[Histonet] Steiner and Steiner method/H. pylori
Steven E. Slap
siksik03 <@t> comcast.net
Wed Dec 10 11:24:52 CST 2003
There certainly is a Wrathin Starry microwave method to demonstrate
spirochetes, and it does not involve anything radioactive.
Here is Joyce Moore's version, as it was taught to me:
Warthin Starry Techniques for Spirochetes
Technique: Sections cut at 4 microns
Control: Tissue containing Spirochetes
Acidulated WaterAcidulate 1 liter distilled water with 0.1 g citric
acid until pH of 3.8-4.4 is reached. A pH of 4.0 is ideal for
staining spirochetes. For demonstrating Donovan Bodies of granuloma
inguinale, a pH of 3.6 is recommended.
2% Silver Nitrate Solution (developer)Silver nitrate C.P. crystals
0.5 gAcidulated water 25.0 ml
1% Silver Nitrate Solution (impregnation)Silver nitrate C.P. crystals
0.5 gAcidulated water 50.0 ml
0.5% Hydroquinone SolutionHydroquinone crystals, photographic quality
0.35 gAcidulated water 25.0 ml
5% Gelatin SolutionGelatin 1.5 gAcidulated water 25.0 ml
1. Attach slides to automatic stainer, deparaffinize, hydrate to water.
2. Rinse in distilled water, 2 changes.
3. Place slides in 1% silver nitrate solution for 45 seconds in
microwave. Let stand for 1 minute at room temperature. (Note:
alternatively, in a laboratory microwave, heat at 80% power, 60 C for
5 minutes, no standing time required).
4. Preheat for 45 seconds in microwave in separate flasks: 2%
silver nitrate, 5% gelatin, 0.15% hydroquinone. Preheat empty flask
in microwave with these.
5. Mix developer solution in order given: Use warm empty flask:
2% silver nitrate 1.5 ml5% gelatin 3.75 ml0.15% hydroqinone 2.0 ml
6. When step 5 is completed, remove slides from silver solution.
Do not rinse. Place slides horizontally on a slide rack and cover
with developer. Allow sections to develop until they are light yellow
to golden brown, approximately 1 minute or less. (Note: developing
step can also be carried out in a laboratory microwave at 80% power
set for 60 C for 20 seconds).
7. Rinse thoroughly in 50 ml tap water which is preheated to
approximately 56 C in the microwave at 450W for 45 seconds.
8. Rinse in distilled water.
9. Attach to automatic stainer, dehydrate and clear. Mount in a
xylene soluble mounting medium.
Spirochetes - blackBackground - pale yellow to light brown
Adapted for the Jefferson Regional Medical Center Histo-Path Laboratory
At 4:30 PM -0500 12/9/03, Derek and/or Lynda Leopold wrote:
>Would anyone care to comment on the Steiner and Steiner (microwave)
>method for spirochetes? I susupect my lab's pathologists are
>interested in it for viewing H. pylori, yet in all the "HP" talk on
>this list I have yet to hear it mentioned. How does it compare from
>a tech safety standpoint? As a brand-new tech, I still do crazy
>things like read the safety warnings on ingredients, and that whole
>uranyl nitrate radioactive thing is sort of bothering me. We use no
>protective equipment when doing this stain (which the doctors still
>call "Warthin Starry", incidentally, but hey---they're making the
>I'll take anyone's 2 cents...
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
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