Re: Brazil's fluid

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

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 

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 

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