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Preparation of Primary Neurons for Visualizing Neurites in a Frozen-hydrated State Using Cryo-Electron Tomography
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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
Chapters
- 00:05Title
- 01:19Preparing Dishes with EM Grids
- 03:43Preparing and Plating Primary Neurons on EM Grids
- 04:45Vitrifying Neurons on EM Grids
- 08:05Results: Visualizing Neurites in a Frozen-hydrated State Using Cryo-Electron Tomography
- 09:32Conclusion
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.
