Cytometry Lab

Technical overview of the cell sorters

Sorter	FACStar	MoFlo	DiVa	Aria
Lasers in Use	1: 488nm* 2: 605nm**	1: 488nm 2: 633nm 3: UV 351-364nm***	1: 488nm* 2: 605nm** 3: UV 351-364nm	1: 488nm 2: 405nm 2: 375nm 3: 633nm
Nozzle Size	70 and 100 microns	70 microns	70 microns	70 and 100 microns
Sort Tubes	Clear Falcon tubes #2003 12x75mm Polystyrene	Opaque Falcon tubes #2005   12x75mm Polypropylene	Clear Falcon tubes #2003 12x75mm Polystyrene	Clear Falcon tubes #2003 12x75mm Polystyrene
Collection Tubes or Plates	12x75mm plastic or glass 1.5ml eppendorf Plates: Terasaki or 6-96 well	12x75mm plastic or glass 1.5ml eppendorf Plates: Terasaki or 6-96 well	12x75mm plastic or glass 1.5ml eppendorf Plates: Terasaki or 6-96 well	12x75mm plastic or glass 15ml yellow cap 1.5ml eppendorf Plates: Terasaki or 6-96 well
Optimum Cell Concentration	5x106 cells/ml	4x107 cells/ml	3x107 cells/ml	3x107 cells/ml
Typical Sort Rate	5,000 cells/sec	10-30,000 cells/sec	10-20,000 cells/sec	10-20,000 cells/sec
Number of Sort Populations	1 or 2 or Plates	1 to 4 or Plates	1 to 4 or Plates	1 to 4 or Plates
Sort Region Type	Rectangular	Polygonal	Polygonal	Polygonal
Running Pressure	7.5 - 12.5psi	55psi	30psi or 55psi	20psi, 35psi or 70psi

• *     May be switched to 458, 476, 515nm.
• **   May be adjusted continuously from 550-650nm.
• *** May be switched to 405nm.
• Allow 30 mins set-up time for any of these changes

For more details about possible choices of fluorochromes, see Fluorochrome Combinations.

Collection Tubes:

1-2mls of neat FCS or media containing FCS (whatever your cells will be happiest in).The tubes can be either glass or plastic (eg Falcon 2003 or 2005)

 Removal of dead cells and debris:

For smooth running of the sort and maximum yield of cells, run your cells through a dead-cell-removal column (glass Pasteur pipette with a plug of cotton wool in the end) or use a 100um filter prior to sorting.

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