BODIAN stain . Need information !

Richard Green green at biology.utah.edu
Mon Sep 25 12:37:21 EST 1995


In article <439e3e$18a2 at jurpool0.rz.uni-frankfurt.de>, 
oelschlaeger at em.uni-frankfurt.de says...
>
>We are working with the Bodian silver impregnation method
>but so far we have no optimal results. After several attempts 
>we need some advice in which direction to go. If you have got 
>some recipe or if you know somebody working with BODIAN, please 
>send us a message.
>Your help is highly appreciated.

The method we use (occasionally) in our lab is a modified form of that 
described in "A rapid silver impregnation method for nervous tissue:  A 
modified protargol-peroxide technic."  Loots, et al 1979, Stain 
Technology 54(2):97-100.  The protocol is as follows:

Solutions:

Impregnating Solution--
To 100ml dist. water add:
0.5 g Protargol (silver protein)--allow to dissolve.
.005-.010g allantoin
1 ml 1% Cu(NO3)2
1 ml 1% AgNO3
1-2 drops 30% H2O2

Dilute to 1/4 strength before using.

Hot Reducing Bath--
To 20 ml dist. water add:
.05 g hydroquinone
.05 ml 35% formalin

Cold Reducing Bath--
To 40 ml dist water add:
.25 g oxalic acid
.25 ml 35% formalin

Protocol:
1. Rinse sections 3 X 10 min in dH2O
2. Soak in impregnation soln for 15 min.
3. Rinse 5 min in dH2O
4. Reduce in hot reducing bath 5 min.
5. Rinse 3 X 5 min in dH2O
6. Tone in .125 % aqueous gold chloride
7. Rinse 2 X 5 min dH2O
8. Reduce in cold reducing bath 5 min.
9. Rinse 3 min in dH2O
10. Fix in 2% sodium thiosulfate
11. Rinse 3 X 5 min in dH2O

mount sections and dehydrate in EtOH series. Clear w/ xylene or 
clearing agents like Histoclear.  Results in brownish-red fiber 
staining.  Hope this helps.

Viel Glueck, Richard




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