[Histonet] Alizarin red staining

Tony Henwood AnthonyH <@t> chw.edu.au
Sun Aug 20 18:10:39 CDT 2006

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:     5=A6=CCm paraffin sections.


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


Tissue containing calcium.

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


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 =

4.	Rinse in 60% Isopropanol, 30 seconds.

5.	Rinse in absolute isopropanol, 30 seconds.

6.	Rinse in xylene and coverslip.


Calcium - orange red. =20
The calcium deposits stained with Alizarin Red S are also birefringent.


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 <@t> lists.utsouthwestern.edu =
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of pex
Sent: Friday, 18 August 2006 8:56 PM
To: Histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Alizarin red staining=20

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


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