[Histonet] problem with brain sections falling off subbed slides

[Douglas M Burns]

Hello,

      I am new to working with brain, and I am having a bad problem with
both thin and thick rat brain sections sliding off of subbed slides.

      We tried commercial treated slides first, but then  moved on to
subbing them. The subbing was done with gelatin according to a printed
protocol, and I can see the film on the dried slide. When kept horizontal
they seem okay, but as soon as they are put into a slide holder for washing
or anything, they begin to slide off or even move around. After a short
while, many/most of mysteriously vanished (into the wash solution).

      Our slices are sliced by cryostat from a rat brain that has been
fixed with 4% paraformaldehyde and 30% sucrose. The slice comes right off
the cryostat, and then I m ove it onto a warm slide. (Perhaps they are not
warm enough?? I have seen others blow on them to warm them.) Slides
destined for Cresyl violet staining were dried for 3 days at room
temperature before being moved.

       It is the Cresyl violet protocol that causes the most trouble, but I
am seeing some problems with any type of large volume wash. At this point,
I don't have enough experience with this specific tissue and our protocols
to be able to tell for sure what is happening. Thus, I am entirely open to
advice - any advice that might help with the problem.

                     Thank  you    -------------    Doug Burns, Kansas City