Simultaneous two-photon optogenetics and imaging of cortical circuits in three dimensions
- Columbia University, United States
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.
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Author details
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].
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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Simultaneous two-photon optogenetics and imaging of cortical circuits in three dimensions
eLife 7:e32671.
https://doi.org/10.7554/eLife.32671
