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)