BD Biosciences FACSMelody Cell Sorter System 3 Lasers: 4Blue/2Red/3Violet Flow Cytometer

Recent PM from BD; Certified and Ready for a Service Contract! Includes:

• BD FACS Melody 3 lasers (4B, 2R, 3V)
• Plate Sorting module
• Computer loaded with FACSChorus Software
• Baker SterilGard Bio Safety Cabinet BD400XD Optional

Technical Specifications:

Optics

• Excitation laser
• 488-nm blue direct diode laser, nominal power: 16 mW
• 640-nm red direct diode laser, nominal power: 36 mW
• 405-nm violet direct diode laser, nominal power: 36 mW
• Beam size
• 9± 3 x 67 ±5 µm
• Laser beam alignment
• Fixed and spatially separated alignment of all lasers with the cuvette flow cell
• Optical coupling
• The quartz cuvette flow cell is gel-coupled by refractive index-matching optical gel to the fluorescence objective lens for optimal light collection efficiency. Numerical aperture: 1.2.
• Detection channels
• Forward scatter (FSC), side scatter (SSC) and up to nine fluorescence. See the BD FACSMelody Filter Guide for laser and detection configurations and optical filter specifications.

Fluidics

• Sample input
• 5.0-mL polystyrene or polypropylene tubes can be used.
• Temperature control: adjustable through software: 4°C, 22°C, 37°C and 42°C or off
• Sample agitation: adjustable through the software to keep the sample constantly suspended
• Sample flow automatically stops when the sample input tube is empty.
• Flow cell
• Quartz cuvette
• Nozzle
• 100-µm nozzle is removable and can be sonicated.
• A registered key-fit position at the bottom of the cuvette provides fixed stream alignment.
• Fluidic tanks
• Autoclavable 10-L stainless steel sheath container
• 10-L polypropylene waste container

Performance

• System operation
• Time from system powerup to running sample, including powerup, fluidic startup, stream optimization and optional performance checks (instrument and drop delay QC): <20 minutes; excluding optional performance checks: <10 minutes Sorting nozzle tip can be removed during operation, replaced and stream optimized: <3 minutes
• Fluorescence sensitivity
• Fluorescence sensitivity was measured using SPHERO™
• Rainbow Calibration Particles according to the manufacturer’s specifications:
• FITC: 80 molecules of equivalent soluble fluorochrome (MESF-FITC)
• PE: 30 molecules of equivalent soluble fluorochrome (MESF-PE)
• Fluorescence detection efficiency
• Qr is the relative fluorescence detection efficiency, used for describing the detection efficiency of a detector for a specific fluorochrome. The units for Qr are photoelectrons generated per assigned BD unit (ABD unit). One ABD unit, for a given fluorochrome, is defined as the fluorescence of one antibody (fluorochrome to protein 1:1) bound to a cell. The higher the Qr value, the better the relative fluorescence detection efficiency per molecule reference fluorochrome measured in that detector.
• FITC Qr (x1000): 40 photoelectrons/ABD
• PE Qr (x1000): 325 photoelectrons/ABD
• CD4 FITC: 1,600 photoelectrons*
• CD4 PE: 13,000 photoelectrons*
• Fluorescence resolution
• Full-peak coefficient of variation (FPCV): <3.0%, G0/G1 peak for propidium iodide (PI)-stained chicken erythrocyte nuclei (CEN)
• Fluorescence linearity
• Doublet/singlet ratio: PI-stained CEN: 1.95–2.05
• Forward and side scatter sensitivity
• Sensitivity enables separation of 0.5-µm beads from noise.
• Forward and side scatter resolution
• Scatter performance is optimized for resolving lymphocytes, monocytes and granulocytes.

Sort performance

• Droplet sorting
• 34,000 drops per second
• Automated setup, optimization and monitoring of droplet breakoff and sort streams
• Automated drop-delay determination with BD FACS™ Accudrop beads
• Automated clog detection and sort tube protection system using
• Sweet Spot technology
• Purity and yield
• At 23 psi and 34 kHz with an average of 10,000 events per second, a two-way sort of 5% target populations achieved a purity of 98% and a yield of >80% of Poisson’s expected yield for both populations. No limitation is imposed on the event rate. However, yield decreases with higher event rates based on Poisson distribution principles.
• Sort collection
• Two-way sorting: 1.5-, 2.0- and 5.0-mL tubes
• One-way sorting: 6-, 24-, 48-, 96- and 384-well plates, 96-well
• PCR tray, microscope slide (optional)
• Temperature control: water recirculation unit to provide heating or cooling for collection into tube holders, multiwell plates and slides (optional)

Signal processing

• Pulse measurement
• Height, area, width
• Acquisition rate
• The maximum throughput rate is 40,000 events per second, independent of the number of parameters.
• Time
• Time can be correlated to any parameter for kinetic experiments or other applications.
• Channel threshold
• Available for any single parameter from any lase

The simple solution for consistent, quality results Truly easy cell sorting is finally available to your lab. Save time with simplified operation and get quality results. The BD FACSMelody™ Cell Sorter with newly added 4-way cell sorting makes the complex world of flow cytometry and sorting accessible to more researchers, enabling deep scientific insights, lab efficiency and cost savings. The BD FACSMelody Cell Sorter combines proven and exclusive BD technology of high-end sorters with automation and simplified software that takes easy-to-use sorting to a new level.Simplify the workflow with smart automationWith BD FACSChorus Software, researchers are guided throughout the entire cell sorting process using advanced automation technology. Designed to eliminate manual steps, this smart automation simplifies the workflow, making cell sorting easy to learn, and the BD FACSMelody Sorter easy to use.With an intuitive user interface, BD FACSChorus Software enables researchers at all skill levels to achieve high-purity sorting 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 sort controls, monitor sort progress and turn off sort controls when sorting is completed
• Simple to read reports
• Automation makes sorting simple and saves time

Minimal training required for effective operation Learning to operate the BD FACSMelody cell sorter is easy, regardless of a user’s skill level or experience. To aid in producing consistent results, BD FACSChorus software prompts novice users with step-by-step instructions. It eliminates the need for setting up the sort streams and 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 amplitude are optimized.
• The sort streams are focused and steered into tubes or plates.

Streamlined workflow

• Instrument settings, gates and sort settings are available and retrievable from saved experiments.
• Compensation is conveniently computed automatically from the results of compensation controls.
• Compensation controls can be labeled capture beads, cells, BD™ FC Beads, or a combination of types.
• Compensation is automatically recalculated when detector settings are adjusted, simplifying operation and saving time.

System anticipates potential issues and identifies root causes

• Filter/mirror units communicate with the software to confirm the optical configuration supports your experiment.
• Stream stability is monitored to detect and correct stream aberrations. If the stream cannot be corrected, sorting will pause and the operator is notified.
• When a sample tube is empty, the sample flow is stopped before 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.

Designed to empower scientific discovery Simplified operation does not mean reduced performance. The BD FACSMelody System features excellent sensitivity for accurate resolution of low density cell markers and high throughput for collecting rare cells. With a BD-pioneered optical system, the BD FACSMelody Cell Sorter maximizes signal generation, collection and detection through lasers with fixed alignment and independent focusing.

Detect and sort dim cells and rare cells

• The patented gel-coupled cuvette with fixed alignment ensures that lasers are precisely focused on the sample stream to generate the greatest signal for optimizing the resolution of dim cells and adjacent populations.
• Spatially separated laser beams ensure minimal optical background noise and improved sensitivity from each laser for resolving dim populations.
• The sorter can process many cells quickly to improve the isolation of rare cells.

Optimized system of reagents, software and hardware

• Instrument configurations are optimized for bright BD Horizon Brilliant and BD OptiBuild Reagents.
• Laser and color configurations facilitate multicolor experiments.
• Sweet Spot technology maintains sort stream stability so yield and purity are maintained.

Reproducible results

• Using BD CS&T Technology for daily QC ensures consistent performance across all detectors and ensures day-to-day and experiment-to-experiment reproducibility.
• Run once every 60 days, spectrally matched BD™ FC Beads determine spillover values for various standard fluorochromes. The values remain independent of experiment settings and are automatically updated and maintained after QC runs.
• Run any new fluorochrome as a single-color control to add its spillover properties to the software’s list of reagents. The fluorochrome’s spillover can be calculated when setting up an experiment even if different detector gains are used.