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[Neuroscience] Re: Golgi stain

Bill via neur-sci%40net.bio.net (by connelly.bill from gmail.com)
Mon Jul 21 03:36:28 EST 2008


Are you sure you fixed the slices properly? i.e. is it possible you
just lysed the cells with the H2O?

On Jul 20, 9:06 pm, Hyunchul Lee <h... from medsci.usyd.edu.au> wrote:
> 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:h... from medsci.usyd.edu.au



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