Our research focuses on creating smart materials that utilize synthetic cell membranes assembled at liquid interfaces for autonomic sensing, actuation, and energy conversion. Unlike single membrane assemblies, systems featuring many membranes have the potential to offer multi-functionality, greater transduction sensitivity, and even emergent behaviors in response to environmental stimuli, similar to living tissue, which utilizes networks of highly packed cells to accomplish tasks. Here, we present for the first time a novel microfluidic platform capable of generating a stream of alternating droplet compositions, i.e. A-B-A-B, and sequentially capturing these droplets in precise locations to enable the spontaneous formation of synthetic lipid bilayers between droplets of different compositions (i.e. A and B) in an enclosed substrate. This platform preserves a key feature of the droplet interface bilayer (DIB) method, which allows asymmetric conditions within and across the membrane to be prescribed by simply using droplets containing different species. In this work, we demonstrate the ability to assemble bilayers consisting of asymmetric lipid compositions and, separately, show that alternating droplets containing the same lipid type can also be used to control the direction of ion channel insertion. In the first study, A and B droplet types contain liposomes comprised of different lipid types, which are used to establish an asymmetric composition of the leaflets that make up the lipid bilayer. This asymmetry results in a dc, non-zero membrane potential, which we measure via membrane capacitance versus bias voltage. In the second study, alamethicin peptides are included in only one of the droplet types, which enable voltage-dependent insertion to occur only at one polarity. Cyclic voltammetry measurements are performed to confirm the direction of insertion of alamethicin channels in bilayers. Also, these results show the ability to perform simultaneously electrical measurements on multiple DIB, which increases the experimental capacity and efficiency of a microfluidic approach. The ability to produce alternating droplets in a high throughput manner with electrical access provides a system to investigate the effects of lipid asymmetry on the function of membrane proteins in a controlled model system.

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