[Histonet] brain sections cut on Cryo Jane apparatus are falling off slides during ethanol series for a cresyl stain

[Douglas M Burns]

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