I am a graduate student and I've been using flow cytometry to study
the cell cycle in Drosophila cells. I dissociate cells from tissue
samples and stain for DNA content with Hoechst 33342 and it works
great. However, I've been trying to also use GFP as a marker in these
assays and I've found that the Hoechst flouresces in the GFP channel and
obscures the GFP signal to the point where compensation can't fix the
problem. We are using a Cytomation Moflo mahcine and we are using the
UV laser for the Hoechst and the FL2 detector for the GFP. We've tried
shutting off the UV laser and the Hoechst background is still picked up
in the GFP chanel, indicating the problem is with the FL2 detector. The
flow technician isn't sure how to fix the problem. Do you guys have any
suggestions on how to look at cell cycle and GFP flourescence in live
cells? Thanks
Renee Read
Department of Molecular Biology and Pharmacology
Washington University School of Medicine
660 S. Euclid Avenue
St. Louis, MO 63110
lab: (314)-362-7797
fax: (314)-362-7058
rdread@artsci.wustl.edu
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