Lipid-mixing assay is now commonly used to study protein, temperature and ion-dependent membrane fusion events. This assay has been crucial to demonstrate the ability of neuronal and non-neuronal soluble NSF attachment receptor (SNARE) to promote spontaneous fusion of liposomes. This lipid-mixing assay is based on the fluorescence resonance energy transfer (FRET) capability between a donor fluorescent lipid and a quenching lipid. When fusion between donor fluorescent liposomes and nonfluorescent acceptor liposome occurred, FRET decreases. This assay allows a real-time reading of SNARE-mediated liposome fusion.