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Packing density changes of supported lipid bilayers observed by fluorescence microscopy and quartz crystal microbalance-dissipation.

Various properties of supported lipid bilayers such as diffusion and lipid partitioning are well characterized. However, little attention has been paid to their molecular packing density. In this work, the adsorption of 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) vesicles on glass and silicon dioxide was investigated using fluorescence microscopy, quartz crystal microbalance-dissipation (QCM-D), and atomic force microscopy. Fluorescence recovery after photobleaching data showed that the adsorption of large unilamellar vesicles (LUVs) on glass yielded supported bilayers with full mobility under alkaline (pH 8.3) and acidic (pH 3-4) conditions. These fluid bilayers exhibited quite different diffusion constants; those at alkaline pH were ~10 times larger than those at acidic pH. The reason for this pH dependence was clarified by investigation of the rupture of giant unilamellar vesicles (GUVs) on glass. Fluorescence data revealed that the area of planar bilayer patches increased at alkaline pH. Thus, we conclude that the rapid diffusion in alkaline solution arises from the decreased molecular density. QCM-D data showed that dissipation increased in a stepwise manner during vesicle fusion on silicon dioxide at alkaline pH. We attribute this behavior to the decrease in packing density of planar bilayers.