Abstract
Networks of droplets, in which aqueous compartments are separated by lipid bilayers, have shown great potential as a model for biological transmembrane communication. We present a microfluidic system which allows for on-demand generation of droplets that are hydrodynamically locked in a trapping structure. As a result, the system enables the formation of a network of four droplets connected via lipid bilayers and the positions of each droplet in the network can be controlled thanks to automation of microfluidic operations. We perform electrophysiological measurements of ionic currents indicating interactions between nanopores and small molecules to prove the potential of the device in screening of the inhibitors acting on membrane proteins. We also demonstrate, for the first time, a microfluidic droplet interface bilayer (DIB) system in which the testing of inhibitors can be performed without direct contact between the tested sample and the electrodes recording picoampere currents.
Highlights
Stable and functional phospholipid bilayers can be formed using the Droplet InterfaceBilayer (DIB) method—a simple experimental approach that recently gained attention due to its broad capabilities in the field of synthetic biology [1,2]
The droplet interface bilayer (DIB) technique relies on the self-organization of phospholipid molecules at the interface of the aqueous droplets and continuous phase and subsequent formation of the bilayer when monolayers of two droplets are brought into intimate contact
Networks of aqueous droplets separated by lipid bilayers have shown great potential as a model for biological transmembrane communication
Summary
Stable and functional phospholipid bilayers can be formed using the Droplet Interface. Recent inventions of dedicated structures and geometries on a chip, such as rails [9] or traps [10,11,12,13,14] allowed for an ordered assembly of nanoliter droplets into networks, in which a chemical signal was propagating via the transport of molecules through lipid bilayers. The electrophysiological measurements require direct contact of an electrode with the measured sample containing tested proteins or inhibitors This problem has been solved by cyclic washing of electrodes with pure buffer [14] or by fabrication of arrays comprising multiple electrodes [21,22,23]. The physical separation of the outer droplets that host the electrodes from the inner (test) droplets allows us to perform a rapid screening of the interactions of small molecules with the single protein molecule, without needing to wash the electrodes between exchanges of the samples
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