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          Institute: MPI für medizinische Forschung     Collection: Abteilung Biomedizinische Optik     Display Documents



ID: 420208.0, MPI für medizinische Forschung / Abteilung Biomedizinische Optik
Targeted patch−clamp recordings and single−cell electroporation of unlabeled neurons in vivo
Translation of Title:Targeted patch−clamp recordings and single−cell electroporation of unlabeled neurons in vivo
Authors:Kitamura, Kazuo; Judkewitz, Benjamin; Kano, Masanobu; Denk, Winfried; Häusser, Michael
Language:English
Date of Publication (YYYY-MM-DD):2008-01-01
Title of Journal:Nat Methods
Journal Abbrev.:Nat Methods
Volume:5
Issue / Number:1
Start Page:61
End Page:67
Review Status:Peer-review
Audience:Experts Only
Intended Educational Use:No
Abstract / Description: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.
Last Change of the Resource (YYYY-MM-DD):--
External Publication Status:published
Document Type:Article
Communicated by:Wulf Kaiser
Affiliations:MPI f�r medizinische Forschung/Abteilung Biomedizinische Optik
MPI f�r medizinische Forschung/Abteilung Zellphysiologie
External Affiliations:Wolfson Institute for Biomedical Research and Department of Physiology, University College London, Gower Street, London WC1E 6BT, UK; Department of Cellular Neuroscience, Graduate School of Medicine, Osaka University, 2−2 Yamadaoka, Suita, Osaka 565−0871, Japan; PRESTO, Japan Science and Technology Agency, 4−1−8 Honcho Kawaguchi, Saitama 332−0012, Japan
Identifiers:LOCALID:7154
URI:http%3A%2F%2Fwww.nature.com%2Fnmeth%2Fjournal%2Fv5...
URI:http%3A%2F%2Fwww.nature.com%2Fnmeth%2Fjournal%2Fv5...
URI:http%3A%2F%2Fwww.nature.com%2Fnmeth%2Fjournal%2Fv5...
DOI:10.1038%2Fnmeth1150%20
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