Video 3: Long-term live-cell microscopy of HeLa Kyoto cells stably expressing H2B-mRFP and MyrPalm-mEGFP, treated with 100 nM SiR-actin.
FromFluorogenic probes for live-cell imaging of the cytoskeleton
- Gražvydas Lukinavičius1, 8,
- Luc Reymond1, 2, 8,
- Elisa D'Este3,
- Anastasiya Masharina1,
- Fabian Göttfert3,
- Haisen Ta3,
- Angelika Güther4,
- Mathias Fournier5,
- Stefano Rizzo6,
- Herbert Waldmann6,
- Claudia Blaukopf7,
- Christoph Sommer7,
- Daniel W Gerlich7,
- Hans-Dieter Arndt4,
- Stefan W Hell3,
- Kai Johnsson1, 2,
- Journal name:
- Nature Methods
- Volume:
- 11,
- Pages:
- 731–733
- Year published:
- DOI:
- doi:10.1038/nmeth.2972
- Received
- Accepted
- Published online
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The movie shows a region-of-interest of 1/16th of the full image frame and the full movie (0 – 23 h), starting 0.5 h after addition of SiR-actin.More videos from this article
Additional data
Primary authors
These authors contributed equally to this work.
- Gražvydas Lukinavičius &
- Luc Reymond
Affiliations
-
Institute of Chemical Sciences and Engineering, École Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.
- Gražvydas Lukinavičius,
- Luc Reymond,
- Anastasiya Masharina &
- Kai Johnsson
-
National Centre of Competence of Research in Chemical Biology, Lausanne, Switzerland.
- Luc Reymond &
- Kai Johnsson
-
Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.
- Elisa D'Este,
- Fabian Göttfert,
- Haisen Ta &
- Stefan W Hell
-
Institute of Organic Chemistry and Macromolecular Chemistry, Friedrich Schiller University, Jena, Germany.
- Angelika Güther &
- Hans-Dieter Arndt
-
Bioimaging and Optics Platform, École Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.
- Mathias Fournier
-
Max Planck Institute of Molecular Physiology, Dortmund, Germany.
- Stefano Rizzo &
- Herbert Waldmann
-
Institute of Molecular Biotechnology of the Austrian Academy of Sciences, Vienna, Austria.
- Claudia Blaukopf,
- Christoph Sommer &
- Daniel W Gerlich
Contributions
G.L., L.R. and K.J. devised this study. All authors except S.R. and A.G. contributed to manuscript writing. G.L., E.D., A.M., C.B., C.S. and D.W.G. characterized the probes. L.R., A.G. and S.R. performed synthesis of probes, supervised by K.J., H.W. and H.-D.A.; G.L., E.D., F.G. and H.T. performed STED microscopy, guided by S.W.H.; G.L. and M.F. performed SIM microscopy.
Competing financial interests
G.L. and K.J. have filed a patent application on SiR derivatives (PCT/EP2011/064750). S.R., H.-D.A. and H.W. have filed a patent application on the use of jasplakinolide derivatives (PCT/EP2012/073002).
Author details
Gražvydas Lukinavičius
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Luc Reymond
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Elisa D'Este
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Anastasiya Masharina
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Fabian Göttfert
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Haisen Ta
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Angelika Güther
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Mathias Fournier
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Stefano Rizzo
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Herbert Waldmann
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Claudia Blaukopf
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Christoph Sommer
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Daniel W Gerlich
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Hans-Dieter Arndt
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Stefan W Hell
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Kai Johnsson
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Video 1: Long-term live-cell microscopy of untreated control HeLa Kyoto cells stably expressing H2B-mRFP and MyrPalm-mEGFP.The movie shows a region-of-interest of 1/16th of the full image frame and the full movie (0 – 23 h).
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Video 2: Long-term live-cell microscopy of HeLa Kyoto cells stably expressing H2B-mRFP and MyrPalm-mEGFP, treated with 100 nM SiR-tubulin.The movie shows a region-of-interest of 1/16th of the full image frame and the full movie (0 – 23 h), starting 0.5 h after addition of SiR-tubulin.
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Video 3: Long-term live-cell microscopy of HeLa Kyoto cells stably expressing H2B-mRFP and MyrPalm-mEGFP, treated with 100 nM SiR-actin.The movie shows a region-of-interest of 1/16th of the full image frame and the full movie (0 – 23 h), starting 0.5 h after addition of SiR-actin.
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Video 4: Long-term live-cell microscopy of HeLa Kyoto cells stably expressing H2B-mRFP and MyrPalm-mEGFP, treated with 1 nM taxol.The movie shows a region-of-interest of 1/16th of the full image frame and the full movie (0 – 23 h), starting 0.5 h after addition of taxol.
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Video 5: Long-term live-cell microscopy of HeLa Kyoto cells stably expressing H2B-mRFP and MyrPalm-mEGFP, treated with 100 nM jasplakinolide.The movie shows a region-of-interest of 1/16th of the full image frame and the full movie (0 – 23 h), starting 0.5 h after addition of jasplakinolide.
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Video 6: 2D SIM time series of microtubule dynamics in human primary dermal fibroblasts.Cells were stained with 2 μM SiR-tubulin in complete DMEM medium for 60 min at 37°C and washed before imaging on a Nikon Eclipse Ti microscope. Time is indicated in min : s format.
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Video 7: 3D SIM time series of actin dynamics in lamellipodia of human primary dermal fibroblasts.Cells were stained with 2 μM SiR-actin in complete DMEM medium for 120 min and imaged directly without washing on Nikon Eclipse Ti microscope. Time is indicated in min : s format.
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Video 8: STED time series imaging of microtubule dynamics in human primary dermal fibroblasts.Cells were stained with 2 μM SiR-tubulin in complete DMEM medium for 60 min at 37°C and washed before imaging on a STED microscope. Time is indicated in min : s format.
