Thank you for the replies.

Eric Kellar also provided the entire procedure and references which I am
forwarding for anyone else who may be interested in it.

Bill Tyler


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From: "Kellar, Eric" <kellarec@MSX.UPMC.EDU>
To: "'wbtyler@webtv.net'" <wbtyler@webtv.net>
Subject: RE: Bosma Steiner stain
Date: Sat, 19 Dec 1998 13:46:34 -0500
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Bosma-Steiner Method:

Solutions:

-Uranium nitrate-mastix solution:
	Uranyl nitrate	2g
	Alcohol 100%	45 ml
	Mastix stock solution (Chroma art. 3D045)	20 ml
-Silver nitrate solution:
	Silver nitrate	0.7g
	DH20			70 ml
-Alcohol-Mastix:
	Alcohol 100%	60 ml
	Mastix stock 	40 ml
-Brenzcatechin solution:
Brenzcatechin	3.5 g
DH20			70 m
-Amylase solution:
	'alpha-amylase' type VI (Sigma #A6880) 0.7 g
	DH20			70 ml

Fixation:	10% formalin

Technique:	Paraffin sections cut at 3um, or plastic at 2um.

Staining procedure:

1.	Deparaffinize and hydrate to DH20.
2.	Place slides in plastic staining jar containing amylase solution at
37 degree C, heat in oven for 1 hour.
3.	Rinse slides briefly in DH20, place slides in alcohol 95% for 1 min
and in 100% for 2 min.
4.	Place slides in uranium nitrate-mastix solution for 5-10 min.
5.	Rinse in DH20 for 1-2 min.
6.	Place slides in a plastic staining jar containing 70 ml of silver
nitrate solution and irradiate at 450 W (*80%), at 90 degree C for 6-10 min.
Allow the sections to remain in the solution for at least 20 min. until the
proper staining intensity is achieved.
7.	Rinse slides briefly in alcohol 80%, rinse in alcohol 95% for 1 min.
8.	Wash in alcohol 100%, 3x2 min.
9.	Place slides in alcohol-mastix solution for 5 min.
10.	Place slides in a plastic staining jar containing 70 ml of
brenzcatechin solution and irradiate at 450 W, at 90 degree C for 6 min.
11.	Rinse slides in DH20 for 1 min.
12.	Dehydrate, clear and mount.

Note:	Uranyl nitrate is radioactive!

Result:	Spirochete:black
		Backround:yellow or brown-yellow

Reference:	Steiner G (1950) Am J Clin Path 20:489;
			Bosma R (1984) Histotechniek 3:161 (in Dutch); Elias
and Bosma (1987).

Eric Kellar
Histology/Immunohistochemistry
University of Pittsburgh Medical Center





	----------
	From:  wbtyler@webtv.net [SMTP:wbtyler@webtv.net]
	Sent:  Saturday, December 19, 1998 1:32 AM
	To:  HistoNet@Pathology.swmed.edu
	Subject:  Bosma Steiner stain

	Does anyone have a reference for the procedure for this modification
of
	the Steiner stain?  Is anyone using this procedure?

	Thank you.

	William B. Tyler MD
	Department of Pathology
	Geisinger Medical Center
	Danville, PA 17821
	Penn State Geisinger Health System
	Email:wtyler@psghs.edu