Single-Channel Single-Molecule Detection (SC-SMD) System.

The combination of single-molecule fluorescence imaging and electrophysiology provides a powerful tool to explore the stoichiometry and functional properties of ionic channels, simultaneously. Here, we describe a typical SC-SMD experiment from the preparation of plasmids containing the genes encoding for the channels of interest fused to fluorescent proteins to the use of the SC-SMD system for simultaneous patch clamping and single-molecule determinations.