Re: mercury

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Elizabeth Hayes wrote:
> Hi,
>     Just wondering if there is a substitute for Zenker's in the Mallory's
> PTAH technique for fibrin and muscle.....that anyone uses or likes...

Two that we have used, each with their own hazard.


This is discussed in Ann Preece's book: "A Manual for Histologic
Technicians" 1972.

Deparaffinize and hydrate tissue to water.
Place slides in Bouin solution in 60 degree C. oven or waterbath
for 1 hour.
Wash in running water 1-10 minutes, until color is gone from
tissue (Time varies as to tissue.)
Stain as usual with PTAH.

Diane Ferber, HTL(ASCP) discovered this method when we were
students, and we had to do the PTAH stain for the ASCP registry
back in 1980. The usual post-mordant in saturated mercuric chloride
or in Zenker solution was giving us uneven staining, as was getting
a fresh piece of tissue and  fixing it in Zenker. We tried the
Bouin post-mordant, and got wonderful consistent results. We showed
the histotechs at William Beaumont Hospital, and they used it for
the next 15 years.

There is still the hazard of reactive/explosive picric acid.
And keep a lid on the coplin jar while in the oven, or
the fumes will be nasty to your eyes when you open the door.


3 g Potassium Dichromate (Cr2K2O7)
100 mL d. water
Dissolve together. Store at room temperature. Stable for 2-3 months.

100 mL 95% Ethanol (reagent)
10 mL Hydrochloric acid, conc. (HCl)
Slowly add hydrochloric acid, drop by drop, to ethanol. Mix
together. Store at room temperature. Stable for 2 months.

30 mL 3% Potassium Dichromate
10 mL Alcoholic Hydrochloroic Acid
Just before use, mix together, slowly adding the Alcoholic
Hydrochloric Acid to the Potassium Dichromate. Good for one day.

Deparaffinize and hydrate slides.
Place slides in Working Post-Mordant Solution at room temperature
for 5 minutes.
Wash in running tap water until all color is gone from tissue,
about 1-5 minutes (depends on type of tissue).
Stain as usual with PTAH.

Around 1995, one of my students, Mary Kalafus, HTL(ASCP)
wanted to get rid of picric acid in all the stains (her own
personal crusade), so was trying out various post-mordants
for trichromes and PTAH.

We tried the acidified potassium dichromate, as written in
an article by Abrams-Engeman, Karyn, HTL(ASCP): "The Milligan
Trichrome: An Easy, Reliable Staining Procedure"; Surgipath
MICROVIEWS, Vol. 7, Issue No. 2, Fall 1991.

We never got it to work on our Masson or Gomori Trichrome,
but it worked beautifully as a post-mordant for PTAH. We've
been using this method now at our hospital (Not that we get
that many requests any more, now with IM.)

There are hazards with the potassium dichromate. It is a
strong oxidizer, and must be stored separate from other
chemicals. It can cause lung cancer. It is poisonous, and
may be fatal if swallowed. It is a heavy metal, and may not
be allowed to be disposed of down the sink in your area.
Check with your local water treatment plant.

Hope these are of help. Like I said, everything still has
it's own hazard.

> thanks,
> Lib

Peggy A. Wenk, HTL (ASCP)
Anatomic Pathology
Wm. Beaumont Hospital
3601 W. 13 Mile Rd.
Royal Oak, MI 48073-6769

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