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Simultaneous all-optical manipulation and recording of neural circuit activity with cellular resolution in vivo

Nature Methods volume 12, pages 140146 (2015) | Download Citation

  • A Corrigendum to this article was published on 30 June 2015

This article has been updated

Abstract

We describe an all-optical strategy for simultaneously manipulating and recording the activity of multiple neurons with cellular resolution in vivo. We performed simultaneous two-photon optogenetic activation and calcium imaging by coexpression of a red-shifted opsin and a genetically encoded calcium indicator. A spatial light modulator allows tens of user-selected neurons to be targeted for spatiotemporally precise concurrent optogenetic activation, while simultaneous fast calcium imaging provides high-resolution network-wide readout of the manipulation with negligible optical cross-talk. Proof-of-principle experiments in mouse barrel cortex demonstrate interrogation of the same neuronal population during different behavioral states and targeting of neuronal ensembles based on their functional signature. This approach extends the optogenetic toolkit beyond the specificity obtained with genetic or viral approaches, enabling high-throughput, flexible and long-term optical interrogation of functionally defined neural circuits with single-cell and single-spike resolution in the mouse brain in vivo.

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Change history

  • 06 February 2015

    In the version of this article initially published, the size of the scale bar reported in the legend of Figure 3a was incorrect. The correct size is 100 μm, not 50 μm. In addition, the volume of injected virus in the Online Methods section "Titration of calcium indicator expression" had the incorrect unit. The correct volume is 100 nl, not 100 μl. The errors have been corrected in the HTML and PDF versions of the article.

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Acknowledgements

We thank M. Pachitariu, C. Stringer, S. Turaga, M. London and N. Pettit for helpful discussion, analysis routines and software; C. Wilms, C. Schmidt-Hieber, A. Roth, B. Clark, I. Bianco, S. Smith, B. Judkewitz and D. Peterka for discussion and comments on the manuscript; the staff at Bruker Corporation (formerly Prairie Technologies) for enabling customization of the microscope; M. Lochrie at the Stanford Neuroscience Gene Vector and Virus Core (grant no. P30 NS069375-01A1) for advice on use of AAVdj; and K. Deisseroth (Stanford University) for plasmids and access to AAVdj virus. This work was supported by grants from the Wellcome Trust, the Gatsby Charitable Foundation, the European Commission (Marie Curie International Incoming Fellowship grant no. 328048), the European Molecular Biology Organization, the Medical Research Council and the European Research Council.

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Affiliations

  1. Wolfson Institute for Biomedical Research, University College London, London, UK.

    • Adam M Packer
    • , Lloyd E Russell
    • , Henry W P Dalgleish
    •  & Michael Häusser
  2. Department of Neuroscience, Physiology and Pharmacology, University College London, London, UK.

    • Adam M Packer
    • , Lloyd E Russell
    • , Henry W P Dalgleish
    •  & Michael Häusser

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Contributions

A.M.P., L.E.R. and H.W.P.D. performed experiments and analyzed data. A.M.P., L.E.R., H.W.P.D. and M.H. designed the study and wrote the paper.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Michael Häusser.

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DOI

https://doi.org/10.1038/nmeth.3217

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