Optical fusion assay based on membrane-coated spheres in a 2D assembly.

A major goal in neurophysiology and research on enveloped viruses is to understand and control the biology and physics of membrane fusion and its inhibition as a function of lipid and protein composition. This poses an experimental challenge in the realization of fast and reliable assays that allow us, with a minimal use of fluorescent or radioactive labels, to identify the different stages of membrane-membrane interaction ranging from docking to complete membrane merging. Here, an optical two-dimensional fusion assay based on monodisperse membrane-coated microspheres is introduced, allowing unequivocal assignment of docking and membrane fusion. The hard-sphere fluid captures and quantifies relevant stages of membrane fusion and its inhibition without interference from aggregation, liposome rupture, extensive fluorescence labeling, and light scattering. The feasibility of the approach is demonstrated by using an established model system based on coiled-coil heterodimers formed between two opposing membrane-coated microspheres.