You did not say whether these are cross sections or mid sagittal? Cross sections are always tougher. Key is to make sure the cortical bone is well processed and infiltrated with a hard paraffin e.g. Tissue Prep 2 (Fisher Scientific ala Thermo Scientific). Try this old bonehead trick is cut tiny V-shaped notches with razor blade or used microtome blade on the sides of block, parallel to the blade. You may have to do this top and bottom too. Take care to NOT make these notches huge. This permits the paraffin of each section in ribbon to expand, relax onto the water bath along with the bone section.
Another trick is try laying section on RT 5 to 10% alcohol, pick up on slide, then go to warm water bath, lower section slowly to flattening. The key here is to NOT let upper part of paraffin of a section totally release from slide while going into warm water (at an angle) in other words, the section flattens while paraffin portion is still attached to slide during flattening. Also, change the blade frequently. Sharpest possible edge helps, and hopefully high profile which is more stable than low profile for decalcified bone microtomy. Good luck Gayle M. Callis HTL/HT/MT(ASCP) -----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Jack Ratliff Sent: Tuesday, May 11, 2010 5:03 AM To: Connolly, Brett M Cc: <histonet@lists.utsouthwestern.edu> Subject: Re: [Histonet] need tips for cross-sectioning of cortical bone Why not embed in resin (MMA) and take thicker sections and then grind/ polish them down? If you went this route, you could then use flourescent labels and quantify mineral apposition rate and bone formation rate. Let me know if you are interested. I can help you get started and direct you to low cost equipment options. Jack On Apr 22, 2010, at 9:58 AM, "Connolly, Brett M" <brett_conno...@merck.com > wrote: > A colleague is having trouble getting wrinkle-free sections of > decalcified, paraffin embedded femur. > > Any tips?? > > Thanks, > > Brett M. Connolly, Ph.D. > Molecular Imaging Team Leader > Merck & Co., Inc. > PO Box 4, WP-44K > West Point, PA 19486 > tel. 215-652-2501 fax. 215-993-6803 > brett_conno...@merck.com > > > > Notice: This e-mail message, together with any attachments, > contains information of Merck & Co., Inc. (One Merck Drive, > Whitehouse Station, New Jersey, USA 08889), and/or its affiliates > Direct contact information for affiliates is available at http://www.merck.com/contact/contacts.html > ) that may be confidential, proprietary copyrighted and/or legally > privileged. It is intended solely for the use of the individual or > entity named on this message. If you are not the intended recipient, > and have received this message in error, please notify us > immediately by reply e-mail and then delete it from your system. > _______________________________________________ > 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 __________ Information from ESET Smart Security, version of virus signature database 5103 (20100510) __________ The message was checked by ESET Smart Security. http://www.eset.com __________ Information from ESET Smart Security, version of virus signature database 5103 (20100510) __________ The message was checked by ESET Smart Security. http://www.eset.com __________ Information from ESET Smart Security, version of virus signature database 5105 (20100511) __________ The message was checked by ESET Smart Security. http://www.eset.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet