TY  - JOUR
A1  - Wäldchen, Sina
A1  - Lehmann, Julian
A1  - Klein, Teresa
A1  - van de Linde, Sebastian
A1  - Sauer, Markus
T1  - Light-induced cell damage in live-cell super-resolution microscopy
T2  - Scientific Reports
N2  - 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. 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.
KW  - optical reconstruction microscopy
KW  - tag fusion proteins
KW  - localization microscopy
KW  - photodynamic therapy
KW  - diffraction limit
KW  - illumination microscopy
KW  - structured illumination
KW  - fluorescent probes
KW  - in vitro
KW  - dynamics
Y1  - 2015
UR  - https://opus.bibliothek.uni-wuerzburg.de/frontdoor/index/index/docId/14520
UR  - https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-145207
VL  - 5
IS  - 15348
ER  -