Droplet bilayer formation using throughput liquid handling techniques
AssigneeThe Regents of the University of California
InventorsJason L. Poulos, Jacob J. Schmidt
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Abstract
The invention relates to methods and devices for forming and measuring lipid bilayers, for example, by joining lipid monolayers that self-assemble at the interface of aqueous and organic phases using sessile aqueous droplets in contact with a measurement electrode. At least one of the aqueous solution, the lower aqueous phase, and/or the upper organic phase comprise lipids. This abstract is intended as a scanning tool for purposes of searching in the particular art and is not intended to be limiting of the present invention.
Claims19
A method for forming a lipid bilayer, comprising the steps of:
- a) providing a droplet electrode with a tip having at least a sessile droplet of aqueous solution;
- b) providing a well comprising:
- i) an upper organic phase,
- ii) a lower aqueous phase, and
- iii) a well electrode extending into the lower aqueous phase,
- wherein at least one of the aqueous solution, the lower aqueous phase, and the upper organic phase comprise lipids, and
- wherein the droplet electrode and the well electrode are in electrical communication;
- and
- c) inserting the droplet electrode into the well so that a lipid bilayer forms between the lower aqueous phase of the well and the aqueous solution of the droplet electrode tip.
A method for measuring current through a bilayer, comprising:
- a) providing a silver pin tool, wherein the tool comprises an electrode and contains an aqueous solution and lipid vesicles;
- b) providing a well, wherein the well comprises a lower aqueous phase and an upper organic phase;
- wherein the lower aqueous phase comprises an electrode and also comprises lipid vesicles;
- c) inserting the silver pin tool into the well so that a bilayer forms between the lower aqueous phase of the well and the aqueous solution of the pin tool;
- and
- d) measuring the current through the bilayer.
A high-throughput lipid bilayer production device comprising:
- a) a plurality of wells, each well having a well electrode positioned within;
- and
- b) a droplet electrode electrically coupled to each well electrode and positioned for insertion into any of the plurality of wells, the droplet electrode having:
- i) a tip capable of extracting a droplet of liquid, or ii) an internal reservoir capable of extruding a droplet of liquid;
- wherein at least one well further comprises:
- a) an upper organic phase, and
- b) a lower aqueous phase in contact with the well electrode, wherein the lower aqueous phase and/or the upper organic phase comprise lipids.
An electrode assembly for use in high-throughput lipid bilayer production, the electrode assembly comprising:
- a) an electrode carriage and a means for positioning the carriage;
- b) a droplet electrode mounted on the carriage and having a tip capable of extracting a droplet of liquid or an internal reservoir capable of extruding a droplet of liquid;
- and
- c) a well electrode electrically coupled to the droplet electrode and mounted on the carriage;
- wherein at least one well further comprises:
- a) an upper organic phase, and
- b) a lower aqueous phase in contact with the well electrode, wherein the lower aqueous phase and/or the upper organic phase comprise lipids.
A well array for use in high-throughput lipid bilayer production, the well array comprising a plurality of wells, each well comprising:
- a) an organic phase chamber;
- b) an aqueous phase chamber spaced apart from, but connected to, the organic phase chamber;
- and
- c) optionally, a membrane mask at the connection between the aqueous phase chamber and the organic phase chamber;
- wherein at least one well further comprises:
- a) an upper organic phase, and
- b) a lower aqueous phase in contact with the well electrode, wherein the lower aqueous phase and/or the upper organic phase comprise lipids.
A high-throughput lipid bilayer production device comprising:
- a) an electrode assembly comprising:
- i) an electrode carriage and a means for positioning the carriage;
- ii) a droplet electrode mounted on the carriage and having a tip capable of extracting a droplet of liquid or an internal reservoir capable of extruding a droplet of liquid;
- and
- iii) a well electrode electrically coupled to the droplet electrode and mounted on the carriage;
- and
- b) a well array comprising a plurality of wells, each well comprising:
- i) an organic phase chamber;
- ii) an aqueous phase chamber spaced apart from, but connected to, the organic phase chamber;
- and
- iii) optionally, a membrane mask at the connection between the aqueous phase chamber and the organic phase chamber.
Dependencies are inferred from the claim text.