Ice crystals formed in tissues during cryosurgery. I. Light microscopy

The size and distribution of ice crystals formed during cryosurgical procedures in intact animals are not clear. In the present experiment oral mucosa was frozen in situ by means of a surface applied cold probe and was excised and freeze substituted while in the frozen state. It was shown that the form of the frozen tissue was preserved during this procedure and the area frozen was divisible into a zone representing the central part of the lesion and a peripheral zone separating this from normal tissue. Ice crystals within the body of the lesion were intracellular in location but varied somewhat in size. Ice crystals in the boundary zone appeared to be intracellular in the epithelium and both intra- and extracellular in the muscle fibres.It is suggested that the intracellular crystals in the body of the frozen area result in cell death while the extracellular ice in the boundary zone results in a less predictable response.