Photoswitchable fluorescent probes for localization-based super-resolution imaging

In recent years, localization-based super-resolution imaging has been developed to overcome the diffraction limit of far-field fluorescence microscopy. Photoswitchable probes are a hallmark of this technique. Their fluorescence can be modulated between an emissive and dark state whereby the sequential, nanoscale measurement of individual fluorophore positions can be used to reconstruct an image at higher spatial resolution. Despite the importance of photoswitchable probes for localization-based super-resolution imaging, both a mechanistic and quantitative understanding of the essential photoswitching properties is lacking for most fluorophores. In this thesis, we begin to address this need. Furthermore, we demonstrate the development of new probes and methodologies for both multicolor and live-cell super-resolution imaging.