Microfluidic Generation of Giant Lipid Vesicles with Membrane Protein Reconstitution

Giant unilamellar lipid vesicles(GUV) are clean experimental models to study the biophysical mechanisms of biomembranes and transmembrane proteins from a quantitative point of view. Traditional generation methods of GUVs like rehydration or electroswelling produce vesicles with a wide size and composition distribution. These drawbacks limit experiment accuracy, and thus hinder the capacity of experiments to validate theoretical models in lipid membrane biophysics. Here we present a microfluidic emulsion template method to generate monodispersed lipid vesicles with transmembrane protein reconstitution. Compared to other emulsion-templated methods, our protocol allows a wider range of lipid choices, eliminates the influence of residual solvents, requires a shorter incubation time for solvent evaporation, and supports fabrication of lipid vesicles with more complicated structures. Our method opens up opportunities for more quantitative lipid membrane-related experiments and are also potential candidates in synthetic biology constructions.