Re: [Histonet] Testes Bouin's fluid

From:Geoff McAuliffe

Dear Dr. Balaji:

A few questions to clarify your problem:

How long were the testes fixed in Bouin?
Was fixation by immersion or perfusion?
How long was the tissue washed in 70% ethanol?
Why was the tissue 'refixed' in buffered formalin? Tissues properly 
fixed in Bouin should not need to be refixed.
Exactly what is it about the morphology/staining that is "not that great"?

Bouin's is an excellent fixative and, assuming it is made correctly and 
the tissues are fixed promptly and for sufficient time, the most likely 
source of your problems is not the fixative.

Geoff 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

Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583; fax: -4029

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