Blinking fluorescent probes for single-molecule localization-based super-resolution imaging

The rapid advancement of single-molecule localization microscopy (SMLM) provides chances for visualizing biological structures and tracking their functions with unprecedented high resolution. The imaging speed and spatial resolution are highly dependent on the photophysical properties of fluorophores. Therefore, great efforts have been paid to develop new photoswitchable probes and improve their photophysical properties for single molecule localization applications. These developments in SMLM have markedly advanced our understanding on many biological processes. In this review, we first describe the basic principles of SMLM and discuss the factors that affect the imaging quality. Then, we summarize recent achievements in the development of fluorescent probes for SMLM and evaluate their photophysical properties. Some interesting achievements in biological applications are also highlighted. Ultimately, the challenges and future advances of SMLM for biological applications are discussed.