They really don't impact on the analysis if they are a true diagnonal and not that high a fluorescence. We see a little of this but not enough to cause problems. Since the question with leukemias and lymphomas is what does the patient have and since this can be answered with a little diagnonal non-specific artifact, we don't care. If this is a significant problem with all cases, something is very wrong. Maryalice >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) Maryalice Stetler-Stevenson Director Flow Cytometry Unit Laboratory of Pathology, NCI, NIH
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