RE: [Histonet] Alternatives to Weigert's Iron Hematoxylin for Mucin andTrichrom e Staining

From:"Tony Henwood"



You could try the celestine blue-haematoxylin sequence:
This method uses a Modified Harris's haematoxylin but any alum haematoxylin (routinely used in your H&E staining) should work.
Document Procedure:	
The Celestine Blue - Alum Haematoxylin Procedure

Principle:	
Celestine blue resists acid decolourisation and the ferric salt included acts as an additional mordant for the Haematoxylin to provide a reasonably acid resistant nuclear stain.	

Fixation:      10% buffered formalin.
Microtomy:     paraffin sections at 5Ám.

Reagents: 	
1.	Celestine Blue Solution:
		0.5% Celestine Blue (CI 51050) in 
		5% iron alum (ferric ammonium sulphate).

2.   Harris's Haematoxylin Solution.

Procedure:
1.	Deparaffinise and take sections to water.
2.	Celestine blue solution, 5 minutes.
3.	Rinse in tap water.
4.	Harris's Haematoxylin solution, 5 minutes.
5.	Wash in water.
6.	Differentiate in acid alcohol, as necessary.
7.	Wash in water.
8.	Blue in blueing solution.
9.	Wash in water.
10.	Rinse in distilled water.
11.	Proceed with required staining technique.
Reference:	 
Bancroft, J.D. and Stevens, A. (ed):  Theory and Practice of Histological Techniques, first ed.  Churchill Livingstone, Edinburgh, 1977, p 88.

Henwood, T., Llewellyn, B., Montgomery, I., Nader, A., Rittman, B.R., (2007) "Celestine blue and hemalum for staining nuclei" Biotechnic & Histochem 82(3):167-8.


Regards

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
Tel: 612 9845 3306
Fax: 612 9845 3318
the children's hospital at westmead 
Cnr Hawkesbury Road and Hainsworth Street, Westmead 
Locked Bag 4001, Westmead NSW 2145 




-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of McKnight, Tanisha
Sent: Saturday, 9 August 2008 3:26 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Alternatives to Weigert's Iron Hematoxylin for Mucin andTrichrom e Staining
Importance: High


 Hello All:

Can anyone suggest an alternative to Weigert's Iron Hematoxylin for nuclei staining with Mucin and Trichrome protocols? I have some standard hematoxylin (7211) that I will be using for my routine staining. However, with the shortage, Weigerts is on backorder until October. 


Tanisha McKnight, HT(ASCP)
AP-Histology/Specimen Management
Covance CLS, Indianapolis


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