30 Ishii

Figure 6 Freeze-fracture electron micrographs of lipid vesicles prepared in various systems: (A) phospholipid-
methanol-water; (B) phospholipid-ethanol-water; (C) phospholipid-1-propanol-water; (D) phospholipid-2-
propanol-water.

vesicles prepared using methanol and ethanol may be larger than those prepared using propanol.

Observation of Outer Surface of Liposomes

The scanning electron microscopy can be used to examine the homogeneity of the surface
topography of the lipid vesicles, in contrast with freeze-fracture electron micrographs.
Previously, we reported a specific fixation technique using malachite green, which adheres to
phospholipid components (10). Using this technique, complete images of lipid vesicles can be obtained
by scanning electron microscopy. Figure 7 shows typical scanning electron micrographs of lipid
vesicles prepared using various low-molecular-weight alcohols as a lipid solvent. All of the vesicles
were spherical particles with smooth surfaces. In particular, as shown in Figure 7, highly
homogeneous