120th General Meeting of the KCS

Type Symposium
Area Physical Chemistry of Bioimaging and Biospectroscopy
Room No. Room 208+209+210
Time FRI 09:40-:
Code PHYS2-3
Subject Super-resolution Fluorescence Microscopy For Visualizing Long-term Cellular Dynamics
Authors Sang-Hee Shim
Department of Chemistry, Korea University, Korea
Abstract Super-resolution fluorescence microscopy opens new windows for visualizing ultrastructural dynamics. When applied to living cells, super-resolution fluorescence microscopy suffers from the limited length of time-lapse series due to high intensities of illumination and photobleaching of fluorophores. Here, we use a fluorogen-binding protein, UnaG for overcoming the photobleaching limit by controlling the switching kinetics and by supplying a large excess of fluorogenes. UnaG fluorescence can be switched off by blue light and then, recovered by replacing the damaged fluorogen with a fresh one in solution for hundreds of cycles. These switching properties enabled long-term super-resolution imaging based on single-molecule localization, with significant improvement (about 10-fold) in the number of independent super-resolution snapshots. By targeting UnaG via genetic incorporation, we demonstrated super-resolution imaging of various organelles (e.g. endoplasmic recticulum, mitochondria, peroxisome, etc) and structural proteins (e.g. vimentin, keratin, lamin, clathrin, etc). These capabilities enable us to observe cellular dynamics that progress in the time scale of minutes to hours.
E-mail sangheeshim@korea.ac.kr