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

Nat Methods. 2020 Jan;17(1):50-54. doi: 10.1038/s41592-019-0630-5. Epub 2019 Nov 18.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Membrane / metabolism
  • Cell Membrane / ultrastructure*
  • Cryoelectron Microscopy / instrumentation*
  • Cryoelectron Microscopy / methods*
  • Green Fluorescent Proteins / metabolism*
  • HeLa Cells
  • Humans
  • Image Processing, Computer-Assisted / methods*
  • Molecular Imaging / methods*
  • Pattern Recognition, Automated
  • Specimen Handling / methods*


  • Green Fluorescent Proteins