My work showcases the application of the two advanced fluorescence microscopy methods — single-molecule FRET (smFRET) and sub-ensemble Sensitized Emission Imaging (SEI) and complementary data analysis pipelines —to unravel molecular interactions and dynamic processes with high spatiotemporal resolution. These methodologies were employed to investigate three diverse biological systems: (a) the interaction between ribosome and Erythromycin resistance methyltransferases (Erm), which contributes to the emergence of antibiotic resistance, using intermolecular FRET scheme (Accepted in Science Advances); (b) conformational dynamics of a protein PurL in presence of its substrate, FGAR, using intramolecular FRET scheme (Mansuscript under preparation) and (c) the surface heterogeneity of  fibrillar form of an intrinsically disordered protein α-synuclein (α-syn), with the help of SEI (ACS Chem. Neurosci. 2024, 15, 1, 108–118). 

Schematic of TIRF based smFRET data acquisition and analysis pipeline.