Cell Sorting

Cell Sorting Guidelines

Choosing a Sorter

Sony MA900 – 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! Selfrun Training Available
  

Beckman Coulter Astrios EQ – 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
  

Beckman Coulter MoFlo XDP-Flex – 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

Beckman Coulter MoFlo XDP100 – 3 Lasers (405nm, 488nm, 561nm), 10 Parameters (No Red Laser as of 02.13.23)

• 100 um Nozzle Tip
• Cell Lines, Large and Clumpy Cells 
• High-Speed; Max Event Rate = 12,000 events/second (40 million events/hour)
• 4-Way Tube Sorting, Plate Sorting
  
  

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.