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Three classes of lipoidal nitroxide spin probes reversibly perturb erythrocyte membranes at low concentrations (10(-10)-10(-5) M). This is manifest in (a) decreased osmotic fragility and (b) alterations of surface topology. At bulk phase nitroxide concentrations providing maximal osmotic stabilization, the erythrocytes exhibit a classic echinocyte morphology. At nitroxide concentrations very slightly higher than those yielding minimal osmotic fragility (10(-5)-10(-4) M), the cellus undergo a sphering reaction and lyse. The morphologic sequence seen in intact cells is not observed in erythrocyte ghosts. We suggest that the spin probes initially concentrate in focal domains, which expand into echinocytic protrusions primarily due to localized weakening of membrane cohesion. We propose that cell lysis involves an irreversible breakdown in membrane domain structure.

Scanning electron microscopy has been used to characterize alterations of lymphocyte surface topology that occur on contact with erythrocytes during the rosetting reaction. Molt-4 cells, a line of leukemic human lymphocytes, defined as T-cells through their ability to form rosettes with sheep erythrocytes, were used for this. Unreacted Molt-4 cells exhibit surfaces that are virtually smooth and carry few microvilli. In contrast, Molt-4 cells in rosettes display a time-dependent modification of surface topology, characterized by the emergence of numerous, long microvilli, particularly in areas of red cell contact.

Temperature-induced variations in the surface morphology of cultured lymphocytes were evaluated by scanning electron microscopy. At 25-37 degrees the cells' surfaces are largely obscured by numerous undulating microvilli of various lengths but uniform diameter. Temperature changes alter the number of microvilli, their lengths, diameters, distribution, branching, and fusing. Typically, chilling to 0-4 degrees markedly reduces the number of microvilli and increases the diameter of the survivors in a reversible process. In contrast, heating the cells to about 45 degrees rapidly and irreversibly transforms the ordinarily smooth membrane surface into one with a "cobblestone" morphology. At the same time most microvilli disappear and the few that remain clump into a cap. The data suggest that the low-temperature effects reflect a change in the physical state of membrane lipids, while the high-temperature alterations represent thermotropic protein transitions.