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