Abstract
Membrane proteins are notoriously difficult to study. While supported lipid bilayers offer stability and allow the application of surface measurement techniques, integral membrane proteins are often not fully functional when close to a solid surface. We have developed a membrane interferometer which allows free standing membranes suspended above an atomically flat silicon surface to be studied by fluorescence interference measurements (Prasad V. Ganesan and Steven G. Boxer, PNAS, 2009,106, 5627-5632). In this platform, a lipid bilayer is suspended across a micron-sized well, allowing the use of Variable Incidence Angle Fluorescence Interference Contrast Microscopy (VIA-FLIC; Caroline Ajo-Franklin, Prasad V. Ganesan, and Steven G. Boxer, Biophys. J., 2005, 89, 2759-2769). The interferometry measurements in VIA-FLIC can be used to determine the height of fluorescent dyes relative to the mirror with an axial resolution of a few nanometers. Moreover, by incorporating electrodes on both sides of the suspended bilayer, we wish to perform concurrent conductance and optical measurements that will allow us to study the function and conformation of single membrane proteins at the same time. Progress towards incorporating ion channels into the interferometer and making electrophysiology measurements will be reported.
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