Re: Isolating dendrites in the hippocampus

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From:"J. A. Kiernan" <>
To:Histonet <>,
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On Wed, 10 May 2000 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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