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

Toro-Nahuelpan, Mauricio; Zagoriy, Ievgeniia; Senger, Fabrice; Blanchoin, Laurent; Théry, Manuel; Mahamid, Julia · Nat Methods · 2020

basic_science · Level V

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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.

Medical subject headings