Our protocol is to: Use a positively charged slide Fix in 10% formalin for 1 minute Rinse in tap (a few dips) Gill 3 for 1 minute Rinse in tap (a few dips) 3-5 dips in bluing Rinse in tap (a few dips) Eosin 2-3 dips 100% ETOH (a few dips) 100% ETOH (a few dips) Xylene (dip until clear) Mount with Richard- Allan Mounting Medium
After the Gill3, Eosin, and last alcohol, the back of the slide is wiped clean of excess solution to prevent carryover. -----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Reynolds,Donna M Sent: Tuesday, August 16, 2011 10:13 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] H&E on Frozen We do this frequently. Fix slides in Methanol (45ml) + formaldehyde (5ml) for 10 minutes. Wash several changes of tap water Stain with Harris Hematoxylin for 3 minutes Proceed as you would for paraffin sections Get great staining. I am sure if you are using a different Hematoxylin you could use it the same way you use it for paraffin Donna Reynolds HT(ASCP) Chief Histology Technician, Core Immuno Lab U. T. M.D. Anderson Cancer Center, Houston, TX 713-792-8106 e-mail dreyn...@mdanderson.org Message: 2 Date: Fri, 12 Aug 2011 08:52:11 -0700 (PDT) From: Jennifer Sipes <jengirl1...@yahoo.com> Subject: [Histonet] H&E for Frozen Tissue To: "histonet@lists.utsouthwestern.edu" <histonet@lists.utsouthwestern.edu> Message-ID: <1313164331.88113.yahoomail...@web125406.mail.ne1.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Does anyone have a good protocol for staining frozen sections with H&E?? Also, any mounting medium suggestions?? This is the first time I've been asked to do this for frozen. ? Thanks a bunch everyone! Jen ? Jennifer K. Sipes, ALAT Sr. Laboratory Technician Johns Hopkins University Ross 933 720 Rutland Avenue Baltimore, MD? 21205 phone:???? 410-614-0131 fax:???????? 410-955-9677 cell:???????? 443-631-6361 e-mail:? jsip...@jhmi.edu ------------------------------ ** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE This message and any included attachments are from Somerset Medical Center and are intended only for the addressee. The information contained in this message is confidential and may contain privileged, confidential, proprietary and/or trade secret information entitled to protection and/or exemption from disclosure under applicable law. Unauthorized forwarding, printing, copying, distribution, or use of such information is strictly prohibited and may be unlawful. If you are not the addressee, please promptly delete this message and notify the sender of the delivery error by e-mail or you may call Somerset Medical Center's computer Help Desk at 908-685-2200, ext. 4050. Be sure to visit Somerset Medical Center's Web site - www.somersetmedicalcenter.com - for the most up-to-date news, event listings, health information and more. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet