Colleen, You can pre-warm your OCT block inside the cryostat @ -20c (~ hr) from freezer before cutting, then gradually increase the temperature to warmer with 1 degree increment (thickness dependant). I also pre-chilled my slides before used.
Another alternative; - Fix with 4% PFA/PBS for ~ 1 hr on ice (you can freezed aliquot 4%PFA @ -20c so you don't need to make them fresh every time) - Wash 3xPBS buffer to remove excess 4% PFA/PBS, ph 7.4 - Transfer pre-fix zebra fish to 15% & 30% sucrose (30 min each or longer, not crucial), or you can leave them in 30% sucrose & store in fridge without freezing in OCT - Embed with OCT on ICE, not liquid nitrogen - Pre-warm -20c before cutting Good Luck! Madeleine Huey, BS, HTL & QIHC (ASCP) Supervisor - Pathology (IPOX & Histology) madelein...@elcaminohospital.org Message: 1 Date: Thu, 10 Jan 2013 15:16:32 -0600 From: Colleen Forster <cfors...@umn.edu> Subject: [Histonet] zebra fish experts...... To: Histonet <histonet@lists.utsouthwestern.edu> Message-ID: <50ef2fb0.6030...@umn.edu> Content-Type: text/plain; charset="iso-8859-1" Anyone out there doing zebra fish work I need your help! We are trying to fix zebra fish kidneys in 10% formalin and then freeze...the samples just seem to crumble when we try to cut...can someone help me out here with fixing/freezing methods for these little buggers!! Thanks in advance!! Colleen Forster U of MN _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
