Bingo, that is exactly what I was thinking. Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. Ste.215 Aurora, CO 80045 720-859-4060 fax 720-859-4110 www.ihctech.net www.ihcrg.org
-----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Amos Brooks Sent: Friday, May 20, 2011 2:57 PM To: amanda.phi...@nationwidechildrens.org; histonet@lists.utsouthwestern.edu Subject: [Histonet] Alizarin Red Troubleshooting Hi, I seldom do this, but I'll bet I know EXACTLY what the problem is. Try it again and stop the rehydration at 95% ETOH then put it directly into the Alizarin Red (double check the pH of course). This will surely fix it. The calcium gets rinsed out when it is rehydrated. The same thing happens with Von Kossa. I also try to abbreviate rinsing after when dehydrating to coverslipping. Let us know if you still have trouble, Amos Message: 18 Date: Thu, 19 May 2011 09:30:27 -0400 From: "Phipps, Amanda" <amanda.phi...@nationwidechildrens.org> Subject: [Histonet] Alizarin Red Troubleshooting To: "'histonet@lists.utsouthwestern.edu'" <histonet@lists.utsouthwestern.edu> Message-ID: < 20CE7246011C12489BA04BEB85CCDA1F0351BF252B@res2k7ms01> Content-Type: text/plain; charset="us-ascii" 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) Histotechnologist Morphology Core Lab The Research Institute at Nationwide Children's Hospital 700 Children's Drive Columbus, Ohio 43205 (614) 355-3474 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet