Instrument Description

Instruments

BD FACSAria Fusion Cell Sorters

Sterile Biosafety Level II Sorting within HEPA filtered safety cabinet

PC: 64-bit Windows 10, BD FACSDiva software ver. 10

 

4-Laser “4L” BD FACSAria Fusion Cell Sorter (16 fluorescent channels)

  • 405nm – 85mW – “Violet laser”
  • 488nm – 50mW – “Blue laser”
  • 561nm – 50mW – “Yellow/Green laser”
  • 640nm – 100mW – “Red laser”

5-Laser “5L” BD FACSAria Fusion Cell Sorter (18 fluorescent channels)

  • 355nm – 15mW – “UV laser”
  • 405nm – 100mW – “Violet laser”
  • 488nm – 50mW – “Blue laser”
  • 561nm – 50mW – “Yellow/Green laser”
  • 640nm – 100mW – “Red laser”

*IMPORTANT: Please note differences between base configuration filter sets of each instrument at the bottom of the page. In some cases, panels will NOT be able to be used on both instruments!

Nozzle & Configuration

Each nozzle, paired with a specific frequency, generates droplets of varying sizes with contain cells of interest that the instrument can isolate and sort out. Please select the nozzle that best suits your cell type and size while also taking into consideration the number of events each configuration can process through per hour. Note: an event is anything that triggers a response on the cytometer, whether it’s a cell, dead cell, RBC, debris, etc. Efficiency of sorting is based on the flow rate (events/second processed by the instrument) in relation to the sorted population’s overall percentage within the sample. A higher percentage of population of interest within the sample = higher efficiency sort.

We offer three different nozzle sizes:

70 µm Nozzle

  • 70 PSI
  • ~40 million events per hour
  • Useful for most cell types
  • 1 million sorted cells = ~1 mL of recovery volume

85 µm Nozzle

  • 45 PSI
  • 30-40 million events per hour
  • Useful for most cell types that require a gentler stream pressure but without losing processing speed
  • 1 million sorted cells = ~2 mL of recovery volume

100 µm Nozzle

  • 20 PSI
  • ~10-20 million events per hour
  • Best for large, fragile, or adherent cells
  • Default config for plate sorting & 10X Single Cell sorting
  • 1 million sorted cells = ~3-4 mL of recovery volume

Sort Collection

Sorted cells can be collected into:​

  • 0.5 – 2 mL Eppendorf Tubes
  • 5 mL FACS Tubes
  • 15 mL Conical Tubes
  • 6, 12, 48, 96, or 384 Well Plates (Culture or PCR)

Please make sure to have buffer or media in your collection tubes, and choose collection tubes based on the number of cells you expect to get back per sample and the nozzle you are using:

70 µm Nozzle

  • < 500,000 cells – Eppendorf tube with 500 uL of buffer/media
  • 500,000 – 3,000,000 cells – 5 mL FACS tube with 1 mL buffer/media
  • > 3,000,000 cells – 15 mL tube with 3 mL buffer/media

85 µm Nozzle

  • < 350,000 cells – Eppendorf tube with 500 uL of buffer/media
  • 350,000 – 2,000,000 cells – 5 mL FACS tube with 1 mL buffer/media
  • > 2,000,000 – 15 ml tube with 3 mL buffer/media

100 µm Nozzle

  • < 250,000 cells – Eppendorf tube with 500 uL of buffer/media
  • 250,000 – 1,000,000 cells – 5 mL FACS tube with 1 mL buffer/media
  • > 1,000,000 cells – 15 mL tube with 3 mL buffer/media

Note: ​The number of cells that are collected into the tube can range significantly, anywhere between 60-90% from what the sorter has recorded, when counted manually by hemocytometer post-sort. This can especially be the case when collected cells are < 10,000 total. Please take this into consideration when planning your experiments. 

4-Laser Filter Layout

5-Laser Filter Layout

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