[Histonet] jones basement membrane stain

[Lee & Peggy Wenk]

It sounds like maybe the kit is bad. Either the periodic acid isn't
oxidizing the carbohydrates on the basement membrane to aldehydes, or the
methenamine silver solution is bad, so the silver won't bind. Possibly there
is something wrong with the way the tissue was fixed or processed - not
likely, but weird things have happened before.

Some thoughts. First, I'm assuming you are using kits for many of your
stains. Try substituting solutions from other kits, to find out what your
problem is.

1. Get the periodic acid from the PASH kit, and use it instead of your
current periodic acid. Then use the methenamine silver solution from the
PASM kit. If the kidney stains now, then the original periodic acid was bad.

2. IF #1 doesn't solve the problem, get the methenamine silver solution from
the GMS (Grocott) kit. If it's like our procedures (we make them up from
scratch), the GMS methenamine silver solution will have about half the
silver nitrate in it as the PASM methenamin silver solution. So use the
periodic acid from the PASM kit, and substitute the methenamine silver from
the GMS kit. It might take longer than your usual time. But if it works,
then the silver solution was bad in the PASM kit.

3. Alternative, stain the section with a reticulin kit. If that doesn't
stain the reticulin and basement membrane, then your tissue is bad.

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

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Bell, Mandy
Sent: Saturday, August 16, 2008 12:41 PM
To: Histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] jones basement membrane stain

Hello.
I'm looking for any help on the Jones stain for basement membranes.  We're
having trouble getting the basement membranes to stain dark enough, if at
all.  Here is our protocol (we are using a kit from American MasterTech):
 
0.5% Periodic Acid  11min
rinse 2 changes distilled water
methenamine silver (in a waterbath @ 65-70 degrees)
   ****we have tried varying the time by 15 minute increments ranging from 1
hour to 2 hours, 15min
   (we're checking the slides microscopically every ten minutes once the
section is brown) rinse in distilled water 0.2% gold chloride 1 min rinse in
distilled water 3% sodium thiosulfate 2 min rinse in distilled water nuclear
fast red 4 min rinse dehydrate and clear
 
Any help/ insight would be much appreciated!   Thanks !
 
 
Mandy M Bell
Histology Department
 
Community Hospital of the Monterey Peninsula
 
831.625.4791
 
 
P Please consider the environment before printing this e-mail 

_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet