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Int J Biochem Cell Biol. 2018 Aug;101:74-79. doi: 10.1016/j.biocel.2018.05.014. Epub 2018 May 28.

SRRF: Universal live-cell super-resolution microscopy.

Author information

1
MRC Laboratory for Molecular Cell Biology, University College London, Gower Street, London WC1E 6BT, UK; Department of Cell and Developmental Biology, University College London, Gower Street, London, WC1E 6BT, UK; The Francis Crick Institute, 1 Midland Road, London, NW1 1AT, UK.
2
MRC Laboratory for Molecular Cell Biology, University College London, Gower Street, London WC1E 6BT, UK; Department of Cell and Developmental Biology, University College London, Gower Street, London, WC1E 6BT, UK.
3
MRC Laboratory for Molecular Cell Biology, University College London, Gower Street, London WC1E 6BT, UK; Department of Cell and Developmental Biology, University College London, Gower Street, London, WC1E 6BT, UK; The Francis Crick Institute, 1 Midland Road, London, NW1 1AT, UK. Electronic address: r.henriques@ucl.ac.uk.

Abstract

Super-resolution microscopy techniques break the diffraction limit of conventional optical microscopy to achieve resolutions approaching tens of nanometres. The major advantage of such techniques is that they provide resolutions close to those obtainable with electron microscopy while maintaining the benefits of light microscopy such as a wide palette of high specificity molecular labels, straightforward sample preparation and live-cell compatibility. Despite this, the application of super-resolution microscopy to dynamic, living samples has thus far been limited and often requires specialised, complex hardware. Here we demonstrate how a novel analytical approach, Super-Resolution Radial Fluctuations (SRRF), is able to make live-cell super-resolution microscopy accessible to a wider range of researchers. We show its applicability to live samples expressing GFP using commercial confocal as well as laser- and LED-based widefield microscopes, with the latter achieving long-term timelapse imaging with minimal photobleaching.

KEYWORDS:

Fluorescence; Image processing; Live-cell imaging; Super-resolution microscopy

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