Renee I do this all the time on our MoFlo. Which laser positions are the UV and 488 lasers in? I would suggest that they must be: 488 #1 and UV #3 or it probably will behave as you describe. Larry At 02:29 PM 4/17/2002 -0500, you wrote: > 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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