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research article

Live-cell multiplane three-dimensional super-resolution optical fluctuation imaging

Geissbuehler, Stefan
•
Sharipov, Azat  
•
Godinat, Aurélien  
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2014
Nature Communications

Super-resolution optical fluctuation imaging (SOFI) provides an elegant way of overcoming the diffraction limit in all three spatial dimensions by computing higher-order cumulants of image sequences of blinking fluorophores acquired with a classical widefield microscope. Previously, three-dimensional (3D) SOFI has been demonstrated by sequential imaging of multiple depth positions. Here we introduce a multiplexed imaging scheme for the simultaneous acquisition of multiple focal planes. Using 3D cross-cumulants, we show that the depth sampling can be increased. The simultaneous acquisition of multiple focal planes significantly reduces the acquisition time and thus the photobleaching. We demonstrate multiplane 3D SOFI by imaging fluorescently labelled cells over an imaged volume of up to 65×65×3.5 μm<sup>3</sup> without depth scanning. In particular, we image the 3D network of mitochondria in fixed C2C12 cells immunostained with Alexa 647 fluorophores and the 3D vimentin structure in living Hela cells expressing the fluorescent protein Dreiklang.

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Type
research article
DOI
10.1038/ncomms6830
Web of Science ID

WOS:000347681600001

Author(s)
Geissbuehler, Stefan
Sharipov, Azat  
Godinat, Aurélien  
Bocchio, Noelia L.
Sandoz, Patrick A.
Huss, Anja
Jensen, Nickels A.
Jakobs, Stefan
Enderlein, Jörg
Gisou Van Der Goot, F.  
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Date Issued

2014

Publisher

Nature Research

Published in
Nature Communications
Volume

5

Article Number

5830

Subjects

fluorescence nanoscopy

•

cumulant imaging

Editorial or Peer reviewed

REVIEWED

Written at

EPFL

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Available on Infoscience
December 19, 2014
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/109441
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