Development of spectroscopic Single-Molecule Localization Microscopy

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PROJECT SUMMARY One of the major recent scientific breakthroughs was the emergence of super-resolution optical nanoscopy. Nanoscale optical imaging currently plays a crucial role in biomedical sciences and has provided invaluable information about the cellular structure and function at the macromolecular scale. However, the potential of this technology can be immensely enhanced. We propose to develop and optimize a new class of spectroscopic super-resolution imaging tool, referred to as spectroscopic single-molecular localization microscope or sSMLM. It will allow biomedical researchers, for the first time, to achieve multi-color imaging with molecular specificity at sub 10-nm spatial resolution with principally unlimited number of distinguishable molecules. We will develope the sSMLM with modular optical designs and open-source data processing software. The unique spectral analysis optical module will be compatible with existing commercial SMLM systems with minimal or no system modification. The new hardware and software development together with optimized experimental protocols will ensure maximal accessibility of sSMLM by the broad biomedical research community. We anticipate that sSMLM will push the envelope of super-resolution imaging in fundamental biological research beyond current static spatial analysis of only few (<4) molecules.