Re: [Histonet] jones basement membrane stain

From:"Bryan Llewellyn"

Make sure the methenamine silver solution is fresh and you have the correct 
amount of borax or buffer in it.  If that is so, try applying 0.5% aqueous 
thiosemicarbazide for ten minutes or so after the wash following periodic 
acid.  Then wash well with water, rinse with distilled water and place in 
the methenamine silver at 60C until done.  This usually speeds up the 
impregnation and increases contrast at the same time due to lighter 
background staining.

Don't let the name of the chemical be a red flag.  It is a semicarbazide not 
a metallic azide, and it is not explosive nor inflammable.  It is safe. 
This procedure is useful for Gomori's methenamine silver as well.

Bryan Llewellyn

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
dehydrate and clear

Any help/ insight would be much appreciated!   Thanks !

Mandy M Bell
Histology Department

Community Hospital of the Monterey Peninsula


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