Cell Sorting

Cell Sorting Guidelines

Choosing a Sorter

The FCSR operates five cell sorters that provide a wealth of opportunities for cell sorting, including both staff-operated and self-run cell sorters.

1. BD FACSDiscover™ S8 (Becton Dickson)

• Spectral- & Imaging-based Cell Sorter (Full Spectral) with 78 detectors
• Optical and Electronic Imaging-Like Analysis (No Camera)
• Available for either Staff-assisted or Trained, self-run operation
• 5 Lasers (349nm, 405nm, 488nm, 561nm, 637nm)
• 85 um, 100 um or 130 um Nozzle Tip
• Max Event Rate =
  • 11,500 events/second (85um Nozzle),
  • 6,800 events/second (100 Nozzle),
  • 3,000 events/second (130um Nozzle)
• Tube & Plate Sorting
  • 6-Way Tube Sorting (85um Nozzle)
  • 4-Way Tube Sorting (100um Nozzle)
  • 2-Way Tube Sorting (130um Nozzle)
  • Plate Sorting with Indexing (with 100um and 130um Nozzle)
    
• Biosafety Cabinet

2. MA900 (Sony)

• 4 Lasers (405nm, 488nm, 561nm, 638nm), 12 Parameters
• 70um, 100um, or 130um Sorting Chip
• All Cell Types including Fragile Cells
• Lower Pressure
• Max Event Rate = 12,000 events/second (70um Chip) and 4000 events/second (100um Chip)
• 4-Way Tube Sorting, Plate Sorting with Indexing
• User-Friendly! Self-Sort Training Available
  

3. Astrios EQ (Beckman Coulter)

• 5 Lasers (405nm, 488nm, 532nm, 561nm, 642nm), 20 Parameters
• 70 um Nozzle Tip
• Primary Human Cells, Primary Mouse Cells, and Small Particles
• High-Speed; Max Event Rate = 25,000 events/second (90 million events/hour)
• Biosafety Cabinet
• 6-Way Tube Sorting, Plate Sorting with Indexing

4. MoFlo XDP-Flex (Beckman Coulter)

• 5 Lasers (355nm, 405nm, 488nm, 552nm, 641nm), 14 Parameters
• 100 um or 70 um Nozzle Tip
  • 100 Tip: Cell lines, Large and Clumpy cells
  • 70 Tip: Primary Mouse cells
• High-Speed; Max Event Rate:
  • 100 tip: 12,000 events/second (40 million events/hour)
  • 70 tip: 25,000 events/second (90 million events/hour)
• 4-Way Tube Sorting, Plate Sorting
  

5. MACSQuant Tyto (Miltenyi Biotec)

• 3 Lasers (405nm, 488nm, 638nm), 8 Parameters
• Valve-Based Cell Sorting Using Self-Contained Cartridge
  • NOT droplet based, no aerosols, no nozzle or chip
• Two Cartridge Options:
  • Regular cartridge:
    • Flow Rate = 4 ml/hour
    • Max Sort Rate = 5000 sorts/second (Max Event Rate = 20,000 events/second)
    • Max Sample Volume = 10 ml
    • Extremely Low Pressure  3-8 psi
  • High-speed cartridge:
    • Flow Rate = 8 ml/ hour
    • Max Sort Rate = 5000 sorts/second (Max Event Rate = 40,000 events/second)
    • Lower pressure  8-18 psi
• 1-Way Sorting with Retention of Unsorted Fraction
• User-Friendly! Self-Sort Training Available

6. MoFlo XDP100 (Beckman Coulter) RETIRED – May 2024

• The XDP100 workload has been shifted to the XDP Flex with the 100 nozzle tip option.
• Please note the XDP100 used a 561nm laser and XDP Flex uses a 552nm laser instead

Optimizing Samples

• ​Sorting Guidelines and Tips
• Add a viability dye to your stain set to eliminate dead cells.
• Use polypropylene test tubes for both your sample and collection tubes for maximum sort yield.
• Cells should be in a single cell suspension. Cell aggregates are the primary cause of poor sort results. We use a doublet discrimination gate to exclude doublets from the sort gate, but it is not 100% effective which leads to poor purity. Aggregates also increase the number of software aborts and may cause instrument clogs, which take to time to clear and create hazardous aerosols.

Biosafety

Regulatory standards require that cell sorting laboratories comply with BSL2 requirements at a minimum. As cell sorting is known to create aerosols, many organisms which can normally be safely handled at the BSL2 level require BSL3 precautions due to the creation of aerosols. The Flow Cytometry Shared Resource facility is able to comply with these standards only at the level of BSL2 with enhanced precautions. The shared resource will not sort samples which require BSL3 containment.