[Histonet] Re: stains for myocardial injury

[histology <@t> gradymem.org]

Here is a copy of the procedure we used to do.
Angie


MYOCARDIAL ISCHEMIA STAIN
(Hematoxylin-Basic Fuchsin-Picric Acid Method)


PRINCIPLE:
The H & E method does not reveal unequivocal morphologic changes in 
cases of myocardial infarction until 15 to 20 hours have elapsed after 
the onset of the injury.  The HBFP technique detects early changes of 
myocardial infarction by coloring affected fibers red.  Normal and 
frankly ischemic myocardium colors yellow.  The differential staining 
results are believed to be caused by an unstable protein complex 
present in the early phases of myocardial ischemia.

SPECIMEN:
Any well fixed tissue (10% neutral buffered formalin) may be used.
Paraffin embedded tissue is sectioned at 4 microns.

REAGENTS:
Solution A   Alum hematoxylin.
			Ammonium aluminum sulfate       6 gm
			Hematoxylin			0.5 gm
			Yellow mercuric oxide		0.25 gm
			Mix these 3 ingredients in 70ml of distilled 
water.  Boil 10 minutes.  Cool and add 30 ml of glycerol and 4 ml of 
glacial acetic acid.  Filter before use.

Solution B   Basic fuchsin
			This is a 0.1% solution in distilled water.

Solution C   Picric acid
			This is a 0.1% solution in absolute acetone.


STAINING PROCEDURE:
1.  Deparaffinize and hydrate to distilled water.
2.  Stain in solution A for 10 seconds.
3.  Wash in running tap water for 5 minutes.
4.  Stain in solution B for 3 minutes.
5.  Rinse briefly (5-10 seconds) in distilled water.
6.  Rinse briefly (5-10 seconds) in absolute acetone.
7.  Differentiate in solution C until the red color ceases to run off 
the section--usually about 20 seconds . 
8.  Rinse briefly (5-10 seconds) in absolute acetone.
9.  Clear in Americlear.
10. Mount with Mounting Medium.






RESULTS:
Acutely ischemia myocardium--crimson red
Normal myocardium--light brown
Nuclei--blue-purple
Red blood cells, fibrin, plasma, proteins, elastic fibers,    
	collagen--red
Note:  The irregular fuchsin staining of myocardial cells along the 
edge of the section is considered an artifact.

REFERENCES:
Sheehan, Dezna C.:  Theory & Practice of Histotechnology.
	Second Edition, 1980, C. B. Mosby Company.






Angie Barnett, HTL(ASCP)
Grady Memorial Hospital
Pathology Department
405/224-2258
histology <@t> gradymem.org


----- Original Message -----
From: RSRICHMOND <@t> aol.com
Date: Wednesday, August 1, 2007 1:37 pm
Subject: [Histonet] Re: stains for myocardial injury
To: histonet <@t> lists.utsouthwestern.edu

> I remember reading articles about the hematoxylin / basic fuchsin 
> / picric 
> acid stain for recently infarcted myocardium, some time late in 
> the Ordovician 
> Period I think. 
> 
> I've never seen one, but there was a question about the stain on 
> the anatomic 
> pathology board examination (American Board of Pathology) when I 
> took it in 
> November 1971. I remember it because it was the only question on 
> anything 
> contemporary that was on the exam.
> 
> Gayle Callis, if you post the procedure in electronic form, I'd 
> like to have 
> a copy. I can't remember whether it's done from neutral buffered 
> formalin or 
> whether it requires a special fixative. The Harris (mercuric 
> oxide) hematoxylin 
> could make a difference - it does in the Engel-Cunningham variant 
> of the 
> Gomori trichrome stain, used for frozen sections of skeletal 
> muscle. 
> 
> Bob Richmond
> Samurai Pathologist
> Knoxville TN (actually in Harlan KY this week)
> 
> 
> 
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