In order to overcome the diffraction limit, new approaches in fluorescence microscopy have been developed in recent years. In particular, single molecule localization microscopy consists of forcing fluorescent molecules to emit in a spatially and temporally shifted manner in order to determine their position with nanometric precision. This approach combining photophysics, optical development and new AI based processing allows us to open new field of applications in biology. Within the internship (potentially followed by a ERC funded PhD), a new concept of localization will be developed, based on a new optical implementation. This will permit to further push not only the spatial resolution but also open the detection of new functional parameter at the nanoscale (lifetime, spectrum), and to test on biological applications on cell migration