cell cycle problems

robert ashcroft cytomat at netcore.com.au
Tue Feb 10 06:50:07 EST 1998


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 at 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 at dartmouth.edu




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