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Three dimensional particle averaging for structural imaging of macromolecular complexes by localization microscopy

Hamidreza Heydarian, Adrian Przybylski, Florian Schueder, Ralf Jungmann, Ben van Werkhoven, Jan Keller-Findeisen, Jonas Ries, Sjoerd Stallinga, Mark Bates, Bernd Rieger
doi: https://doi.org/10.1101/837575
Hamidreza Heydarian
1Department of Imaging Physics, Delft University of Technology, Delft, the Netherlands
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Adrian Przybylski
2Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany
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Florian Schueder
3Department of Physics and Center for Nanoscience, Ludwig Maximilian University, Munich, Germany
4Max Planck Institute of Biochemistry, Martinsried, Germany
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Ralf Jungmann
3Department of Physics and Center for Nanoscience, Ludwig Maximilian University, Munich, Germany
4Max Planck Institute of Biochemistry, Martinsried, Germany
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Ben van Werkhoven
5Netherlands eScience Center, Amsterdam, the Netherlands
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Jan Keller-Findeisen
2Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany
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Jonas Ries
6Cell Biology and Biophysics Unit, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany
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Sjoerd Stallinga
1Department of Imaging Physics, Delft University of Technology, Delft, the Netherlands
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Mark Bates
2Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany
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  • For correspondence: mark.bates@mpibpc.mpg.de b.rieger@tudelft.nl
Bernd Rieger
1Department of Imaging Physics, Delft University of Technology, Delft, the Netherlands
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  • For correspondence: mark.bates@mpibpc.mpg.de b.rieger@tudelft.nl
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Abstract

We present an approach for 3D particle fusion in localization microscopy which dramatically increases signal-to-noise ratio and resolution in single particle analysis. Our method does not require a structural template, and properly handles anisotropic localization uncertainties. We demonstrate 3D particle reconstructions of the Nup107 subcomplex of the nuclear pore complex (NPC), cross-validated using multiple localization microscopy techniques, as well as two-color 3D reconstructions of the NPC, and reconstructions of DNA-origami tetrahedrons.

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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 10, 2019.
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Three dimensional particle averaging for structural imaging of macromolecular complexes by localization microscopy
Hamidreza Heydarian, Adrian Przybylski, Florian Schueder, Ralf Jungmann, Ben van Werkhoven, Jan Keller-Findeisen, Jonas Ries, Sjoerd Stallinga, Mark Bates, Bernd Rieger
bioRxiv 837575; doi: https://doi.org/10.1101/837575
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Three dimensional particle averaging for structural imaging of macromolecular complexes by localization microscopy
Hamidreza Heydarian, Adrian Przybylski, Florian Schueder, Ralf Jungmann, Ben van Werkhoven, Jan Keller-Findeisen, Jonas Ries, Sjoerd Stallinga, Mark Bates, Bernd Rieger
bioRxiv 837575; doi: https://doi.org/10.1101/837575

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