Gustafsson, N;
Culley, S;
Ashdown, G;
Owen, DM;
Pereira, PM;
Henriques, R;
(2016)
Fast live-cell conventional fluorophore nanoscopy with ImageJ through super-resolution radial fluctuations.
Nature Communications
, 7
, Article 12471. 10.1038/ncomms12471.
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Abstract
Despite significant progress, high-speed live-cell super-resolution studies remain limited to specialized optical setups, generally requiring intense phototoxic illumination. Here, we describe a new analytical approach, super-resolution radial fluctuations (SRRF), provided as a fast graphics processing unit-enabled ImageJ plugin. In the most challenging data sets for super-resolution, such as those obtained in low-illumination live-cell imaging with GFP, we show that SRRF is generally capable of achieving resolutions better than 150 nm. Meanwhile, for data sets similar to those obtained in PALM or STORM imaging, SRRF achieves resolutions approaching those of standard single-molecule localization analysis. The broad applicability of SRRF and its performance at low signal-to-noise ratios allows super-resolution using modern widefield, confocal or TIRF microscopes with illumination orders of magnitude lower than methods such as PALM, STORM or STED. We demonstrate this by super-resolution live-cell imaging over timescales ranging from minutes to hours.
| Type: | Article |
|---|---|
| Title: | Fast live-cell conventional fluorophore nanoscopy with ImageJ through super-resolution radial fluctuations |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1038/ncomms12471 |
| Publisher version: | http://doi.org/10.1038/ncomms12471 |
| Language: | English |
| Additional information: | © The Author(s) 2016. This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
| Keywords: | Science & Technology, Multidisciplinary Sciences, Science & Technology - Other Topics, SINGLE-MOLECULE LOCALIZATION, STRUCTURED ILLUMINATION MICROSCOPY, FLUORESCENCE MICROSCOPY, DIFFRACTION-LIMIT, RESOLUTION, RECONSTRUCTION, ALGORITHM, DYNAMICS, SYMMETRY, STORM |
| UCL classification: | UCL 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 UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Lab for Molecular Cell Bio MRC-UCL UCL > Provost and Vice Provost Offices > UCL BEAMS UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Maths and Physical Sciences |
| URI: | https://discovery.ucl.ac.uk/id/eprint/1508261 |
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