Re: H. Pylori /Alcian Yellow

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From:amos brooks <>
To:a i d a n s c h u r r <>
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    I dug this out of the histonet archives. I knew it was there because I
posted it :-) The procedure is pretty good, It stains the bugs blue on a
yellow background rather than blue on blue like the diff quick. I cut and
pasted the procedure from the archives below 'cause I didnt want to type it
all over again with all the talk of repetitive motion and all :-)
Amos Brooks
    1) Periodic Acid 1%
    2) Sodium Metabisulfite 5%
    3) Alcian Yellow 1% (filter this, its kind of gooey)
                --Poly Scientific item # s2423
    4) Toluidine Blue Stock Solution
                --0.1% Toluidine Blue (Poly Scientific #s2424)
    5) Sodium Hydroxide 3%
    6) Toluidine Blue Working Solution <--- this needs to be prepared fresh

                Toluidine Blue Stock Solution --- 0.5 ml
                Distilled Water ------------------ 50 ml
                Sodium Hydroxide -------------- 2 drops

    1) Dearaffinize and hydrate slides to distilled water
    2) Oxidize in 1% Periodic Acid for 10 min.
    3) Rinse well in tap water
    4) Place slides in Sodium Metabisulfite 5% for 5 min.
    5) Rinse in tap water for 2 min.
    6) Stain in Alcian Yellow 1% for 5 min.
    7) Rinse well in tap water
    8) Stain in freshly prepared Toluidine Blue Working Solution for 3 min.

    9) Rinse well in tap water
    10) Dehydrate QUICKLY then clear and mount as routine

    Helicobacter pylori --- Blue
    Mucin ---------------- Yellow
    Background ---------- Pale Blue

    This Alcian Yellow is yucky stuff dont forget to filter it before using

it. Whatever you use to stain the Alcian Yellow step in will be stained
forever so
choose your glassware accordingly. The bugs should have a sharp contrast of

blue on yellow epithelium you should be pleased. Common screw ups are using

Periodic acid from the PAS stain which is a different concentration
usually. Also too much or too little Sodium Hydroxide in the Toluidine Blue


good luck
Amos Brooks

a i d a n s c h u r r wrote:

> Hi all,
> We have a new pathologist, and she has asked for an Alcian
> Yellow for H. pylori.   I cannot find any references for this in any of
> my textbooks.  Does anyone have a protocol for this, and if you do,
> is it actually any good?!  What are other folks out there in
> Histoland using to demonstrate H. pylori?  Our other pathologists
> just use an H&E.
> Thanks in advance...
> Aidan
> ___________________________________________________
> a i d a n   c   s c h u r r
>      mlso,  histology department
>       hutt valley health
>        lower hutt, new zealand
>      ph.  ++64 4 5709173
>      fax  ++64 4 5709214
> ___________________________________________________

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