Fast, Multi-modal Nanoscopy with SRRF-Stream
Super-resolution radial fluctuations (SRRF) is a synthesis of temporal fluctuation analysis and localization microscopy. One of the key differences between SRRF and other super-resolution methods is its applicability to live-cell dynamics because it functions across a very wide range of fluorophore densities and excitation powers.
SRRF is applied to data from imaging modes which include widefield, TIRF and confocal, where short frame bursts (e.g. 50 frames) can be processed to deliver spatial resolution similar to or better than structured illumination microscopy (SIM). With sparse data e.g. direct STORM, SRRF provides resolution similar to Gaussian fitting localization methods. Thus SRRF could provide a route to super-resolution without the need for specialized optical hardware, exotic probes or very high power densities.
We present a fast GPU-based algorithm, “SRRF-Stream” and apply it to imagery from our new multimodal imaging platform, Dragonfly. SRRF-Stream runs on an NVidia GPU, and streams images direct from camera to GPU for real time processing. We present preliminary data from EMCCD and sCMOS cameras, and various fluorophores including fluorescent proteins and organic dyes.
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