Live-cell nanoscopy enabled with transient labeling and the control of fluorophore blinking

Abstract

Live-cell super-resolution of proteins labeled with genetically encoded fluorescent tags is a challenging task because of the imperfect labeling and the inevitable deterioration of the signal in the course of the experiment. Incomplete maturation of the covalently attached fluorescent tags, inefficient photoconversion, and photobleaching further complicate prolonged live-cell nanoscopy. We have implemented two strategies for lowering the photodamage: ensuring the dynamic replacement of damaged molecules and establishing conditions for the robust intrinsic blinking of the tags at lower illumination powers.