Sequential production of various types of asymmetric lipid vesicles using pulse jet flow

This paper describes a method of sequential formation of asymmetric lipid vesicles, which applies pulse jet flow to an asymmetric planar lipid bilayer with multiple lipid components formed by Droplet Split-and-Contact Method. In our previous work, we generated asymmetric lipid vesicles by deforming an asymmetric planar lipid bilayer using a pulsed jet flow. However, it was difficult to create various types or large amounts of the asymmetric lipid vesicles in a single double-well device. In this study, asymmetric planar lipid bilayers with multiple lipid components were formed by contacting various types of lipid monolayers using movable wells on a revolving table. We successfully produced two types of the asymmetric lipid vesicles containing red or green fluorescent lipids on the outer leaflet with a single device in a high-throughput manner using the proposed method. The asymmetricity of the generated vesicles was evaluated by the distribution of the fluorescence intensities. The proposed device will be applicable for generating artificial cell models with multiple asymmetric lipid components.