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Super-resolution microscopy encompasses optical microscopy techniques that produce images in which structures are laterally resolved beyond the diffraction limit of light (250nm). These techniques have revolutionized fluorescence microscopy and enabled probing cellular structure in unprecedented detail. Since Eric Betzig, Stefan Hell and William Moerner were awarded the 2014 Nobel prize in Chemistry for developing super resolved fluorescence microscopy, the methodologies have continuously evolved and are becoming increasingly amenable to real time and quantitative imaging. Super resolution imaging is now also being used in conjunction with other microscopy techniques to further extend their abilities to uncover nanoscale detail. This Collection showcases the latest research presenting applications of super resolution microscopy—to characterize cellular ultrastructure and dynamics, in medicine—as well as technological improvements and quantitative methods applied to the techniques.