From: william nostrom (slickwilli@hotmail.com)
Date: Mon Feb 09 1998 - 16:46:23 EST
ON Feb 9th, you wrote: >Can someone point me to the state-of-the-art for cell cycle analysis with: >1) cells fixed in formaldehyde >2) live cells > >We have GFP-fusion-protein-transfected cells and, in the first instance, need >to fix in formaldehyde (not ethanol) to retain the GFP but want to get good DNA >profiles. Is there a stain that works better than PI with formaldehyde-fixed >cells or is there a method for opening up the fixed histones/DNA so as to >improve the PI profile without losing the GFP? > >In the second instance, we would like to keep the cells alive during this >procedure. Is there a staining method with one of the Hoechst dyes that will >give us histograms sharp enough to allow cell cycle analysis or is there any >better vital alternative? > > >Thanks. >Alice Interestingly enough, I just got an article on that. It is in Cytometry 29:286-291 (1997). "Use of a Membrane-Localized GFP Allows Simultaneous Identification of Transfected Cells and Cell Cycle Analysis by FC" by RF Kalejta. Hope this helps. Bill Nostrom Trudeau Institute Saranac Lake, NY ______________________________________________________ Get Your Private, Free Email at http://www.hotmail.com
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