Re: Rat testis paraffin embedding for in situ hyb.

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From:Kevin R Gray <grayk@bms.com> (by way of histonet)
To:histonet <histonet@magicnet.net>
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Joe,
We use a saturated solution of lithium carbonate in 70% ethanol.

Kevin Gray
Bristol-Myers Squibb, NJ

Saby, Joseph wrote:
>
> Kevin-
>
> Sorry for the repeat, by my first missive did not make it to the HistoNet.
> What percentage lithium carbonate do you use in your 70% EtOH?
>
> Joe Saby, BA HT
> Parke-Davis, Ann Arbor, MI
>
> -----Original Message-----
> From: Kevin R Gray [mailto:grayk@bms.com]
> Sent: Thursday, February 18, 1999 8:10 AM
> To: alexander.brands@uni-tuebingen.de
> Cc: HistoNet@Pathology.swmed.edu
> Subject: Re: Rat testis paraffinembedding for in situ hyb.
>
> Alexander,
>
> Our lab fixes rat testis in Bouin's fixative.  It is removed in 70%
> ethanol containing lithium carbonate during the course of a workday with
> the ethanol Li/Carbonate mixture being replaced as the Bouin's comes out
> of the tissue.  The testis are then processed overnight under routine
> conditions to paraffin.  They are sectioned at 3 um for PAS staining by
> our histotechnician.
>
> Kevin Gray
> Bristol-Myers Squibb
> New Jersey, USA
>
> Alexander Brands wrote:
> >
> > 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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