Fast live-cell conventional fluorophore nanoscopy with ImageJ through super-resolution radial fluctuations

Research output: Contribution to journalArticlepeer-review


  • Nils Gustafsson
  • Siân Culley
  • George Ashdown
  • Pedro Matos Pereira
  • Ricardo Henriques

Colleges, School and Institutes

External organisations

  • Centre for Mathematics and Physics in Life Sciences and Experimental Biology (CoMPLEX), University College London, Gower Street, London WC1E 6BT, UK
  • Quantitative Imaging and Nanobiophysics Group, MRC Laboratory for Molecular Cell Biology, Department of Cell and Developmental Biology, University College London, Gower Street, London WC1E 6BT, UK
  • Department of Physics and Randall Division of Cell and Molecular Biophysics, King's College, London


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.


Original languageEnglish
Article number12471
Number of pages9
JournalNature Communications
Publication statusPublished - 12 Aug 2016