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Advances in High Resolution, High Content Optical Biomolecular Imaging

Peter T. C. So
Department of Mechanical and Biological Engineering, MIT

A central challenge of modern biology is the need to understand how the interactions of protein machines affect cellular physiology and the pathology. Optical imaging and spectroscopy afford unprecedented opportunities in studying these dynamical processes in vivo. In this lecture, I will focus on two microscopy technologies. First, time-resolved fluorescence resonance energy transfer microscopy has been developed to study mechanotransduction processes. Specifically, we demonstrate for the first time the quantification of association constants of focal adhesion proteins in vivo. Second, the resolution of optical imaging has been limited by the diffraction of light. I will describe standing-wave total internal reflection microscopy, a wide-field, high speed optical imaging technique with demonstrated 80 nm resolution. The possibility of extending the resolution of this technique to sub-50 nm based on surface plasmon excitation will be examined.