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Hi,
How do people out there measure the CD34+ population by flow, do you
gate on CD45+ cells or around the MNC fractions?
Let me know.
Many thanks.
Ed.
We use the sequential gating method of Sutherland, et al.
Experimental Hematology 22:1003-1010 (1994).
Briefly, the method involves gating on CD45 (all nucleated WBC's),
then on CD34+ cells. Use both of these gates to generate a CD45 x
side scatter histogram. The CD34 cells form a discrete cluster of dim
CD45 and low side scatter. Gate this region and use all three gates
to look at forward angle x side scatter. You can then use another
region for a final count on the CD34+ cells.We use this method to
enumerate CD34+ cells in pheresis products and whole blood specimens
from patients undergoing mobilization for transplant.
K. Helm, University of Colorado Health Sciences Center
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