1. Nuclear magnetic resonance, light scattering and freeze fracturing electron microscopic techniques were used to characterize the size of unilamellar phospholipid vesicles of 1,2-dimyristoyl-sn-glycero-3-phosphocholine. 2. Differential scanning calorimetric and light scattering analyses showed that very small unilamellar vesicles obtained by the sonication method exhibit a downward shifted, largely broadened phase transition with a slightly decreased enthalpy change when compared with multilayered liposomes. 3. The phase transition of vesicles with variable diameter as obtained by injection methods resembled the pattern of multilayered liposomes the more the diameter was increased. 4. Repeated cycling through the lipid phase transition was shown to have a progressive effect on a fusion process. This effect was strongly increased when the osmolarity of the medium was enhanced (e.g. by the addition of cryoprotectors). Furthermore it was shown that ice-water of the systems caused abrupt fusion of the lipid structures. 5. Controversial results in the literature on the thermotropic behavior of vesicles could be explained in terms of these fusion processes.
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