Re: [Histonet] Steiner and Steiner method/H. pylori
Re: [Histonet] Steiner and Steiner method/H.
There certainly is a Wrathin Starry microwave method to
demonstrate spirochetes, and it does not involve anything
Here is Joyce Moore's version, as it was taught to me:
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
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
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 big bucks....)
I'll take anyone's 2 cents...
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