TruCTotal Events: 10068ParameterLymph EventsBead EventsCD3 CD3 CD8 CD3 CD4 CD3 CD4 CD8 CD16 CD56 CD19 CD45 4/8 Ratio10CD19 APC-ACD3 54.683.64CD16856CD19 CD16856 CCD19 CD19 APC-ACD3-CD4-CD8CD45 PerCP-Cy5.5-A000633C028.001.fcsCD4 CD8 CD4 PE-Cy7-ABeadsSSC-ASSC-ASSC-ACD4 CD8-CD4-CD8 CD8 APC-Cy7-ACD16856-CD19CD16856 CD19-CD16 56 PE-AReagent Lot ID: 39825Monitoring HIV—Identification and enumerationof lymphocyte subsetsThe BD Multitest 6-color TBNK kit is the firstand only 6-color In Vitro Diagnostic applicationto provide a complete immune panel in a singletube, saving valuable time and resources forsample processing.
APPLICATIONSCountTotal Events: 17083SSC-AThe BD HLA-B27 module for the BD FACSCantoII system provides a complete system for rapiddetection of HLA-B27 antigen, which is clinicallyrelevant to the evaluation of ction of HLA-B27 AntigenFSC-HCD3 PE-AAnti-HLA-B27 FITC-AReagent Lot ID: 37681:145308001.001.fcsGated EventsPreset HLA-B27 MarkerSample HLA-B27 MedianConclusion2015145166HLA-B27 positive sample11
Versatile, easy-to-useA Powerful Platform for Research UseThe BD FACSCanto II system brings BD flowcytometry and multicolor analysis to a new levelto help researchers meet complex challengesand advance their research.BD FACSDiva Software for Setup, Acquisition, and AnalysisBD FACSDiva software efficiently controls the setup,acquisition, and analysis of flow cytometry data from theBD FACSCanto II workstation. BD FACSDiva operatingsoftware is common across many BD cell analyzers and cellsorters, including BD LSR and BD FACSAria systems, givingresearchers application flexibility.BD FACSDiva software enables researchers to preview andrecord data from multiple samples with an automatedacquisition process. Acquisition templates, user-definableexperiment layouts, and simple compensation procedures alsomanaged by the software further facilitate acquisition.For analysis, the software includes powerful features suchas hierarchical snap-to gating, a variety of plots, and batchanalysis. Recorded data can be analyzed by creating plots,gates, population hierarchies, and statistical views on aBD FACSDiva global worksheet. The global worksheet canbe used to analyze multiple sample tubes from the sameexperiment, to save time. User-definable batch analysisand automated capabilities such as gate resizing, pausingbetween data files, exporting statistics, and printing beforeproceeding to the next data file are additional BD FACSDivafeatures.Lysed whole human blood12104CD45 AmCyan-A105-5030104105-959103CD2 PE-Cy7-A-503103104CD2 PE-Cy7-A105105105104CD8 APC-A10410310300CD41 PerCP-Cy5-5-A02-106 0 10103104CD3 Pacific Blue-A105-236103-6930103103CD56 PE-A104104CD57 FITC-A102-263050M 1100105105SSC-A (x 1,000)100 150 200 250Eight-color immunophenotyping panel using FITC,PE, PerCP-Cy 5.5, PE-Cy 7, APC, APC-Cy7, AmCyan,and Pacific Blue fluorophores.-5030103104CD4 APC-Cy7-A105
RESEARCH APPLICATIONS105250104-5000103CD2 PE-Cy7-A15050100SSC-A15050100SSC-APanel #15-45/3/57/16/4/2/56/8200(x 1,000)250Panel #15-45/3/57/16/4/2/56/8200(x 1,000)Panel #15-45/3/57/16/4/2/56/8150200250102(x 1,000)FSC-A105103104105CD4 PerCP-Cy5.5-A103105105105-6850103104105CD3 AmCyan-ABD FACSDiva Worksheet104Lysed whole human blood sample in an 8-colorpanel using V450, AmCyan, FITC, PE, PerCP-Cy5.5,PE-Cy7, APC and APC-H7 fluorophores.-294-1970103CD56 APC-A104104CD2 PE-Cy7-APanel #15-45/3/57/16/4/2/56/810310400Panel #15-45/3/57/16/4/2/56/80 102103CD3 AmCyan-A-647-500CD3 AmCyan-A01051051040-1,370-2630105-685-685Panel #15-45/3/57/16/4/2/56/8103CD57 FITC-A1041030CD8 APC-H7-A104Panel #15-45/3/57/16/4/2/56/8105Panel #15-45/3/57/16/4/2/56/8CD16 PE-A103CD45 V450-A104100103500-685103104105CD3 AmCyan-A0-685103104105CD3 AmCyan-ACytometer Setup and TrackingBD Cytometer Setup and Tracking (CS&T) beadsprovide, in a single vial reagent format, automateddetermination of BD FACSCanto II configurationbaselines and monitoring of performance over time,using BD FACSDiva software.BD CompBeads Compensation Particle Sets aredesigned to optimize compensation settings inmulticolor flow cytometry. Particularly helpfulwhen using tandem dye conjugates, these particlesets simplify establishing compensation correctionsfor spectral overlap for any combination offluorochrome-labeled antibodies.13
SERVICETechnical expertise from BDCommitted to Customer SuccessBD Biosciences is fully committed to the successand satisfaction of its customers. Supportingflow cytometry applications for over 35 years,BD training, support, and field service teams arededicated to helping customers achieve optimalinstrument performance, ease of use, andstreamlined workflow. With unmatched flowcytometry experience, this world-class serviceorganization is available to help with yourBD FACSCanto II product installation, futureupgrades, and application support.TrainingHands-on training is included with each BD FACSCanto IIproduct. Training courses are held at BD training centersworldwide. BD flow cytometry training courses combinetheory and practice to provide participants with the skillsand experience they need to take full advantage of thecapabilities of their BD FACSCanto II system.Technical Application SupportBD Biosciences technical applications support specialists areavailable to provide field- or phone-based assistance andadvice. Expert in all aspects of flow cytometry, BD technicalapplication specialists are well equipped to address customerneeds in both instruments and applications support.Field ServiceWhen instrument installation or service is required, aBD Biosciences Technical Field Service Engineer can bedispatched to the customer site. BD Biosciences field serviceengineers are located across the world. On-site service andmaintenance agreements are available to provide long-termsupport for BD FACSCanto II systems.14
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Built on more than 25 years of BD experience and leadership in flow cytometry and multicolor analysis, the BD FACSCanto II system is an easy-to-use benchtop analyzer that delivers proven performance, accuracy, and high quality results. . At the heart of the cytometer, for example, the fluidics system features a fixed alignment flow cell to .
