Parallel and Automated Formation of Lipid Bilayers on Microstructured Chips for Ion Channel and Nanopore Recordings

Abstract

Bilayer recording is a well-established technique for in-depth studies of biophysical properties of ion channels and is particularly suited for functional studies on proteins residing in intracellular membranes. Moreover, this technique supports a host of powerful emerging analytical techniques using biological nanopores as molecular sensors. Despite its proven value, bilayer recording can be frustrating due to the capricious nature of lipid bilayers, which have to be formed manually one by one and which often lack stability. We here show a new approach and device, which speeds up the entire process by the rapid and simultaneous formation of 16, highly stable micrometer-sized bilayers using Micro-Electrode-Cavity-Array (MECA)-Chips. A study will be presented showing that the MECA supports high-resolution polymer sizing with a single biological nanopore in a parallel format (Fig.1). Additionally, data on a variety of channel proteins recorded from proteoliposomes will be shown. using a surface containing micron-sized apertures in glass substrates, the fusion of vesicles on the surface becomes an attractive method for electrophysiology and then to reconstitute membrane proteins into the lipid bilayer.Fig. 1MonoPEG-28-mediated blockages of HL-nanopore(s) on a MECA chip.View Large Image | View Hi-Res Image | Download PowerPoint Slide