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Superresolution microscopy for bioimaging at the nanoscale: from concepts to applications in the nucleus

Authors Georgieva M, Nöllmann M

Received 5 May 2015

Accepted for publication 1 July 2015

Published 28 September 2015 Volume 2015:6 Pages 157—169

DOI https://doi.org/10.2147/RRB.S60501

Checked for plagiarism Yes

Review by Single-blind

Peer reviewer comments 3

Editor who approved publication: Professor Zvi Kelman

Mariya Georgieva,1 Marcelo Nöllmann1,2

1Centre de Biochimie Structurale, Centre National de la Recherche Scientifique (UMR5048), Institut National de la Santé et la Recherche Médicale (U1054), 2University of Montpellier, Montpellier, France

Abstract: Superresolution microscopy breaks the diffraction limit of light, making it possible to visualize a broad range of subcellular components with nearly molecular scale detail. The potential of this powerful tool is continuously growing since the implementation of optical configurations and data analyses compatible with the technically challenging, yet frequent in biology, thick and crowded samples. We review the principles underlying stimulated emission depletion, structured illumination, and single-molecule localization microscopy approaches, and their technical developments, with an emphasis on three-dimensional and live-cell imaging. Special attention is brought to the new requirements for probe efficiency, namely their size and their photophysical properties. Finally, recent applications exploring the interphase nucleus are described to illustrate the performance of superresolution techniques.

Keywords: fluorescence microscopy, superresolution techniques, nucleus, single-molecule localization microscopy, structured-illumination microscopy

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