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
