This is a copy of the Warthin-Starry procedure we use in our laboratory.
Our pathologists prefer it over the Steiner. We have modified the
original procedure to include microwaving. This makes the staining much
better and a whole lot quicker!


STAIN:      Warthin-Starry
            
PURPOSE:    To stain spirochetes
           
TECHNIQUE:  Cut paraffin sections 4 to 5 microns.   Use control slides.
              

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 .................... 100.00 ml
                       
     1% Silver Nitrate
       Silver nitrate .....................   1.00 gm
       Acidulated Water .................... 100.00 ml
                         
     0.15% Hydroquinone
       Hydroquinone .......................   0.15 gm
       Acidulated water..................... 100.00 ml
                         
     5% Gelatin
       Gelatin ............................   5.00 gm
       Acidulated water..................... 100.00 ml
                         
     Developer Solutions - Mix when ready to use.
                                   
                                   
       2% silver nitrate ........  9.0 ml
       5% gelatin ...............  22.5ml
       0.15% hydroquinone .......  12.0ml
        

PROCEDURE:  (USE ACID-CLEANED GLASSWARE)
            
     1.  Deparaffinize and hydrate to distilled water.

     2.  Place in 1% silver nitrate, microwave 20     seconds. Standing
time 1 min..      
Mix developer.(mix immediatly before use)

     3.  Place slides in developer. Microwave on high for 15-20 seconds.
Immediatly 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 56oC).

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

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


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

Kathy Wortham
USDA, Food Safety Inspection Service
950 College Station Road
Athens, Ga 30605

Phone# 706-546-3556
E-mail  kjwortham@yahoo.com



--- "Dr. Naseem" <nasimas@khi.compol.com> wrote:
> Hi colleagues..
>      can someone tell me about the procedure &
> inference of warthin starry
> stain. we are not doin this stain in our lab. i wanna
> establish the WS stain
> in our lab.
> any suggestion is also welcomed
> Dr. Naseem Ahmed
> Karachi Paksitan
> 
> 
> 
> 




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