Betsy, I would dissolve the OCT in Hanks solution (or similar cell culture medium - not saline). Place in dilute OCT (50/50 with distilled water), place on stirrer until the lung settles in the OCT 30-60min. Blot with absorbent paper, refreeze in fresh OCT. It should cut quite well.
I have used this previously on some tissue that was received the same way. It seems that the OCT had "freeze-dried" prior to receipt in my lab. 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 Molinari, Betsy Sent: Thursday, 8 April 2010 12:46 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] frozen lung sections Hi all, I have received some lung samples on OCT. These lungs have not been in PBS or any fixative. They went directly from dissection into OCT and frozen. I cannot get a section. The temp of the cryostat is -20. I have done some reading and it seems the best results come from immersion in PBS, then 4% paraformaldehyde then into sucrose. Can anyone clarify the procedure or offer advice? I am unfamiliar with this technique since most of my frozen sectioning is heart tissue and it does not present the same problem as the lung. Thanks, Betsy Betsy Molinari HT(ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave MC1-283 Houston,TX 77030-2607 832-355-6524 832-355-6812 _______________________________________________ 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