>>> HI Bill,
>>> > I need to do some cerebellar slices. I'm trying to
> > figure out the best
> > age band and the best orientation of the slice.
>> > I see some people using very young animals
> > for cerebellar
> > slices for no apparent reason. Though others use
> > months old animals.
> > Apart from the general, "younger animals make
> > slices" rule of
> > brain slices, is there any specific cut off I
> > be aware of? (I'm
> > after granule cells)
>> Age would depend on which developmental stage you're
> trying to target ..... people chose the those post
> natal ages because of the beginning of synaptic
> maturation that begins mostly around p10 (that's to
> the best of my understanding).
>> also, I would say, having done some cerebellar
> (though not for physiology), the sagittal sections
> the easiest ..... you can use a regular slicer for
> p10-p14 and then vibratome for >p15 ....
> Now, I suppose if you wish to target the parallel
> fibres of graule cells, the frontal sections are
> you'll be after .... although, I must admit that
> frintal slices are far more difficult that sagittal
> > Also, what the the best plane of section to cut
> > slices in. The one
> > time I tried making cerebellar slices, it reminded
> > me of trying to
> > shave a cauliflower, little tiny bits fell of,
> > rather than big proper
> > slices. I would imagine you would either glue it
> > the stage of the
> > vibrotome via the sagital mid-line, or along the
> > ventral surface of
> > the brain stem.
>> srangely enough they don't fall apart like
> bits, (if you stick to ~300 um sections for live
> and you can go upt 30um for fixed/frozen) ...that's
> from personal experience ... I suppose one can go
> lower depending on the blade and apparatus one is
> using ....
>> hope that helps,
> > Any tips would be appreciated,
> > Thanks in advance.
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