Lattice micropatterning for cryo-electron tomography studies of cell-cell contacts

J Struct Biol. 2021 Dec;213(4):107791. doi: 10.1016/j.jsb.2021.107791. Epub 2021 Sep 11.

Abstract

Cryo-electron tomography is the highest resolution tool available for structural analysis of macromolecular complexes within their native cellular environments. At present, data acquisition suffers from low throughput, in part due to the low probability of positioning a cell such that the subcellular structure of interest is on a region of the electron microscopy (EM) grid that is suitable for imaging. Here, we photo-micropatterned EM grids to optimally position endothelial cells so as to enable high-throughput imaging of cell-cell contacts. Lattice micropatterned grids increased the average distance between intercellular contacts and thicker cell nuclei such that the regions of interest were sufficiently thin for direct imaging. We observed a diverse array of membranous and cytoskeletal structures at intercellular contacts, demonstrating the utility of this technique in enhancing the rate of data acquisition for cellular cryo-electron tomography studies.

Keywords: bioengineering; cryo-EM; cryo-ET; micropatterning.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cadherins / metabolism
  • Cell Communication*
  • Cell Nucleus / metabolism
  • Cell Nucleus / ultrastructure
  • Cells, Cultured
  • Cryoelectron Microscopy / methods*
  • Electron Microscope Tomography / methods*
  • Endothelial Cells / cytology
  • Endothelial Cells / metabolism
  • Endothelial Cells / ultrastructure
  • Extracellular Matrix / metabolism
  • Extracellular Matrix / ultrastructure
  • Humans
  • Imaging, Three-Dimensional / methods*
  • Intercellular Junctions / metabolism
  • Intercellular Junctions / ultrastructure*
  • Microscopy, Atomic Force / methods
  • Microscopy, Confocal / methods
  • Microscopy, Electron, Transmission / methods*
  • Reproducibility of Results

Substances

  • Cadherins