RE: [Histonet] Orcein

From:Margaret Blount

Dear Wen,

Here is the method from my old favourite staining book, reference; Manual of
Histological Demonstration Techniques H.C. Cook, Butterworths, 1974, ISBN 0
407 74700 1.

Note that Orcein dye batches vary in quality and as a general rule,
synthetic orcein is better than natural orcein.

Dissolve 1g orceing in 100ml of 70% ethanol with the aid of gentle heat
(waterbath!). Cool, filter and add 1ml of concentrated hydrochloric acid.

1. take sections to 70% ethanol
2. Stain with orcein for 1 to 2 housr at 37C
3. Rinse with 70% ethanol and differentiate in 1% acid alcoholif necessary
then wash well in water
4. Counterstain as required, suitable counterstains are Haematoxylin, 0.1%
aqueous azure A or methylene blue for 1 to 2 minutes. 
5. Dehydrate, clear and mount.
Elastin dark brown
Background according to the counterstain.

NOTE: when I used this method the elastin gave a weak pale brown stain and I
prefer the method in the following reference:

Roman et al. (1967) Orcein-Haematoxyoin in iodides ferric chloride as a
stain for elastic fibres with metanil yellow counterstaining. Stain
Technology 42: 199.

I ave attached a copy of my nethod card below. This is a nice method and
works very well, picking up very fine elastic fibres.

Good luck

-----Original Message-----
From: wen eng []
Sent: Tuesday, March 29, 2005 10:50 PM
To: histonet
Subject: [Histonet] Orcein

 I was asked to do Orcein stain for elastic fibres. I couldn't find method.
Could anybody tell me where I can find it or send me a copy? I really
apperciate any input.

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