Re: RE H pylori

From:Tara Miller <>

The TB/AY stain is also known as the "rapid staining method". We buy the kit 
from Poly Scientific (and I know that they will send you a copy of the 
staining method of you call them.) A quick run down of the stain below is 
done by flooding the slides on a rack except for the last part.

1.Periodic acid 1% aqueous - 10 min.
2.Rinse in tap water
3.Sodium metabisulfite 5% aqueous - 5 min.
4.Rinse in tap water for 2 min.
5.Alcian yellow 1% - 5 min.
6.Rinse with tap water
7.Stain in a coplin jar in freshly prepared Tol. blue working solution (see 
below) for 3 min.
8.Rinse well in tap water
9.Blot slides dry, dehydrate quickly, clear and mount

Working Toluidine Blue:
50 ml distilled water
0.5 ml of stock 1% Toluidine blue aqueous
2 drops 3% Sodium hydroxide aqueous

Heliobacter pylori - blue
Mucin - yellow
Background - pale blue

A personal note: I don't know what the Alcian yellow from Poly Scientific is 
mixed with - water???  We were for a time making up most of our solutions, 
except for the alcian yellow and the toluidine blue, but the kit isn't that 
expensive and you really don't go through too much solution when you're 
dropping it directly on the slide.  Also, during dehyration we do 
95%,100%,100% just several dips each as this is where differentiation takes 

In the April 1999 issue of "Histo-Logic" several methods of H. pylori 
staining technics were compared and the "Rapid" method came out on top.

Poly Scientific

Tara Oakes, HT
Central Dupage Hospital

>From: (Flores, Teresa)
>To: Tara Miller <>
>Subject: Re: RE H pylori
>Date: Thu, 10 May 2001 09:21:38 -0500
>Tara, what is the procedure for preparing the TB/AB Stain?
>Thanks, Teresa
> >We use the toluidine blue/alcian yellow method. Our pathologists like how
> >the HP stand out blue against the yellow stained mucin. (And, it makes it
> >easier for us techs to check how well our control slide stained too.) 
> >are a few other labs that have started using this method after seeing our
> >slides. :)
> >
> >Tara Oakes, HT
> >Central Dupage Hospital
> >
> >
> >>To:
> >>Subject: RE H pylori
> >>Date: Tue, 08 May 2001 20:52:02 -0600
> >>
> >>Hello,
> >>The pathologist at our hospital would like to know what stain method the
> >>labs are doing for H pylori.
> >>
> >>Thanks for the feedback I'm sure to receive.
> >>
> >>Louise Yarrow MLT
> >>Foothills Hospital
> >>Calgary,Alberta, Canada
> >>
> >>
> >
> >_________________________________________________________________
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