Fixing Cells for FACS

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• Grow 5 ml of cells
• Spin and wash in water
• Spin and resuspend in 1.5 ml H2O
• While slowly vortexing add 3.5 ml of ice cold 95% ethanol
• Store at 4C until the day before the FACS analysis
• On the day before the FACS analysis wash the cells in 50 mM Na-citrate (prepared from a 0.5M stock)
• Resuspend in 1 ml Na-citrate, 100 ug/ml RNAse A (boiled)
• Incubate at 37C for 2h
• One h before the FACS analysis add 1 ml Na-citrate containing 4 ug/ml PI
• Transfer 0.5 ml into a FACS tube (6 ml plastic tube)
• Store samples in the dark (wrap with foil) until measurement