Re: Brazil's fluid
This was my thought exactly when I saw this formula, just a variation of Bouins.
"J. A. Kiernan" wrote:
> I think of all these picric-alcohol-formalin-acetic mixtures
> as "alcoholic Bouin." The idea of adding the acetic acid to the
> stock solution immediately before using is probably to avoid the
> slow reversible reaction with ethyl alcohol, which forms ethyl
> acetate (fruity odour) and depletes the acetic acid. BUT for
> some purposes, especially insect histology, aged alcoholic Bouin
> is claimed to be better than the fresh stuff. R.E.Gregory et al
> published two thorough studies of this in 1980, in
> Stain Technology 55:143-148 and 55:151-160. Gregory's optimal
> fixative (which does not need to be aged) contains ethyl acetate.
> John A. Kiernan
> Department of Anatomy and Cell Biology
> The University of Western Ontario
> London, Canada N6A 5C1
> RSRICHMOND@aol.com wrote:
> > Tony Henwood has posted some information and a request for a reference
> > concerning Brazil's fixative. I append his entire post below.
> > Eleven years ago I corresponded with Arthur H. Cohen, M.D. in the Department
> > of Pathology, Division of Renal Pathology LAC-Harbor-UCLA Medical Center in
> > Torrance CA, and I still have the correspondence on file. I asked him:
> > I just read your article "Frozen-section analysis of allograft renal biopsy
> > specimens: reliable histopathologic data for rapid decision making" in Arch
> > Pathol Lab Med 1991;115:386-389; April 1991. You refer to your article
> > "Masson's trichrome stain in the evaluation of renal biopsies: an appraisal"
> > in Am J Clin Pathol 1976;65:631-643 which apparently contains your formula
> > for alcoholic Bouin's solution, in which you fixed renal biopsy specimens for
> > at least an hour before frozen section.
> > He replied: I am enclosing the recipe for alcoholic Bouin's fixative which is
> > also known as Dubosq-Brazil [Duboscq-Brazil] fixative. Stains commonly
> > employed in renal pathology (Masson's trichrome, periodic acid-methenamine
> > silver) come out especially well.
> > 80% ethanol 150 mL
> > 37% formaldehyde 60 mL
> > picric acid 1 g
> > Add 1 mL glacial acetic acid to 14 mL stock solution before use.
> > Leave in the fixative from 6 to 24 hours depending on the size of the tissue.
> > Transfer to 70% alcohol to await processing for light microscopy.
> > Reference: Histopathology Laboratory Procedures, National Cancer Institute
> > Samurai pathologist's note: To avoid handling solid picric acid, since
> > saturated aqueous picric acid contains about 21 g/L of picric acid, 30 mL of
> > it would contain about 600 mg. If one amended this formula to:
> > saturated aqueous picric acid 30 mL
> > absolute ethanol 120 mL
> > 37% formaldehyde 60 mL
> > one would get a mixture that contained about 60% of the amount of picric acid
> > present in the previous formula.
> > The only other reference that I [the Samurai Pathologist, that is] could
> > locate was in The microtomist's vade-mecum (Bolles Lee): a handbook of the
> > methods of animal and plant microscopic technique. 11th ed. Gatenby JB and
> > Beams HW, eds. Blakiston, Philadelphia, 1950. p. 325. This venerable
> > reference spell it Duboscq-Brazil.
> > Bob Richmond
> > Samurai Pathologist
> > Knoxville TN
> > ***************
> > My laboratory has used a fixative called BRAZIL'S FLUID for fixing tissues
> > where small amounts of glycogen are being sought. It is supposedly a good
> > fixative for glycogen. Unfortunately I have no reference for this fixative
> > nor its uses. The formula is as follows:
> > Absolute alcohol 1650ml
> > Formalin 600ml
> > Trichloracetic acid 7.5g
> > Picric acid 10g
> > Dissolve picric acid, then Trichloracetic acid in the alcohol, then add the
> > formalin.
> > Fixation time:-
> > Normal block size 3-4 mm thick 24 hours
> > Needle biopsy 3-4 hours
> > The only one I know of is Dubosq-Brasil fluid which is similar (an alcoholic
> > Bouin's fluid):
> > ethyl alcohol 80% 150ml
> > picric acid 1g
> > immediately before use add:
> > Formol 35% 60ml
> > acetic acid, pure 15ml.
> > From: Bock, P.: "Romeis Mikroskopische Technik", 17th. ed., p. 98.
> > Would anyone have a reference for BRAZIL'S FLUID that they could share with
> > me?
> > Thanks muchly,
> > Tony Henwood JP, BappSc, GradDipSysAnalys, CT(ASC)
> > Laboratory Manager
> > The Children's Hospital at Westmead,
> > Locked Bag 4001, Westmead, 2145, AUSTRALIA.
> > Tel: (02) 9845 3306
> > Fax: (02) 9845 3318
> > http://www.histosearch.com/homepages/TonyHenwood/default.html
> > http://us.geocities.com/tonyhenwoodau/index.html
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