Free-Standing Lipid Bilayers Based on Nanopore Array and Ion Channel Formation

Area of Science:

• Nanotechnology
• Biophysics
• Materials Science

Background:

• Integrated nanopores are promising for label-free biopolymer detection but require stable lipid bilayers for protein function.
• Existing methods for creating lipid bilayers can be unstable, limiting nanopore sensor applications.

Purpose of the Study:

• To describe a novel method for producing stable lipid bilayers across nanopore arrays on silicon nitride substrates.
• To demonstrate the utility of these integrated nanopores for ion channel studies and biopolymer characterization.

Main Methods:

• A painting technique was used to embed alpha-hemolysin (α-HL) into lipid bilayers across nanopore arrays.
• Real-time monitoring of membrane formation, stability, and ion channel recordings was performed using patch clamp.
• Electrical recordings assessed bilayer conductance, resistance, and capacitance.

Main Results:

• Stable lipid bilayers were formed across nanopore arrays, exhibiting low conductance (<10 pS) and high resistance (>1.0 GΩ).
• The silicon nitride substrate provided excellent membrane stability, with membrane lifetimes ranging from 5 to 24 hours.
• Single alpha-hemolysin (α-HL) ion channels were successfully incorporated and recorded, showing characteristic current amplitudes (~100±10 pA).

Conclusions:

• The developed method enables robust and stable lipid bilayer formation on integrated nanopore devices.
• These integrated nanopore systems facilitate the analysis of ion channel functions under diverse conditions.
• This technology holds potential for high-throughput medical screening and diagnostics.