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