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Real time multi-modal super-resolution microscopy through Super-Resolution Radial Fluctuations (SRRF-Stream)

Cooper, J; Browne, M; Gribben, H; Catney, M; Coates, C; Mullan, A; Wilde, G; (2019) Real time multi-modal super-resolution microscopy through Super-Resolution Radial Fluctuations (SRRF-Stream). In: Proceedings of SPIE: Single Molecule Spectroscopy and Superresolution Imaging XII. SPIE Green open access

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Abstract

Super-resolution radial fluctuations (SRRF) is a combination 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 enhancements similar to or better than structured illumination microscopy (SIM). On the other hand, with sparse data e.g. stochastic optical reconstruction microscopy (STORM), SRRF can deliver 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 GPUbased SRRF algorithm termed “SRRF-Stream” and apply it to imagery from an iXon EMCCD coupled to a multi-modal imaging platform, Dragonfly. The new implementation is <300 times faster than the standard CPU version running on an Intel Xeon 3.5GHz 4 core processor, and < 20 times faster than the NanoJ GPU implementation, while also being integrated with acquisition for real time use. In this paper we explore the image resolution and quality with EMCCD and sCMOS cameras and various fluorophores including fluorescent proteins and organic dyes.

Type: Proceedings paper
Title: Real time multi-modal super-resolution microscopy through Super-Resolution Radial Fluctuations (SRRF-Stream)
Event: Single Molecule Spectroscopy and Superresolution Imaging XII
Location: San Francisco (CA), USA
Dates: 2nd-7th February 2019
ISBN-13: 9781510624108
Open access status: An open access version is available from UCL Discovery
DOI: 10.1117/12.2510761
Publisher version: https://doi.org/10.1117/12.2510761
Language: English
Additional information: This version is the version of record. For information on re-use, please refer to the publisher’s terms and conditions.
UCL classification: UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences
URI: https://discovery.ucl.ac.uk/id/eprint/10076905
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