General Lab Rules and collection tubes
Food and Drinks
Food and Drinks are not allowed in any of the Flow Cytometry Core Facility laboratories.
Logging in Instruments’ Computers
Enter your ONYEN and Password. Be sure you have selected to login to the AD domain.
Logging off Instruments’ Computers
Please log off the machine when you are finished. You may be charged for extra time if you do not.
Sample Tubes Requirements
- CyAn ADP – 12x75mm (5mL) polystyrene or polypropylene (cloudy) plastic tubes. Note: some types of irradiated polypropylene bullet tubes (0.5 mL) can cause erroneous autofluorescence in the 450/50 channel. Avoid USA Scientific #1412-1410.
- LSR II / LSRFortessa – 12x75mm (5mL) polystyrene plastic tubes
- ImageStreamX MkII – 1.5mL Eppendorf tubes
- MoFlo – 12x75mm (5mL) polystyrene or polypropylene plastic tubes to 50ml tubes
- MoFlo XDP – 12x75mm (5mL) polystyrene or polypropylene plastic tubes to 50ml tubes
- FACSAria II – 12x75mm (5mL) polystyrene or polypropylene plastic tubes to 50ml tubes and 1mL microtube size
Sort Collection Tubes
Use either 12x75mm (5mL) polypropylene plastic tubes or 12x120mm (15mL) polypropylene.
Note: Polystyrene tubes are not ideal as they accumulate charges during sorting which can result in losses of sorted cells.
- For the 12x75mm tubes place ~1ml of media in the tube.
- For the 12x120mm tubes place ~2ml of media in the tube.
- When using the 70µm tip we can collect ~3×106 cells per tube.
- When using the 100µm tip we can place about ~1×106 cells per tube.
- When using the 12x120mm tubes increase the numbers above by 3 fold.
Filter your samples prior to running on any instrument (Filter Options).
DNA from lysed cells in the medium can cause cells to clump. DNAse I (Sigma Cat. No. D-4513) in the presence of magnesium chloride (Sigma Cat. No. M-2670) will help reduce cellular aggregation. Treat cells for 15 to 30 minutes in a solution of 100 µg/mL DNAse and 5 mM MgCl2 in HBSS at room temperature. Wash the cells once in the presence of 5 mM MgCl2 in HBSS. Gently suspend the cells in Stain Buffer (BSA) containing MgCl2 and 25-50 µg/mL DNAse (as a maintenance dose). DNAse I requires a concentration of at least 1 mM magnesium to work effectively, although 5 mM is optimal. Note: It is important to minimize the presence of dead cells during this procedure, since actin released from dead cells irreversibly inhibits DNAse I.
Follow the cleaning listed in the Quick Reference or affixed to the instrument. Please clean any spills and throw any used pipet tips, tube, tube caps, aluminum foil…in the trash.
Make sure you stop your run before the end of your scheduled time to perform cleaning and in order not to delay the next user.