Keith, Interesting information! About the whole blood, ours (my samples anyway) are never refrigerated. For some reason, the FACS Lyse did not provide consistent lysing for us. NOW, that's not to say that it just needed a whirl on the vortex to mix everything nicely JUST prior to adding the lysing solution, as I found was needed to bring the effectiveness of the Gentrak and Coulter product to 100% effectiveness. I think that the Gebtrak and Coulter products "in my opinion" are better due to speed of the procedure....BTW, thanks for the info on the NH4CL....I'll have to try it again to see if a sample is indeed "readable" with a small RBC pellet present. Patricia Echeagaray Flow Cytometry Services Southern Research Frederick Frederick MD USA Keith Bahjat wrote: > > With regard to lysing solutions. DO NOT refrigerate whole > blood and expect the red cells to lyse properly. Snip > I only mention this as I have used BD's lysing solution for > a very long time, and I have NEVER had it not adequately > lyse 100 uL of peripheral anticoagulated (EDTA, Na Heparin, > or ACD) whole blood when 2 mL of lysing solution is used > UNLESS the sample was refrigerated. > > As for NH4Cl, I have always seen CONSISTENT performance with > this in non-refrigerated blood. The quirk is that it > CONSISTENTLY leaves a pellet of rbc's. BUT, as NH4Cl does > not shrink the cytoplasm of the cell (as lysing solutions > with fixatives do), the lymphocytes are quite easy to > resolve from the rbc's.
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