NBT/BCIP in situ detection problem

Paul Klosen klosen at neurochem.u-strasbg.fr
Fri Oct 20 11:33:50 EST 2000

Peter Ashby wrote:

> Are you sure it is a problem with the in situ rather than your
> microscopy which could produce these symptoms? Are you doing in situs on
> sections or are you sectioning whole mounts? Either way it could
> indicate that your staining is simply stronger than it used to be, you
> could try incubation for less time.

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.
Paul Klosen, PhD
CNRS UMR 7518 Neurobiologie des Fonctions Rythmiques et Saisonnieres
Universite Louis Pasteur  12, rue de l'Universite F-67000 Strasbourg,
Tel.  Fax.  klosen at neurochem.u-strasbg.fr

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