I second what Gayle said.

Kidney biopsies from different patients develop the silver at different
rates.
So each kidney needs to be monitored for development. We don't use a
control. The patient's kidney IS the control (built in).

We find this individuality also happens with the trichrome stain. Some
patient's kidneys really stain red, others don't want to pick up any red.
And the same with blue. So each patient's kidney is individually
checked when we do the Masson trichrome.

(For the anticipated question - no, we've never had a problem
with CAP. We have a sheet in the front of our staining manual,
listing those stains in which the patient's tissue is the built in control,
and where to look at in the patient's tissue. Such as PAMS, look
at the glomeruli; Trichrome - blood vessel; PASH, if kidney look
at the glomerular loops, etc.)

One other thought for problems with silver staining PAMS on kidney
biopsies. We have found, if they let the kidney biopsies dry out, even
a little, before it is placed in the fixative, the silver staining will be
impaired. The H&E, PASH and Trichromes all look fine, but the
silver will NOT pick up, particularly around the edges (which seems
to be where many of the gloms want to hang out, for some reason).
So, when they are dividing the biopsy up into parts for EM, IM and
histology, they MUST keep the biopsy moist in a drop of buffer, such
as FA or PBS. Even a couple of minutes on a gauze pad will cause
problems with the PAMS stain. If the drying is minimal, than only the
outside edges  will not pick up silver. If the drying is more prolonged,
then the entire kidney will not pick up silver.

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



----- Original Message -----
From: "Gayle Callis" <uvsgc@montana.edu>
To: "Teri Johnson" <terij@prlnet.com>; <histonet@pathology.swmed.edu>
Sent: Wednesday, May 09, 2001 1:14 PM
Subject: Re: Jones PAMS


> Questions:
>
> Are you doing microwave heating to perform the silver staining?  Is the
> thickness of your section 1 - 2 um?
>
> My renal pathologist, when I first started doing these, said to develop
the
> stain for THE glomerular basement membrane, and you may have to do this
> carefully with microscopic examination.  If you are not in the habit of
> using a microscope for this stain, it is a good thing to do faithfully.
>
> Stain sections in hot methenamine silver solution for prescribed time per
> your protocol, rinse them in cold water, examine on microscope (keep them
> wet) and if the glomerular basement membrane is not dark enough, rinse the
> slide with HOT distilled water to equilibrate back to temp of methenamine
> silver heated solution, return to same, and develop a few minutes longer.
> I examined one slide from EVERY kidney biopsy slide I did, to insure
proper
> staining, and looked at the PATIENT'S section, not just the normal control
> - there can be differences in diseased kidney versus normal kidney, and
you
> need to see the patients glomerular basement membrane. Once this became
> practice, staining was consistent.
> Worked fine with either MW heated or waterbath heated MS solution.
>
> Make sure the silver solution is not funky, mirror like silver deposits on
> side of brown bottle indicate methenamine silver solution has gone bad.
> This must be stored in refrig, and made up less to have better turnover of
> silver solution to maintain fresh reagent. I also store silver nitrate
> (stock) in the refrig as it is a bit hygroscopic.
>
> I never reused periodic acid, made it up FRESH each time to insure best
> oxidation of the membranes. Its cheap and easy to weigh out, Culling
> advised this even for PASH.
>
> At 09:59 AM 5/9/01 -0500, you wrote:
> >Hi all!
> >
> >We have a pathologist who specializes in renal pathology, and our current
> >arsenal/panel consists of H&E, PAS, and Jones PAMS.  The Jones is a
tricky
> >little bugger. One day it will stain properly and correctly, the next day
it
> >stains only the elastic and tubules in the biopsy, not the glomerular
> >basement membrane. We've tried troubleshooting this and can find no
> >correlation to the periodic acid, methenamine, or the silver. Can anyone
out
> >there help us with their experience?
> >
> >-Teri Johnson
> >Physicians Reference Laboratory
> >Overland Park, KS
> >
> >
> >
> >
> Gayle Callis
> MT,HT,HTL(ASCP)
> Histopathology Supervisor
> Veterinary Molecular Biology
> Montana State University - Bozeman
> Bozeman MT 59717-3610
>
> 406 994-6367
> 404 994-4303 (FAX)
>
>
>