From: robert ashcroft (cytomat@netcore.com.au)
Date: Tue Feb 10 1998 - 06:50:07 EST
For the live stuff Alice, the Ho33342 is hard to beat. See Cytometry Supplement 3:85ff, 1988 figure two and the text. We sorted and got mRNA from cells in different parts of the cycle. Cheers Bob -----Original Message----- From: Alice L. Givan [SMTP:Alice.L.Givan@dartmouth.edu] Sent: Friday, February 06, 1998 2:36 AM To: Cytometry Mailing List Cc: Anne K. Warner Subject: cell cycle problems Hello Flowers, 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 Alice L. Givan Englert Cell Analysis Laboratory Dartmouth Medical School Lebanon, New Hampshire NH 03756 USA tel 603-650-7661 fax 603-650-6130 e-mail givan@dartmouth.edu
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