[Histonet] FETAL BRAIN FIXATION

Thu Oct 21 22:57:36 CDT 2004

Tony,

You are not alone in being confused by the nomenclature of fixatives.
I share your confusion and so do most authors of textbooks!!!
Anyone making a slight modification to a fixative mixture seems to have put
his name on it!!!
There are numerous examples in the texts.
There can be little doubt that Clarke (1851) was the originator of using a
combination of alcohol and acetic acid for fixation,
yet many authors attribute Carnoy (1887). Indeed, Carnoy did use it (Carnoy
A), but only his contribution of adding chloroform to the mixture (Carnoy B)
should be acknowledged (see Baker and Lillie).
    (Reference: Clarke J L.(1851) Researches into the structure of the
spinal cord, Phil. Trans. R.S. 141: 601-622)

Following the introduction of formaldehyde (Blum 1893) a great number of
people utilized it in various combinations with other fixatives.
Most notably Bouin (1897), who combined it with picric and acetic acids the
result of which is still in wide use today.
Brasil (1905) also experimented with added formaldehyde in several
mixtures.He used the combination of formaldehyde and Clarke's
fluid for general histological work (the one I gave) and an alcoholic
version of Bouin's fluid for protozoa. It is the latter which is commonly
called Brasil's fluid!!
(Reference: Brasil, L.(1905) Nouvelles recherches sur la reproduction des
gregarines monocystidees, Arch. Zool. Exp. 4: 69-99)

Brasil's combination of formaldehyde, acetic acid and alcohol as a fixative
mixture is known by many names. Lillie lists Tellyesniczky, Fekete, Opie &
Lavin, Bodian and Lillie's FAA as the same fixative with only minor
variations in the individual component concentrations! None the less Brasil
was the originator.

Incidentally, I can find no reference to the substitution of trichloracetic
for the acetic acid in the original Brasil's fluid you are using!
The formula I have gives:
80% alcohol        1500 mL
Formalin                600 mL
Glacial acetic acid  150 mL
Picric acid                10 g
Your formula is no doubt another of those minor modifications that actually
went unnamed!!

You will find a reference to the so-called good fixation of glycogen by
Brasil and similar fixatives in Kiernan's textbook under Gendre's fluid, yet
another name for alcoholic Bouin!!!!!

Yours in confusion,

Bryan

----- Original Message -----
From: "Tony Henwood" <AnthonyH <@t> chw.edu.au>
To: "'Bryan Hewlett'" <bhewlett <@t> cogeco.ca>; <lpwenk <@t> sbcglobal.net>;
"Histonet" <histonet <@t> lists.utsouthwestern.edu>
Sent: Thursday, October 21, 2004 7:41 PM
Subject: RE: [Histonet] FETAL BRAIN FIXATION

>
> 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
> >
> >
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>
>
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