[Histonet] Question concerning H. pylori staining

Gudrun Lang gu.lang at gmx.at
Sun Apr 12 03:38:29 CDT 2020

I found this instruction for a Hp- stain, that sounds similiar to yours.
They want the slides to be airdried after water-rinsing and before xylen.
But the result should be blue bacteria, not purple.
I would try to let the slides air-dry for about half a minute, then rinse
very-very short in absolute ethanol (one dip). The colour should turn blue
and some dye will be extracted to give a clearer result. Then fast into
xylen, dip a few times, then coverslip.

I assume, that the periodic acid should render some mucins stainable with
Alcian yellow, to give a more contrasted result.

Gudrun Lang

-----Ursprüngliche Nachricht-----
Von: Bob Richmond via Histonet [mailto:histonet at lists.utsouthwestern.edu] 
Gesendet: Donnerstag, 9. April 2020 20:05
An: Histonet at lists.utsouthwestern.edu
Betreff: Re: [Histonet] Question concerning H. pylori staining

Michelle (where) asksi: >>We have a question about staining for H-Pylori
Using Quick Stain (Periodic acid 1%, Alician Yellow, Sodium Metabisulfate,
Toluidine Blue stock, Sodium Hydroxide) we notice what clearly  looks like
the H-Pylori purple stained clusters, but after dehydration in 100% alcohol
the purple clusters seem to disappear. Should we just dehydrate using a
slide oven instead of the alcohol? For how long at what temp? Could the
alcohol be affecting the purple color making it too light to see?<<

You identify Helicobacter pylori by its morphology - curved, angled, or
"gull wing" bacilli. Is that what you're seeing? If you do this stain, you
should know how it's interpreted.

The Alcian yellow (correct spelling) method is needlessly complex. What
does the periodic acid do? - A solution of toluidine blue (Diff-Quik 2 or a
generic equivalent) suffices - don't use Diff-Quik 1 with it - dehydrate
through alcohols rapidly.

Bob Richmond
Samurai Pathologist
Maryville TN
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