Micropatterning of electron microscopy grids for improved cellular cryo-electron tomography throughput

  • Engel L
  • Vasquez C
  • Montabana E
  • et al.
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Abstract

Cryo-electron tomography is the highest resolution tool available for structural analysis of macromolecular complexes within their native cellular environment. 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 leverage photo-micropatterning of EM grids to optimally position endothelial cells to enable high-throughput imaging of cell-cell contacts. This method increases the distance between contacts and the thicker cell nuclei such that the regions of interest are sufficiently thin for imaging. The resulting pipeline enables the observation of a diverse array of structures at endothelial cell-cell contacts, demonstrating the utility of this technique in enhancing the rate of data acquisition for cellular cr o-electron tomo ra h studies

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Engel, L., Vasquez, C., Montabana, E., Sow, B., Walkiewicz, M., Weis, W., & Dunn, A. (2021). Micropatterning of electron microscopy grids for improved cellular cryo-electron tomography throughput. Microscopy and Microanalysis, 27(S1), 2570–2573. https://doi.org/10.1017/s1431927621009132

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