Multiphoton imaging of neural structure and activity in Drosophila through the intact cuticle
- Princeton University, United States
- Cornell University, United States
- 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
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
- Upinder Singh Bhalla, Tata Institute of Fundamental Research, India
Version history
- Preprint posted: October 9, 2019 (view preprint)
- Received: April 4, 2021
- Accepted: January 23, 2022
- Accepted Manuscript published: January 24, 2022 (version 1)
- Accepted Manuscript updated: January 25, 2022 (version 2)
- 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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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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