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

[John A. Kiernan]

The first thing you need to find out is whst's in
the kit. 

Is the method the one traditionally known as the
rapid Golgi method? This involves fixation in an
osmium tetroxide-potassium dichromate mixture, for
2 to 7 days (the time affects the outcome, but
unpredictably) followed by immersion in aqueous
silver nitrate for a few days, and preparing thick
celloidin or paraffin sections. There are plenty
of later variants, having in common a silver
chromate end product, but these should not be
called "the rapid Golgi method". The sections are
mounted in thick Canada balsam (the real stuff)
without coverslips. If a coverslip is applied, the
preparations fade with time. There are various
chemical tricks to prevent such fading.

The other major member of this group of methods is
Cox's, with mercuric chloride and potassium
dichromate, but no silver. The dark deposit in
this case is thought to be a mixture of mercurous
oxide and colloidal mercury. These methods are
often called Golgi-Cox, even though Golgi didn't
invent or use them. Ideally Golgi-Cox preparations
should also be mounted without coverslips, but
they will keep for a few years in DPX with
coverslips.

John Kiernan
Anatomy,  UWO
London,  Canada
____________________
John Fernandez wrote:
> 
> 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>
> 
> --------------------------------------------------------------------------------
> 
> 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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