Re: controlling autofluorescence red cells/whole blood

From: G Robin Barclay (Robin_Barclay@compuserve.com)
Date: Wed Feb 09 2000 - 19:09:41 EST


Dear Steve
I do not normally see any autofluorescence when looking at red cells in
whole blood - are you diluting your blood with anything (usual culprits are
phenol red or foetal calf serum) which might introduce "autofluorescence"? 
Other autofluorescent events could be eosinophils - but I wouldn't expect
many.  What is your context?  I am counting cells labelled with a human
monoclonal anti-Rh, visualised with FITC-anti-hu-IgG, and experience very
little background.  However, I have introduced PE-anti-glycophorin-A
(anti-red cell) and positively select (gate on) the GPA-positive cells
before counting the (small) numbers of different Rh cells in the mixtures,
to avoid inclusion of any leucocytes in my counts.
Robin Barclay
SNBTS Edinburgh



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