Live cell super resolution imaging by radial fluctuations using fluorogen binding tags

Nanoscale. 2019 Feb 21;11(8):3626-3632. doi: 10.1039/c8nr07809b.


Fluorescence-Activating and absorption-Shifting Tag (FAST) is a novel genetically encoded optical highlighter probe. Since the fluorescence of FAST originates from the stochastic and reversible diffusive association of a fluorogenic ligand, we investigate the application of FAST using Super-Resolution Radial Fluctuations (SRRF) to achieve routine imaging below the diffraction limit in a widefield epifluorescence microscope. We show that intensity fluctuation analysis like SRRF allows the imaging of FAST-tagged proteins with sub - 100 nm resolution in live cells. FAST co-labeled with conventional fluorophores enables real time multicolour 2D and 3D super-resolution imaging, indicating that FAST can be used for the observation of sub-diffraction limited structures in both living and fixed samples.

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Fluorescent Dyes / chemistry*
  • Green Fluorescent Proteins / chemistry*
  • Green Fluorescent Proteins / metabolism
  • Mice
  • Microscopy, Fluorescence
  • Photobleaching


  • Fluorescent Dyes
  • Green Fluorescent Proteins