Re: Fixation of rat testes
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From: | "J. A. Kiernan" <jkiernan@julian.uwo.ca> |
To: | "RODRIGUEZ, RICHARD" <RICHARD.RODRIGUEZ@spcorp.com> |
Reply-To: | |
Date: | Tue, 22 Jun 1999 13:13:27 -0400 (EDT) |
Content-Type: | TEXT/PLAIN; charset=US-ASCII |
On Tue, 22 Jun 1999, RODRIGUEZ, RICHARD wrote:
> We are looking for expertise in fixation or post fixation of rat
> testes in fixatives other than formalin. We are currently post-fixing
> our 3-month rat study testes in Bouin's, but we would like to
> eliminate the waste and rinsing in alcohols for this method. We have
> had suggestions to switch to Karnovsky's fixative (please correct the
> spelling if needed). We know nothing of this fixative and would like
> to find out more.
Karnovsky's fixative is a buffered formaldehyde & glutaraldehyde
mixture intended for electron microscopy. Glutaraldehyde does
not prepare specimens well for paraffin embedding but it would
be OK if you embed in plastic or if you cut froxen sections.
It interferes with some stains.
Both Bouin and Karnovsky contain plenty of formaldehyde. If you
want a fixative without any you might try Clarke's, which is
1 volume of glacial acetic acid plus 3 volumes of ethanol, or
Carnoy, which is ethanol 60, chloroform 30, acetic acid 10 parts
by volume. Both work very rapidly. You could expect a whole rat's
testis to be penetrated and fixed in 6 hours or so. You can
then take the specimens straight into 100% alcohol. For most
organs structural preservation and response to common stains
are better than in formaldehyde-fixed paraffin embedded material,
and much better than in objects that have been only partly
fixed (e.g. overnight) in formaldehyde.
John A. Kiernan,
Department of Anatomy & Cell Biology,
The University of Western Ontario,
LONDON, Canada N6A 5C1
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