Alcian Blue stains for Barrett's esophagus
OK, since our dear Samurai Pathologist dragged me into this, here's
my 2 cents' worth (thanks for the plug, Bob!).
The basis of any procedure for Barrett's esophagus is to
differentiate neutral (gastric-type) from acidic (intestinal-type)
mucus. As John Kiernan pointed out, AB + PAS is the standard method.
The typical sequence using Alcian blue first is possible today, at
least with Anatech's certified dye powder or stain solution. Color
differentiation is fine (for a color photomicrograph, see our
website: select Product Literature, then Innovators, then "Searching
for Helicobacter". The last page contains the picture;it is not
Barrett's esophagus, but it shows neutral vs. acidic mucus).
Actually, my favorite was Maxwell's procedure until Alcian yellow
disappeared. For a look at that, see our Innovator titled "Three
Dyes, Three Dilemmas". Maxwell stained first with AB (the example in
the Innovator used pH 1.0, but for Barrett's I would use 2.5-2.8).
This is followed with periodic acid then sodium metabisulfite then
Alcian yellow. We finished it off with a red nulear counterstain,
Brazilliant!. The color contrat between emerald green acidic mucus
and bright yellow neutral mucus is spectacular.
You can obtain the same results without Alcian yellow by using our AB
at 2.5-2.8; followed by 1% periodic acid for 10 min, then our Hp
Yellow stain. Hp Yellow is a yellow Schiff-like reagent (technically
pseudo-Schiff, but don't worry about that here). AB stains only the
acidic mucus. Periodic acid oxidizes nearly all mucus to their
aldehyde forms, which then react very specifically with Hp Yellow (or
regular Schiff's Reagent). Follow the Hp Yellow with a 10% aqueous
acetic acid rinse to remove ionically bound yellow dye. Counterstain
nuclei red for best contrast. Brazilliant! provides a Harris-type
crispness to the chromatin, which really enhances the picture.
Complete directions are included in the package insert for Hp Yellow.
Naturally the specimen must be properly fixed and processed for best
results.
If anyone has a control block of Barrett's esophagus I would love to have it.
Dick
--
Richard W. Dapson, Ph.D.
Anatech Ltd.
Battle Creek, MI
800-ANATECH (800-262-8324)
269-964-6450
email@anatechltdusa.com
Web address: anatechltdusa.com
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