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Nanoscale subcellular architecture revealed by multicolor 3D salvaged fluorescence imaging

View ORCID ProfileYongdeng Zhang, View ORCID ProfileLena K. Schroeder, View ORCID ProfileMark D. Lessard, Phylicia Kidd, Jeeyun Chung, Yuanbin Song, Lorena Benedetti, View ORCID ProfileYiming Li, Jonas Ries, Pietro De Camilli, James E. Rothman, David Baddeley, View ORCID ProfileJoerg Bewersdorf
doi: https://doi.org/10.1101/613174
Yongdeng Zhang
1Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06510, USA
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Lena K. Schroeder
1Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06510, USA
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  • ORCID record for Lena K. Schroeder
Mark D. Lessard
1Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06510, USA
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Phylicia Kidd
1Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06510, USA
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Jeeyun Chung
1Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06510, USA
2Department of Neuroscience, Yale University School of Medicine, New Haven, CT 06510 USA
3Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, CT 06510 USA
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Yuanbin Song
4Section of Hematology, Department of Internal Medicine, Yale School of Medicine, New Haven, CT 06510, USA
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Lorena Benedetti
1Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06510, USA
2Department of Neuroscience, Yale University School of Medicine, New Haven, CT 06510 USA
3Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, CT 06510 USA
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Yiming Li
6Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg 69117, Germany
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  • ORCID record for Yiming Li
Jonas Ries
6Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg 69117, Germany
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Pietro De Camilli
1Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06510, USA
2Department of Neuroscience, Yale University School of Medicine, New Haven, CT 06510 USA
3Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, CT 06510 USA
5Kavli Institute for Neuroscience, Yale University School of Medicine, New Haven, CT 06510 USA
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James E. Rothman
1Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06510, USA
7Nanobiology Institute, Yale University, West Haven, CT 06516, USA
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David Baddeley
1Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06510, USA
7Nanobiology Institute, Yale University, West Haven, CT 06516, USA
8Auckland Bioengineering Institute, University of Auckland, Auckland 1010, New Zealand
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Joerg Bewersdorf
1Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06510, USA
5Kavli Institute for Neuroscience, Yale University School of Medicine, New Haven, CT 06510 USA
7Nanobiology Institute, Yale University, West Haven, CT 06516, USA
9Department of Biomedical Engineering, Yale University, New Haven, CT 06520, USA
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  • ORCID record for Joerg Bewersdorf
  • For correspondence: joerg.bewersdorf@yale.edu
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Abstract

Combining the molecular specificity of fluorescent probes with three-dimensional (3D) imaging at nanoscale resolution is critical for investigating the spatial organization and interactions of cellular organelles and protein complexes. We present a super-resolution light microscope that enables simultaneous multicolor imaging of whole mammalian cells at ~20 nm 3D resolution. We show its power for cell biology research with fluorescence images that resolved the highly convoluted Golgi apparatus and the close contacts between the endoplasmic reticulum and the plasma membrane, structures that have traditionally been the imaging realm of electron microscopy.

One Sentence Summary Complex cellular structures previously only resolved by electron microscopy can now be imaged in multiple colors by 4Pi-SMS.

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Posted April 18, 2019.
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Nanoscale subcellular architecture revealed by multicolor 3D salvaged fluorescence imaging
Yongdeng Zhang, Lena K. Schroeder, Mark D. Lessard, Phylicia Kidd, Jeeyun Chung, Yuanbin Song, Lorena Benedetti, Yiming Li, Jonas Ries, Pietro De Camilli, James E. Rothman, David Baddeley, Joerg Bewersdorf
bioRxiv 613174; doi: https://doi.org/10.1101/613174
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Nanoscale subcellular architecture revealed by multicolor 3D salvaged fluorescence imaging
Yongdeng Zhang, Lena K. Schroeder, Mark D. Lessard, Phylicia Kidd, Jeeyun Chung, Yuanbin Song, Lorena Benedetti, Yiming Li, Jonas Ries, Pietro De Camilli, James E. Rothman, David Baddeley, Joerg Bewersdorf
bioRxiv 613174; doi: https://doi.org/10.1101/613174

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