RE: [Histonet] Testes Bouin's fluid

From:"Barry R Rittman"

An important point here is that during fixation in Bouin's some of the
protein picrates that are formed are soluble in water but are rendered
insoluble when the tissue is placed in 70% or higher percentage alcohol.
Placing tissue in water (and therefore presumably in aqueous formalin)
allows some of the picrates to leach out of the tissue. 
I like Bouin's for fixing tissues such as testes (animal testes that is
- we don't get too many from patients here in the dental school!!) as
the coarse precipitate allows nuclear details to be clearer.
Barry

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Craig
Squire
Sent: Monday, May 03, 2004 11:31 AM
To: balajimr@drreddys.com
Cc: Histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Testes Bouin's fluid


I work in a contract research lab and we keep almost all of our testes,
and sometimes epididymides, in 70% alcohol permanently.  We rinse with
distilled water and then store in 70%.  All of  the slides are then
stained with H&E and they turn out really nice. When we place the tissue
in 70% COH we do not trim the tissue right away, we wait a few days.
Hope this helps.

Heather Squire
----- Original Message ----- 
From: "John Kiernan" 
To: 
Cc: 
Sent: Monday, May 03, 2004 9:40 AM
Subject: Re: [Histonet] Testes Bouin's fluid


> It does not make sense to put specimens in neutral
> buffered formaldehyde after fixing them in Bouin's
> fluid. Bouin's is, for many purposes, a better
> fixative than NBF because it confers characteristic
> patterns of chromatin in nuclei of different cell
> types and it also permits brighter staining of
> collagen and cytoplasm with anionic dyes. Bouin's
> fluid extracts cytoplasmic RNA and it can also
> make DNA Schiff-positive (Feulgen hydrolysis), and
> can damage red blood cells.
>
> Staining by both haemalum and eosin is stronger
> after Bouin's than after NBF, and in many tissues
> the microanatomy, as seen in paraffin sections, is
> more lifelike. There is less differential shrinkage
> of cells, tubules etc than you see in paraffin
> sections of specimens fixed in neutral formaldehyde.
> --
> -------------------------------
> John A. Kiernan
> Department of Anatomy and Cell Biology
> The University of Western Ontario
> London,   Canada   N6A 5C1
>    kiernan[AT]uwo.ca
>    http://publish.uwo.ca/~jkiernan/
>    http://instruct.uwo.ca/anatomy/530/index.htm
> _______________________________
> balajimr@drreddys.com wrote:
> >
> > Dear Histonetter,
> >
> > We have been preserving testes (of rats and mouse) in bouin's 
> > solution
and
> > after 24 hrs washing them with 70% alcohol and fixing it back in 
> > 10%NBF. But staining later with H & E is not that great. Can anybody

> > suggest me the best protocol for fixing,processing and staining  of 
> > testes.
> >
> > Dr. Balaji
> > Dept. Pre clinical safety evaluation,
> > Discovery research,
> > Dr. Reddys Laboratories ltd. Bollaram Road,
> > Miyapur, Hyderabad, 500 049
> >  Andhra Pradseh, INDIA 
> > _______________________________________________
> > Histonet mailing list
> > Histonet@lists.utsouthwestern.edu 
> > http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
> _______________________________________________
> Histonet mailing list
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>


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