Investigation of biomolecular interactions at single-molecule level

Numerous biomolecules and their interactions possess physiologically relevant heterogeneities that can be unraveled only by means of single-molecule tools. All current single-molecule imaging methods, however, require fluorescent labelling or immobilization onto a surface, which might alter the biomolecule’s natural behavior. Recently, a ground-breaking optical microscopy method has been developed – Nanofluidic Scattering Microscopy (NSM) [1] – whose unprecedented resolution enabled to bypass those limitations and to image individual proteins in free motion without any label. Monitoring of individual biomolecules diffusing in solution represents a yet unexplored opportunity for studies of molecular transport, interactions between individual biomolecules, or analysis of complex biofluids. With further advances in this novel technology, NSM is expected to have a significant impact in the field of single-molecule sciences and to inspire new strategies for early diagnosis and therapeutic intervention.

The dissertation will be focused on further development of this novel method and its application in investigation of biomolecular interactions.

[1] B. Špačková, et al: Nanofluidic Scattering Microscopy for label-free weight and size screening of single diffusing biomolecules. Nature Methods 19, 751-758 (2022) https://doi.org/10.1038/s41592-022-01491-6