Nick, NBF may not work for Delia. She needs to do IF on the frozen sections and NBF fixation will require her to do AR on the sections (esp if she is localising immunoglobulins and complement).
Possibly warming the frozen tissue, using a thumb, may help. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Nicholas David Evans Sent: Saturday, 1 August 2009 5:37 AM To: 'DELIA GARCIA'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Please Help! We fix skin by sandwiching it between two sponges soaked in either 4% PFA or 0.4%PFA (for paraffin or cryo respectively) and compressed in a cassette. This flattens the skin. Following several hours fixation the skin is usually flat and rigid enough to handle and embed. It can then either by dehydrated for paraffin or soaked in sucrose for cryo. -----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of DELIA GARCIA Sent: Friday, July 31, 2009 11:42 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Please Help! Hello, I am in dire need of some suggestions. I work for a Derm lab and we occasionally get fresh skin punches that we freeze to do FITC (immunoflouresence). I have a really hard tiime keeping the tissue from curling makeing almost impossible to get a decent section on the slide. Anyone out there that might have experienced the same thing I would love to learn your techniques that overcame this problem. I have tried lightly blowing, freeze spraying, and lightly brushing acetone on the cold plate (not all at the same time). Nothing seems to ease it up. PLEASE HELP! I have one in the cryostat now. Thank you so very much. Delia _______________________________________________ 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 ********************************************************************* This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet