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Tailoring cryo-electron microscopy grids by photo-micropatterning for in-cell structural studies

View ORCID ProfileMauricio Toro-Nahuelpan, Ievgeniia Zagoriy, Fabrice Senger, Laurent Blanchoin, View ORCID ProfileManuel Théry, Julia Mahamid
doi: https://doi.org/10.1101/676189
Mauricio Toro-Nahuelpan
1Structural and Computational Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany
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  • ORCID record for Mauricio Toro-Nahuelpan
Ievgeniia Zagoriy
1Structural and Computational Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany
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Fabrice Senger
2CytomorphoLab, Interdisciplinary Research Institute of Grenoble, Laboratoire de Physiologie Cellulaire et Végétale, Université Grenoble-Alpes/CEA/ CNRS/INRA, Grenoble, France
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Laurent Blanchoin
2CytomorphoLab, Interdisciplinary Research Institute of Grenoble, Laboratoire de Physiologie Cellulaire et Végétale, Université Grenoble-Alpes/CEA/ CNRS/INRA, Grenoble, France
3CytomorphoLab, Hôpital Saint Louis, Institut Universitaire d’Hématologie, Université Paris Diderot, Paris, France
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Manuel Théry
2CytomorphoLab, Interdisciplinary Research Institute of Grenoble, Laboratoire de Physiologie Cellulaire et Végétale, Université Grenoble-Alpes/CEA/ CNRS/INRA, Grenoble, France
3CytomorphoLab, Hôpital Saint Louis, Institut Universitaire d’Hématologie, Université Paris Diderot, Paris, France
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Julia Mahamid
1Structural and Computational Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany
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  • For correspondence: julia.mahamid@embl.de
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Abstract

Spatially-controlled cell adhesion on electron microscopy (EM) supports remains a bottleneck in specimen preparation for cellular cryo-electron tomography. Here, we describe contactless and mask-free photo-micropatterning of EM grids for site-specific deposition of extracellular matrix-related proteins. We attained refined cell positioning for micromachining by cryo-focused ion beam milling. Complex patterns generated predictable intracellular organization, allowing direct correlation between cell architecture and in-cell 3D-structural characterization of the underlying molecular machinery.

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Posted June 19, 2019.
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Tailoring cryo-electron microscopy grids by photo-micropatterning for in-cell structural studies
Mauricio Toro-Nahuelpan, Ievgeniia Zagoriy, Fabrice Senger, Laurent Blanchoin, Manuel Théry, Julia Mahamid
bioRxiv 676189; doi: https://doi.org/10.1101/676189
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Tailoring cryo-electron microscopy grids by photo-micropatterning for in-cell structural studies
Mauricio Toro-Nahuelpan, Ievgeniia Zagoriy, Fabrice Senger, Laurent Blanchoin, Manuel Théry, Julia Mahamid
bioRxiv 676189; doi: https://doi.org/10.1101/676189

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