Re: Jones PAMS
|From:||firstname.lastname@example.org (Teri Johnson)|
Yes, our sections are 2 microns (more or less, you know). And we always use
the microscope to determine endpoint. I was given a suggestion to only make
up 100 ml of the methenamine, and we might try that. I also think it's a
good idea to make up the periodic acid fresh every time.
I'll try some of the techniques sent here, and I thank you all for your
Physicians Reference Laboratory
Overland Park, KS
----- Original Message -----
From: Gayle Callis <email@example.com>
To: Teri Johnson <firstname.lastname@example.org>; <email@example.com>
Sent: Wednesday, May 09, 2001 12:14 PM
Subject: Re: Jones PAMS
> 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
> 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
> staining, and looked at the PATIENT'S section, not just the normal control
> - there can be differences in diseased kidney versus normal kidney, and
> 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
> >little bugger. One day it will stain properly and correctly, the next day
> >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
> >there help us with their experience?
> >-Teri Johnson
> >Physicians Reference Laboratory
> >Overland Park, KS
> Gayle Callis
> Histopathology Supervisor
> Veterinary Molecular Biology
> Montana State University - Bozeman
> Bozeman MT 59717-3610
> 406 994-6367
> 404 994-4303 (FAX)
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