Re: NBT/BCIP in situ detection problem

From:Heike Grabsch <>

Dear Paul,

I do have the same problems with NBT/BCIP since long time and I tried a 
couple of things - no success so far, I gave up (I have to admit). In 
case you get a good problem solution, please let me know.

Heike Grabsch,
Dept of Patholgoy
University of Duesseldorf
Moorenstrasse 5
40225 Duesseldorf

Paul Klosen wrote:
> Hi,
> Recently we have run into a nasty problem with our NBT/BCIP in situ
> detections. The in situ part seems to work fine and the alk phos detections
> is sensitive.
> However, under the microscope, the precipitate is completely granular and
> seems to be located above the tissue, which is absolutely unsuitable for
> photomicrography. We have tried varying several parameters (pH, salt and
> MgCl2 conc., ..) and different lots and sources of both NBT and BCIP, but
> nothing seems to bring us back to our previous results, where we had a nice
> cell-filling precipitate, albeit a bit diffuse as expected with NBT/BCIP.
> Has anyone encountered a similar problem, and how did you get rid of it ?
> Thanks
>                                                                          -=-
>                                                                         (o -) O
> ===============================oOo==(_)==OOo=============================> Paul Klosen, PhD
> CNRS UMR 7518 Neurobiologie des Fonctions Rythmiques et Saisonnieres
> Universite Louis Pasteur  12, rue de l'Universite
> F-67000 Strasbourg, FRANCE
> Tel.  Fax.

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