Re: Autofluorescence in bone marrow samples

From: Maryalice Stetler-Stevenson (stetler@box-s.nih.gov)
Date: Tue Oct 05 1999 - 12:53:56 EST


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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