I have a problem with sectioning brains on a Leica cyrostat. We are trying to take 20 um coronal sections (which we have had no problem doing in the past) but it looks like the cryostat is taking extra thick sections (even though it is set at 20 um).
Any tips to get the correct thickness of our tissue slices?
What model is it? Does it section one thick then the next one thinner? My guess is you have something loose on the knife stage or the objective holder. Or the wrong "degree" of knife/blade inclination.. It should be about 3-4 degrees, there is a scale bar on the left hand side of the knife base on most Leica models..
Yes we have the Leica CM3050S and the CM1950's, as well as having had a variety of other Leica models.. And Matthews comments about "over extension" are spot on..... Sometimes with mouse or rat brain sectioning, you can get the sample "flexing" between the cerebrum and the cerebellum. I always trim off the cerebellum and fix the cerebrum direct to the aluminium objective/specimen holder, to avoid that problem. (depends which bit of the brain you are working on;-)
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