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> A graduate student in our lab would like to compare the relative antigen
> density of a marker on cells from a particular patient population
> versus that of normal individuals. The patient and normal samples have
> to be run on different days. I have explained to her the problems
> inherent in this type of experiment: differences in instrument settings,
> variabililty in staining conditions, the "relative" nature of
> fluorescence intensity, etc. However, her mentor insists that this
> type of experiment can and must be done.
>
> Can anyone advise us on the best method for approaching this problem
> by flow cytometry? Doesn't someone sell beads which are standardized
> as to the amount of fluorescence on them? Can we use these as
> internal standards to control for experiment-to-experiment
> variability in fluorscence intensity of the antigen density of cells?
>
> Steve Shivers
> Bone Marrow Transplant Program
> Moffitt Cancer Center
> at the University of South Florida
>
> shivers@aarlo.moffitt.usf.edu
Dear Steve,
You can do this by routinely setting up your instruments with the same
standard beads or dump some of the beads in with the samples, setting the
peak for the beads on the same channel each time. We use Flow
Cytometry Standards for the bead source. This worked fine for us over
years. If your instrument (you don't say what kind) can't do this you
have a problem.
Deb Berglund
MSU
Bozeman, MT 59717
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