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[Neuroscience] Re: Cerebellar slices, advice?

Christian Wilms via neur-sci%40net.bio.net (by usenet02 from out-of-phase.de)
Wed Mar 21 05:53:36 EST 2007


We work with cerebellar slices on a regualr basis. The plane you decide
to cut in depends on what you are looking for. Sagital slices will have
intact climbing fibers and good Purkinje cell dendritic trees, but the
parallel fibers will be severed. Transversal slices will leave long
parts of the parallel fibers intact, but Purkinje cell dendrites will
often be severed. I've also found that climbing fibers tend to be
severely damaged in transversal slices. It seems to be a good idea to
use fairly thick slices if you are cutting in the transversal plane (400
microns), whereas thin slices (200 microns) work fine for the sagital
plane.

People using sagital slices usually stick to a basic preparation
technique. We normally only use the Vermis. Hence, after removing the
hind section of the skull I use a scapel to severe the hemispheres from
the vermis as parallel to the midline as possible. Next I cut the Vermis
free from the visual cortex with a clean cut perpendicular to the
midline. For slicing I glue the explant down on one of the two sagital
surfaces.

We tend to use animals between P18 and 24. At this age mice and rats
have fully developed parallel fibers and Purkinje cells. A collegue
working on granule cells tends to use P60 animals - that would fit to
Nadia's comments.

Let me know if you have any specific questions.

h2h, Christian


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