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Preparation of Cells for Flow Cytometry
For the preparation of single cells derived from tissue culture cell lines.
Single cells must be suspended at a density of 105-107 cells/ml to keep the narrow bores of the flow cytometer and its tubing from clogging up. The concentration also influences the rate of flow sorting, which typically progresses at 2,000-20,000 cells/second. Phosphate Buffered Saline (PBS) is a common suspension buffer.
The most straight forward samples for flow cytometry are non-adherent cells from tissue cell culture. Here we describe methods for both tissue culture cell lines and adherent tissue culture cell lines. However analysis may be required from cells derived from other sources, Bio-Rad has a protocol (FC3) for the preparations of tissues from a variety of sources including; peritoneal macrophages, thymus cells, spleen cells and bone marrow. Further to this a protocol (FC2) is also available for the sample preparation of human peripheral blood mononuclear cells.
This method provides a general procedure for use with tissue culture cells in suspension.
This method provides a general procedure for use with adherent tissue culture cells