Hemifusion of giant unilamellar vesicles with planar hydrophobic surfaces: a fluorescence microscopy study.

Vesicle adhesion and fusion to interfaces are frequently used for the construction of biomimetic surfaces in biosensors and drug delivery. Ubiquitous in cell biology, vesicle fusion involves the transformation of two separate membranes into one contiguous lipid bilayer. In distinction, the deposition of vesicle membranes to hydrophobic surfaces requires the transformation of a lipidic bilayer into a monomolecular layer - a topologically distinct process termed hemifusion. Here, we used hydrophobically terminated self-assembled monolayers (SAMs) on solid surfaces to track the hemifusion of fluorescently labeled giant unilamellar vesicles (GUVs) at the single vesicle level with video time resolution (≈53 ms). We observed that a dilute monolayer, consisting of lipid extracted from the outer GUV leaflet, spreads outward across the hydrophobic surface from the vesicle adhesion site. Subsequently, bilayer hemifusion occurs by vesicle rupture near the hydrophobic surface, followed by spreading of lipid in a dense monolayer. GUV lipids thus transfer to the SAM surface in two concentric zones: an outer hemifusion zone comprises lipids drawn from the outer GUV leaflet and an inner hemifusion zone comprises lipids from both the inner and outer GUV leaflets and grows at a rate of ≈1000 µm