LAUSR

Abstract The diffraction limit is no longer a concept that stands as a true constant in imaging, with fluorescence switching–based methods having made the breakthrough to circumvent this limit. Multiple ingenious solutions have been presented over the last decades and continue to be explored. The techniques used today have undergone constant development both conceptually and technically, which has enabled an increased number of biological studies at the molecular scale. Here we review recent developments to stimulated emission depletion microscopy, reversible saturable optical fluorescence transitions microscopy, single-molecule localization microscopy, and MINFLUX and mention key applications of these methods. Finally, we present our view on what the future holds for super-resolution imaging, especially in terms of even more challenging live-cell imaging in different biological model systems.