I have tried this method as well and it works just as described below. Sarah E. Lewis HT, ASCP The Research Institute at Nationwide Children's Hospital Center for Gene Therapy Neuromuscular Division Rm WA3110 (614)722-2204
-----Original Message----- From: Tony Reilly [mailto:tony_rei...@health.qld.gov.au] Sent: Tuesday, October 01, 2013 7:15 PM To: Histonet; Timothy Morken Cc: jmitch...@neurology.wisc.edu Subject: Re: [Histonet] thawing and refreezing muscle? Hi Tim A guy in our histology group gave a presentation debunking the theory that ice crystal artefact is irreversible. His lab receives frozen specimens from thousands of kilometres away from labs with no histology staff. If there was ice crystal artefact he defrosts the specimen at room temperature. Mop the specimen with tissues to remove as much OCT and moisture as possible. Allow the specimen to remain at room temperature on the bench for 20-30 minutes depending on the size of the specimen to allow evaporation of more moisture. Refreeze the specimen with OCT and cut sections. The result was absence of ice crystal artefact but some separation of muscle bundles which does not interfere with staining. It appears the ice crystals do not damage the fibres just cause separation. So removing the moisture eliminates the problem. If you have a lot of muscle perhaps you could separate a small amount to trial the procedure. regards Tony Tony Reilly B.App.Sc. , M.Sc. Chief Scientist, Anatomical Pathology Pathology Queensland-PA Laboratory ________________________________________________ Health Services Support Agency | Department of Health Level 1, Building 15,Princess Alexandra Hospital Ipswich Road,WOOLLOONGABBA Qld4102 Ph: 07 3176 2412 Mob: 0402 139411 Fax: 07 3176 2930 Email: tony_rei...@health.qld.gov.au Web: www.health.qld.gov.au/qhcss/ >>> "Morken, Timothy" <timothy.mor...@ucsfmedctr.org> 10/2/2013 2:07 am >>> >>> Histonetters, Does anyone have a good method for thawing muscle and refreezing for histochemistry? With good results? We have some bulk-frozen muscle (centimeter thick) and that is what they want to try. Tim Morken Supervisor, Electron Microscopy/Neuromuscular Special Studies Department of Pathology UC San Francisco Medical Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ******************************************************************************** This email, including any attachments sent with it, is confidential and for the sole use of the intended recipient(s). This confidentiality is not waived or lost, if you receive it and you are not the intended recipient(s), or if it is transmitted/received in error. Any unauthorised use, alteration, disclosure, distribution or review of this email is strictly prohibited. The information contained in this email, including any attachment sent with it, may be subject to a statutory duty of confidentiality if it relates to health service matters. If you are not the intended recipient(s), or if you have received this email in error, you are asked to immediately notify the sender by telephone collect on Australia +61 1800 198 175 or by return email. You should also delete this email, and any copies, from your computer system network and destroy any hard copies produced. If not an intended recipient of this email, you must not copy, distribute or take any action(s) that relies on it; any form of disclosure, modification, distribution and/or publication of this email is also prohibited. Although Queensland Health takes all reasonable steps to ensure this email does not contain malicious software, Queensland Health does not accept responsibility for the consequences if any person's computer inadvertently suffers any disruption to services, loss of information, harm or is infected with a virus, other malicious computer programme or code that may occur as a consequence of receiving this email. Unless stated otherwise, this email represents only the views of the sender and not the views of the Queensland Government. ********************************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
