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From:"McCollough, Carol" <> (by way of histonet)
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This is a previous response from Bob.  Hope it helps.  Welcome to Histonet,
Regards -
Carol B. McCollough, HT(ASCP)
Diagnostics & Histology Laboratory Manager
Maryland Department of Natural Resources
Fisheries Service
Cooperative Oxford Laboratory
904 S. Morris Street
Oxford, MD 21654

Subject:        Re: Genta Stain
Date:           Wednesday, November  6, 1996 21:57:46 EST

Here it is - I copied it out for somebody who wanted it - certainly haven't
tried it and don't intend to - of academic interest only, in the worst sense
of that abused word.

Bob Richmond
Samurai Pathologist
Knoxville TN

1. Sections of formalin fixed tissue are cut in the usual fashion,
deparaffinized, and rehydrated to 	distilled water.
2: Sensitize: 1% aqueous uranyl nitrate (uranium nitrate) for 3 minutes.
3. Rinse thoroughly in distilled water.
4. Silver stain: 1% aqueous silver nitrate in distilled water, beginning at
room temperature, and 	heating in the microwave oven to just below the
boiling point (do not boil).
5. Rinse in three changes of distilled water.
6. Rinse in two changes of 95% alcohol.
7. Rinse in two changes of 100% alcohol.
8. Place in 2.5% gum mastic in distilled water for 5 minutes.
9. Air dry for one minute.
10. Rinse in two changes of distilled water.
11. Reduce in REDUCING SOLUTION in a 45oC. water bath for 10 to 15 minutes,
until sections 	develop satisfactorily, with black or brown bacteria on a
light yellow background. (Check 	development with a microscope.)
12. Rinse thoroughly in distilled water.
13. Stain in ALCIAN BLUE pH 2.5 for 10 minutes.
14. Rinse in running tap water.
15. Stain in Harris hematoxylin for 5 minutes.
16. Rinse in running tap water.
17. Dip quickly in acid alcohol.
18. Rinse in running tap water.
19. Blue in 0.25% ammonia water.
20. Rinse in running tap water.
21. Stain in eosin up to 5 minutes.
22. Rinse quickly in tap water.
23. Dehydrate, clear, and mount in resin.

REDUCING SOLUTION: 10 mL of 2.5% gum mastic, 25 mL of 2% hydroquinone, 5 mL
of absolute alcohol, and 2.5 mL of 0.04% silver nitrate. Do not filter.

Criteria used for the semiquantitative scoring of the density of
Helicobacter pylori:

0. None visible.
1. Rare scattered isolated organisms, or one or two small clusters in
surface mucus or in a pit.
2. Few scattered organisms. Bacteria can be found without difficulty, and
many 10x fields 	contain some. They are not crowded or in large
3. Organisms uniformly distributed on surface and in pits. Organisms usually
can be seen with no 	difficulty at 4x, and almost every 10x field
contains some. They are not crowded and do 	not form large clumps.
4. Very large numbers of organisms on most mucosal surfaces and pits, with
bacteria forming an 	almost continuous lining on mucous cells.
5. Organisms on entire mucosa, with no free areas.

Robert M. Genta MD. George O. Robason HT(ASCP). David Y. Graham MD. (Baylor
in Houston, at VA center there). Simultaneous visualization of Helicobacter
pylori and gastric morphology: a new stain. Human Pathology, March 1994:

-----Original Message-----
From: Decarli, Terri []
Sent: Thursday, November 04, 1999 12:59 PM
To: ''
Cc: Decarli, Terri

I am looking for a procedure for the Genta Stain sometimes used for H.
pylori.  I attended the Culling lecture at Providence and it was discussed
but the procedure was not in the reference material.  Thanks for your reply.

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