Journal
NEURON
Volume 95, Issue 5, Pages 1048-+Publisher
CELL PRESS
DOI: 10.1016/j.neuron.2017.08.018
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Funding
- BBSRC [BB/K001817/1]
- Scientifica
- European Union FP7 Marie Curie Initial Training Network [289146]
- Royal Society [2011/R2]
- Biotechnology and Biological Sciences Research Council [BB/K001817/1] Funding Source: researchfish
- Medical Research Council [G1000512] Funding Source: researchfish
- BBSRC [BB/K001817/1] Funding Source: UKRI
- MRC [G1000512] Funding Source: UKRI
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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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