Nanopore Platforms for Ion Channel Recordings and Single Molecule Detection and Analysis
First Claim
1. A nanopore device comprising:
- a membrane having a thickness with a first, exterior, side and a second, interior, side, the first side being opposite the second side, wherein the membrane is made from a material selected from the group consisting of Si, SiO2, Si3N4, alumina, nitrides, metals, polymers, and combinations thereof;
at least one nanopore extending through the membrane, thus forming at least one channel connecting the first side and the second side of the membrane, wherein the nanopore has a first opening, a second opening, and a depth, wherein the first opening opens to the first side of the membrane, and the second opening opens to the second side of the membrane, and wherein surface portions of the first side and the at least one channel connecting the first side and the second side of the membrane include modifying molecules defining hydrophobic surfaces; and
means for spanning across the first opening of the nanopore, wherein the means for spanning across the first opening of the nanopore is a supported lipid bilayer structure or a spanning lipid bilayer structure.
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Abstract
A nanopore device includes a membrane having a nanopore extending there through forming a channel from a first side of the membrane to a second side of the membrane. The surface of the channel and first side of the membrane are modified with a hydrophobic coating. A first lipid monolayer is deposited on the first side of the membrane, and a second lipid monolayer is deposited on the second side of the membrane, wherein the hydrophobic coating causes spontaneous generation of a lipid bilayer across the nanopore orifice. Sensing entities, such as a protein ion channel, can be inserted and removed from the bilayer by adjusting transmembrane pressure, and adapter molecules can be electrostatically trapped in the ion channel by applying high transmembrane voltages, while resistance or current flow through the sensing entity can be measured electrically.
53 Citations
17 Claims
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1. A nanopore device comprising:
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a membrane having a thickness with a first, exterior, side and a second, interior, side, the first side being opposite the second side, wherein the membrane is made from a material selected from the group consisting of Si, SiO2, Si3N4, alumina, nitrides, metals, polymers, and combinations thereof; at least one nanopore extending through the membrane, thus forming at least one channel connecting the first side and the second side of the membrane, wherein the nanopore has a first opening, a second opening, and a depth, wherein the first opening opens to the first side of the membrane, and the second opening opens to the second side of the membrane, and wherein surface portions of the first side and the at least one channel connecting the first side and the second side of the membrane include modifying molecules defining hydrophobic surfaces; and means for spanning across the first opening of the nanopore, wherein the means for spanning across the first opening of the nanopore is a supported lipid bilayer structure or a spanning lipid bilayer structure. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14)
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15. A method for single molecule detection of an analyte using a nanopore device including a membrane having a thickness, with a first, exterior, side and a second, interior, side, the first side being opposite the second side, wherein the membrane is made from a material selected from the group consisting of glass, silicate, ceramic, Si, SiO2, Si3N4, alumina, nitrides, metals, polymers, and combinations thereof;
- at least one nanopore extending through the membrane, thus forming at least one channel connecting the first side and the second side of the membrane, wherein the nanopore has a first opening, a second opening, and a depth, wherein the first opening opens to the first side of the membrane, and the second opening opens to the second side of the membrane, and wherein surface portions of the first side and the at least one channel connecting the first side and the second side of the membrane include modifying molecules defining hydrophobic surfaces; and
means for spanning across the first opening of the nanopore, wherein the means for spanning across the first opening of the nanopore is a supported lipid bilayer structure or a spanning lipid bilayer structure, wherein a sensing entity is embedded in the lipid bilayer structure spanning across the first opening of the nanopore the method comprising;providing a sample solution containing an analyte of interest; contacting the nanopore device with the sample solution such that the first, exterior, side of the membrane is immersed in the sample solution and the at least one nanopore is filled with the sample solution; applying an appropriate voltage across the first and second side of the membrane; measuring the resistance or current flow through the at least one nanopore using either alternating current (AC) or direct current (DC) measurements; and analyzing the observed resistance or current flow to produce a useful output. - View Dependent Claims (16, 17)
- at least one nanopore extending through the membrane, thus forming at least one channel connecting the first side and the second side of the membrane, wherein the nanopore has a first opening, a second opening, and a depth, wherein the first opening opens to the first side of the membrane, and the second opening opens to the second side of the membrane, and wherein surface portions of the first side and the at least one channel connecting the first side and the second side of the membrane include modifying molecules defining hydrophobic surfaces; and
Specification