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From: Linda Pilarski Professor
Immunology
University of Alberta(403) 492-2275
Henry Wotris asks about doing absolute cell counts on the Facscan. We do this
by taking an aliquot of known volume from a sample in a known volume, diluting
it to a convenient cvolume, e.g. 0.2 ml, and running the whole aliquot through
the instrument. The number of events in the file is the number of cells in
your original aliquot and from there you can calculate the number of cells in
your original sample. You can also get counts for large and small cells in the
sample using this is needed by looking at FALS vs SSC and calculating back to
your sample. We also use this method to compare samples by looking at the
number of cells passing through the probe per second from samples in equal
volumes. Then the flow rates can be directly compared to determine the
relative number of cells in each sample. This is quicker than running the
whole thing.
Hope this helps.
Linda Pilarski
Regards,
Linda
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