Re: Isolating dendrites in the hippocampus
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From: | "J. A. Kiernan" <jkiernan@julian.uwo.ca> |
To: | Histonet <histonet@pathology.swmed.edu>, WPDyckman@lbl.gov |
Reply-To: | |
Content-Type: | TEXT/PLAIN; charset=US-ASCII |
On Wed, 10 May 2000 WPDyckman@lbl.gov wrote:
> I would like to know how to best go about isolating and examining
> dendrite spines for atrophy (what stain is most appropriate, etc.)? We
> are cutting sections of perfused mouse brains, and are interested in
> shifting our focus from brain region volumes to the condition of the
> dendrites in the hippocampal region.
To look at dendritic spines you need a Golgi-Cox method, which
stains only a small proportion of the neurons, but shows the
stained cells in their entirity: black against a clear background.
There are many variants. A modern one that I can recommend is that
of R Gibb & B Kolb (1998) J. Neurosci. Methods 79(1): 1-4. The
thick sections are cut with a vibratome, and the lengthy
business of nitrocellulose embedding is avoided.
If your specimens have already been fixed in formaldehyde, you
will need a modification of the Golgi (as distinct from Cox)
technique, such as that of Gonzalez-Burgos & 2 others (1992)
Biotech. & Histochem. 5: 288-296. This ends up with thick
paraffin sections. For a very detailed account of a Golgi
method that involves plastic embedding, followed by light and
electron microscopy, see Alan Peters (1981) Ch.5 in Current
Trends in Morphological Techniques, ed. JE Johnson, Vol 1,
pp 187-212. Boca Raton: CRC Press. The original account of
this LM/EM method also gave very full technical instructions:
A. Fairen & 2 others (1977) J. Neurocytol. 6: 311-337.
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