[Histonet] Alizarin red staining
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 =
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% =
2. Absolute Isopropanol
3. 60% Isopropanol
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
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 ?
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