Re: Basal bodies
|From:||"J. A. Kiernan" <email@example.com>|
On Thu, 18 Jan 2001, Karen Pawlowski wrote:
> I was wondering if there are any stains/methods that would allow the
> identification of basal bodies or centromeres in a non-dividing cell.
> I would like to determine the basal body position in a cell without
> resorting to serial TEM. My thinking is that I might be able to do
> it in whole cells with the right marker. Any comments?
This is classical cytology, so you need a classical method! Here
are a few suggestions.
1. Osmium tetroxide is a must, either in the primary fixative
or as a post-fix. A glutaraldehyde-osmium sequence, as for EM,
2. Plastic embedding followed by staining 1um sections with a
basic dye may be all you need. Otherwise, cut paraffin sections
as thinly as possible (The 19th century methods for centrosomes
were, according to McClung's Handbook, done this way, after
3. Try treating paraffin sections of postosmicated tissue with
ethyl gallate, which makes a more darkly coloured product at
sites of osmium binding. This method was used with great elegance
in a study of the hypothalamus and neurohypophysis by
D. G. Montemurro - J. Endocrinol. 35: 271-279 (1966).
4. Try Heidenhain's iron-haematoxylin method. With careful
differentiation this can make almost anything visible.
5. When you have found a good method let us all know about
it, by way of Histonet.
John A. Kiernan,
Department of Anatomy & Cell Biology,
The University of Western Ontario,
LONDON, Canada N6A 5C1
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