Assessing photodamage in live-cell STED microscopy

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Fig. 1: Short- and long-term effects of live-cell STED imaging on COS7 and HeLa cells.

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Acknowledgements

We thank S. Baguley, F. Bottanelli, and L.K. Schroeder for generating and providing plasmids; A.E.S. Barentine for determining the STED microscope resolution; E.B. Kromann for support with the custom-built STED microscope; M. Germino and Y. Zhang for helpful discussions regarding data analysis; A. Kumar (Enzo Life Sciences, Inc.) for support regarding the use of FluoForte; and B.G. Rash for comments on the manuscript. This work was supported by grants from the Wellcome Trust (095927/A/11/Z and 203285/B/16/Z to D.T., J.E.R., and J.B.) and the G. Harold & Leila Y. Mathers Foundation (to J.B.). The used Leica SP8 STED microscope was supported by NIH (S10OD020142). M.D.L. and J.B. acknowledge support by the NIH-funded Yale Center of Excellence in Hematology (U54DK106857). D.T. and J.B. were supported by the Yale Diabetes Research Center (NIH P30DK045735).

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Authors

Contributions

N.K., A.G., and J.B. conceived and designed the experiments. N.K., A.G., and M.D.L. performed the experiments. N.K., A.G., M.D.L., G.H., D.T., J.E.R., and J.B. analyzed and discussed the data. G.H. performed the statistical analysis. N.K., A.G., and J.B. wrote the manuscript.

Corresponding author

Correspondence to Joerg Bewersdorf.

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Competing interests

J.B. has financial interests in Bruker Corp. and Hamamatsu Photonics. The other authors declare no competing interests.

Supplementary information

41592_2018_145_MOESM3_ESM.avi

STED time-lapse video of a representative COS7 cell

41592_2018_145_MOESM4_ESM.avi

Video of a COS7 cell surviving for at least 24 h after STED irradiation

41592_2018_145_MOESM5_ESM.avi

Video of a COS7 cell dying a few hours after STED irradiation

Supplementary Text and Figures

Supplementary Methods and Supplementary Notes

Reporting Summary

Supplementary Video 1

STED time-lapse video of a representative COS7 cell

Supplementary Video 2

Video of a COS7 cell surviving for at least 24 h after STED irradiation

Supplementary Video 3

Video of a COS7 cell dying a few hours after STED irradiation

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Kilian, N., Goryaynov, A., Lessard, M.D. et al. Assessing photodamage in live-cell STED microscopy. Nat Methods 15, 755–756 (2018). https://doi.org/10.1038/s41592-018-0145-5

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