Intramolecular optical microscopy with sub-nm spatial resolution in single biomolecules and single-protein sequencing

Fluorescence microscopy has witnessed a true resolution revolution in the past decades.

The invention of methods circumventing the classical diffraction limit of ≈200 nm has allowed researchers – for the first time – to reach sub-diffraction resolution with optical fluorescence microscopy.

With these so-called super-resolution techniques, processes within cells can be observed at thus far unprecedented spatial resolutions.

However, while incumbent super-resolution approaches enable researchers to resolve intermolecular distances between e.g. protein molecules in small protein clusters, the final frontier in optical bioimaging to resolve intramolecular distances in the sub-nm regime for single molecules is still elusive.

With this research proposal, I want to develop and apply a paradigm-shifting imaging technology that allows intramolecular optical microscopy with sub-nm spatial resolution in single biomolecules.To achieve this goal, I will combine DNA-Point Accumulation Imaging Nanoscale Topography (DNA-PAINT) with DNA Exchange to overcome the current limit of optical nanoscopy in terms of spatial resolution.

To do so, I will build a microscope with ultra-high stability and sensitivity.

The project is conceived as an integral approach including the design of optics, instrumentation, labelling probes and data analysis.Such a capability provided by the project IntraMol would be a game changer in fluorescence microscopy, as it could – for the first time – allow scientists to resolve intramolecular distances e.g. within single proteins, thus enabling structural biology studies and provide insights into conformations of proteins even enable single-molecule protein sequencing using optical microscopy in situ.