Two-photon targeted patching (TPTP) in vivo

Komai, S., Denk, W., Osten, P., Brecht, M., Margrie, T. W. (2006) Two-photon targeted patching (TPTP) in vivo. Nature Protocols, 1 (2). pp. 647-52. ISSN 1745-2473

URL: http://www.ncbi.nlm.nih.gov/pubmed/17406293
DOI: 10.1038/nprot.2006.100

Abstract

Two-photon-excited fluorescence laser-scanning microscopy (2PLSM) has provided a wealth of information about the spatiotemporal properties of biological processes at the single cell and population level. Because such nonlinear optical methods allow for imaging deep within biological tissue, 2PLSM can be combined with patch-clamp techniques to obtain electrophysiological recordings from specific fluorescently labeled cells in vivo. Here a protocol referred to as two-photon targeted patching (TPTP) describes a method that may be used to record from cells in the intact animal labeled by virtually any type of fluorophore. We target neurons that have been optically and genetically identified using green fluorescent protein (GFP) expressed under the control of a specific promoter. TPTP when combined with genetic approaches therefore permits electrophysiological recordings from specified neurons and their compartments, including dendrites. This technique may be repeated in the same preparation many times over the course of several hours and is equally applicable to non-neuronal cell types.

Item Type: Paper
Uncontrolled Keywords: Animals Electrophysiology methods Gene Expression Regulation Green Fluorescent Proteins genetics metabolism Mice Mice Inbred C57BL Microscopy Confocal instrumentation methods Neurons metabolism Patch-Clamp Techniques instrumentation methods Photons
Subjects: Investigative techniques and equipment > microscopy > flourescence microscopy
Investigative techniques and equipment > optical devices > lasers
organs, tissues, organelles, cell types and functions > cell types and functions > cell types > neurons
organs, tissues, organelles, cell types and functions > cell types and functions > cell types > neurons
organs, tissues, organelles, cell types and functions > cell types and functions > cell types > neurons

Investigative techniques and equipment > patch-clamp recording
Investigative techniques and equipment > recording devices > patch-clamp recording
CSHL Authors:
Communities: CSHL labs > Osten lab
Depositing User: CSHL Librarian
Date Deposited: 13 Dec 2011 16:28
Last Modified: 08 Apr 2013 15:10
Related URLs:
URI: http://repository.cshl.edu/id/eprint/22842

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