RE: Alcian Yellow for Helicobacter

From:"Jon C. Kerr" <> (by way of histonet)

Included is a protocol my institution uses.  The protocol was obtained from
the Stainsfile website, as referenced from the Journal of Histotechnology.
We actually use three stains for H. B. pylori: Giemsa, Alcian yellow, and
any of the thiazine dyes.  Each of our pathologist has his own flavor that
he likes, so we cut, slice, dice, and stain as per their desires.
Therefore, each is pleased with the stain he requests.  The Alcian yellow
does a fine job.

Jon Kerr
Reference Pathology Services
Sandy, Utah

Helicobacter pylori  - Alcian Yellow Toluidine Blue Stain (Leung and Gibbon)

Buffered formalin is satisfactory
Other fixatives were not tested.
Paraffin sections at 5 micrometers are suitable.

Step #
Deparaffinize and hydrate to distilled water
1	Xylene
2	Xylene
3	Xylene
4	Alcohol
5	Alcohol
6	Alcohol
7	Alcohol
8	Distilled water rinse for about 30 dips
9	Oxidize in solution A for 10 minutes
10 Wash well with running tap water
11 Treat with solution B for 5 minutes
12 Wash in running tap water for 2 minutes
13 Stain in solution C for 5 minutes
14 Wash well with running tap water
15 Stain in freshly prepared solution D for 3 minutes
16 Wash well with running tap water
17 Blot dry
Dehydrate in graded alcohol
11	Alcohol
12	Alcohol
13	Alcohol
14 Alcohol
Clear in clearing agent
15	Xylene
16	Xylene
17 Xylene
18 Mount

Helicobacter pylori: 	Blue
Mucin:			Yellow
Background:		Pale blue

Solution A
1. 1.0 gr. Periodic acid
2.	100 ml distilled water

Solution B
1. 5.0 gr. Sodium Metabisulphate
2. 100 ml distilled water
3. 1 ml 1 N hydrochloric acid

Solution C
1.	1.0 gr. Alcian Yellow
2.	100 ml 50% Ethanol
4. 3 ml glacial acetic acid

Solution D
1. 0.5 ml 1% aqueous Toluidine Blue
2. 50 ml Distilled water
3. 2 drops 3% Sodium hydroxide


This is most useful as a screening method.
Alcian yellow is a monoazo dye that stains similarly to alcian blue. After
oxidation gastric mucin is stained yellow. This cannot later be replaced by
toluidine blue. The final result is blue bacteria on a yellow background.


Leung, K., & Gibbon, K.J.
A rapid staining method for He

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