Stain specific for DNA (no RNA) and excitable with ArKr-Laser

Steffen Dietzel (dietzel@sun0.urz.uni-heidelberg.de)
Fri, 29 Nov 1996 16:02:36 +0100

I need some help about DNA-staining. I want to do confocal microscopy with
stained (mitotic) chromosomes in Drosophila tissue preparations.
Since I have not yet found an appropriate DNA stain, I thought that the
flow-cytometry community might be able to provide help due to the
experience with all sorts of fluorescent stains.

I am searching a stain for DNA that does not stain RNA - like
Dapi or the Hoechst dyes. But since I want to use it with a confocal only
equipped with an ArKr-Laser (488, 568, 647nm and a weaker line at 476nm) I
can not use these. I also would like to use GFP-Fluorescence, so a red
stain would be preferable.
I have tried already the following without success (probably because of
RNA-staining): Propidium Iodide, Bobo-3, Hexidium Iodide, LDS-751, Ethidium
Bromide, Dihydroethidium.
As one might guess from the set of dyes, originally I wanted to do the
staining in living cells, but... :-(

So if anybody should know a stain that could work, please let me know.

I assume that chromomycin A3 could be helpfull, but until now I did not
find a spectrum of the excitation or emission.
Also, can somebody comment on 7-Aminoactinomycin (7-AAD)? Does it stain
also RNA? I have found contradictory opinions so far.

I am also thinking whether a classical, non-fluorescent stain could be
helpfull, that could be recorded with DIC or Phase Contrast on the confocal
in parallel to the fluorescent dyes. Has anyone suggestions what could be
tried or experience with this?

Please send a copy of your reply directly to my e-mail address. I have
searched the Web-Site at Purdue Univ quite long, but I did not find how I
can subscribe to the list. I can check the answers in the Web Archive, but
loading times are rather long.

Steffen Dietzel

-- -------------------------------------------------------------
Dr. Steffen Dietzel
Zentrum fuer molekulare Biologie Heidelberg (ZMBH)
University of Heidelberg
Im Neuenheimer Feld 282
69120 Heidelberg, Germany
Phone: +49/6221/54-6836 or -6879. Fax: -5893
listservmail:dietzel@sun0.urz.uni-heidelberg.de
personal mail: dietzel@wotan.iwr.uni-heidelberg.de

-- -------------------------------------------------------------
Dr. Steffen Dietzel
Zentrum fuer molekulare Biologie Heidelberg (ZMBH)
University of Heidelberg
Im Neuenheimer Feld 282
69120 Heidelberg, Germany
Phone: +49/6221/54-6836 or -6879. Fax: -5893
listservmail:dietzel@sun0.urz.uni-heidelberg.de
personal mail: dietzel@wotan.iwr.uni-heidelberg.de