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Membrane Transport Processes Analyzed by a Highly Parallel Nanopore Chip System at Single Protein Resolution
Published on: August 16, 2016
Multichannel simultaneous measurements of single-molecule translocation in alpha-hemolysin nanopore array
Toshihisa Osaki1, Hiroaki Suzuki, Bruno Le Pioufle
1Institute of Industrial Science, The University of Tokyo, Japan.
Analytical Chemistry
|December 17, 2009
Summary
This study introduces a novel microarray system for simultaneously monitoring ionic currents through alpha-hemolysin nanopores in lipid bilayer membranes. The system demonstrates efficient bilayer formation and detects nucleic acid translocation events for high-throughput applications.
Area of Science:
- Biophysics
- Nanotechnology
- Microfluidics
Background:
- Simultaneous monitoring of ionic currents through nanopores is crucial for molecular analysis.
- Forming stable and reproducible lipid bilayer membranes in microfluidic devices presents challenges.
Purpose of the Study:
- To develop a microarray system for simultaneous monitoring of multiple ionic currents through alpha-hemolysin nanopores.
- To optimize lipid bilayer formation in a microfluidic device for enhanced nanopore sensing.
Main Methods:
- Utilized self-assembling lipid molecules at the interface of aqueous and organic solvents for bilayer formation.
- Employed a microfluidic device with a hydrophobic polymer film to create and stabilize lipid bilayers across micrometer-sized apertures.
- Controlled solvent composition (n-decane and 1-hexanol) for rapid and reproducible bilayer formation and simultaneous alpha-hemolysin nanopore incorporation.
Main Results:
- Successfully formed stable lipid bilayer membranes within the microfluidic device.
- Achieved simultaneous monitoring of ionic currents through an array of alpha-hemolysin nanopores.
- Obtained up to four synchronous ionic current signals from eight monitored wells.
- Detected translocation events of nucleic acid molecules by observing blocked current profiles.
Conclusions:
- The developed microarray system enables efficient and reproducible lipid bilayer formation with integrated nanopores.
- The system shows promise for high-throughput analysis, including the detection of nucleic acid molecules.
- This technology advances nanopore sensing capabilities for molecular diagnostics and research.
