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

ABSTRACT: 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.

SUBMITTER: Toro-Nahuelpan M

PROVIDER: S-EPMC6949126 | biostudies-literature | 2020 Jan

REPOSITORIES: biostudies-literature

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

Toro-Nahuelpan Mauricio M Zagoriy Ievgeniia I Senger Fabrice F Blanchoin Laurent L Théry Manuel M Mahamid Julia J

Nature methods 20191118 1

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 corre ...[more]

PMID: 31740821

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