DNA staining in living embryos!?
CHAD A. RAPPLEYE
rappleye at biomail.ucsd.edu
Wed Jun 14 14:50:39 EST 2000
The eggshell surrounding the embryo is the main obstacle as it will not
let most substances into the embryo. The solution of course is to
permeabilize the egg shell so that your DNA stains can get into the
blastomeres. Some options to permeabilizing the egg shell are:
perforation of the egg shell with a laser (Schierenberg and Junkersdorf,
1992 Roux's Arch Dev Biol 202:10-16), chemical and enzymatic treatments to
remove the egg shell layers (Edgar, 1995 Methods in Cell Biology
48:303-321), or use of a mutant strain that has compromised egg shells
(Rappleye, et al., 1999 Genes & Dev 13:2838-2851). You can contact me if
you need more information on the above. Alternatively you can consider
using a live tag for following the DNA such as a histone:GFP.
UC, San Diego
On Tue, 13 Jun 2000, Nobody wrote:
> Dear all,
> I am in the process of trying to staining the DNA in the living embryos,
> but I was curious to find out whether anyone else has ever had a similar
> experience, or can offer any suggestions as to what might be happening.
> I was really wandering if, there is a simple recipe for this staining
> procedure that one can simply prepare. I appreciate any inputs, ideas or
> suggestions for my purpose.
> Many thanks in advance,
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