Super-resolution fluorescence microscopy-based single-molecule spectroscopy

Abstract

The rise of super-resolution fluorescence microscopy (SRM) has revolutionized our understanding of the nanoscale world. SRM has recently been correlated with spectroscopy techniques to obtain rich spectral information of individual molecules in addition to localization information. This technique ultimately provides multidimensional and functional information of single molecules, which is generally masked in the ensemble averages of previous bulk measurements and creates exciting new opportunities and challenges for single-molecule spectroscopic imaging. We review the recent developments in single-molecule spectroscopic imaging technique and how the challenges are addressed to effectively obtain single-molecule spectroscopic information. Particular attention has been devoted to the applications of this method, including single-molecule chemical kinetic studies and single-molecule polarity sensing. Finally, we address the possible future enhancements and applications. Thus, we demonstrate how a single-molecule spectroscopic imaging technique adds new dimensions of information to the SRM, providing new opportunities in a wide range of research fields.