[Histonet] Alizarin Red staining

From:"Tamara Franz-Odendaal"

I routinely do Alizarin red staining for bone in embryos. The protocol is very simple.  Some steps are only required if you need to bleach pigment (skin or eye pigment) or digest surrounding tissue from your specimens.  Essentially, after fixing, place samples in water overnight, then into saturated tetraborate (Borax) for 12 hours, then into 1% KOH with alizarin red (1mg/ml) overnight, then water to rinse out excess staining (30min).  Then into a graded series of 1% KOH with glycerol (80:20), (50:50), (20:80) until pure glycerol.
 If you need to bleach or digest tissue you need to add a hydrogen peroxide and trypsin step. I presume you are doing you staining on whole tissue pieces and not sections.  


Tamara Franz-Odendaal, PhD.
Post-doctoral Researcher
Biology Dept., Dalhousie University
Halifax, NS, B3H 4J1
902-494 3335
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