Cytokine increases autofluorescence!?

From: Jason Gush (jgush@malaghan.org.nz)
Date: Tue Apr 16 2002 - 14:58:18 EST


Hello out there flow-folk,

I am working with transformed haemopoietic cell lines, treating them with
cytokine, and measuring the expression of surface markers by single colour
flow.  I would appreciate advise on how to analyze the following situation:

My problem is that in response to cytokine, a number of the lines increase
in size, granularity, as well as showing a significant increase in
autofluorescence (e.g. with the same settings FL1, FL2 medians go from
approximately 6 and 9 to 11 and 27 respectively).  How should I interpret
positive markers which show a similar log increase?

I believe a sensible thing to do is to use a spare fluorescence channel to
compensate for autofluorescence.  However is compensation applied to just
the treated data set/samples, or is this bad behavior and the same
compensation should be applied to both conditions?

Thanks in advance,
Jason.

Jason Gush
Malaghan Institute of Medical Research
P.O. Box 7060.
Wellington, NEW ZEALAND.



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