Live cell super resolution imaging by radial fluctuations using fluorogen binding tags. Issue 8 (8th February 2019)

• Record Type: Journal Article
• Authors: Venkatachalapathy, Muthukumaran; Belapurkar, Vivek; Jose, Mini; Gautier, Arnaud; Nair, Deepak
• Is Part Of: Nanoscale. Volume 11:Issue 8(2019)
• Page Start: 3626
• Page End: 3632
• Publication Date: 2019-02-08
• Subjects: Nanoscience -- Periodicals; Nanotechnology -- Periodicals; 620.505
• DOI: 10.1039/c8nr07809b
• Languages: English
• ISSNs: 2040-3364

Abstract

Abstract : Combination of SRRF and stochastic labeling based on FAST:Fluorogen complexes to achieve super-resolution in 2D, 3D and in time-lapse. Abstract : 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.