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Biochemistry

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JoVE Journal Biochemistry
Micropatterning Transmission Electron Microscopy Grids to Direct Cell Positioning within Whole-Cell Cryo-Electron Tomography Workflows
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Micropatterning Transmission Electron Microscopy Grids to Direct Cell Positioning within Whole-Cell Cryo-Electron Tomography Workflows

Article DOI: 10.3791/62992-v • 09:53 min • September 13th, 2021
September 13th, 2021 •
*1,2,3, *1,4, 1,2,3, 1,2,3,5
1Department of Biochemistry, University of Wisconsin, Madison, 2Cryo-Electron Microscopy Research Center, Department of Biochemistry, University of Wisconsin, Madison, 3Midwest Center for Cryo-Electron Tomography, Department of Biochemistry, University of Wisconsin, Madison, 4Department of Chemistry, University of Wisconsin, Madison, 5Morgridge Institute for Research
* These authors contributed equally

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Summary
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The goal of this protocol is to direct cell adhesion and growth to targeted areas of grids for cryo-electron microscopy. This is achieved by applying an anti-fouling layer that is ablated in user-specified patterns followed by deposition of extra-cellular matrix proteins in the patterned areas prior to cell seeding.

Tags

Micropatterning Transmission Electron Microscopy TEM Grids Cell Positioning Cryo-electron Tomography Nanometer-level Resolution Macromolecular Complexes Native Frozen Hydrated State Cryo-correlative Light Electron Microscopy Cryo-focused-ion-beam Milling Throughput EM Grids Rapid Reproducible Broadly Adaptable Method Pipettes Tweezers Cell Culture Techniques Fluorescence Microscopy Transmission Electron Microscopy Carbon Grids
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