Democratising live-cell high-speed super-resolution microscopy

The extension of PALM and STORM based methods to live-cell dynamics is limited due to their reliance on intense phototoxic illumination and long acquisition times. In this talk I will describe a new approach, Super-Resolution Radial Fluctuations (SRRF), capable of achieving super-resolution with illumination orders of magnitude lower than methods such as SMLM or STED. It also enables live-cell imaging with conventional fluorophores using modern widefield, confocal or TIRF microscopes, achieving resolutions better than 150nm at 1 frame per second. Meanwhile, in datasets suitable for SMLM analysis SRRF achieves resolutions matching standard analysis techniques (20nm). We demonstrate, using SRRF, live-cell super-resolution images of microtubule, mitochondrial dynamics, the dynamic nanoscale reorganisation of HIV-1 host receptors, as well as extensive cortical actin remodelling during the formation of the immunological T-cell synapse.