[Neuroscience] Golgi stain

Hyunchul Lee via neur-sci%40net.bio.net (by hlee from medsci.usyd.edu.au)
Sun Jul 20 04:06:50 EST 2008

Hi, I need help... 

I've been trying out a Golgi impregnation protocol- it was my first try and it wasn't great, but it worked...
in a tiny corner of my brain anyways... I'd cut it on a freezing microtome.  After having a quick look and taking
a couple of photos, I decided to leave them in distilled water overnight.

I came back the next day, and mounted them, only to find that the staining had vanished with only brown blobs dotting
where the cell profiles used to be... so here's a question- does Golgi staining dissolve in water?  I was thinking of
double staining with immunohistochemistry, but obviously, if the Golgi reaction product dissolves in water in a space of
only a few hours, this is impossible- yet I HAVE heard of others doing immunocytochemistry on Golgi-labeled sections...

Also- a reddish brown precipitate (which I guess is the silver chromate reaction product of the Golgi stain) forms a coat
around my brain, and little label happens deep within the brain (I'm staining tiny black six mouse brains)... is this normal?
I am washing my brain for 30 min in distilled water after the chromation step.

BTW, here's the protocol I'm using:
Gonzalez-Burgos et al., 1992. Golgi method without osmium tetroxide for the study of the central nervous system. Biotechnic & Histochemistry 67 (5):288-296

Thanks for any helpful suggestions! 
Hyunchul Lee
PhD student

SystemsNeuroscience Laboratory
N121 Anderson Stuart Bldg.(F13)
The University of Sydney , NSW, 2006
Email:hlee from medsci.usyd.edu.au


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