We use the same procedure listed below except that I dilute ALL of the
reagents (including the Silver solutions) in acidulated water.  I use
Citric acid to make the DI water acidic.  This is not done very
scientifically --- I put a few grains of citric acid in about 1 liter of
water.  Check the pH and add citric acid until I get pH 4.0.  After
deparaffinizing, sections are well rinsed in DI water straight from the
tap, then held in the acidulated water for 5 - 10 minutes.  If I'm too
busy to do the stain right away, I like to hold them in the acidulated
water until I'm ready.  I have found that using acidulated water is the
key to a good WS stain... at least in this lab it is. 
Also, keep your solutions fresh.  I like to prepare mine every 6 months.


Happy staining *g*
Connie McManus
Utah Veterinary Diagnostics Laboratory
Utah State University
Logan, UT
Phone:  435/797-1891
fax: 435/797-2805
email: convmcm@cc.usu.edu


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Mildred
Fail
Sent: Thursday, September 02, 2004 4:48 AM
To: histonet@lists.utsouthwestern.edu; Sarah.Blachford@ngh.nhs.uk
Subject: Re: [Histonet] Warthin Starey

Sarah,
      We use the following method with good results though the Steiner
is used more often some of our pathologists prefer the Warthin-Starry.
Solutions needed
1% Silver Nitrate 
2% Silver Nitrate 
0.15% Hydroquinone
5% Gelatin (use gentle heat to mix) 
All of the above are  stored at 2-8 degrees Celsius for up to 6 months.
!. Preheat a coplin jar with 1% silver nitrate in a 54-60 degree water
bath for 30 minutes. Place your gelatin bottle in the water bath at the
same time.
 2. Deparaffinize and hydrate slides to distilled water
3. Place slides in the preheated 1% Silver Nitrate and incubate for 30
minutes in a 54-60 degree water bath. At the same time place a vial of
1.5 mls of 2% silver nitrate, a vial of 3.75 5% gelatin, and a vial of 2
mls of 0.15% hydroquinone into the 54-60 degree waterbath to preheat.for
30 minutes.
4. Prepare the developer solution by mixing the solutions in the
following order. 1.5 mls of 2% silver Nitrate
          3.75mls of 5 % gelatin and 2.0 mls of 0.15% hydroquinone
5. Remove slides from heated silver (DO NOT RINSE) Place slides
horizontally on a paper towel and cover with developer. Allow sections
to develop until they are a light golden brown. Check microscopically
for end point
6. Rinse thoroughly in hot tap water
7. Dehydrate, clear and mount
 Rena Fail
 Medical University of SC
 


>>> "Blachford, Sarah - Histopath Main" 
09/02/04 05:27AM >>>

Can anyone help us.	
	
Has anyone got a reliable warthin starey method.  We have tried many
methods
but cannot get them to work.

Also what is the use of gelatin in the method


Sarah


Northampton General Hospital NHS Trust 
Cliftonville, Northampton NN1 5BD 

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