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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