AW: [Histonet] jones basement membrane stain

[Gudrun Lang]

Hi Bell,
You can try to prolong the incubation time untill the coplin jar turns
black. Also the slides will get a shade of silver. With our protocol this is
usually about 60-75 min in 60°C oven. (Best practice is to catch the moment
just before the blackening of the glass begins)
If this "breaking" of the silver-solution doesn't occur, I would think, that
it is ineffective.
Then don't wash the slides too excessive in water. Cleaning the slides back
with paper towel is helpful.
Goldchlorid turns the brown silver into black. Our protocol says 10 min in
0,1% Goldchloride.

Hope this helps
Gudrun Lang
 
Biomed. Analytikerin
Histolabor
Akh Linz
Krankenhausstr. 9
4020 Linz
+43(0)732/7806-6754
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Von: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] Im Auftrag von Bell,
Mandy
Gesendet: Samstag, 16. August 2008 18:41
An: Histonet <@t> lists.utsouthwestern.edu
Betreff: [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
 
 
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