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TitleA cost-effective method for preparing, maintaining, and transfecting neurons in organotypic slices.
Publication TypeJournal Article
Year of Publication2014
AuthorsSoares, Cary, Kevin F. H. Lee, Denise Cook, and Jean-Claude Béïque
JournalMethods Mol Biol
Volume1183
Pagination205-19
Date Published2014
ISSN1940-6029
KeywordsAnimals, Biolistics, Brain, Equipment Design, Mice, Neurons, Organ Culture Techniques, Plasmids, Rats, Transfection
Abstract

The cellular and molecular mechanisms that underlie brain function are challenging to study in the living brain. The development of organotypic slices has provided a welcomed addition to our arsenal of experimental brain preparations by allowing both genetic and prolonged pharmacological manipulations in a system that, much like the acute slice preparation, retains several core features of the cellular and network architecture found in situ. Neurons in organotypic slices can survive in culture for several weeks, can be molecularly manipulated by transfection procedures and their function can be interrogated by traditional cellular electrophysiological or imaging techniques. Here, we describe a cost-effective protocol for the preparation and maintenance of organotypic slices and also describe a protocol for biolistic transfection that can be used to introduce plasmids in a small subset of neurons living in an otherwise molecularly unperturbed network. The implementation of these techniques offers a flexible experimental paradigm that can be used to study a multitude of neuronal mechanisms.

DOI10.1007/978-1-4939-1096-0_13
Alternate JournalMethods Mol. Biol.
PubMed ID25023311