The extension of PALM and STORM based methods to live-cell dynamics is limited due to their reliance on the sparse activation of non-conventional photo-switchable probes, generally requiring intense phototoxic illumination and long acquisition times. In this talk I will describe a new approach, Super-Resolution Radial Fluctuations (SRRF), based on a novel image analysis technique, and provided as a fast GPU-enabled ImageJ plugin. The requirement for sparse activation of fluorophores is circumvented in SRRF by calculating the temporal correlations present after applying a simple and fast image transform that quantifies the radial symmetries in short image sequences. The applicability of SRRF to non-photoswitching datasets makes super-resolution possible 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. We demonstrate, using SRRF, live-cell super-resolution images of microtubule and mitochondrial dynamics, as well as extensive cortical actin remodelling during the formation of the immunological T-cell synapse.


Seminar, July 19, 2017, 12:00. Seminar Room

Hosted by Prof. Pablo Loza-Álvarez