H Pylori Alcian Yellow

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From:"A G DU TOIT" <AGDT@GERGA.SUN.AC.ZA>
To:HistoNet Server <HistoNet@pathology.swmed.edu>
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The method is on the StainsFile web page:
http://netbistro.com/~bryand
Look in the staining techniques under Alcian yellow.


ubject: Re: H. Pylori /Alcian Yellow

Hi,
    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
**********************************************************
REAGENTS
    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

PROCEDURE
    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

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

COMMENTS
    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

the
Periodic acid from the PAS stain which is a different concentration
usually. Also too much or too little Sodium Hydroxide in the 
Toluidine Blue
Hope that's what you are looking for.
Regards.
Andre du Toit




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