title: Photofunctional surfaces for quantitative fluorescence microscopy: monitoring the effects of photogenerated reactive oxygen species at single cell level with spatiotemporal resolution.
abstract: Herein, we report on the implementation of photofunctional surfaces for the investigation of cellular responses by means of quantitative fluorescence microscopy. The developed substrates are able to produce reactive oxygen species under the fluorescence microscope upon irradiation with visible light, and the behavior of cells grown on these surfaces can be consequently investigated in situ and in real time. Moreover, a suitable methodology is presented to simultaneously monitor phototriggered morphological changes and the associated molecular pathways with spatiotemporal resolution employing time-resolved fluorescence anisotropy at the single cell level. The results showed that morphological changes can be complemented with a quantitative evaluation of the associated molecular signaling cascades for the unambiguous assignment of reactive oxygen species-related photoinduced apoptosis. Indeed, similar phenotypes are associated with different cellular processes. Our methodology facilitates the in vitro design and evaluation of photosensitizers for the treatment of cancer and infectious diseases with the aid of functional fluorescence microscopy.
authors.affiliation: †Physikalisches Institut and Center for Nanotechnology (CeNTech), Westfälische Wilhelms-Universität Münster, Heisenbergstraße 11, D-48149 Münster, Germany.
authors.forename: Linda
authors.initials: L
authors.lastname: Stegemann
authors.affiliation: ‡Department of Systemic Cell Biology, Max Planck Institute of Molecular Physiology, Otto-Hahn Straße 11, D-44227 Dortmund, Germany.
authors.forename: Klaus C
authors.initials: KC
authors.lastname: Schuermann
authors.affiliation: †Physikalisches Institut and Center for Nanotechnology (CeNTech), Westfälische Wilhelms-Universität Münster, Heisenbergstraße 11, D-48149 Münster, Germany.
authors.forename: Cristian A
authors.initials: CA
authors.lastname: Strassert
authors.affiliation: ‡Department of Systemic Cell Biology, Max Planck Institute of Molecular Physiology, Otto-Hahn Straße 11, D-44227 Dortmund, Germany.
authors.forename: Hernán E
authors.initials: HE
authors.lastname: Grecco
mesh_terms: D006367:HeLa Cells; D006801:Humans; D008027:Light; D008856:Microscopy, Fluorescence / Q000379:methods*; D057054:Molecular Imaging / Q000379:methods*; D053758:Nanoparticles / Q000737:chemistry* / Q000528:radiation effects; D010777:Photochemistry / Q000379:methods; D017382:Reactive Oxygen Species / Q000378:metabolism*; D062211:Spatio-Temporal Analysis
chemical_list: D017382:Reactive Oxygen Species
keywords: apoptosis; caspase activity; cell death; necrosis; photoactive surfaces; phototherapy; quantitative functional microscopy; reactive oxygen species