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  Targeted patch-clamp recordings and single-cell electroporation of unlabeled neurons in vivo

Kitamura, K., Judkewitz, B., Kano, M., Denk, W., & Häusser, M. (2008). Targeted patch-clamp recordings and single-cell electroporation of unlabeled neurons in vivo. Nature methods, 5(1), 61-67. doi:10.1038/nmeth1150.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-002C-063E-3 Version Permalink: http://hdl.handle.net/21.11116/0000-0000-DBEE-E
Genre: Journal Article

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 Creators:
Kitamura, Kazuo, Author
Judkewitz, Benjamin1, Author              
Kano, Masanobu, Author
Denk, Winfried1, Author              
Häusser, Michael2, Author              
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1Department of Biomedical Optics, Max Planck Institute for Medical Research, Max Planck Society, ou_1497699              
2Department of Cell Physiology, Max Planck Institute for Medical Research, Max Planck Society, ou_1497701              

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 Abstract: Here we describe an approach for making targeted patch-clamp recordings from single neurons in vivo, visualized by two-photon microscopy. A patch electrode is used to perfuse the extracellular space surrounding the neuron of interest with a fluorescent dye, thus enabling the neuron to be visualized as a negative image ('shadow') and identified on the basis of its somatodendritic structure. The same electrode is then placed on the neuron under visual control to allow formation of a gigaseal ('shadowpatching'). We demonstrate the reliability and versatility of shadowpatching by performing whole-cell recordings from visually identified neurons in the neocortex and cerebellum of rat and mouse. We also show that the method can be used for targeted in vivo single-cell electroporation of plasmid DNA into identified cell types, leading to stable transgene expression. This approach facilitates the recording, labeling and genetic manipulation of single neurons in the intact native mammalian brain without the need to pre-label neuronal populations.

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Language(s): eng - English
 Dates: 2007-08-242007-11-262007-12-232008-01
 Publication Status: Published in print
 Pages: 7
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 Table of Contents: -
 Rev. Type: Peer
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Title: Nature methods
  Other : Nature methods
Source Genre: Journal
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Publ. Info: New York, NY : Nature Pub. Group
Pages: - Volume / Issue: 5 (1) Sequence Number: - Start / End Page: 61 - 67 Identifier: ISSN: 1548-7091
CoNE: https://pure.mpg.de/cone/journals/resource/111088195279556