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BD FACSMelody Cell Sorter The simple solution for consistent, quality results

The right instrumentis needed for thebest resultsSimplicity, affordability andquality are keyCell sorting is fast becoming an essential step on the path to deeper understandingand new discoveries in the life sciences. Although cell sorting often is performed oninstruments owned by core labs, there are issues associated with transportingsamples and timely access.A readily available on-site sorter that provides consistent, reliable results couldbenefit researchers in many fields of investigation. And, researchers need a solutionthat is easy to learn, use and maintain. That solution is here.2

The BD FACSMelody cell sorter makes sorting easyTruly easy cell sorting is finally available to your lab. Now you can save time with simplified operation and get qualityreproducible results. The BD FACSMelody cell sorter makes the complex world of flow cytometry and sorting accessible to moreresearchers, enabling deep scientific insights, lab efficiency and cost savings.Incorporating technology exclusive to BD, the BD FACSMelody cell sorter is based on the proven BD FACSAria platform of cellsorters. With fixed alignment and gel-coupled cuvettes, BD FACSAria sorters have a reputation for driving superior performanceand reliability.Easy to use: Automation of complex tasks and guidedprocedures facilitate consistent results, even when skill levelsvary among users, to enable deep scientific insights for moreresearchers.Lab efficiency and productivity: Save time and money withan affordable cell sorting solution based on a streamlinedworkflow to improve throughput, minimize downtime andpromote experimental success.High performance: A complete system of reagents, softwareand hardware works together to detect and sort low densitycell markers and rare cells. The system delivers reproducibleresults while protecting the operator and the sample, andenables sorting into tubes or a range of standard plates.Trusted brand: Rely on highly responsive field service supportand expertise in flow cytometry instrumentation andapplications from BD, backed by more than 40 years ofexperience and innovation in flow cytometry and thousandsof peer-reviewed publications citing BD flow cytometryinstruments, software and reagents.3

Easy to learn and useSimplify the workflow with smart automationWith BD FACSChorus software, researchers are guided throughout the entire cellsorting process using advanced automation technology. Designed to eliminatemanual steps, this smart automation simplifies the workflow, making cell sortingeasy to learn, and the BD FACSMelody sorter easy to use.With an intuitive user interface, BD FACSChorus softwareenables researchers at all skill levels to achieve high-puritysorting with: Screens that are dedicated to each single step On-screen instructions and tips that display when needed Preset data tools to save the user clicks and time Instrument management behind the scenes to activate sortcontrols, monitor sort progress and turn off sort controlswhen sorting is completed Simple to read reportsAutomation makes sorting simple and saves time100%55%Typical sorter Power up system Start up fluidics Wait for fluidics to stabilize Select nozzle size and appropriatesheath pressure Adjust drop drive amplitude andfrequency for optimaldroplet breakoff Focus side streams Calculate drop delay Turn on stream monitoring Aim sort streams Run sample Define sort criteriareductionin stepsBD FACSMelody cell sorter Power up system Run fluidic startup Confirm drop delay Run sample Define sort criteria0%Sort setup protocol4

Minimal training required for effective operationLearning to operate the BD FACSMelody cell sorter is easy, regardless of a user’s skill level orexperience. To aid in producing consistent results, BD FACSChorus software prompts noviceusers with step-by-step instructions. It eliminates the need for setting up the sort streamsand monitoring the sort, reducing the risk of poor sort outcomes.Automation of routine tasks Typically the system is ready in less than 17 minutes,maximizing uptime. Fluidic startup automatically debubbles the stream. Sheath pressure, drop drive frequency and amplitudeare optimized. The sort streams are focused and steered into tubes or plates.Streamlined workflow Instrument settings, gates and sort settings are available andretrievable from saved experiments. C ompensation is conveniently computed automatically fromthe results of compensation controls.System anticipates potential issues andidentifies root causes Filter/mirror units communicate with the software to confirmthe optical configuration supports your experiment. S tream stability is monitored to detect and correct streamaberrations. If the stream cannot be corrected, sorting willpause and the operator is notified. W hen a sample tube is empty, the sample flow is stoppedbefore bubbles can be introduced into the nozzle. Alerts remind the operator to perform maintenance. Sorting is stopped when the target number of cells is reached,inhibiting overflowing of collection tubes. Compensation controls can be labeled capture beads, cells,BD FC Beads, or a combination of types. Compensation is automatically recalculated when detectorsettings are adjusted, simplifying operation and saving time.5

Lab efficiency and productivitySave time with simplified operation and maintenanceThe BD FACSMelody cell sorter allows researchers to work moreefficiently and improves throughput. A streamlined workfloweliminates manual setup of the stream and monitoring of thesort, which minimizes hands-on time and provides walk-awaysorting capability.Fast results to improve lab throughput Fixed alignment means there is no lengthy procedure withalignment beads required at startup. Only perform instrument QC when needed. It is not requiredeach time the sorting nozzle is installed. Even with instrumentQC, you can typically run your sample in less than 17 minutescompared to less than 8 minutes without QC. Automated stream setup and steering into tubes or platessaves valuable minutes.Minimal downtime Instrument QC continuously monitors the system to trackperformance and predict problems such as laser failuresbefore they occur. The sorter design facilitates fast troubleshooting, such aseasy and quick replacement of the sorting nozzle, with noneed to re-align the laser and stream or run QC beads. Youcan resume sorting within 3 minutes because the nozzle isseparate from the fixed-alignment flow cell, which is wherethe sample is interrogated.Cost savings The BD FACSMelody cell sorter features a fixed flow cell andreusable nozzle, eliminating the need to purchase dailyconsumables. This affordable solution allows for in-house cell sorting, whichcan save money compared to outsourcing.6Quick resultsTypically run your sample in less than 17 minutes.1. System powerup3 minutes2. Laser and stream alignment0 minutes3. Fluidics startup3 minutes4. Stream optimization2 minutes5. Daily performance checks with beads9 minutes6. Experiment settings andcompensation normalizedfor consistent results0 minutes17

250x103x10320010398.8%GFP BrightGFP 50GFP Bright200250x103250x103200GFP 02103GFP-A103104105GFP-A104105Transfected GFP cell sortingHEK-293 cells (ATCC CRL-1573 ) were transfected over 24 hours with AcGFP. Untransfected cells were usedas a negative control to set the instrument and draw the gates (not shown). Two populations of GFP cells,dim and bright, could be discriminated. GFP bright cells were identified and sorted at 1,000 total events/second in purity mode. Post-sort analysis revealed a purity of 98.8%.7

High performanceDesigned to empower scientific discoverySimplified operation does not mean reduced performance. The BD FACSMelodysystem features excellent sensitivity for accurate resolution of low density cellmarkers and high throughput for collecting rare cells.With a BD-pioneered optical system, the BD FACSMelody cell sorter maximizessignal generation, collection and detection through lasers with fixed alignment andindependent focusing.Detect and sort dim cells and rare cells The patented gel-coupled cuvette with fixed alignmentensures that lasers are precisely focused on the sample streamto generate the greatest signal for optimizing the resolutionof dim cells and adjacent populations.Optimized system of reagents, software and hardware Instrument configurations are optimized for brightBD Horizon Brilliant and BD OptiBuild reagents. Spatially separated laser beams ensure minimal opticalbackground noise and improved sensitivity from each laser forresolving dim populations. Sweet Spot technology maintains sort stream stability so yieldand purity are maintained. The sorter can process many cells quickly to improve theisolation of rare cells.8 Laser and color configurations facilitate multicolorexperiments.

Reproducible results Using BD CS&T technology for daily QC ensures consistentperformance across all detectors and ensures day-to-day andexperiment-to-experiment reproducibility. Run once every 60 days, spectrally matched BD FC Beadsdetermine spillover values for various standard fluorochromes.The values remain independent of experiment settings andare automatically updated and maintained after QC runs.Operator and sample protection A custom Class II biological safety cabinet, verified to meetpersonnel and product protection standards with theinstrument installed in the work area, is available. It includes abuilt-in aerosol management system that evacuates aerosolsand operates independently of the cabinet for an extra levelof protection. Run any new fluorochrome as a single-color control to add itsspillover properties to the software’s list of reagents. Thefluorochrome’s spillover can be calculated when setting up anexperiment even if different detector gains are used.1031004CD4 FITC-A102 1051031031004CD25 BV421-A102105103104CD25 521030104Tregs3.7%99.7%CD127 Alexa Fluor 647-A105CD127 Alexa Fluor 647-A105104103102CD127 Alexa Fluor 647-A1010520105104104 0CD4 FITC-ATregs3.7%Tregs post-sort-102102Tregs post-sort-1020Tregs pre-sort-102005050100CD4 103SSC-ASSC-A100CD4 102CD127 Alexa Fluor 647-ATregs pre-sort0250200CD4 pre-sort150150250x103-102CD4 pre-sort200x1031004CD25 BV421-A102105103104CD25 BV421-A105Regulatory T-cell (Treg) sorting using a panel with minimal spectral overlapPeripheral blood mononuclear cells (PBMCs) were isolated from human whole blood and stained with an antibodycocktail for the identification and sorting of Tregs. Lymphocytes were first gated based on light scatter, and followingdoublet discrimination (not shown), helper T cells were defined based on CD4 expression. From the CD4 gate, Tregswere then identified as CD25 CD127dim/- cells and sorted at 3,500 total events/second in purity mode. Post-sortanalysis revealed a cell purity of 99.7%. With each fluorochrome excited by a different laser and cross-laser excitationaccounted for, spectral overlap between channels was minimal ( 1%).9

Trusted partnerRely on an industry leader for experimental successResearchers expect the best when it comes to BD, a leader in flow cytometrysorting, and the BD FACSMelody cell sorter hits the mark. With more than 40 years of experience and innovation in flow cytometry,BD continues to develop and improve instrumentation, software andreagent solutions. Tens of thousands of peer-reviewed publications cite BD flow cytometryinstruments, software and reagents. BD offers world-class technical application support and highly responsive fieldservice support. The BD FACSMelody cell sorter was developed from proven BD FACS technology.10