[Histonet] Fixation using Modified Bouins and NBF

[Tony Henwood]

Why use NBF (which I assume is neutral buffered formalin) when there are two
acids present?

Regards

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
The Children's Hospital at Westmead,
Locked Bag 4001, Westmead, 2145, AUSTRALIA.
Tel: 612 9845 3306
Fax: 612 9845 3318




-----Original Message-----

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From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Ahmed, T
(Tanni)
Sent: Friday, 7 October 2005 5:41 PM
To: John A. Kiernan
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Fixation using Modified Bouins and NBF



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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