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Simultaneous two-photon optogenetics and imaging of cortical circuits in three dimensions

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Cite as: eLife 2018;7:e32671 doi: 10.7554/eLife.32671

Abstract

The simultaneously imaging and manipulating of neural activity in three-dimensions could enable the functional dissection of neural circuits. Here we have combined two-photon optogenetics with simultaneous volumetric two-photon calcium imaging to manipulate neural activity in mouse neocortex in vivo in 3D, while maintaining cellular resolution. Using a hybrid holographic approach, we simultaneously photostimulate more than 80 neurons over 150 μm in depth in cortical layer 2/3 from mouse visual cortex. We validate the usefulness of the microscope by photoactivating in 3D selected groups of interneurons, suppressing the response of nearby pyramidal neurons to visual stimuli. Our all-optical method could be used as a general platform to read and write activity of neural circuits.

Article and author information

Author details

  1. Weijian Yang

    Department of Biological Sciences, Columbia University, New York, United States
    For correspondence
    wejyang@ucdavis.edu
    Competing interests
    No competing interests declared.
    ORCID icon 0000-0003-0941-3496
  2. Luis Carrillo-Reid

    Department of Biological Sciences, Columbia University, New York, United States
    Competing interests
    No competing interests declared.
  3. Yuki Bando

    Department of Biological Sciences, Columbia University, New York, United States
    Competing interests
    No competing interests declared.
  4. Darcy S Peterka

    Department of Biological Sciences, Columbia University, New York, United States
    Competing interests
    Darcy S Peterka, is listed as an inventor of the following patent: Devices.
    ORCID icon 0000-0001-7351-5820
  5. Rafael Yuste

    Department of Biological Sciences, Columbia University, New York, United States
    For correspondence
    rmy5@columbia.edu
    Competing interests
    Rafael Yuste, is listed as an inventor of the following patent: Devices.
    ORCID icon 0000-0003-4206-497X

Funding

National Eye Institute (DP1EY024503)

  • Rafael Yuste

National Institute of Mental Health (R44MH109187)

  • Darcy S Peterka

Defense Advanced Research Projects Agency (N66001-15-C-4032)

  • Rafael Yuste

National Institute of Mental Health (R01MH100561)

  • Rafael Yuste

National Eye Institute (R21EY027592)

  • Darcy S Peterka

National Institute of Mental Health (R01MH101218)

  • Rafael Yuste

Defense Advanced Research Projects Agency (W91NF-14-1-0269)

  • Rafael Yuste

Army Research Laboratory (W911NF-12-1-0594)

  • Rafael Yuste

Army Research Office (W911NF-12-1-0594)

  • Rafael Yuste

Burroughs Wellcome Fund (1015761)

  • Weijian Yang

Uehara Memorial Foundation

  • Yuki Bando

National Eye Institute (R01EY011787)

  • Rafael Yuste

National Institute of Mental Health (R41MH100895)

  • Rafael Yuste

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: This study was performed in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. All of the animals were handled according to approved institutional animal care and use committee (IACUC) protocols of Columbia University [protocol ID: AC-AAAM5100, AC-AAAM7951].

Reviewing Editor

  1. Karel Svoboda, Reviewing Editor, Janelia Research Campus, Howard Hughes Medical Institute, United States

Publication history

  1. Received: October 10, 2017
  2. Accepted: February 5, 2018
  3. Accepted Manuscript published: February 7, 2018 (version 1)

Copyright

© 2018, Yang et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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