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JoVE Journal Neuroscience
Preparation of Primary Neurons for Visualizing Neurites in a Frozen-hydrated State Using Cryo-Electron Tomography
 

Preparation of Primary Neurons for Visualizing Neurites in a Frozen-hydrated State Using Cryo-Electron Tomography

Article DOI: 10.3791/50783-v • 09:59 min • February 12th, 2014
February 12th, 2014 •
1, 2, 3, 4, 2, 3, 4
1Department of Molecular Physiology and Biophysics, Baylor College of Medicine, 2Department of Neuroscience, Baylor College of Medicine, 3Department of Neuroscience, University of California at San Diego, 4National Center for Macromolecular Imaging, Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine

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요약
Automatic Translation

To preserve neuronal processes for ultrastructural analysis, we describe a protocol for plating of primary neurons on electron microscopy grids followed by flash freezing, yielding samples suspended in a layer of vitreous ice. These samples can be examined with a cryo-electron microscope to visualize structures at the nanometer scale.

Tags

Primary Neurons Visualizing Neurites Frozen-hydrated State Cryo-electron Tomography Dendrites Axons Electrical Impulses Synaptic Communication Ultrastructural Features Organic Solvent Dehydration Resin Embedding Artifacts Growing Neurites Flash-freezing Electron Microscopy Grids Cryo-electron Tomography (cryo-ET) Nanometer Resolution Morphological Differences Dorsal Root Ganglion (DRG) Neurites Hippocampal Neurites Near-native State Neurological Disorders
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