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Light-induced cell damage in live-cell super-resolution microscopy
Open accessAbstract
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 ~1 kW cm−2 at 640 nm for several minutes, the maximum dose at 405 nm is only ~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.
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Cites
Tetrazolium dyes as tools in cell biology: new insights into their cellular reduction
Berridge, M. V., Herst, P. M., & Tan, A. S. (2005) · Biotechnology Annual Review · doi:10.1016/s1387-2656(05)11004-7 · Cited by 2,222
Cited by
Fluorescence nanoscopy in cell biology
Sahl, S. J., Hell, S. W., & Jakobs, S. (2017) · Nature Reviews Molecular Cell Biology · doi:10.1038/nrm.2017.71 · Cited by 990
Plasmonic Biosensing: Focus Review
Mejía-Salazar, J. R., & Oliveira, O. N. (2018) · Chemical Reviews · doi:10.1021/acs.chemrev.8b00359 · Cited by 544
Tay, N. E. S., Lehnherr, D., & Rovis, T. (2022) · Chemical Reviews · doi:10.1021/acs.chemrev.1c00384 · Cited by 518
Switchable Fluorophores for Single-Molecule Localization Microscopy
Li, H., & Vaughan, J. C. (2018) · Chemical Reviews · doi:10.1021/acs.chemrev.7b00767 · Cited by 319
Seeing beyond the limit: A guide to choosing the right super-resolution microscopy technique
Valli, J., Garcia-Burgos, A., Rooney, L. M., Vale de Melo e Oliveira, B., Duncan, R. R., & Rickman, C. (2021) · Journal of Biological Chemistry · doi:10.1016/j.jbc.2021.100791 · Cited by 159
Metadata
- Authors
- Wäldchen, S., Lehmann, J., Klein, T., van de Linde, S., & Sauer, M.
- Venue
- Scientific Reports
- Published
- 2015-10-20
- Volume
- 5
- Issue
- 1
- Pages
- 15348-15348
- ISSN
- 2045-2322 (ISSN-L)
- OpenAlex
- W1782900133
- Cited by
- 536
- References
- 1
- FWCI
- 23.75
- Type
- journal article
- Language
- en
- Open access
- gold
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- Wäldchen, S., Lehmann, J., Klein, T., van de Linde, S., & Sauer, M. (2015). Light-induced cell damage in live-cell super-resolution microscopy. Scientific Reports. https://doi.org/10.1038/srep15348
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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
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