I was having similar issues with frozen rat and human cartilage with the cryojane. After a lot of trial and error; I have found that the 4X slides and air-drying overnight has worked extremely well for us. The tissues never fall off any more! Good luck, Kim
Kim Merriam, MA, HT(ASCP)QIHC Cambridge, MA ________________________________ From: "Connolly, Brett M" <brett_conno...@merck.com> To: Douglas M Burns <dmbur...@gmail.com>; "histonet@lists.utsouthwestern.edu" <histonet@lists.utsouthwestern.edu> Sent: Thursday, December 8, 2011 9:46 PM Subject: RE: [Histonet] brain sections cut on Cryo Jane apparatus are falling off slides during ethanol series for a cresyl stain Doug, Hopefully you are using the CryoJane slides (not the subbed slides) and the UV glue activator and keeping both slides and tape cold in the cryostat. Be aware that you can get CryoJane slides with different amounts of adhesive (0.5-4x). I would suggest going to the 4x slides. When peeling the tape off I pull it slowly and evenly almost back on itself at an acute angle- this greatly reduces the incidence of the section coming off with the tape... Haven't had a problem with section detachment in ethanols. Brett Brett Connolly, Phd Merck Research Labs West Point, PA ________________________________________ From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Douglas M Burns [dmbur...@gmail.com] Sent: Thursday, December 08, 2011 5:29 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] brain sections cut on Cryo Jane apparatus are falling off slides during ethanol series for a cresyl stain Hello, Histo-users, Follow-up to our original posting about fixed-brain cryosections falling off of subbed slides. We fished out our Cryo-Jane apparatus, and using this apparatus has resulted in a great improvement in sections and getting sections stuck onto the slides. This works pretty well until we start using the ethanol series to dehydrate and delipidate before a Crystal violet stain for Nissl bodies. (That is, for acqueous steps everything seems to stay glued to the slide; however, once ethanols are involved pieces start to come off.) To summarize, we are trying to go from cut blocks of paraformaldehyde-fixed brain infused with sucrose and then frozen at -80. We take them out, unfrozen, and cut them on a Leica cryostat. Early attempts with subbed slides did not work reliably, so we have turned to 'Jane. Jane is slightly slower than a brush, and seems to work fairly well. However, we have had trouble with the tape pulling parts of sections off of the glued slides. When we introduce the nonacqeous solvents, our problems become more obvious. We have not so far gone to a post-fix, because we wanted to simplify the overall process. Could this be the mistake? We want to use IHC for c-Fox and possibly orexin - and similar - but we want cresyl-stain to locate positively the area of interest in the cut block. Any suggestions? Also, are there any 'Jane enthusiasts out there? thank you in advance --------------- Doug Burns, Kansas City _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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