Re: Mounting media woes

From:klosen@neurochem.u-strasbg.fr (Paul Klosen)

>Try one of the newer liquid mounting medias for chromogens sensitive to
>solvents, like BCIP/NBT, or AEC, it is supposed to work for fluorescence
>but I have not used it for that application.

I can only concur. These new drying media relt in perfectly transparent
preparations just as organic solvent-based media. And they work for some
fluorescent applications like the fast red fluorescence or tyramide
deposited fluorochromes. I don't know about standard immunofluorescence.

As Gayle indicated, a critical detail with these media is to leave them to
dry BEFORE coverslipping with Eukitt, DEPEX or whatever organic medium you
use. During this drying, the refractive index of the medium increases,
which is critical for the transparency of the preparation. Furthermore,
some stains will DIFFUSE or even wash out completely (!!! seen this with
Vectors NOVARED) if the sections are immediately coverslipped. After
immediate coverslipping, only a small rim of the medium around the
coverslip will dry, the remainder will remain liquid.

Paul Klosen


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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@neurochem.u-strasbg.fr==============================





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