Forming and loading giant unilamellar vesicles with acoustic jetting

Giant unilamellar vesicles (GUVs) are a useful platform for reconstituting and studying membrane-bound biological systems, offering reduced complexity compared to living cells. Several techniques exist to form GUVs and populate them with biomolecules of interest. However, a persistent challenge is the ability to efficiently and reliably load solutions of biological macromolecules, organelle-like membranes, and/or micrometer-scale particles with controlled stoichiometry in the encapsulated volume of GUVs. Here, we demonstrate the use of acoustic streaming from high-intensity focused ultrasound to make and load GUVs from bulk solutions, without the need for nozzles that can become clogged or otherwise alter the solution composition. In this method, a compact acoustic lens is focused on a planar lipid bilayer formed between two aqueous solutions. The actuation of a planar piezoelectric material coupled to the lens accelerates a small volume of liquid, deforming the bilayer and forming a GUV containing the solution on the transducer side of the bilayer. As demonstrated here, acoustic jetting offers an alternative method for the generation of GUVs for biological and biophysical studies.