Here's what worked for us - Out of the old Ann Preece book.

Fix as usual in formalin, and process as you would for similar size tissue.

Section 4-5 um (I always vote for thinner, when working on registry
material) and place on a subbed slide. (Charged, poly-L-lysine, etc.) Helps
to hold it on through the rest of the steps. Dry in 60 degree oven for about
an hour (helps to hold the tissue on the slide during the rest of the
steps).

After deparaffinization and running down to water, post-fix in Bouin for 1
hour in a 60 degree C. oven or waterbath. Then wash in slowly running cold
water for about 10 minutes, or until all the yellow is gone. (Yes, just like
post-fixing to do a trichrome.)

Then, stain as usual with the PTAH.

Should work great, nice intense colors with very little splotching, and
sectioning should be your usual formalin quality. No mercury disposal to
worry about.

This is assuming, of course, that the PTAH solution is still good and the
procedure was followed correctly.

Now - as far as any PTAH questions on the written/computer test - the
correct answer to fixation/post-fixation for PTAH is "Zenker". As that's
what is written in all the rest of the histo books.

Hope that helps.

Peggy A. Wenk, HTL(ASCP)SLS
William Beaumont Hospital
Royal Oak, MI 48073

----- Original Message -----
From: "UniPath IHC" 
To: 
Sent: Thursday, August 12, 2004 2:13 PM
Subject: [Histonet] Mallory's PTAH


I actually already tried to use formalin-fixed muscle.  You're exactly
right, it sectioned great, but I could never get any decent staining.  I
tried post-fixing for 24 hrs. in Zenker's vs. post-fixing for 3 Hrs in
Zenker's at 56 degrees.  I tried all three protocols in Sheehan and the
protocol in Carson.  I'm at a loss for what to do next if I can't make the
Zenker fixed muscle work.  At least I've learned a lot about the stain.  :-)



Brianna Jackson, BS, QIHC

UniPath, LLC

Denver, CO



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