Lattice micropatterning for cryo-electron tomography studies of cell-cell contacts
Autor: | Belle M. Sow, Claudia G. Vasquez, Marcin P. Walkiewicz, William I. Weis, Leeya Engel, Alexander R. Dunn, Elizabeth A. Montabana |
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Rok vydání: | 2020 |
Předmět: |
Electron Microscope Tomography
Materials science Cryo-electron microscopy Cell Communication Microscopy Atomic Force Article law.invention Imaging Three-Dimensional Microscopy Electron Transmission Structural Biology law Humans Cytoskeleton Cells Cultured Cell Nucleus Microscopy Confocal Cell cell contact Cryoelectron Microscopy Resolution (electron density) Endothelial Cells Reproducibility of Results Cadherins Extracellular Matrix Lattice (module) Intercellular Junctions Biophysics Cryo-electron tomography Tomography Electron microscope Micropatterning |
Zdroj: | J Struct Biol |
Popis: | 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 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 the 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. |
Databáze: | OpenAIRE |
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