4.7 Article

A chip-based array for high-resolution fluorescence characterization of free-standing horizontal lipid membranes under voltage clamp

Journal

LAB ON A CHIP
Volume 22, Issue 15, Pages 2902-2910

Publisher

ROYAL SOC CHEMISTRY
DOI: 10.1039/d2lc00357k

Keywords

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Funding

  1. International Graduate College 1642 Soft Matter Science: Concepts for the Design of Functional Materials of the Deutsche Forschungsgemeinschaft (DFG)
  2. German Ministry for Research and Education through Project Management PTJ [FKZ 031B0864A]
  3. BW-Foundation through Project Management VDI (MSDS-BioMem)
  4. Ministry of Commerce of the State of Baden-Wurttemberg
  5. DFG [290424854]
  6. BWFoundation

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This study presents a novel chip-based array device that allows high-resolution optical measurements and voltage clamp electrical recordings for studying membranes and membrane proteins. The device enables optical access to standing lipid membranes, while controlling membrane voltage and recording electrical signals using micropatterned electrodes. This device provides a powerful tool for combined electrical-optical studies of membranes and membrane proteins.
Optical techniques, such as fluorescence microscopy, are of great value in characterizing the structural dynamics of membranes and membrane proteins. A particular challenge is to combine high-resolution optical measurements with high-resolution voltage clamp electrical recordings providing direct information on e.g. single ion channel gating and/or membrane capacitance. Here, we report on a novel chip-based array device which facilitates optical access with water or oil-immersion objectives of high numerical aperture to horizontal free-standing lipid membranes while controlling membrane voltage and recording currents using individual micropatterned Ag/AgCl-electrodes. Wide-field and confocal imaging, as well as time-resolved single photon counting on free-standing membranes spanning sub-nanoliter cavities are demonstrated while electrical signals, including single channel activity, are simultaneously acquired. This optically addressable microelectrode cavity array will allow combined electrical-optical studies of membranes and membrane proteins to be performed as a routine experiment.

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