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Article Dans Une Revue Nature Methods Année : 2020

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

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Micropatterning of cryo-EM grids enables controlled adhesion of mammalian cells for cryo-ET-based structural studies. This approach leads to reproducible cellular morphology and improves focused ion beam thinning of cells for in-cell structural analyses. Spatially controlled cell adhesion on electron microscopy supports remains a bottleneck in specimen preparation for cellular cryo-electron tomography. Here, we describe contactless and mask-free photo-micropatterning of electron microscopy grids for site-specific deposition of extracellular matrix-related proteins. We attained refined cell positioning for micromachining by cryo-focused ion beam milling. Complex micropatterns generated predictable intracellular organization, allowing direct correlation between cell architecture and in-cell three-dimensional structural characterization of the underlying molecular machinery.
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hal-02379109 , version 1 (15-10-2020)

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Mauricio Toro-Nahuelpan, Ievgeniia Zagoriy, Fabrice Senger, Laurent Blanchoin, Manuel Thery, et al.. Tailoring cryo-electron microscopy grids by photo-micropatterning for in-cell structural studies. Nature Methods, 2020, 17 (1), pp.50-54. ⟨10.1038/s41592-019-0630-5⟩. ⟨hal-02379109⟩
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