From: Arnold Pizzey (a.pizzey@ucl.ac.uk)
Date: Wed Nov 18 1998 - 05:44:29 EST
>X-Sender: marina_polonskaia@hms.harvard.edu
>Mime-Version: 1.0
>Date: Thu, 12 Nov 1998 13:49:24 +0100
>To: Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu>
>From: Marina Polonskaia <mpolonskaia@hms.harvard.edu>
>Subject: Cell Cycle of alive cells
>
>
>We are looking for working reliable methods for DNA content staining for
>alive cells. I know that few of them are exist but I heard that people
>always have problems with them.
>
>We'll appreciate any references, advices.
>
>Thank you. Marina
Greetings Maria,
I have recently tried 'Syto16' from molecular probes, it's a
cell-permeable Nucleic acid stain that is green fluorescent, the big
problem with it is that it has some affinity for RNA and, in some cell
types may also stain cytosolic components, I know this sounds as though
Syto16 is totaly unsuitable for DNA analysis but, it all depends on what
you want to do.... Differentiation of G0/G1, S, G2/M would be
difficult using Syto16 alone, as the cell cycle peaks are convolved with
the RNA/cytosolic distribution. If you simply want to analyse/seperate
cycling/noncycling cells however, combining Syto16 with forward scatter
gives a good correlation when sorted cells are fixed,restained with
Propidium and reanalysed.
best wishes
Arnold.
--------------------------------
Arnold Richard Pizzey
Department of Haematology
98 Chenies Mews
London WC1E 6HX
a.pizzey@ucl.ac.uk
(044) 0171 209 6234
fax: 0171 209 6222
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