[Histonet] FETAL BRAIN FIXATION

Tony Henwood AnthonyH <@t> chw.edu.au
Thu Oct 21 18:41:46 CDT 2004


We have also used a modified Bouin's Fixative for our fetal brains. 
Bryan do you have a reference for the modified Brasil's Fixative?
The Brasil's fixative that we have used for liver biopsies where a metabolic
disease is suspected is as follows:

Absolute alcohol 		1650ml
Formalin 			600ml
Trichloracetic acid 		7.5g
Picric acid 			10g

It has been suggested as a good fixative for glycogen, though I can't find a
reference for this. The above solution has been used in my lab for years,
before I arrived here, hence my confusion. In fact one of our recent papers
used this fixative and it had to be published with out a reference, so if
anyone can shed some light I would be most appreciative:

V-M. Mangan, V. Farago, M. Kelly, and A. F. Henwood (2002) " An Amylase
Reagent with a Long Shelf Life for the Removal of Glycogen from Tissue
Sections" J Histotechnol. 25(3): 153-4.

You will note that our Brasil's fixative contains picric acid. 
The nomenclature seems very confused!
Any insights?

Regards,

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

-----Original Message-----
From: Bryan Hewlett [mailto:bhewlett <@t> cogeco.ca] 
Sent: Thursday, 21 October 2004 11:33 PM
To: lpwenk <@t> sbcglobal.net; Histonet
Subject: Re: [Histonet] FETAL BRAIN FIXATION


Peggy,

We faced the same issue a number of years ago.
Our solution was to use a modified Brasil fixative.
The pediatric pathologist was delighted with the results!
The formula follows;

Modified Brasil Fluid.

Ethanol............................................. 700 mL
40% w/v Formaldehyde.....................100 mL
Glacial Acetic acid..............................50 mL
add D.water to make up to.................1000 mL

Fetal brains are left to fix for 1-3 weeks, rinsed in 70% ETOH, then sliced
and photographed.
Selected blocks are processed using the same reagents and times as for NBF
fixed material.

Hope this helps,

Bryan

----- Original Message -----
From: <lpwenk <@t> sbcglobal.net>
To: "Histonet" <histonet <@t> lists.utsouthwestern.edu>
Sent: Thursday, October 21, 2004 4:56 AM
Subject: [Histonet] FETAL BRAIN FIXATION


> Anyone have a good fixative for the very soft fetal brains?
>
> Our neuropathologist does not want to use Bouin, as the tissue turns
yellow
> and photographs do not look the way they are supposed to.
>
> Another one of our pathologists remembers using an
alcohol-acetone-formalin
> mixture years ago as a resident in another institution, but doesn't know
the
> percentage of any of the reagents.
>
> I've tried several search engines and PubMed, and can't find anything.
>
> Thanks.
>
> Peggy A. Wenk, HTL(ASCP)SLS
> William Beaumont Hospital
> Royal Oak, MI 48073
>
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>



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