Fast live-cell conventional fluorophore nanoscopy with ImageJ through super-resolution radial fluctuations.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 27514992.
- Also identified by DOI 10.1038/ncomms12471 and PMC identifier 4990649.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
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.
Medical subject headings
- Fluorescent Dyes
- Microscopy, Confocal
- Microscopy, Fluorescence
- Nanotechnology
- Optical Imaging