Hi Laurie, No, the tissue hasn't been fixed before freezing. My prof doesn't prefer the method, but if nothing else works, we might give it a try.
Any suggested protocols for fixation? Thanks! Dakshna ----- Original Message ----- From: "Laurie J King" <king.lau...@marshfieldclinic.org> To: "Dakshnapriya Balasubbramanian" <dakshnapr...@neo.tamu.edu> Sent: Tuesday, May 20, 2014 10:37:35 AM Subject: RE: [Histonet] HELP- Cryosectioning FAT! Dakshna, Has this tissue been fixed before freezing by any chance? laurie -----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Balasubbramanian, Dakshnapriya Sent: Tuesday, May 20, 2014 10:23 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] HELP- Cryosectioning FAT! Hi all, I've been having problems with cryosectioning fatty tissue for a long time now. The section leaves a hole in the middle. I know this is probably a much discussed topic, but I've tried a lot of strategies with no luck. The tissue is of mouse origin and has micro-lesions buried inside fat. So the fat needs to be cut in order to get to the lesion. I started with a temperature of -17C and tried upto -25C and also at -50C; didn't work at any of these temperatures. I tried rubbing the block, blade and anti-roll plate with dry ice-again,no luck. I rapid-freeze the tissue with OCT in LN2 and then store the block at -80C. Could I try sectioning the block directly from -80C? (never done that). Any other suggestions on how to tackle this? Any advice is much appreciated!! Thanks, Dakshna _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ______________________________________________________________________ The contents of this message may contain private, protected and/or privileged information. If you received this message in error, you should destroy the e-mail message and any attachments or copies, and you are prohibited from retaining, distributing, disclosing or using any information contained within. Please contact the sender and advise of the erroneous delivery by return e-mail or telephone. Thank you for your cooperation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
