Enhanced neuroimaging with a calcium sensor in ex-vivo Drosophila melanogaster brains using closed-loop adaptive optics light-sheet fluorescence microscopy

Antoine Hubert; Georges Farkouh; Fabrice Harms; Cynthia Veilly; Sophia Imperato; Mathias Mercier; Vincent Loriette; François Rouyer; Alexandra Fragola

doi:10.1117/1.JBO.28.6.066501

16 June 2023 Enhanced neuroimaging with a calcium sensor in ex-vivo Drosophila melanogaster brains using closed-loop adaptive optics light-sheet fluorescence microscopy

Antoine Hubert, Georges Farkouh, Fabrice Harms, Cynthia Veilly, Sophia Imperato, Mathias Mercier, Vincent Loriette, François Rouyer, Alexandra Fragola

Author Affiliations +

Journal of Biomedical Optics, Vol. 28, Issue 6, 066501 (June 2023). https://doi.org/10.1117/1.JBO.28.6.066501

Abstract

Significance

Adaptive optics (AO) has been implemented on several microscopy setups and has proven its ability to increase both signal and resolution. However, reported configurations are not suited for fast imaging of live samples or are based on an invasive or complex implementation method.

Aim

Provide a fast aberration correction method with an easy to implement AO module compatible with light-sheet fluorescence microscopy (LSFM) for enhanced imaging of live samples.

Approach

Development of an AO add-on module for LSFM based on direct wavefront sensing without requiring a guide star using an extended-scene Shack-Hartmann wavefront sensor. The enhanced setup uses a two-color sample labeling strategy to optimize the photon budget.

Results

Fast AO correction of in-depth aberrations in an ex-vivo adult Drosophila brain enables doubling the contrast when imaging with either cell reporters or calcium sensors for functional imaging. We quantify the gain in terms of image quality on different functional domains of sleep neurons in the Drosophila brain at various depths and discuss the optimization of key parameters driving AO.

Conclusion

We developed a compact AO module that can be integrated into most of the reported light-sheet microscopy setups, provides significant improvement of image quality and is compatible with fast imaging requirements such as calcium imaging.

CC BY: © The Authors. Published by SPIE under a Creative Commons Attribution 4.0 International License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.

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Antoine Hubert, Georges Farkouh, Fabrice Harms, Cynthia Veilly, Sophia Imperato, Mathias Mercier, Vincent Loriette, François Rouyer, and Alexandra Fragola "Enhanced neuroimaging with a calcium sensor in ex-vivo Drosophila melanogaster brains using closed-loop adaptive optics light-sheet fluorescence microscopy," Journal of Biomedical Optics 28(6), 066501 (16 June 2023). https://doi.org/10.1117/1.JBO.28.6.066501

Received: 20 February 2023; Accepted: 22 May 2023; Published: 16 June 2023

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