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research article

Complementarity of PALM and SOFI for super-resolution live-cell imaging of focal adhesions

Deschout, Hendrik  
•
Lukes, Tomas  
•
Sharipov, Azat  
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2016
Nature Communications

Live-cell imaging of focal adhesions requires a sufficiently high temporal resolution, which remains a challenge for super-resolution microscopy. Here we address this important issue by combining photoactivated localization microscopy (PALM) with super-resolution optical fluctuation imaging (SOFI). Using simulations and fixed-cell focal adhesion images, we investigate the complementarity between PALM and SOFI in terms of spatial and temporal resolution. This PALM-SOFI framework is used to image focal adhesions in living cells, while obtaining a temporal resolution below 10 s. We visualize the dynamics of focal adhesions, and reveal local mean velocities around 190 nmmin (-1). The complementarity of PALM and SOFI is assessed in detail with a methodology that integrates a resolution and signal-to-noise metric. This PALM and SOFI concept provides an enlarged quantitative imaging framework, allowing unprecedented functional exploration of focal adhesions through the estimation of molecular parameters such as fluorophore densities and photoactivation or photoswitching kinetics.

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Type
research article
DOI
10.1038/ncomms13693
Web of Science ID

WOS:000389878100001

Author(s)
Deschout, Hendrik  
Lukes, Tomas  
Sharipov, Azat  
Szlag, Daniel
Feletti, Lely  
Vandenberg, Wim
Dedecker, Peter
Hofkens, Johan
Leutenegger, Marcel  
Lasser, Theo  
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Date Issued

2016

Publisher

Nature Publishing Group

Published in
Nature Communications
Volume

7

Article Number

13693

Editorial or Peer reviewed

REVIEWED

Written at

EPFL

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LBEN  
LOB  
Available on Infoscience
December 19, 2016
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/132055
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