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Annual Review of Physical Chemistry

Volume 75, 2024

Single-Molecule Spectroscopy and Super-Resolution Mapping of Physicochemical Parameters in Living Cells

Abstract

By superlocalizing the positions of millions of single molecules over many camera frames, a class of super-resolution fluorescence microscopy methods known as single-molecule localization microscopy (SMLM) has revolutionized how we understand subcellular structures over the past decade. In this review, we highlight emerging studies that transcend the outstanding structural (shape) information offered by SMLM to extract and map physicochemical parameters in living mammalian cells at single-molecule and super-resolution levels. By encoding/decoding high-dimensional information—such as emission and excitation spectra, motion, polarization, fluorescence lifetime, and beyond—for every molecule, and mass accumulating these measurements for millions of molecules, such multidimensional and multifunctional super-resolution approaches open new windows into intracellular architectures and dynamics, as well as their underlying biophysical rules, far beyond the diffraction limit.

Keyword(s): functional imaging, intracellular physicochemical parameters, multidimensional super-resolution microscopy, single-molecule spectroscopy, spatial mapping, spectral microscopy
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/content/journals/10.1146/annurev-physchem-070623-034225
2024-06-28
2025-04-22
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/content/journals/10.1146/annurev-physchem-070623-034225
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Single-Molecule Spectroscopy and Super-Resolution Mapping of Physicochemical Parameters in Living Cells
Annual Review of Physical Chemistry 75, 163 (2024); https://doi.org/10.1146/annurev-physchem-070623-034225
/content/journals/10.1146/annurev-physchem-070623-034225
/content/journals/10.1146/annurev-physchem-070623-034225
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