[Histonet] RE: Warthin Starry Help

[Roberta Horner]

I am also in a small lab and this is how I do our Warthin Starry.  I usually only have 1 or 2 slides to stain at a time and I stain them in a (brand new never before used for anything) plastic 5-slide mailer.  I mix the reagents in disposable 20 ml beakers.  I stain the slides using my flotation bath turned all the way up. 

I put the acidulated water in the water bath.  Then I measure out the Silver Nitrate in the mailer and mix it with the acidulated water then I measure the hydroquonine, gelatin, and the silver nitrate for the developer in the 20 ml beakers.  I add water to the gelatin only at this point an put in between the glass rods on the water bath so it will warm up and dissolve the gelatin.  When the impregnation with the silver nitrate is finished then I mix the hydroquinone and the developer silver nitrate.  I lay the slides on the rods over the water bath then flood the sections with the developer.  Rinse in hot water and dehydrate etc.

The only time I have had problems is in the winter.  My lab is poorly insulated and the water bath sets by the window so I take slide folders and make a wind break.

I hope this helps.  I have tried the microwave method and could never get it to work.

Roberta Horner HT/HTL
Animal Diagnostic Lab
Penn State University


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Campbell, Tasha M.
Sent: Tuesday, January 14, 2014 9:55 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Warthin Starry Help

I need help with warthin starry stain please.  I am a very small lab trying to save money so I want to make the reagents myself.  I have sigma Aldrich dry reagents and have been making the reagents and following the instructions correctly and I did get the stain to work wonderfully when I first started.  Then suddenly it quit working and I cannot get it to work again.  I have made fresh reagents day after day and I have a pH of 3.8 for the acidulated water.  The tissue will stain normally with the yellow to brown background and I the developing solution turns black when it is left on paper but the h.pylori organisms are not staining.  I am getting very frustrated and need some help please.

 

Thank you.

 

Tasha Campbell, HTL

tmcampbell <@t> fmh.org

 

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