Re: pyramidal cell staining
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From: | "J. A. Kiernan" <jkiernan@julian.uwo.ca> (by way of Marvin Hanna) |
To: | histonet@histosearch.com |
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Content-Type: | text/plain; charset="us-ascii" |
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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