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Robotic Automation of In Vivo Two-Photon Targeted Whole-Cell Patch-Clamp Electrophysiology

Research output: Contribution to journalArticlepeer-review

Luca A Annecchino, Alexander R Morris, Caroline S. Copeland, Oshiorenoya E. Agabi, Paul Chadderton, Simon R Schultz

Original languageEnglish
Pages (from-to)1048-1055.e3
JournalNeuron
Volume95
Issue number5
Early online date30 Aug 2017
DOIs
Accepted/In press11 Aug 2017
E-pub ahead of print30 Aug 2017
Published30 Aug 2017

Bibliographical note

Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

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Abstract

Whole-cell patch-clamp electrophysiological recording is a powerful technique for studying cellular function. While in vivo patch-clamp recording has recently benefited from automation, it is normally performed "blind," meaning that throughput for sampling some genetically or morphologically defined cell types is unacceptably low. One solution to this problem is to use two-photon microscopy to target fluorescently labeled neurons. Combining this with robotic automation is difficult, however, as micropipette penetration induces tissue deformation, moving target cells from their initial location. Here we describe a platform for automated two-photon targeted patch-clamp recording, which solves this problem by making use of a closed loop visual servo algorithm. Our system keeps the target cell in focus while iteratively adjusting the pipette approach trajectory to compensate for tissue motion. We demonstrate platform validation with patch-clamp recordings from a variety of cells in the mouse neocortex and cerebellum.

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