Schematic illustration of the experimental strategy. We immobilized vesicles labeled with DiD and biotinylated lipids on the PEGylated quartz surface via specific binding of biotin and neutravidin, and removed nonimmobilized vesicles by flow washing. We then introduced another group of vesicles, having the same lipid composition but labeled with DiI, along with C2AB into the flow chamber. When we injected Ca2+ ions, the DiI-labeled vesicles rapidly bound to the surface-immobilized vesicles using the Ca2+-bound C2AB molecules as the molecular glue. If there was any degree of lipid mixing as a result of vesicle bridging, the FRET efficiency correspondingly increased.