Single-Molecule Localization Microscopy Propelled by Small Organic Fluorophores with Blinking Properties

Super-resolution fluorescence imaging by single-molecule localization microscopy (SMLM) has paved the way for a better understanding of cellular structures and processes at the molecular scale. Since SMLM relies on the fluorescence switching and detection of single molecules of fluorescent probes, the probe performance is of crucial importance. Small organic fluorophores have contributed greatly to SMLM, owing to their brightness, photostability, and potential for chemical modification and functionalization. Here, we provide an overview of the techniques of SMLM, including a discussion of the characteristics of existing small organic fluorophores. In particular, we highlight a novel class of spontaneously blinking fluorophores that we have recently developed, describing some of their applications, together with details of the experimental methods.