An hypothesis for survival of spermatozoa via encapsulation during plane front freezing

Plane front freezing presents the possibility of encapsulating individual cells in the ice phase. The cells may also be pushed ahead of the plane front ice interface, as is always the case for conventional dendritic freezing, where the cells are pushed ahead of the thickening dendrite arms. Cells which are encapsulated during freezing are exposed to hypotonic liquid (pure water) initially upon thawing, while cells which are pushed into the last liquid to freeze are exposed to hypertonic liquid upon thawing. Some exposure to hypertonic intercellular liquid prior to freezing may be required to build up the salt and CPA content in the intracellular liquid and thereby avoid intracellular ice formation at the given cooling rate. Encapsulation of cells by a plane front ice interface should result in three regions of cell survival in the sample: an initial region of cell death due to intracellular ice formation, a final region of cell death due to overexposure to hypertonic intercellular liquid, and an intermediate region of cell survival, where neither damage mechanism has operated to a lethal level. An advantage of plane front freezing over dendritic freezing is that the regions of cell survival and death should be geometrically separate in the sample, rather than mixed at the dendritic microstructural level, as is the case for dendritic freezing. Samples containing populations with very high or very low survival rates for spermatozoa could be obtained by simply cutting up the frozen sample.