Understanding the underlying mechanisms of vesicle diffusion near membranes can provide a comprehensive interpretation of exocytosis and endocytosis. For the first time, we experimentally and theoretically characterize the dynamics of small vesicles (~50 nm in diameter) that are diffusing near target supported planar bilayers. The population distributions and diffusion kinetics of the vesicles are affected by changing the ionic strength and pH of the buffer, as well as the lipid composition of the target membrane. Using total internal reflection-fluorescence correlation spectroscopy, we examine the validity of theoretical analyses of vesicle–membrane interactions. Effective surface charges on neutral bilayers are also analyzed by comparing experimental and theoretical data, and we demonstrate the possibility that vesicle dynamics can be modified by surface charge redistribution of the planar bilayer. Based on these results, we hypothesize that small vesicles, diffusing close to target biomembranes are spatially restricted, which may represent an addition mechanism for controlling fusion or fission dynamics.