Katie L. Holland, Sarah E. Plutkis, Brian P. English, Timothy A. Daugird, Abhishek Sau, Jonathan B. Grimm, Qinsi Zheng, Ankith Sharma, Sandeep Dave, Anja Schmidt, Fariha Rahman, Liangqi Xie, Peng Dong, Ariana N. Tkachuk, Timothy A. Brown, Robert H. Singer, Zhe Liu, Siegfried M. Musser, Wesley R. Legant, Catherine G. Galbraith, Luke D. Lavis
Spontaneously blinking fluorophores toggle between nonfluorescent and fluorescent forms without caging groups or redox buffers, enabling super-resolution imaging. The intrinsic blinking of such dyes is governed by molecular structure and modulated by environment; there is no one-size-fits-all fluorophore suitable for every imaging context. We report dyes with tuned on:off ratios that enable single-molecule localization microscopy and super-resolution optical fluctuation imaging of biomolecular structures in vitro and in cells.