A Method For Orienting Cryostat Sections For Three-Dimensional Reconstructions

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Abstract
Mouse cerebellum for cryostat sectioning was embedded together with reference tissue [liver] impaled on the needles of a specially constructed trident. To maintain spatial orientation, reference and experimental tissue are frozen in Tissue-Tek OCT compound in the manner usual for enzyme histochemistry using a simply constructed carrier. The trident is removed by heating its handle, thus leaving holes in the reference tissue. These, in turn, are used as reference points to orient successive sections with respect to each other.