NBT/BCIP in situ detection problem

Paul Grimm pgrimm at ucsd.edu
Fri Oct 20 20:29:47 EST 2000


Is it possible that the mounting medium or technique is different and affecting
the NBT/BCIP deposits? I used glycerol gel to mount the cover slip when I was
using NBT/BCIP for in situ's. Sometimes a batch of glycerol gel would not work
as well as another.
Suggest that you try temporarily mounting one of your slides with water or
saline and a coverslip to see how the deposit looks under the microscope before
formal mounting.

The other issue is whether you use polyvinyl alcohol in your NBT development. I
think PVA can give a more "granular" appearance.

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, but when we observe our slides under the
> microscope, the precipitate is completely granular and seems to be
> located above the tissue. We have tried varying several parameter (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
> ?
> Thanx
>
> --
> 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. 03.88.35.85.04  Fax. 03.88.24.04.61  klosen at neurochem.u-strasbg.fr

--
Paul C. Grimm
Associate Professor
Pediatric Nephrology
University of California at San Diego

Email pgrimm at ucsd.edu
Phone 619-543-5218
Fax   619-543-3575
Snail mail
UCSD Pediatrics
Mail Stop 0831
9500 Gilman Drive,
La Jolla California
92093-0831

Rome did not create a great empire by having meetings...
they did it by killing all those who opposed them.
------
Where mediocrity is the norm, it is not long before mediocrity becomes the
ideal" A. N. Wilson







More information about the Methods mailing list