Super-resolution microscopy techniques have revolutionized imaging and found numerous applications in probing and understanding biological processes on nanometer length scales, due to their unprecedented ability to break the optical diffraction limit by several orders of magnitude. However, most current super-resolution microscopies require fluorophore-labeled samples, which are highly subject to degradation and potentially perturbative, limiting the scope and time resolution of super-resolution measurements.<br/><br/>Our lab has developed an imaging technique capable of achieving similar spatial resolution without the need for external fluorescent labels, by combining aspects of stimulated Raman microscopy with stimulated emission depletion microscopy. This microscope provides far-field Raman images with resolution well below the diffraction limit. In addition to providing nanoscale information on chemical composition, the technique also enables interrogation of dynamics on the ultrafast timescale, providing information on both structure and function in deliberately nanostructured systems such as photovoltaic devices. This talk will discuss the photophysics behind our super-resolution Raman technique, our efforts to determine the practical resolution limit, as well as emphasizing its current applications to membrane protein biophysics. Current research focuses on using this technique to determine the effect of local cellular environments on membrane protein dynamics, and to investigate localized cooperative effects in protein interactions. Measurements on lipid dynamics and the mechanism of endocytosis of drug delivery vehicles will be discussed. The use of a label-free, super-resolution Raman imaging technique is a powerful new approach in determining how localized membrane environments affect chemical reaction mechanisms and dynamics.

Towards label-free super-resolution Raman imaging

256th National Meeting (2018)

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