Modified Triple Stain (Carbol Fuchsin/Alcian Blue/Hematoxylin-Eosin) for the Identification of Helicobacter pylori.
[Figure ILLUSTRATION OMITTED]
The stain is performed in our laboratory with a Sakura autostainer.
1. Deparaffinize and rehydrate to distilled water.
2. Place in carbol fuchsin (with 0.25% hydrogen peroxide) for 30 seconds to 2 minutes, as described above.
3. Rinse thoroughly in tap water for 2 minutes. Increase the wash time if the tissue is too dark.
4. Place in 1% Alcian blue in 3% acetic acid, pH 2.5, for 1 minute.
5. Rinse with tap water for 5 minutes.
6. Stain with Gill's hematoxylin for 7 minutes.
7. Rinse with tap water for 5 minutes.
8. Rinse with 0.25% ammonia water for 30 seconds.
9. Stain with eosin for 30 seconds.
10. Rinse with tap water.
11. Dehydrate to absolute alcohol, clear in xylene, and coverslip.
It has become increasingly recognized that evaluating sections stained only with H&E has a reduced accuracy for the detection of H pylori, and special stains have been recommended.24 Factors influencing the choice of a special stain include the time and expense of processing. The described triple stain uses commonly available reagents and can be performed on an autostainer, which has its advantage in general laboratories.
HALA M. T. EL-ZIMAITY, MD Gastrointestinal Mucosa Pathology Laboratory, Departments of Medicine and Pathology
Veterans Affairs Medical Center and Baylor College of Medicine
Houston, TX 77030
[1.] El-Zimaity HMT, Ota H, Scott S, Killen DE, Graham DY. A new triple stain for Helicobacter pylori suitable for the autostainer. Arch Pathol Lab Med. 1998;122:732-736.
[2.] Deltenre M, Glupczynski Y, De Prez C, et al. The reliability of urease tests, histology and culture in the diagnosis of Campylobacter priori infection. Scand J Gastroenterol Suppl. 1989;160:19-24.
[3.] Dixon MF, Genta RM, Yardley JH, Correa P. Classification and grading of gastritis. The updated Sydney System. International Workshop on the Histopathology of Gastritis, Houston 1994. Am J Surg Pathol. 1996;20:1161-1181.
[4.] Kolts BE, Joseph B, Achem SR, Bianchi T, Monteiro C. Helicobacter pylori detection: a quality and cost analysis. Am J Gastroenterol. 1993;88:650-655.
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|Author:||EL-ZIMAITY, HALA M. T.|
|Publication:||Archives of Pathology & Laboratory Medicine|
|Date:||Oct 1, 2000|
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