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Correlative cryogenic montage electron tomography for comprehensive in-situ whole-cell structural studies

Jie E. Yang, Matthew R. Larson, Bryan S. Sibert, Joseph Y. Kim, Daniel Parrell, Juan C. Sanchez, Victoria Pappas, Anil Kumar, Kai Cai, Keith Thompson, View ORCID ProfileElizabeth R. Wright
doi: https://doi.org/10.1101/2021.12.31.474669
Jie E. Yang
1Department of Biochemistry, University of Wisconsin, Madison, WI USA
2Cryo-Electron Microscopy Research Center, Department of Biochemistry, University of Wisconsin, Madison, WI USA
3Midwest Center for Cryo-Electron Tomography, Department of Biochemistry, University of Wisconsin, Madison, WI USA
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Matthew R. Larson
1Department of Biochemistry, University of Wisconsin, Madison, WI USA
2Cryo-Electron Microscopy Research Center, Department of Biochemistry, University of Wisconsin, Madison, WI USA
3Midwest Center for Cryo-Electron Tomography, Department of Biochemistry, University of Wisconsin, Madison, WI USA
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Bryan S. Sibert
1Department of Biochemistry, University of Wisconsin, Madison, WI USA
2Cryo-Electron Microscopy Research Center, Department of Biochemistry, University of Wisconsin, Madison, WI USA
3Midwest Center for Cryo-Electron Tomography, Department of Biochemistry, University of Wisconsin, Madison, WI USA
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Joseph Y. Kim
1Department of Biochemistry, University of Wisconsin, Madison, WI USA
4Department of Chemistry, University of Wisconsin, Madison, WI USA
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Daniel Parrell
1Department of Biochemistry, University of Wisconsin, Madison, WI USA
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Juan C. Sanchez
1Department of Biochemistry, University of Wisconsin, Madison, WI USA
5Biophysics Graduate Program, University of Wisconsin, Madison, WI USA
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Victoria Pappas
1Department of Biochemistry, University of Wisconsin, Madison, WI USA
5Biophysics Graduate Program, University of Wisconsin, Madison, WI USA
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Anil Kumar
1Department of Biochemistry, University of Wisconsin, Madison, WI USA
2Cryo-Electron Microscopy Research Center, Department of Biochemistry, University of Wisconsin, Madison, WI USA
3Midwest Center for Cryo-Electron Tomography, Department of Biochemistry, University of Wisconsin, Madison, WI USA
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Kai Cai
1Department of Biochemistry, University of Wisconsin, Madison, WI USA
2Cryo-Electron Microscopy Research Center, Department of Biochemistry, University of Wisconsin, Madison, WI USA
3Midwest Center for Cryo-Electron Tomography, Department of Biochemistry, University of Wisconsin, Madison, WI USA
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Keith Thompson
1Department of Biochemistry, University of Wisconsin, Madison, WI USA
2Cryo-Electron Microscopy Research Center, Department of Biochemistry, University of Wisconsin, Madison, WI USA
3Midwest Center for Cryo-Electron Tomography, Department of Biochemistry, University of Wisconsin, Madison, WI USA
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Elizabeth R. Wright
1Department of Biochemistry, University of Wisconsin, Madison, WI USA
2Cryo-Electron Microscopy Research Center, Department of Biochemistry, University of Wisconsin, Madison, WI USA
3Midwest Center for Cryo-Electron Tomography, Department of Biochemistry, University of Wisconsin, Madison, WI USA
6Morgridge Institute for Research, Madison, WI, USA
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  • ORCID record for Elizabeth R. Wright
  • For correspondence: [email protected]
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Abstract

Imaging large fields of view while preserving high-resolution structural information remains a challenge in low-dose cryo-electron tomography. Here, we present robust tools for montage electron tomography tailored for vitrified specimens. The integration of correlative cryo-fluorescence microscopy, focused-ion beam milling, and micropatterning produces contextual three-dimensional architecture of cells. Montage tilt series may be processed in their entirety or as individual tiles suitable for sub-tomogram averaging, enabling efficient data processing and analysis.

Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • https://github.com/wright-cemrc-projects/cryoet-montage

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 January 02, 2022.
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Correlative cryogenic montage electron tomography for comprehensive in-situ whole-cell structural studies
Jie E. Yang, Matthew R. Larson, Bryan S. Sibert, Joseph Y. Kim, Daniel Parrell, Juan C. Sanchez, Victoria Pappas, Anil Kumar, Kai Cai, Keith Thompson, Elizabeth R. Wright
bioRxiv 2021.12.31.474669; doi: https://doi.org/10.1101/2021.12.31.474669
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Correlative cryogenic montage electron tomography for comprehensive in-situ whole-cell structural studies
Jie E. Yang, Matthew R. Larson, Bryan S. Sibert, Joseph Y. Kim, Daniel Parrell, Juan C. Sanchez, Victoria Pappas, Anil Kumar, Kai Cai, Keith Thompson, Elizabeth R. Wright
bioRxiv 2021.12.31.474669; doi: https://doi.org/10.1101/2021.12.31.474669

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