Rehydration of Freeze Substituted Brain Tissue for Pre-embedding Immunoelectron Microscopy.

Autor:	Pérez-Garza J; Department of Physiology and Neurobiology, University of Connecticut, 75 North Eagleville Rd. Unit 3156, Storrs, CT 06269-3156, USA., Parrish-Mulliken E; Department of Physiology and Neurobiology, University of Connecticut, 75 North Eagleville Rd. Unit 3156, Storrs, CT 06269-3156, USA., Deane Z; Department of Physiology and Neurobiology, University of Connecticut, 75 North Eagleville Rd. Unit 3156, Storrs, CT 06269-3156, USA., Ostroff LE; Department of Physiology and Neurobiology, University of Connecticut, 75 North Eagleville Rd. Unit 3156, Storrs, CT 06269-3156, USA.; Connecticut Institute for the Brain and Cognitive Sciences, University of Connecticut, 337 Mansfield Rd. Unit 1272, Storrs, CT 06269-1272, USA.; Institute of Materials Science, University of Connecticut, 25 King Hill Rd. Unit 3136, Storrs, CT 06269-3136, USA.
Jazyk:	angličtina
Zdroj:	Microscopy and microanalysis : the official journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada [Microsc Microanal] 2023 Sep 29; Vol. 29 (5), pp. 1694-1704.
DOI:	10.1093/micmic/ozad077
Abstrakt:	Electron microscopy (EM) volume reconstruction is a powerful tool for investigating the fundamental structure of brain circuits, but the full potential of this technique is limited by the difficulty of integrating molecular information. High quality ultrastructural preservation is necessary for EM reconstruction, and intact, highly contrasted cell membranes are essential for following small neuronal processes through serial sections. Unfortunately, the antibody labeling methods used to identify most endogenous molecules result in compromised morphology, especially of membranes. Cryofixation can produce superior morphological preservation and has the additional advantage of allowing indefinite storage of valuable samples. We have developed a method based on cryofixation that allows sensitive immunolabeling of endogenous molecules, preserves excellent ultrastructure, and is compatible with high-contrast staining for serial EM reconstruction. Competing Interests: Conflict of Interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. (© The Author(s) 2023. Published by Oxford University Press on behalf of the Microscopy Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
Databáze:	MEDLINE
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