[Histonet] Re: Golgi stain question/Rapid Golgi Stain (FD Neurotech)

[John Fernandez]

Hi all,

our lab has recently purchased and used the Rapid Golgi Stain Kit from FD
Neurotech, resulting in very impressive staining down to dendritic spine
level, however as Julie Heinrich found, within 24 hours this staining
becomes very granular with loss of distinct morphology that was otherwise
seen previously. It progressively worsens over time.

Has anyone successfully used the FD Neurotech Rapid Golgi kit? Or does
anyone know of a mounting medium that can be used to adequately stop the
reaction that is taking place? Any ideas on what could be causing this
progressive deterioration of staining would be very much appreciated.

Thanks in advance,

John


John Fernandez
Research Assistant
Department of Medicine
University of Melbourne
Austin & Repatriation Medical Centre
Heidelberg, VIC 3084
Ph: (03) 9496 3257



Re: Golgi stain question
From: "J. A. Kiernan" <jkiernan <@t> uwo.ca>

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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 <@t> 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 <@t> uwo.ca
   http://publish.uwo.ca/~jkiernan





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