Light-induced cell damage in live-cell super-resolution microscopy
Zitieren Sie bitte immer diese URN: urn:nbn:de:bvb:20-opus-145207
- Super-resolution microscopy can unravel previously hidden details of cellular structures but requires high irradiation intensities to use the limited photon budget efficiently. Such high photon densities are likely to induce cellular damage in live-cell experiments. We applied single-molecule localization microscopy conditions and tested the influence of irradiation intensity, illumination-mode, wavelength, light-dose, temperature and fluorescence labeling on the survival probability of different cell lines 20-24 hours after irradiation. InSuper-resolution microscopy can unravel previously hidden details of cellular structures but requires high irradiation intensities to use the limited photon budget efficiently. Such high photon densities are likely to induce cellular damage in live-cell experiments. We applied single-molecule localization microscopy conditions and tested the influence of irradiation intensity, illumination-mode, wavelength, light-dose, temperature and fluorescence labeling on the survival probability of different cell lines 20-24 hours after irradiation. In addition, we measured the microtubule growth speed after irradiation. The photo-sensitivity is dramatically increased at lower irradiation wavelength. We observed fixation, plasma membrane permeabilization and cytoskeleton destruction upon irradiation with shorter wavelengths. While cells stand light intensities of similar to 1 kW cm\(^{-2}\) at 640 nm for several minutes, the maximum dose at 405 nm is only similar to 50 J cm\(^{-2}\), emphasizing red fluorophores for live-cell localization microscopy. We also present strategies to minimize phototoxic factors and maximize the cells ability to cope with higher irradiation intensities.…
Autor(en): | Sina Wäldchen, Julian Lehmann, Teresa Klein, Sebastian van de Linde, Markus Sauer |
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URN: | urn:nbn:de:bvb:20-opus-145207 |
Dokumentart: | Artikel / Aufsatz in einer Zeitschrift |
Institute der Universität: | Fakultät für Biologie / Theodor-Boveri-Institut für Biowissenschaften |
Sprache der Veröffentlichung: | Englisch |
Titel des übergeordneten Werkes / der Zeitschrift (Englisch): | Scientific Reports |
Erscheinungsjahr: | 2015 |
Band / Jahrgang: | 5 |
Heft / Ausgabe: | 15348 |
Originalveröffentlichung / Quelle: | Scientific Reports 5:15348 (2015). DOI: 10.1038/srep15348 |
DOI: | https://doi.org/10.1038/srep15348 |
Allgemeine fachliche Zuordnung (DDC-Klassifikation): | 5 Naturwissenschaften und Mathematik / 53 Physik / 530 Physik |
Freie Schlagwort(e): | diffraction limit; dynamics; fluorescent probes; illumination microscopy; in vitro; localization microscopy; optical reconstruction microscopy; photodynamic therapy; structured illumination; tag fusion proteins |
Datum der Freischaltung: | 06.11.2018 |
Lizenz (Deutsch): | CC BY: Creative-Commons-Lizenz: Namensnennung 4.0 International |