Hi Tim, Try fixing for 45 min at 50oC. I think it possible that the tissue is not adequately fixed.
Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Laboratory Manager & Senior Scientist, the Children's Hospital at Westmead Adjunct Fellow, School of Medicine, University of Western Sydney Tel: 612 9845 3306 Fax: 612 9845 3318 Pathology Department 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 Morken, Timothy Sent: Thursday, 5 December 2013 4:31 AM To: Histonet Subject: [Histonet] Rush bx schedule? For tissue, what step most contributes to subsequent swelling of tissue when the block is soaked? I'm thinking the 100% ETOH, xylene clearing and paraffin infiltration all contribute, but at what percentage? Here is the current schedule, which apparently was meant to mimic a microwave schedule that was used before the MW quit working. The tissue - kidney and liver bx for the most part, up to 10 per day, is coming out soft and swells a bit when soaked. So sectioning is difficult. We need to have a rapid schedule but need good sections. Obviously we are pushing the limits on this so need a bit of leeway for variation in tissue So, I'm considering which of these steps is the most critical to lengthen. This is on a VIP5 but we can also use a peloris for this processing Formalin 2 min, 50degC (this is not necessary at all, IMHO) 80% ETOH 5 min, 50degC 95%ETOH 5 min, 50degC 100% ETOH 8 min, 50degC Xylene, 9 min 50degC Paraffin, 4 min, Paraffin, 5 min Thanks for your insight! Tim Morken Supervisor, Electron Microscopy and Neuromuscular Special Studies UC San Francisco Medical Center Box 1656 505 Parnassus Ave San Francisco, CA 94143 USA 415.353.1266 (office) tim.mor...@ucsfmedctr.org<mailto:tim.mor...@ucsfmedctr.org> _______________________________________________ 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 Sydney Children's Hospitals Network. This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Sydney Childrens Hospital's Network 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
