Re: Cell Culture after DNA Ploidy Staining

Gregor Rothe (gregor.rothe@klinik.uni-regensburg.de)
Fri, 18 Jul 1997 08:35:33 +0200

On 17 Jul 1997 16:33:06 Gerhard Nebe-von-Caron wrote:

>
> Depending on how rough an estimation you can get away with,
> you could try dihydroethidium (or Hydroethidine as it was
> originally called) or LDS751. They do stain live cells but
> will not give you the profiles you get with the A-T specific
> dyes. I am not sure whether the new syto stains from
> Molecular Probes offer any higher DNA specificity. You could
> also try to contact Terry Hoy at Cardiff if you could get
> some of their 'magic red' to try out.
>

Dihydroethidium staining does NOT lead to a fluorescence distribution
which correlates to cell cycle. The cellular red fluorescence levels
achieved following intracellular oxidation rather resemble different
levels of metabolic activity of these cells far below saturation of
DNA binding sites (J Leukoc Biol 47:440,'90).

Gregor Rothe

> ______________________________ Reply Separator _________________________________
> Subject: Cell Culture after DNA Ploidy Staining
> Author: l.gaudry@unsw.edu.au at INTERNET
> Date: 16/07/97 21:01
>
>
> Hello,
> I would like to ask if anyone has a method for estimating DNA Ploidy using
> 488nm or 633nm Laser so that the cells are alive and sortable before
> culturing.If you use Hoechst and UV the above can be done,I am told.My
> problem is no UV capabilities hence I am looking for some other way.
> Hope someone out there can help me.
> Thank you,Leonie
>
*************************************************************************
Institute for Clinical Chemistry and Laboratory Medicine
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Tel. +49 (941) 944-6204
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Internet: Gregor.Rothe@klinik.uni-regensburg.de
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