Multiphoton imaging of neural structure and activity in Drosophila through the intact cuticle

  1. Max Jameson Aragon
  2. Aaron T Mok
  3. Jamien Shea
  4. Mengran Wang
  5. Haein Kim
  6. Nathan Barkdull
  7. Chris Xu
  8. Nilay Yapici  Is a corresponding author
  1. Princeton University, United States
  2. Cornell University, United States
  3. University of Florida, United States

Abstract

We developed a multiphoton imaging method to capture neural structure and activity in behaving flies through the intact cuticles. Our measurements show that the fly head cuticle has surprisingly high transmission at wavelengths > 900 nm, and the difficulty of through-cuticle imaging is due to the air sacs and/or fat tissue underneath the head cuticle. By compressing or removing the air sacs, we performed multiphoton imaging of the fly brain through the intact cuticle. Our anatomical and functional imaging results show that 2- and 3-photon imaging are comparable in superficial regions such as the mushroom body, but 3-photon imaging is superior in deeper regions such as the central complex and beyond. We further demonstrated 2-photon through-cuticle functional imaging of odor-evoked calcium responses from the mushroom body g-lobes in behaving flies short-term and long-term. The through-cuticle imaging method developed here extends the time limits of in vivo imaging in flies and opens new ways to capture neural structure and activity from the fly brain.

Data availability

All data generated or analyzed during this study are included in the manuscript and supporting file; Source Data files have been provided.

Article and author information

Author details

  1. Max Jameson Aragon

    Princeton Neuroscience Institute, Princeton University, Princeton, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Aaron T Mok

    School of Applied and Engineering Physics, Cornell University, Ithaca, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Jamien Shea

    Department of Neurobiology and Behavior, Cornell University, Ithaca, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Mengran Wang

    School of Applied and Engineering Physics, Cornell University, Ithaca, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Haein Kim

    Department of Neurobiology and Behavior, Cornell University, Ithaca, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Nathan Barkdull

    Department of Physics, University of Florida, Gainesville, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1174-5046
  7. Chris Xu

    School of Applied and Engineering Physics, Cornell University, Ithaca, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3493-6427
  8. Nilay Yapici

    Department of Neurobiology and Behavior, Cornell University, Ithaca, United States
    For correspondence
    ny96@cornell.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1130-5083

Funding

National Science Foundation (DBI-1707312)

  • Nilay Yapici

National Institute of General Medical Sciences (R35 GM133698)

  • Nilay Yapici

Pew Charitable Trusts (Scholars Award)

  • Nilay Yapici

Alfred P. Sloan Foundation (Scholars Award)

  • Nilay Yapici

American Federation for Aging Research (Grants for Junior Faculty)

  • Nilay Yapici

National Science Foundation (DBI-1707312)

  • Chris Xu

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

Reviewing Editor

  1. Upinder Singh Bhalla, Tata Institute of Fundamental Research, India

Publication history

  1. Preprint posted: October 9, 2019 (view preprint)
  2. Received: April 4, 2021
  3. Accepted: January 23, 2022
  4. Accepted Manuscript published: January 24, 2022 (version 1)
  5. Accepted Manuscript updated: January 25, 2022 (version 2)
  6. Version of Record published: February 15, 2022 (version 3)

Copyright

© 2022, Aragon 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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  1. Max Jameson Aragon
  2. Aaron T Mok
  3. Jamien Shea
  4. Mengran Wang
  5. Haein Kim
  6. Nathan Barkdull
  7. Chris Xu
  8. Nilay Yapici
(2022)
Multiphoton imaging of neural structure and activity in Drosophila through the intact cuticle
eLife 11:e69094.
https://doi.org/10.7554/eLife.69094
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