Re: pyramidal cell staining

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From:"J. A. Kiernan" <> (by way of Marvin Hanna)
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On Wed, 26 Jan 2000, Mary Georger wrote:

> We have been having difficulty with a very old technique used for
> staining pyramidal cells, Golgi Cox. We have used this technique for
> over 50 years and I have to guess that there must be a better way to go.

    If you have been unhappy with it for 50 years, it's surprising
    that you didn't give up and try something else in 1951!

> Does anyone have a suggestion for a stain to demonstrate pyramidal cells
> in the cerebral cortex that can be used with formalin fixed tissue that
> has already been processed in paraffin?

   Briefly, either No or Yes.  No if you want to see the dendritic
   branching details.  Yes if you need to see only the cell bodies
   of the neurons.

   The original (1873) and "rapid" (?1880s) Golgi methods and Cox's
   important variant (?1890s) can be done only on whole pieces of
   CNS tissue. There are more recent techniques that don't need
   special primary fixatives; they will work after neutral formaldehyde,
   for example. I can commend the technique of Gibb & Kolb, 1998
   (J. Neurosci. Methods 79(1): 1-4), with which a graduate student
   working in my lab generated thousands of well impregnated sections
   in 1977-1999. It's a great improvement on the earlier methods that
   required nitrocellulose embedding.

   Bear in mind that a Golgi preparation shows only a small proportion
   (perhaps 1%) of the neurons, but with all their dendritic branches.
   Axons are generally not impregnated, especially with Golgi-Cox.

   If all you need to do is distinguish cortical pyramidal neurons
   from interneurons and glial cells, this is easy. Stain sections with
   a basic (cationic) dye. Toluidine blue, thionine and neutral red
   are all good; many people like cresyl violet, and there are lots
   of others. A blood stain (Giemsa or Wright) does a pretty good
   job. H & E doesn't. With these methods, pyramidal cell bodies (all
   of them) appear as triangular objects, but the dendrites and their
   branchings are invisible.

 John A. Kiernan,
 Department of Anatomy & Cell Biology,
 The University of Western Ontario,
 LONDON,  Canada  N6A 5C1

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