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Begin with 5 x 105 to 1 x 106 cells per ml to be fixed and examined. Place the cells in a 6-ml polystyrene Falcon tube (#2058).
Pellet the cells and pour off the supernatant leaving a small amount of liquid in the bottom of the tube.
Gently vortex the cells and slowly (drop-wise) add 70% cold EtOH to approximately 2 ml (approximately 1 pasteur pipet worth of EtOH). This is to avoid clumping of the cells during fixation.
Add cold 70% EtOH to 5 ml and store the cells at 4°C overnight. The cells can be stored at this point for up to 2 weeks at 4°C or 6 months at -20°C, however it is best to at least let the cells fix overnight in EtOH.
To prepare the cells for flow cytometry, pellet the EtOH fixed cells and wash once in approximately 3-ml of cold 1X PBS.
