General Sorting Information
| Machine |
Aria II/IIu |
FACS Melody |
Sony SH800 |
BSL-3 FACS Aria IIu |
|
Lasers |
(BSL-2 only) 5B 2V 2R "S" Aria (BL2+) 2B 6V 3R 5YG "2" Aria |
4B 5V 3R |
488/633/405/561 |
5B 3V 4R 2UV |
| Nozzle sizes (µm) | 70/85/100/130 |
100 | 70/100 | 70/85/100 |
| Cell Concentration | 1-2 x107 | 1-2 x107 | 1-2 x107 | 1-2 x107 |
| Sort Rate | up to 17,000 cells/sec. (70µm) | up to 6,000 cells/sec. | up to 5,000 cells/sec. | up to 17,000 cells/sec. (70µm) |
| Operating Pressure (psi) | 70/45/20/10 |
20 | 20/40 | 70/45/20 |
Nozzle choice - The diameter of your cells should be less than 30% of the diameter of the nozzle. Otherwise cells can be damaged and side streams can be diffused.
Sort Rate - Each nozzle size has a limitation for the rate at which cells can be sorted efficiently. As a rule-of-thumb, divide the diameter of the nozzle by 5 to see the maximum rate at which the nozzle can sort.
Running media - 1% FBS in any phenol-red-free media. HEPES can be added at a final concentration of 25mM to media to keep cells well buffered for all-day sorts, and/or to collection media.
Collection media - This is usually 20% FBS in whichever media the cells are happiest. This is diluted 4-5x and ends up being approximately 5% FBS.
Collection plates: Bring plates for collection with media pre-loaded in the wells. Do not fill wells above 1/2 maximum capacity to avoid spilling.
Filtering : All samples are filtered through 37µ mesh prior to running by core facility personnel, however if you wish to filter the samples yourself refer to the ordering information below. Do not bring samples in tubes with filter caps, as these are not sealed. Replace filter cap with an impermeable cap before transporting your samples!
Sterile 12x75 mm tubes with filter caps are available; by Corning/Falcon #352235 - Fisher Cat. number 08-771-23
Nitex Nylon Filtration Mesh order from: ELKO Filtering Co, LLC. (elkofiltering.com) Nylon mesh roll 37µ opening size, 40" wide, sold by the yard) Item#: 03-37/24