in situ pigment problem

SKucenas stahlhut at
Fri Mar 25 06:56:00 EST 2005


I've never seen embryos develop pigment during an in situ, but I know that
if your PTU is not good to begin with, it may appear that the embryos are
OK at the start of the procedure, but by coloration, you do get
"non-specific" labeling of what appears to be melanocytes.  We
affectionately call it leopard spots!  I don't know why this occurs at the
very end, but we found that making fresh PTU every week avoids this issue
in our hands.  Hope this helps!

alanmayer at wrote:
> Periodically (and always at the worst times) PTU'ed embryos will develop
pigment (melanin) at the end of an in situ, upon exposure to
> alkaline Tris buffer (NTMT).  Has anybody experienced this and know
how to
> avoid it?
> Thanks!
> ---


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