Rat testis paraffinembedding for in situ hyb.

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From:Alexander Brands <alexander.brands@uni-tuebingen.de> (by way of histonet)
To:histonet <histonet@magicnet.net>
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Has anyone experience with fixation of (rat) testis and
paraffinembedding for subsequent staging of the germcells?

Problems
It seems to be impossible to cut the Isopropanol dehydrated tissue
thinner than 5Ám, which is nessecary for staging the germcells.

The 5Ám sections float apart as soon as they come into the water bassin,
before I can "catch" them with the slide.

When I try to differrentiate the Toluidinblue stained (5Ám) sections
with 1% HCl-EtOH the color gets lost cmpletely.

General
I am trying nonradioactive mRNA in situ hybridization especially on the
tubuli seminiferi.
I want to asign the detected mRNA to the different stages of the
germcells, in particular distinguish between the two compartments made
by the blood-testis barrier.

Thanks for your help

Alexander Brands, M.D.
Institute of Toxicology
University of Tuebingen
Germany




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