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Issue 8, 2019
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Live cell super resolution imaging by radial fluctuations using fluorogen binding tags

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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.

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

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Publication details

The article was received on 25 Sep 2018, accepted on 17 Jan 2019 and first published on 24 Jan 2019


Article type: Paper
DOI: 10.1039/C8NR07809B
Citation: Nanoscale, 2019,11, 3626-3632

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

    M. Venkatachalapathy, V. Belapurkar, M. Jose, A. Gautier and D. Nair, Nanoscale, 2019, 11, 3626
    DOI: 10.1039/C8NR07809B

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