Dear 'Netters,

	Here is the Warthin-Starry procedure we use, as posted by Kathy Wortham
on April 2, 1999.


STAIN: Warthin-Starry	

PURPOSE: To stain spirochetes

TECHNIQUE: Cut paraffin sections 4 to 5 microns.  Use control slide.

SOLUTIONS:   (USE ACID CLEANED GLASSWARE)

Acidulated Water

	One liter of triple distilled water acidulated with a weak solution of
citric acid (1% or less) to bring solution to a pH of 3.8 to 4.4.

2% Silver Nitrate	

	Silver Nitrate.................   2.00 gm
	Acidulated Water...............  100 ml

1% Silver Nitrate

	Silver Nitrate.................    1.00 gm
	Acidulated Water...............   100 ml

0.15% Hydroquinone

	Hydroquinone...................    0.15 gm
	Acidulated Water...............    100 ml

5% Gelatin
	
	Gelatin........................    5.00 gm
	Acidulated Water...............    100 ml

Developer Solution - Mix when ready to use

	2% Silver Nitrate..............   9.0 ml
	5% Gelatin.....................   22.5 ml
	0.15% Hydroquinone.............   12.0 ml

PROCEDURE:  (USE ACID CLEANED GLASSWARE)

	1. Depariffinize and hydrate to distilled water.

	2. Place in 1% Silver Nitrate, microwave 20 seconds on high. Let 
           stand 1 min. (Mix developer)

	3. Place slides in developer. Microwave on high for 15-20 
	   seconds. Immediately remove from microwave and stir           
constantly using a disposable transfer pipette. Remove from           
developer when tissues are light brown. Time of development           
10-30 seconds.

	4. Rinse quickly in HOT tap water (about 60 seconds).

	5. Dehydrate, clear, and coverslip with a synthetic mounting           
medium.

RESULTS:

Spirochetes..................black
Background...................pale yellow to light brown


REFERENCES:   AFIP, pp. 238-240; Sheehan, 2nd Edition, p. 240; Preece,
pp. 270-272; Thompson, pp. 1048-1049




Rae Ann Staskiewicz
Galesburg Animal Disease Lab
Galesburg, IL

	3.