Re: Elastic stains and Myelin
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From: | Tony Henwood <henwood@mail.one.net.au> |
To: | HistoNet <HistoNet@Pathology.swmed.edu>, "Yanosy, Bonnie" <lbyano@bpthosp.org> |
Reply-To: | |
Date: | Tue, 23 Mar 1999 21:27:16 +0000 |
Content-Type: | text/plain; charset=US-ASCII |
> Date: Wed, 17 Mar 1999 11:44:00 -0500
> From: "Yanosy, Bonnie" <lbyano@bpthosp.org>
> Subject: Elastic stains and Myelin
>
> Hi Histonetters,
> Would any of you know of a published reference regarding the staining
> of Myelin by any Elastic tissue staining method. My Pathologist would
> appreciate any info. Thanks, Bonnie Yanosy
>
The following method may be useful:
WEIL'S MYELIN STAIN
NOTES:
Weil's stain is a modification for paraffin sections of the
Weigert-Pal-Kulschitsky technique for tissue blocks. The principle of
these methods involves reduction of chrome salt to chromium dioxide by
myelin, the chromium subsequently acting as a mordant for the
Haematoxylin.
FIXATION: 10% buffered formalin
MICROTOMY: 8um paraffin sections
SOLUTIONS:
1. Staining Solution:
a) Haematoxylin Solution:
Haematoxylin stock solution: 10g haematoxylin in 100ml
absolute alcohol. Allow to ripen naturally for 6
weeks.
Dilute haematoxylin solution 1 part with 4 parts of
distilled water for use.
b) Iron Alum Solution:
4% aqueous ferric ammonium sulphate.
Mix equal volumes of preheated (56-60 oC) solutions a) and b),
immediately prior to use.
2. 2% aqueous ferric ammonium sulphate (differentiating solution).
3. Weigert's Differentiator:
Borax (sodium tetraborate) 2g
Potassium ferricyanide 2.5g
Distilled water 200ml
CONTROL: Spinal cord.
STAINING PROCEDURE:
1. Dewax and hydrate sections to distilled water.
2. Freshly prepared staining solution at 56-60 oC (slide drying
oven), 30 minutes.
3. Wash well in water.
4. Partially differentiate in iron alum differentiating solution,
until myelin sheaths stand out blueish-black on a pale grey
background, approx, 5 minutes. (If unsure, check sections at
1 minute intervals).
5. Wash in tap water, 10 minutes.
6. Complete differentiation in Weigert's differentiator, 1 to 2
minutes. Control this differentiation step carefully,
microscopically, until the myelin is an intense deep blue against
a creamy background.
7. Wash well in tap water.
8. Dehydrate, clear and mount.
RESULTS:
Myelin blue-black
Red blood cells black
Nuclei blue
Background cream
Tony Henwood
www2.one.net.au/~henwood
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