Re: staining (acid-alcohol after haemalum)
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From: | "J. A. Kiernan" <jkiernan@julian.uwo.ca> |
To: | Edna_J_Gonzalez/Powderject@powderject.com |
Reply-To: | |
Date: | Tue, 28 Sep 1999 11:40:28 -0400 (EDT) |
Content-Type: | TEXT/PLAIN; charset=US-ASCII |
On Tue, 28 Sep 1999 Edna_J_Gonzalez/Powderject@powderject.com wrote:
> in running tap water, I dip the slides two times in 1% HCl in 70% ethanol.
> At this point the slides will turn light red or pink, but now they are not
> turning to light red, they stay purple.
This means that either (a) your acid-alcohol is not acidic
enough, or (b) 2 dips is not enough to allow the acid-
alcohol to permeate the sections, change the colour, and
extract some of the alum-haematein. If your acid-alcohol
was freshly made from 70% (or 95%) alcohol and concentrated
hydrochloric acid explanation (a) is unlikely, so the first
thing to try is a few more dips in the acid-alcohol. You
should see some colour (red) being extracted. Then wash
in tap water (made slightly alkaline if necessary, to
change the alum-haematein colour to blue), and check with
a microscope to see if you have a pure nuclear stain. If it
is overstained, repeat the acid-alcohol treatment. If you
have extracted too much, go back to the alum-haematein and
try again.
Many people prefer a progressive nuclear staining, controlled
by the time in alum-haematoxylin, so that there is then no
need to overstain and remove the excess with acid-alcohol.
John A. Kiernan,
Department of Anatomy & Cell Biology,
The University of Western Ontario,
LONDON, Canada N6A 5C1
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