Droplet Split-and-Contact Method for High-Throughput Transmembrane Electrical Recording

• Published in: Analytical chemistry (Washington) Vol. 85; no. 22; pp. 10913 - 10919
• Main Authors: Tsuji, Yutaro, Kawano, Ryuji, Osaki, Toshihisa, Kamiya, Koki, Miki, Norihisa, Takeuchi, Shoji
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 19.11.2013
• Subjects: Analytical chemistry; Aqueous chemistry; Cell Membrane - metabolism; Channels; Conductance; Contact; Droplets; Electrophysiology - methods; Formations; Hemolysin Proteins - chemistry; Hemolysin Proteins - metabolism; High-Throughput Screening Assays - methods; Humans; Ion channels; Ion Channels - metabolism; Ions; Lipid Bilayers - chemistry; Lipids; Membranes; Nanopores; Oils - chemistry; Recording
• ISSN: 0003-2700; 1520-6882
• DOI: 10.1021/ac402299z

This paper describes the rapid and repetitive formation of planar lipid bilayers via a mechanical droplet contact method for high-throughput ion channel analysis. In this method, first, an aqueous droplet delivered in a lipid-in-oil solution is mechanically divided into two small droplets. Second, the two small droplets contact each other, resulting in the lipid bilayer formation. Third, an ion channel is immediately reconstituted into the bilayer and the transmembrane current signals are measured. By repeating this procedure, massive data sets of the channel signals can be obtained. This method allowed us to perform statistical analysis of α-hemolysin conductance (n = 256 within 30 min) and channel inhibition experiments by contacting different types of the droplets in a short time frame.