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. 2018 Oct;15(10):755-756.
doi: 10.1038/s41592-018-0145-5.

Assessing photodamage in live-cell STED microscopy

Nicole Kilian  1 , Alexander Goryaynov  1 , Mark D Lessard  1 , Giles Hooker  2   3 , Derek Toomre  1   4 , James E Rothman  1   4 , Joerg Bewersdorf  5   6   7
Affiliations

Assessing photodamage in live-cell STED microscopy

Nicole Kilian et al. Nat Methods. 2018 Oct.
No abstract available

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Figures

Figure 1 |
Figure 1 |. Short and long-term effects of live-cell STED imaging on COS7 and HeLa cells.
(a) Cytoplasmic Ca2+-level response of SNAP-Sec61β expressing SiR-labeled cells under negative control conditions (no excitation or STED illumination). (b) Positive control using Ionomycin. (c) STED-irradiated cells using an 8-kHz resonant scanner. (d) STED-irradiated cells with ROS scavenging buffer added. (e-l) Representative fluorescence (e,g-i,k,l) and brightfield (f,j) images of a HeLa cell before and after STED irradiation in ROS scavenging buffer visualizing cell viability via cell morphology and ER movement. Scale bars: 10 μm (j), 5 μm (l). Confocal images (e,i), STED images (g,h,k,l). (m) Long-term viability of STED-irradiated and control cells. Cells are categorized in alive, dead and indeterminable (labeled as “?”; see Supplementary Methods) after 24 h. Statistical information (N = total number of cells; M = number of independent experiments): (a) HeLa: N=17, M=3; COS7: N=18, M=4; (b) HeLa: N=15, M=3; COS7: N=15, M=3; (c) HeLa: N=30, M=3; COS7: N=30, M=4; (d) HeLa: N=32, M=4; COS7: N=30, M=5; (e-l) N=10, M=2; (m) HeLa: N=15, M=3; COS7: N=15, M=3; control HeLa: N=20, M=3; COS7: N=28, M=4.
See this image and copyright information in PMC

References

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