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Preparation of Primary Neurons for Visualizing Neurites in a Frozen-hydrated State Using Cryo-Electron Tomography
Published on: February 12, 2014
Cryo-electron microscopy of nervous tissue. A review
1Department of Cytology, Anatomical Institute, University of Bochum, Germany.
Abstract:
The present work attempts to demonstrate that cryofixation is a valuable method for the study of the nervous tissue. The use of the newly developed methods of cryofixation and freeze-etching without fixatives or cryoprotectants allows new exciting perspectives for the electron microscopical observation of cellular components, emphasizing their three-dimensional morphological structures. Significant contributions have been made on the fine structure of the cytoskeleton, cell membranes and cell organelles. The components of the cytoskeleton are distributed in different composition through the perikarya, dendrites and axon. The ubiquitous presence of the cytoskeleton suggests a crucial role in the functional activities of the neurons, especially in relation to the intracellular communication and to developmental and regeneration processes. Vitrified cellular membranes of myelin sheaths and rod outer segments have been observed in hydrated state by using cryofixation and cryotransfer techniques. These procedures allow new insights into the supramolecular structure and an approximation of morphological data to the present biophysical membrane model including a critical comparison with the current descriptions gained by conventional electron microscopy.

