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

Deschout, H., Lukes, T., Sharipov, A., Szlag, D., Feletti, L., Vandenberg, W., et al. (2016). Complementarity of PALM and SOFI for super-resolution live-cell imaging of focal adhesions. Nature Communications, 7: 13693. doi:10.1038/ncomms13693.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-002C-31A5-7 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-002C-7D22-2
Genre: Journal Article

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 Creators:
Deschout, H., Author
Lukes, T., Author
Sharipov, A., Author
Szlag, D., Author
Feletti, L., Author
Vandenberg, W., Author
Dedecker, P., Author
Hofkens, J., Author
Leutenegger, M.1, Author              
Lasser, T., Author
Radenovic, A., Author
Affiliations:
1Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society, ou_578627              

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 Abstract: 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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Language(s): eng - English
 Dates: 2016-12-19
 Publication Status: Published online
 Pages: -
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 Table of Contents: -
 Rev. Method: Peer
 Identifiers: DOI: 10.1038/ncomms13693
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Title: Nature Communications
Source Genre: Journal
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Pages: 11 Volume / Issue: 7 Sequence Number: 13693 Start / End Page: - Identifier: -