Department of Chemistry, University of Cincinnati, Cincinnati, Ohio 45221, United States.
Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, Koganei-shi, Tokyo 184-8588, Japan.
Anal Chem. 2020 Aug 4;92(15):10856-10862. doi: 10.1021/acs.analchem.0c02720. Epub 2020 Jul 13.
Biological nanopores reconstituted into supported lipid bilayer membranes are widely used as a platform for stochastic nanopore sensing with the ability to detect single molecules including, for example, single-stranded DNA (ssDNA) and miRNA. A main thrust in this area of research has been to improve overall bilayer stability and ease of measurements. These improvements are achieved through a variety of clever strategies including droplet-based techniques; however, they typically require specific microfabrication techniques to prepare devices or special manipulation techniques for microdroplets. Here, we describe a new method to prepare lipid bilayers using a recessed-in-glass Ag/AgCl microelectrode as a support structure. The lipid bilayer is formed at the tip of the microelectrode by immersing the microelectrode into a layered bath solution consisting of an oil/lipid mixture and an aqueous electrolyte solution. In this paper, we demonstrate this stable, supported lipid bilayer structure for channel current measurements of pore-forming toxins and single-molecule detection of ssDNA. This Ag/AgCl-supported lipid bilayer can potentially be widely adopted as a lipid membrane platform for nanopore sensing because of its simple and easy procedure needed to prepare lipid bilayers.
生物纳米孔重新构成的支撑脂质双层膜被广泛用作随机纳米孔传感的平台，具有检测单分子的能力，例如单链 DNA（ssDNA）和 miRNA。该研究领域的一个主要重点是提高双层稳定性和测量的便利性。这些改进是通过各种巧妙的策略实现的，包括基于液滴的技术；然而，它们通常需要特定的微制造技术来制备器件或特殊的微滴操纵技术。在这里，我们描述了一种使用凹陷玻璃 Ag/AgCl 微电极作为支撑结构来制备脂质双层的新方法。脂质双层在微电极的尖端形成，方法是将微电极浸入由油/脂质混合物和水性电解质溶液组成的分层浴溶液中。在本文中，我们展示了这种稳定的、支撑的脂质双层结构，用于孔形成毒素的通道电流测量和 ssDNA 的单分子检测。由于制备脂质双层所需的简单易行的步骤，这种 Ag/AgCl 支撑的脂质双层有可能被广泛应用于纳米孔传感的脂质膜平台。
