[Histonet] Alizarin Red Troubleshooting

Phipps, Amanda Amanda.Phipps <@t> nationwidechildrens.org
Thu May 19 08:30:27 CDT 2011

Hi All

We are currently trying to stain for calcium deposits using Alizarin Red.  A few months ago our protocol worked fine, but this time we are getting zero staining.  Reagent has been made fresh, and pH is between 4.1 and 4.2.

Here is our protocol.

Deparaffinize to distilled H20
Stain slides with Alizarin Red S for 30 seconds to 5 minutes and observe reaction microscopically
Shake off excess dye and blot sections
Dehydrate in acetone 10 seconds to 20 seconds, then acetone-xylene (1:1) solution 10-20 seconds
Clear in xylene and mount in synthetic mounting medium.

Any help is greatly appreciated!!

Amanda Phipps, HTL (ASCP)
Morphology Core Lab
The Research Institute at Nationwide Children's Hospital
700 Children's Drive
Columbus, Ohio 43205
(614) 355-3474

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