Ultrastructure of red cells frozen with hydroxyethyl starch

The freeze fracture appearance of red cells frozen in the presence of varying concentrations of hydroxyethyl starch (HES) is described. A technique is used which allows examination of a small portion of cells from a larger unit. The frozen cells appear distorted probably as a result of osmotic dehydration but indicate no evidence of intracellular ice. The frozen mixture with HES has three phases--a particulate phase consisting of the concentrated HES (and other salts), a sculptured ice phase and the red cells. When the concentration of HES is increased, the particulate phase becomes more prominent and at 14% HES appears to surround nearly all cells. In cells frozen in saline alone and 4% HES, the cytoplasm in a majority of cells has numerous cavities and depressions. Since such units haemolyse badly when thawed, it is possible that these regions indicate structural damage. In contrast, those units frozen with 14% HES (in which nearly 85% of the cells survive freeze-thaw) possess cells which only infrequently have such regions in the cytoplasm.