[Histonet] jones basement membrane stain

[Bell, Mandy]

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
 
 
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