Video 5: Imaging of HI-Virus uptake in living cells
FromUltrafast, temporally stochastic STED nanoscopy of millisecond dynamics
- Jale Schneider1, 2,
- Jasmin Zahn1, 2,
- Marta Maglione3,
- Stephan J Sigrist3,
- Jonas Marquard1, 2,
- Jakub Chojnacki4,
- Hans-Georg Kräusslich4,
- Steffen J Sahl5,
- Johann Engelhardt1, 2,
- Stefan W Hell1, 2, 5,
- Journal name:
- Nature Methods
- Volume:
- 12,
- Pages:
- 827–830
- Year published:
- DOI:
- doi:10.1038/nmeth.3481
- Received
- Accepted
- Published online
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Video acquisition of dynamics of EGFP-labeled HI-Viruses taken up by living HeLa cells. The effective frame rate of the acquisition is 125 fps. The data shown are laterally smoothed with a Gaussian of 100 nm and temporally smoothed over five frames.More videos from this article
Additional data
Affiliations
-
German Cancer Research Center (DKFZ), Optical Nanoscopy Division, Heidelberg, Germany.
- Jale Schneider,
- Jasmin Zahn,
- Jonas Marquard,
- Johann Engelhardt &
- Stefan W Hell
-
Bioquant Center, Heidelberg, Germany.
- Jale Schneider,
- Jasmin Zahn,
- Jonas Marquard,
- Johann Engelhardt &
- Stefan W Hell
-
Institute for Biology and Genetics, Free University of Berlin, Berlin, Germany.
- Marta Maglione &
- Stephan J Sigrist
-
Department of Infectious Diseases, Virology, University of Heidelberg, Heidelberg, Germany.
- Jakub Chojnacki &
- Hans-Georg Kräusslich
-
Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.
- Steffen J Sahl &
- Stefan W Hell
Contributions
J.S. and J.E. developed the EOD scanning system and set up the microscope. J.S. and J.Z. performed the experiments and analyzed the data. M.M., S.J. Sigrist, J.C. and H.-G.K. provided samples. J.M. performed single-antibody imaging. J.S., J.E., S.J. Sahl and S.W.H. wrote the manuscript. The latter also conceived and guided the project and is responsible for its main points. All authors discussed the data and commented on the manuscript.
Competing financial interests
The authors declare no competing financial interests.
Author details
Jale Schneider
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Jasmin Zahn
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Marta Maglione
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Stephan J Sigrist
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Jonas Marquard
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Jakub Chojnacki
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Hans-Georg Kräusslich
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Steffen J Sahl
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Johann Engelhardt
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Stefan W Hell
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Supplementary Figure 1: Increased photon yield for several fluorophores by ultrafast scanning compared to the conventional slow-scanning mode.Hover over figure to zoom
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Supplementary Figure 2: Resolution in ultrafast scanning STED nanoscopy.Hover over figure to zoom
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Supplementary Figure 3: Temporally stochastic STED imaging of single antibodies conjugated to Atto 647N.Hover over figure to zoom
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Video 1: Image build-up with ultra-fast scanning
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Video 2: Imaging of diffusing beads
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Video 3: Imaging of vesicle dynamics in living Drosophila larvae
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Video 4: Imaging of vesicle dynamics in living Drosophila larvae (laterally smoothed)
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Video 5: Imaging of HI-Virus uptake in living cells