Processing

• The sample is received by the laboratory in saline or a tissue culture media (ie. RPMI) so that the cells are kept alive.
• The cell suspension is washed, the red cells are lysed and the white cells are counted.
• If the sample is a tissue, it may need disaggregation prior to counting (the tissue is mechanically separated and pushed through a cell strainer).
• A slide is made from the sample to look at the morphology of the cells.

Staining

• Once the cell count is determined, the cells are resuspended in a buffer at an optimal concentration for staining (ie. 1 x 10E6/100ul).
• Fluorochrome-conjugated antibodies are either added to the cells individually or a cocktail of antibodies is made (many antibodies in the same tube).
• The staining is done in the dark due to sensitivity of the fluorochromes to light.

Acquisition

• Once the cells are stained, washed and fixed they are ready to be run on the cytometer.
• Running the sample is called Acquisition because you are acquiring the cellular events that you are later going to analyze.