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Abstract

Closed lipid bilayers in the form of giant unilamellar vesicles (GUVs) are commonly used membrane models. Various methods have been developed to prepare GUVs, however it is unknown if all approaches yield membranes with the same elastic, electric, and rheological properties. Here, we combine flickering spectroscopy and electrodefomation of GUVs to measure, at identical conditions, membrane capacitance, bending rigidity and shear surface viscosity of palmitoyloleoylphosphatidylcholine (POPC) membranes formed by several commonly used preparation methods: thin film hydration (spontaneous swelling), electroformation, gel-assisted swelling using poly(vinyl alcohol) (PVA) or agarose, and phase-transfer. We find relatively similar bending rigidity value across all the methods except for the agarose hydration method. In addition, the capacitance values are similar except for vesicles prepared via PVA gel hydration. Intriguingly, membranes prepared by the gel-assisted and phase-transfer methods exhibit much higher shear viscosity compared to electroformation and spontaneous swelling, likely due to remnants of polymers (PVA and agarose) and oils (hexadecane and mineral) in the lipid bilayer structure.