Multiplexed Parallel Single Transport Recordings on Nanopore Arrays

Abstract

We introduce a nanofabricated silicon chip for massively multiplexed analysis of membrane channels and transporters in suspended lipid membranes that does not require any surface modification or organic solvent. Transport processes through single membrane complexes are monitored by fluorescence. The chip consists of an array of well-defined nanopores, addressing an individual pyramidal back-reflecting 30-fL compartment. The setup allows simultaneous analyses of ∼1,000 single transmembrane events in one field of view, observing translocation kinetics of transmembrane complexes.

Membrane-on-a-Chip: Microstructured Silicon/Silicon-Dioxide Chips for High-Throughput Screening of Membrane Transport and Viral Membrane Fusion

“…The sharp transitions to the import stop were previously interpreted by diffusion of membrane pores out of the suspended bilayer into the solid-supported bilayer part next to the cavities. 4 However, at low ionic strength or other conditions with low insertion efficiency we did not observe such interrupted transport signatures, and in the previous study the experiments were performed only at high ionic strength. Instability or even disassembly of RHL heptamers has not been described so far, but single-channel recordings of RHL were previously conducted only at favorable insertion conditions.…”

“…Herein, nanopore arrays were fabricated by electron-beam lithography, while stable lipid bilayers could be formed on top of the <100 nm pores. 4 Leakage of a fluorophore entrapped in bilayer-sealed cavities was used to detect membrane pore formation. Although very accurate and sophisticated nanopore arrays can be created by electron-beam lithography, the rather difficult manufacturing process leads to low piece counts and high production costs.…”

Through a Window, Brightly: A Review of Selected Nanofabricated Thin-Film Platforms for Spectroscopy, Imaging, and Detection

“…Fluorescence is the most likely optical technique to be used in conjunction with a nanopore, but one could imagine that a suitably structured gold film, separated by a well-defined nanopore gap containing an analyte, might also be used for surface-enhanced Raman spectroscopy. 20,27,46,61,90,[267][268][269][305][306][307][308][309][310][311][312][313][314][315] Extensions to Nanopore Arrays. We conclude our discussion of the merging of physical-and optical-based nanopore sensing and manipulation approaches, with the consideration of nanopore arrays.…”

Multiplex-Einzelmolekülanalyse von Transmembranprozessen

“…Reproduziert aus[4], © American Chemical Society. Präparation lösemittelfreier freitragender Lipidmembranen auf Nanoporen.…”

“…Maßbalken: 10 μm. Reproduziert aus[4], © American Chemical Society.Abb. 3: Parallele Analyse vonMembrantransport am Beispiel von α-Hämolysin.…”