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

By Mauricio Toro-Nahuelpan, Ievgeniia Zagoriy, Fabrice Senger, Laurent Blanchoin, Manuel Théry, Julia Mahamid

Posted 19 Jun 2019
bioRxiv DOI: 10.1101/676189 (published DOI: 10.1038/s41592-019-0630-5)

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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