Micro- and Nanostructured Devices for the Investigation of Biomolecular Interactions

Abstract

Cell membrane receptors and ion channels are essential in many different cellular processes. To analyze the activity of membrane proteins in vesicles and biological cells, we fabricated micro-nano structured chips, enabling the application of electro physiology and fluorescence-based techniques. A SU-8 biochip was developed to simultaneously micromanipulate and investigate optically and electrically individual vesicles in a microfluidic channel. Lipid vesicles were transported, positioned by electrophoretic movement on a micrometer sized aperture, and fused to form a planar suspended membrane, which is suited to study ion channel activity. Aiming to investigate G protein coupled receptor signaling pathways in native-like environment, we developed a method for producing well-Oriented planar cell membrane sheets on silicon films containing nanoaperture arrays. The accessibility of extracellular and cytosolic surfaces was demonstrated by targeting membrane constituents side-specifically with fluorescent markers. Our approach can be applied for studying membrane proteins from a large variety of cells and cellular or ganelles using chip-based screening assays.

Keywords

• Electrophysiology
• Fluorescence techniques
• Membrane proteins
• Micro-nanoapertures