[Histonet] Fixation using Modified Bouins and NBF

[Ahmed, T (Tanni)]

John,

Composition of Modified Bouins we use is - 
36gms picric acid
200mls glacial acetic acid
400mls 10% NBF
3.6L Distilled water

Tissues are fixed for at least 24 hrs. Bouins is changed to 70% IMS
after the recommended time - i.e 24 hrs however this period can be
extended up to a maximum of 3 days.

We are fixing avian bursa specimens in 10% NBF at the moment and
performing IHC to stain for IBD (Infectious Bursal Disease) using the
Vector ABC Elite kit. Our pathologist would like to go back to the days
and try using Modified Bouins to fix bursal tissues then perform the
same IHC. I have never used toxic MD fixative before and trying to
convince him that 10% NBF works just as well...I will have a go though!
no harm in trying old techniques.

Thanks,

Tanni


-----Original Message-----
From: John A. Kiernan [mailto:jkiernan <@t> uwo.ca] 
Sent: 6 October 2005 16:52
To: Ahmed, T (Tanni)
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] Fixation using Modified Bouins and NBF


What is the composition of the "modified Bouin's"
fixative, and how long did the specimens remain in
it? Back in the old days Bouin's was considered a
good fixative for some immunofluorescent applications, especially
localizing protein and peptide hormones. The picric acid in the solution
subdues much of the autofluorescence.
-- 
-------------------------------
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
_______________________________
"Ahmed, T (Tanni)" wrote:
> 
> 
> Dear Histonetters,
> 
> I would like to know if fixation of avian bursa tissue with Modified 
> Bouins fixative over 10% Neutral Buffered Formalin has any advantagous

> effect on immuno precipitation?
> 
> Thanks in advance,
> 
> Tanni
> 
> 
> 
> 
> 
> Tanni  S Ahmed
> 
> Scientific Officer - Histopathology, R&D
> 
> 
> 
> Intervet UK Ltd.
> 
> Walton Manor,
> 
> Walton, Milton Keynes,
> 
> Buckinghamshire,
> 
> MK7 7AJ, UK.
> 
> 
> 
> Tel. +44(0)1908 685552/685543
> 
> Fax +44(0)1908 685614
> 
> E-mail: tanni.ahmed <@t> intervet.com
> 
> 
> 
> 
> 
> 
> 
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