Dear All, I'm posting this for a colleague, so don't have all the details, but hope that someone may have some suggestions. They are using "whole blood" bone marrow preparations in analysis of various leukaemias/lymphomas, and consistently find a problem with autofluorescent cells that give signals in both the FL1 and FL2 channels (they have that typical non-specific diagonal look on a dot-plot of FL1-FL2). The non-specific signals appear even in the absence of any antibody, so this seems to be a true autofluorescence rather than non-specific antibody binding. They are reluctant to gate out on FSC/SSC after back-gating as they feel that they risk losing important cells. Does anyone have a simple way of overcoming this problem, or even know for certain what the cells are likely to be. They are not seen after ficoll separation, and scatter suggests that they are of myeloid origin Andy (andy@serotec.co.uk)
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