Mariano,I have been doing IHC on frozen sections and one thing I can tell you --- and I stress this point emphatically --- do not thaw the tissues, then freeze them again and try to do IHC on them. This causes the cells to lyse which in turn will destroy epitopes. Keep the tissue frozen. I would section them as they came. Connie McManus, HTUniverisity of UtahSchool of MedicineDept of Dermatology----- Original Message ----From: Mariano S. Viapiano To: histonet@lists.utsouthwestern.eduSent: Friday, July 6, 2007 4:40:10 PMSubject: [Histonet] Flash-freezing samples in OCT?Hi everyone, We just received some frozen brain tumor samples toprocess for IHC but are a bit confused about how theywere prepared. We fix our samples in 4% PFA, change=0Athem to 30% sucrose until they sink, embed in OCT andfreeze them at -80C until cryostat sectioning. On theother hand, the samples we received were freshspecimens (just out of the OR) dipped in OCT andsnap-frozen in liquid nitrogen. What would be the bestthing to do? Section them as they came, or thaw them,remove the OCT and do our fixation procedure beforesectioning? I am a bit concerned about the OCT havingacted as a =93retardant” for the snap-freezing of thosesamples when they were submerged in nitrogen (or am Ijust imagining things?)Thank you!MarianoMariano S. Viapiano, PhDAssistant ProfessorCenter for Molecular Neurobiology andDepartment of Neurological SurgeryThe Ohio State University226B Rightmire Hall1060 Carmack Rd., Columbus OH 43210=0ATel (614) 292-4362Fax (614) 292-5379 ____________________________________________________________________________________Need a vacation? Get great dealsto amazing places on Yahoo! Travel.http://travel.yahoo.com/_______________________________________________=0AHistonet mailing listHistonet@lists.utsouthwestern.eduhttp://lists.utsouthwestern.edu/mailman/listinfo/histonet ____________________________________________________________________________________No need to miss a message. Get email on-the-go with Yahoo! Mail for Mobile. Get started.http://mobile.yahoo.com/mail
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