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Isotropic 3D electron microscopy reference library of whole cells and tissues

View ORCID ProfileC. Shan Xu, View ORCID ProfileSong Pang, View ORCID ProfileGleb Shtengel, View ORCID ProfileAndreas Müller, View ORCID ProfileAlex T. Ritter, View ORCID ProfileHuxley K. Hoffman, View ORCID ProfileShin-ya Takemura, View ORCID ProfileZhiyuan Lu, View ORCID ProfileH. Amalia Pasolli, View ORCID ProfileNirmala Iyer, View ORCID ProfileJeeyun Chung, View ORCID ProfileDavis Bennett, View ORCID ProfileAubrey V. Weigel, View ORCID ProfileTobias C. Walther, View ORCID ProfileRobert V. Farese Jr., View ORCID ProfileSchuyler B. van Engelenburg, View ORCID ProfileIra Mellman, View ORCID ProfileMichele Solimena, View ORCID ProfileHarald F. Hess
doi: https://doi.org/10.1101/2020.11.13.382457
C. Shan Xu
1Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA
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  • For correspondence: xuc@janelia.hhmi.org hessh@janelia.hhmi.org
Song Pang
1Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA
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Gleb Shtengel
1Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA
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Andreas Müller
2Molecular Diabetology, University Hospital and Faculty of Medicine Carl Gustav Carus, TU Dresden, Dresden, Germany
3Paul Langerhans Institute Dresden (PLID) of the Helmholtz Center Munich at the University Hospital Carl Gustav Carus and Faculty of Medicine of the TU Dresden, Dresden, Germany
4German Center for Diabetes Research (DZD e.V.), Neuherberg, Germany
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Alex T. Ritter
5Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, USA
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  • ORCID record for Alex T. Ritter
Huxley K. Hoffman
6Molecular and Cellular Biophysics Program Department of Biological Sciences, University of Denver, Denver, CO 80210, USA
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  • ORCID record for Huxley K. Hoffman
Shin-ya Takemura
1Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA
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  • ORCID record for Shin-ya Takemura
Zhiyuan Lu
1Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA
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  • ORCID record for Zhiyuan Lu
H. Amalia Pasolli
1Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA
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  • ORCID record for H. Amalia Pasolli
Nirmala Iyer
1Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA
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Jeeyun Chung
7Department of Molecular Metabolism, Harvard T.H. Chan School of Public Health, Boston, MA 02115, USA
8Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA
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Davis Bennett
1Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA
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Aubrey V. Weigel
1Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA
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Tobias C. Walther
7Department of Molecular Metabolism, Harvard T.H. Chan School of Public Health, Boston, MA 02115, USA
8Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA
9Howard Hughes Medical Institute, Boston, MA 02492, USA; Broad Institute of Harvard and MIT, Cambridge, MA 02124, USA
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Robert V. Farese Jr.
7Department of Molecular Metabolism, Harvard T.H. Chan School of Public Health, Boston, MA 02115, USA
8Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA
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Schuyler B. van Engelenburg
6Molecular and Cellular Biophysics Program Department of Biological Sciences, University of Denver, Denver, CO 80210, USA
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Ira Mellman
5Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, USA
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  • ORCID record for Ira Mellman
Michele Solimena
2Molecular Diabetology, University Hospital and Faculty of Medicine Carl Gustav Carus, TU Dresden, Dresden, Germany
3Paul Langerhans Institute Dresden (PLID) of the Helmholtz Center Munich at the University Hospital Carl Gustav Carus and Faculty of Medicine of the TU Dresden, Dresden, Germany
4German Center for Diabetes Research (DZD e.V.), Neuherberg, Germany
10Max-Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany
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  • ORCID record for Michele Solimena
Harald F. Hess
1Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA
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  • ORCID record for Harald F. Hess
  • For correspondence: xuc@janelia.hhmi.org hessh@janelia.hhmi.org
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Summary

Understanding cellular architecture is essential for understanding biology. Electron microscopy (EM) uniquely visualizes cellular structure with nanometer resolution. However, traditional methods, such as thin-section EM or EM tomography, have limitations inasmuch as they only visualize a single slice or a relatively small volume of the cell, respectively. Here, we overcome these limitations by imaging whole cells and tissues with enhanced Focus Ion Beam Scanning Electron Microscopy (FIB-SEM) in high resolution with month-long acquisition duration. We use this approach to generate reference 3D image datasets at 4-nm isotropic voxels for ten different examples, including cultured cells (cancer, macrophages, and T-cells) as well as tissues (mouse pancreatic islets and the Drosophila fan-shaped body). We open access to all datasets in OpenOrganelle, an interactive web platform that allows accessing both the original 3D EM data, and subsequent organelle segmentation. Together, these data will serve as a reference library to explore comprehensive quantification of whole cells and their constituents, thus addressing questions related to cell identities, cell morphologies, cell-cell interactions, as well as intracellular organelle organization and structure.

Competing Interest Statement

Portions of the technology described herein are covered by U.S. Patent 10,600,615 issued to C.S.X. and H.F.H., and assigned to HHMI.

Footnotes

  • https://openorganelle.janelia.org

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted November 14, 2020.
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Isotropic 3D electron microscopy reference library of whole cells and tissues
C. Shan Xu, Song Pang, Gleb Shtengel, Andreas Müller, Alex T. Ritter, Huxley K. Hoffman, Shin-ya Takemura, Zhiyuan Lu, H. Amalia Pasolli, Nirmala Iyer, Jeeyun Chung, Davis Bennett, Aubrey V. Weigel, Tobias C. Walther, Robert V. Farese Jr., Schuyler B. van Engelenburg, Ira Mellman, Michele Solimena, Harald F. Hess
bioRxiv 2020.11.13.382457; doi: https://doi.org/10.1101/2020.11.13.382457
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Isotropic 3D electron microscopy reference library of whole cells and tissues
C. Shan Xu, Song Pang, Gleb Shtengel, Andreas Müller, Alex T. Ritter, Huxley K. Hoffman, Shin-ya Takemura, Zhiyuan Lu, H. Amalia Pasolli, Nirmala Iyer, Jeeyun Chung, Davis Bennett, Aubrey V. Weigel, Tobias C. Walther, Robert V. Farese Jr., Schuyler B. van Engelenburg, Ira Mellman, Michele Solimena, Harald F. Hess
bioRxiv 2020.11.13.382457; doi: https://doi.org/10.1101/2020.11.13.382457

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