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MODIFIED WARTHIN-STARRY METHOD (pH 4.0) FOR SPIROCHETES AND OTHER MICROORGANISMS FIXATION: 10% buffered neutral
MODIFIED WARTHIN-STARRY METHOD (pH 4.0)
FOR SPIROCHETES AND OTHER MICROORGANISMS
FIXATION: 10% buffered neutral formalin.
SECTIONS: Paraffin, 6 micrometers.
SOLUTIONS
1% CITRIC ACID SOLUTION
Citric acid
Distilled water
1 .0 gm
100.0 ml
ACIDULATED WATER SOLUTION
Sterile water
Add enough 1% citric acid solution to reach a pH 4.0.
1 000.0 ml
1% SILVER IMPREGNATING SOLUTION
Silver nitrate
Acidulated water
1 .0 gm
100.0 ml
2% SILVER NITRATE SOLUTION
4
Silver nitrate
2.0 gm
Acidulated water
Place flask in a flotation bath set at 54°C.
100.0 ml
5% GELATIN SOLUTION
Gelatin
Acidulated water
Place flask in a flotation bath set at 54°C.
5.0 gm
1 00.0 ml
0.15% PYROCATECHOL SOLUTION
Pyrocatechol
Acidulated water
Place flask in a flotation bath set at 54°C.
0.15 gm
100.0 ml
DEVELOPING SOLUTION
Silver
nitrate solution, 2%
1 .5
ml
Gelatin solution, 5%
Pyrocatechol solution, 0.15%
3.75 ml
2.0 ml
Just before use combine in a small beaker in the order given. Mix the silver
nitrate solution and the gelatin solution thoroughly before adding the
pyrocatechol solution.
214
AFIP Laboratory Methods in Histotechnology
PROCEDURE 1. Deparaffinize and hydrate to distilled water. 2. Impregnate in 1% silver nitrate solution
PROCEDURE
1.
Deparaffinize and hydrate to distilled water.
2. Impregnate in 1% silver nitrate solution at 43°C for 30 minutes.
3. Place slides on staining rack using glass rods. Flood with the developing
solution. Allow color to develop to yellow brown.
4. Wash quickly and thoroughly in hot tap water to stop reaction.
5. Dehydrate and clear through 95% ethyl alcohol, absolute ethyl alcohol, and
xylene, 2 changes each, 2 minutes each.
6. Mount with resinous medium.
RESULTS
Background
Spirochetes and other microorganisms
tan to yellow
black
REFERENCES
Allen TC, Luna LG, Wear DJ. Modified Warthin-Starry technique. AFIP Letter.
1986;144:3.
Kerr DA. Improved Warthin-Starry method of staining spirochetes in tissue sections.
J
Am
Clin Pathol. 1938;8:63.
Bacteria, Fungi, and Other Microorganisms
215