We use 10% for 10 minutes as well, but after bleaching in sodium bisulphite
we place in 0.5% thiosemicarbazide for 5 minutes.  This has an aldehyde
blocking group at one end of its molecule and a very strong reducing agent
at the other, stronger than the aldehyde.  The result is a faster and more
contrasty impregnation in fungus staining as well as BMs.

Hayashi, I., Tome, Y. and Shimosato, Y., 1989.
Thiosemicarbazide used after periodic acid makes methenamine silver staining
of renal glomerular basement membranes faster and cleaner.
Stain Technology 64: 185-190.

Bryan Llewellyn

----- Original Message -----
From: "Lee & Peggy Wenk" <lpwenk@mail.netquest.com>
To: <Kimcatk@aol.com>; <histonet@pathology.swmed.edu>
Sent: December 8, 2000 2:26 AM
Subject: Re: GMS & Steiner Chapman / Re: Seeking Staining Procedures


> We use 10% chromic acid (aqueous) for 10 minutes, room temperature.
> We used to use 4% for 1 hour, room temp. But the 10% for 10 min. gave
> us the same results and saved us 50 minutes. This solution can be reused
> for months. When it starts to turn brown, instead of being orange, it's
> time to remake the solution.
>
> We tried the 1 minute microwave with the 4%, but we never liked it. The
> tissues tended to fall off the slides, and we found that the solution
would
> turn brown after microwaving just a couple of times so we were making
> new solution every week or so.
>
> Peggy A. Wenk, HTL(ASCP)
> William Beaumont Hospital
> Royal Oak, MI
>
> ----- Original Message -----
> From: <Kimcatk@aol.com>
> To: <histonet@pathology.swmed.edu>
> Sent: Thursday, December 07, 2000 1:47 PM
> Subject: GMS & Steiner Chapman / Re: Seeking Staining Procedures
>
>
> > Dear HistoNet colleagues,
> >      It is great to have HistoNet working again!  Thank you to those of
> you who responded to my posting.  I got some leads on staining for fungi
and
> spirochetes, but I still have some questions.
> >      Has anyone done Grocott's Methenamine Silver (GMS) without heating
> the chromic acid?  I heard that it is possible to oxidize in the chromic
> acid for an hour instead of heating it, but I do not have a procedure.
> >      Also, I am looking for a procedure for Steiner Chapman because that
> replaces the uranium/uranyl nitrate with some type of zinc solution.
> >      I would be very grateful for any help you could provide.
> >
> > Sincerely,
> >
> > Kimberly Atkin HT (ASCP)
> > Boston, MA
> >
> >
> > In a message dated Tue, 5 Dec 2000  8:52:00 AM Eastern Standard Time,
> Kimcatk writes:
> >
> > << Dear HistoNet colleagues,
> >
> >      I am seeking to replace some staining procedures in my laboratory
> with NON-microwave methods.  Optimally, I am hoping to find stains that
are
> free of heavy metals or any particularly toxic or dangerous components.  I
> can not use mercury.
> >
> >      I need to find staining protocols or kits to demonstrate the
> following:
> >
> > Fungal organisms
> > Spirochetes, Campylobacter, and Legionella organisms
> >
> >      In addition, the gram stain procedure I use is very cumbersome and
I
> would be extremely grateful for a simpler method.
> >
> >      Thank you all very much in advance for your help.
> >
> > Sincerely,
> >
> > Kimberly Atkin HT (ASCP)
> >
> > Boston, Massachusetts
> >
> >  >>
> >
> >
> >
> >
>
>
>