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Maryalice
Maryalice Stetler-Stevenson
Director Flow Cytometry Unit
Laboratory of Pathology, NCI, NIH
We try very hard to run the cells (lymphomas, leukemias, breast
cancers) the same day they were stained and fixed. However, because of
the times they are delivered to us, we sometimes have to stain and fix
around 5 or 6 p.m. and then acquire on the FACScan the next morning.
Under these conditions, we see no significant increase in background
(auto-?) fluorescence in the controls. When specimens are delivered
late on a Friday and acquired the following Monday (Tuesday in the
case of a holiday) we begin to see significant background fluorescence
on the controls. This fluorescence increases when the cells sit in the
refrigerator longer, say a week or two. This throws doubt upon the
validity of the percent positive calculations. Our conclusion from
this experience is to run the cells *as soon as possible* after
staining and fixation, but no longer than *one day* later, even if it
incurs overtime at the hospital. Also, we encourage our sources to get
samples to us no later than 3 p.m.
Steve Micko
Emory University Hospital
Atlanta, GA
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