Hi Liz,
I inherited the following protocol for ACL samples. It works quite well, but times probably could be reduced - the optimisation is on my to-do list. 70% Alcohol - 1 hour 90% alcohol - 1 hour 100% Alcohol -2 hours 100% alcohol - 3 hours 100% alcohol - 4 hours Xylene - 1.5 hours Xylene - 1.5 hours Xylene - 3 hours Wax - 3 hours Wax - 3 hours Wax - 4 hours I cut the sections at 8um so they hold together better. Takes a while for all the wrinkles to disappear when floating out on water-bath so be patient Hope this helps. Andrew Andrew Prior Histologist Tissue Regenix Group E-mail: a.pr...@tissueregenix.com<mailto:a.pr...@tissueregenix.com> Website: www.tissueregenix.com<http://www.tissueregenix.com/> ------------------------------ Message: 3 Date: Wed, 12 Jun 2013 16:59:52 -0400 From: Elizabeth Ronan <lizro...@umich.edu<mailto:lizro...@umich.edu>> Subject: [Histonet] Paraffin processing native sheep ACL To: histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu> Message-ID: <51b8e148.3020...@umich.edu<mailto:51b8e148.3020...@umich.edu>> Content-Type: text/plain; charset=ISO-8859-1; format=flowed Hello, I need to paraffin process native sheep anterior cruciate ligament (ACL) that has been fixed in 10% neutral buffered formalin for 7 days. I was wondering if anyone with more expertise on this subject could guide me with the best lengths of times in the alcohols, xylenes, and paraffin to fix ACL. I tried a 12 hour program but the sections crumbled in the middle and it appeared that the paraffin had not fully perfused the ligament. I have access to the following program, and can alter the lengths of the steps for as long as desired: Program: 70% 80% 95% 95% 100% 100% Xylene Xylene Paraffin Paraffin Paraffin Any advice is much appreciated. Thanks for your time, Liz The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential and/or privileged material. Any review, retransmission, dissemination or other use of, or taking of any action in reliance upon, this information by persons or entities other than the intended recipient is prohibited. If you received this in error, please contact the sender and delete the material from any computer. Although we routinely screen for viruses, addressees should check this e-mail and any attachment for viruses. We make no warranty as to absence of viruses in this e-mail or any attachments. Registered Office: The Biocentre, Innovation Way, Heslington, York, YO10 5NY Registered No. 05969271 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
