On Fri, 30 Mar 2001, Julie Heinrich wrote:

> I am terribly sorry to bother you- I have been looking through the 
> histonet web page and have found several helpful replies from you 
> about the Gibb & Kolb Golgi stain method.  I haven't managed to 
> figure out how to properly post a question there - 

You can't. It's a collection of old (archived) communications.
To ask or answer questions you have to subscribe to the
listserver. This is done by sending an email to
 histonet@pathology.swmed.edu  with the one word  subscribe
in the Subject line. Nothing else. You'll then get an automatic
reply from the listserver telling you all about it.

> I'm attempting to use the method on avian tissue. I have obtained 
> some decent looking tissue so far (though the stain is a dark 
> tan/brown, rather than the preferable black that I had expected) yet 
> after time the stain turns very 'grainy'. Within a matter of just 24 
> hours, the dendrites/spines look like collections of dots rather than 
> complete structures, and it gets worse with time.

> Do you have any idea why this might be happening? (I'm following 
> their protocol to the best of my knowledge, and use fresh solutions 
> each time I run tissue)

A graduate student here called Tim Ho did great numbers of Kolb
Golgis on rat brains a few years ago. He went to Kolb's lab in
Lethbridge, Alta for guidance. His initial problem was that the
unstained spaces between the black cells were pale green and
a bit granular. If I remember rightly, the washing after the
Golgi-Cox solution needed to be more thorough. 

Your problem is different, and may relate to the mounting medium.
Traditionally the sections were mounted in thick Canada balsam
without coverslips. A modern synthetic medium + coverslip can
be followed by fading, but I haven't heard of this happening
in 24 hours. 6 months, yes. Are you cutting vibratome sections 
of adequate thickness? I think you must be doing something wrong 
at or after the sectioning step, but don't know what. Sorry I 
can't be more helpful.  

----------------------------------------
John A. Kiernan
Department of Anatomy & Cell Biology
The University of Western Ontario
London,  Canada   N6A 5C1
   kiernan@uwo.ca
   http://publish.uwo.ca/~jkiernan