A copper block method for freezing non-cryoprotected tissue to produce ice-crystal-free regions for electron microscopy
1976; Wiley; Volume: 106; Issue: 3 Linguagem: Inglês
10.1111/j.1365-2818.1976.tb02406.x
ISSN1365-2818
Autores Tópico(s)Nanopore and Nanochannel Transport Studies
ResumoSUMMARY Two methods are described for fracturing unfixed, uncryoprotected mouse liver that had been frozen by contact with a copper block at liquid nitrogen temperature. The best method involved precise fracturing of a few microns of surface tissue with a cooled glass knife, using the Cryokit attachment of the LKB III ultramicrotome. The resulting replicas were free from electron microscopically‐visible ice crystals. A noticeable feature of replicas from unfixed, uncryoprotected tissue was the extent of plastic deformation of certain cellular structures. Cytoplasmic macromolecules, and to a lesser extent intramembranous particles, often appeared to be ‘stretched’ to form fibrils.
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