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Recently we have started looking at retics for the first time and have
observed something that seems unusual that I hope you can help us with. We
are actually doing 2-color FACS (we have a 440) with thiazole orange (TO) to
detect retics and a
phycoerythrin conjugated antibody to look at another marker. The filters
are a 530\30 for TO and 585/42 for PE. The problem we are having is that
sometimes the TO-only stained cells sometimes appear in the lower right
(530/30) quadrant as expected but sometimes they appear in the two-color
quadrant as if they were doubly stained. And it seems to be one or the
other, i.e., one sample will have essentially all the retics in the
"retic" quadrant and another will have all the retics in the 2-color
quadrant. One clue as to what might be the cause is that these samples
have remained in the Vacutainer tube with anti coagulant for
several weeks. We
are trying to get some fresh patient samples to determine if this
"incubation" might be the cause. Until we do, can anyone tell me if this
is what you might expect - "two-color" retics after sitting in coagulant?
Thanks.
Ray
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Raymond B. Hester, Ph.D.
Flow Cytometry Lab
The University of South Alabama Each day is a gift;
Mobile, AL 36688-0002 that's why it's called,
Email: rhester@jaguar1.usouthal.edu "the present".
Voice: (334) 460-6029 - Anonymous
FAX: (334) 460-6073
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