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[Neuroscience] Re: Golgi stain

Hyunchul Lee via neur-sci%40net.bio.net (by hlee from medsci.usyd.edu.au)
Tue Jul 22 05:07:53 EST 2008


Dear Bill, thanks for your suggestion.

As embarrassing as it may be to think that it was something so simple- I think you're right.  I left another batch of sections in 
30% sucrose overnight, and they seem fine.

Sorry for causing a ruckus everyone... but I would like to know why people seem so worried about the Golgi product deteriorating...
and why many groups (though not all) go to the trouble of doing single-section Golgi staining on immunolabeled sections instead of
performing immunolabeling on Golgi impregnated sections.  Also, many investigators choose to section paraffin-embedded brains
and some papers seem to imply that this is important to maintain the integrity of labeling.  The number of different chemical tricks
used to preserve Golgi labeling, as well as techniques such as gold-toning and deimpregnation makes me wonder... what makes 
them necessary?

At least for now, I'm one happy dude!

 
Hyunchul Lee
PhD student

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




----- Original Message ----
From: Bill <connelly.bill from gmail.com>
To: neur-sci from magpie.bio.indiana.edu
Sent: Monday, July 21, 2008 6:36:28 PM
Subject: [Neuroscience] Re: Golgi stain

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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