title: Complementarity of PALM and SOFI for super-resolution live-cell imaging of focal adhesions.
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 nm min-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.
authors.affiliation: Laboratory of Nanoscale Biology &Laboratoire d'Optique Biomédicale, STI - IBI, Ecole Polytechnique Fédérale de Lausanne, Station 17, CH-1015 Lausanne, Switzerland.
authors.forename: Hendrik
authors.initials: H
authors.lastname: Deschout
authors.affiliation: Laboratory of Nanoscale Biology &Laboratoire d'Optique Biomédicale, STI - IBI, Ecole Polytechnique Fédérale de Lausanne, Station 17, CH-1015 Lausanne, Switzerland.|Department of Radioelectronics, FEE, Czech Technical University in Prague, Technická 2, 166 27 Prague 6, Czech Republic.
authors.forename: Tomas
authors.initials: T
authors.lastname: Lukes
authors.affiliation: Laboratory of Nanoscale Biology &Laboratoire d'Optique Biomédicale, STI - IBI, Ecole Polytechnique Fédérale de Lausanne, Station 17, CH-1015 Lausanne, Switzerland.
authors.forename: Azat
authors.initials: A
authors.lastname: Sharipov
authors.affiliation: Laboratory of Nanoscale Biology &Laboratoire d'Optique Biomédicale, STI - IBI, Ecole Polytechnique Fédérale de Lausanne, Station 17, CH-1015 Lausanne, Switzerland.
authors.forename: Daniel
authors.initials: D
authors.lastname: Szlag
authors.affiliation: Laboratory of Nanoscale Biology &Laboratoire d'Optique Biomédicale, STI - IBI, Ecole Polytechnique Fédérale de Lausanne, Station 17, CH-1015 Lausanne, Switzerland.
authors.forename: Lely
authors.initials: L
authors.lastname: Feletti
authors.affiliation: Department of Chemistry, University of Leuven, Celestijnenlaan 200F, B-3001 Heverlee, Belgium.
authors.forename: Wim
authors.initials: W
authors.lastname: Vandenberg
authors.affiliation: Department of Chemistry, University of Leuven, Celestijnenlaan 200F, B-3001 Heverlee, Belgium.
authors.forename: Peter
authors.initials: P
authors.lastname: Dedecker
authors.affiliation: Department of Chemistry, University of Leuven, Celestijnenlaan 200F, B-3001 Heverlee, Belgium.
authors.forename: Johan
authors.identifier: 0000-0002-9101-0567
authors.initials: J
authors.lastname: Hofkens
authors.affiliation: Abteilung NanoBiophotonik, Max-Planck-Institut für biophysikalische Chemie, Am Fassberg 11, 37077 Göttingen, Germany.
authors.forename: Marcel
authors.initials: M
authors.lastname: Leutenegger
authors.affiliation: Laboratory of Nanoscale Biology &Laboratoire d'Optique Biomédicale, STI - IBI, Ecole Polytechnique Fédérale de Lausanne, Station 17, CH-1015 Lausanne, Switzerland.
authors.forename: Theo
authors.identifier: 0000-0002-4948-7580
authors.initials: T
authors.lastname: Lasser
authors.affiliation: Laboratory of Nanoscale Biology &Laboratoire d'Optique Biomédicale, STI - IBI, Ecole Polytechnique Fédérale de Lausanne, Station 17, CH-1015 Lausanne, Switzerland.
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Radenovic
mesh_terms: D000818:Animals; D002448:Cell Adhesion / Q000502:physiology*; D005347:Fibroblasts / Q000502:physiology*; D051379:Mice; D008853:Microscopy / Q000379:methods*; D051419:Paxillin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D051381:Rats; D013194:Staining and Labeling; D013997:Time Factors
chemical_list: D051419:Paxillin