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Light-induced cell damage in live-cell super-resolution microscopy

Please always quote using this 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.show moreshow less

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Metadaten
Author: Sina Wäldchen, Julian Lehmann, Teresa Klein, Sebastian van de Linde, Markus Sauer
URN:urn:nbn:de:bvb:20-opus-145207
Document Type:Journal article
Faculties:Fakultät für Biologie / Theodor-Boveri-Institut für Biowissenschaften
Language:English
Parent Title (English):Scientific Reports
Year of Completion:2015
Volume:5
Issue:15348
Source:Scientific Reports 5:15348 (2015). DOI: 10.1038/srep15348
DOI:https://doi.org/10.1038/srep15348
Dewey Decimal Classification:5 Naturwissenschaften und Mathematik / 53 Physik / 530 Physik
Tag:diffraction limit; dynamics; fluorescent probes; illumination microscopy; in vitro; localization microscopy; optical reconstruction microscopy; photodynamic therapy; structured illumination; tag fusion proteins
Release Date:2018/11/06
Licence (German):License LogoCC BY: Creative-Commons-Lizenz: Namensnennung 4.0 International