in situ pigment problem

Thomas Hall thomasehall at hotmail.com
Sun Mar 27 04:28:00 EST 2005


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This is a longer process, but I tend to cross my lines onto Golden.

Tom Hall
Currie Lab

From: ""SKucenas"" <stahlhut at slu.edu>
To: zbrafish at net.bio.net
Subject: Re: in situ pigment problem
Date: Fri, March 25, 2005 6:56 am

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

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