ratbrain slices 2

From:Frouwke Kuijpers

I also like to have information about fixation, processing and imbedding in
paraffin of rat brains.
We have a good protocol for frozen sections (40 um) and do imuno using the
Free Floating Method.  Disadvantage of this method is that not used sections
cannot preserve for a long time, we do add Na-azide to the vial but after a
we don't trust it anymore . Because we use special breeded rats which are
not so
often available I want to look if the procedure also can be done using
paraffin techniques.
I presume preservation of the sections will be better, sections can be
thinner and less antibody can be used ($$$). We have  paraffin-experience
with all kind of fish and 'xenopus' (a kind of frog), the story on our
department is that doing this on ratbrain is much more difficult but nobody
can explain why.
So, is it worth trying this or is it better using  my own freezing-protocols
and accept the disadvantages of the FF method?

What kind of fixative (does anyone has experience with Zn-formaline, I
already read omething about post fixation with it ?)
Processing schedule (Times used for a whole ratbrain)
How long paraffin sections can be stored and a good immune-staining is still

I hope somebody can make me wise,
thanks in advance.

Dept. Cellular Animal Physiology
University of Nijmegen
Toernooiveld 1
6525 ED Nijmegen
the Netherlands

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