[Histonet] Rapid Golgi

[Hyunchul Lee]

Hello,

I've been trying out this 'osmium-free' rapid Golgi protocol (not Golgi-Cox), and it seems to work-
However, I've been having a couple of problems...

After impregnation with silver nitrate (0.75%) solution, I immersed my brain in 30% sucrose for 24 hrs,
then sectioned it on a freezing microtome.  Out of my eagerness, I mounted a section and viewed it
under a microscope- I was quite impressed!  As it was getting late, I returned my sections to distilled water
so that I could mount them the follwing day.

Upon returning the next day, I mounted my sections.  I hadn't coverslipped them yet...
I viewed these again under the microscope and found that the staining I had seen earlier was nowhere to be seen!
All I could see were brown blotches roughly corresponding to where the perikarya were.
Google gave me the impression that others have been having similar problems regarding the longevity of
Golgi-stained sections.  Is there a way to stop this decomposition of rapid-Golgi product?
We don't have Canada-balsam though...

I had been hoping to perform immunohistochemistry on these same sections, and I have heard of people
immunolabeling sections AFTER Golgi labeling, yet if the Golgi product has such a short life-span in water,
this would be impossible.  Is there a way to stop this decay in rapid Golgi material?

Thank you in advance,

 
Hyunchul Lee
PhD student

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