NBT/BCIP in situ detection problem

Peter Ashby p.r.ashby at dundee.MAPS.ac.uk
Sat Oct 21 06:14:40 EST 2000


In article <39F073EE.DC083AFE at neurochem.u-strasbg.fr>, Paul Klosen 
<klosen at neurochem.u-strasbg.fr> wrote:

> The problem is not with the in situ. We perform ISH on sections. When we
> follow the reaction development under a scope, it develops normally.
> Just, when we later on look at the slides on the microscope, we have a
> granular deposit instead of the more diffuse typical deposit. This is
> very annoying, because it seems to be located above the section, and
> does not outline the cells. We have the problem with several probes and
> only with alk phos detection. With TSA and perox detection the result is
> the same as usual, but we need the alk phos for our double in situs.

Your description confirms that it is most likely a problem with your 
mounting medium as outlined by someone else. I have seen this effect 
with a number of different enzyme stain products including LacZ from 
transgenic embryos. Have a look at your mounting medium as that is the 
most likely culprit unless you have suddenly increased the magnification 
you are using to view your sections.


Peter

-- 
Peter Ashby
Wellcome Trust Biocentre
University of Dundee
Dundee, Scotland
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