Staining of Whole Blood. -Reply

From: Antony Bakke (bakkea@ohsu.edu)
Date: Thu Oct 16 1997 - 17:55:53 EST


BD has developed a procedure for no wash, no lyse staining of whole blood.  It
depends on multiple color staining in which one of the antibodies stains all of
the leukocytes.  This is usually CD45.  You set up the flow cytometer to trigger
on the fluorescence of CD45 for leukocytes, thus eliminating all of the red
cells, since they do not fluoresce and, therefore, do not trigger the
instrument.  The leukocytes can then be displayed and gated as usual.

I have only tried this a few times and it is difficult.  The amount of blood
used, the flow rate and the trigger threshold seem to be critical to prevent
overwhelming the detectors and having a high coincidence of red cells and
leukocytes, which alters the scatter pattern.  I would suggest talking to
someone at BD.

Hope this helps.  Good flowing,
Tony Bakke



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