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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
Para preservar los procesos neuronales para el análisis ultraestructural, se describe un protocolo para la siembra de las neuronas primarias en rejillas de microscopía electrónica, seguido de congelación instantánea, dando muestras en suspensión en una capa de hielo vítreo. Estas muestras pueden ser examinados con un crio-microscopio electrónico para visualizar estructuras a escala nanométrica.
