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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).
J.B. has financial interests in Bruker Corp. and Hamamatsu Photonics. The other authors declare no competing interests.
Supplementary Methods and Supplementary Notes
STED time-lapse video of a representative COS7 cell
Video of a COS7 cell surviving for at least 24 h after STED irradiation
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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