RE: [Histonet] Alizarin red staining

From:"Tony Henwood"

The following works well:

Alizarin Red S for Calcium

This technique allows the direct demonstration of insoluble calcium cations where Alizarin Red S forms an insoluble orange red lake.  The technique is made specific by pH control (Garvey et al 1989).

Alizarin Red S also forms lakes with other metals such as barium, aluminium, mercury and magnesium. Fortunately, the bright red colouration of calcium can be easily differentiated from the dark red of other metal-lakes.

Fixation:      10% buffered formalin.

Microtomy:     5m paraffin sections.

Solutions:

1.	1% aqueous Alizarin Red S (CI 58005) adjust pH (to 4.2) with 10% ammonium hydroxide.

2.	Absolute Isopropanol

3.	60% Isopropanol

4.	Xylene

Controls:

Tissue containing calcium.

Negative Control - place for 20 minutes in citrate buffer pH 3-4 and wash well before staining.

Technique:

1.	Deparrafinise sections and hydrate.

2.	Rinse slides in distilled water.

3.	Place in Alizarin Red S solution 1-5 minutes, controlling with the light microscope until the deposits are orange-red in colour but not diffuse.

4.	Rinse in 60% Isopropanol, 30 seconds.

5.	Rinse in absolute isopropanol, 30 seconds.

6.	Rinse in xylene and coverslip.

Results:

Calcium - orange red.  
The calcium deposits stained with Alizarin Red S are also birefringent.

Regards

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
The Children's Hospital at Westmead,
Locked Bag 4001, Westmead, 2145, AUSTRALIA.
Tel: 612 9845 3306
Fax: 612 9845 3318




-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of pex
Sent: Friday, 18 August 2006 8:56 PM
To: Histonet@lists.utsouthwestern.edu
Subject: [Histonet] Alizarin red staining 


Hello,
   
  I would like to perform Alizarin Red staining for mineralization nodules.
  Can someone do me a favor to show your protocol about it ?
   
  Thank you.
   
  Guofeng 

 		
---------------------------------
 =D1Ż-3.5G20M
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