University of Cambridge > > Biophysical Seminars > Probing intracellular physicochemical environments at the nanoscale, one molecule at a time

Probing intracellular physicochemical environments at the nanoscale, one molecule at a time

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Recent advances in super-resolution microscopy methods based on single-molecule imaging (single-molecule localization microscopy; SMLM ) have led to ~10 nm spatial resolution and exciting new biology. We are advancing beyond the structural (shape) information offered by existing methods, to reveal (intracellular) functional parameters, including chemical polarity, diffusivity, and reactivity, with nanoscale resolution and single-molecule sensitivity. To this end, we have been developing new strategies to perform high-throughput, multidimensional single-molecule spectroscopy in the wide-field. In particular, with spectrally resolved SMLM , we encoded functional parameters into the emission spectra of single probe molecules, and so unveiled rich, nanoscale functional and compositional heterogeneities in the membranes of live mammalian cells. With single-molecule displacement/diffusivity mapping (SMdM), we mapped out intracellular diffusivity with unprecedented spatial resolution and fidelity, and hence discovered that diffusion in the mammalian cytoplasm and nucleus are both spatially heterogeneous at the nanoscale, and identified the net charge of the diffuser as a previously overlooked, key determinant of diffusion rate. By adding rich functional dimensions to the already powerful super-resolution microscopy, we thus open up new ways to reveal fascinating local heterogeneities in both live cells and chemical systems.

This talk is part of the Biophysical Seminars series.

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