Random-access two-photon holographic optogenetic stimulation combined with brain-wide functional light-sheet imaging in larval zebrafish

Antoine Hubert; Mattéo Dommanget-Kott; Sébastien Wolf; Thomas Panier; Georges Debrégeas; Volker Bormuth

doi:10.1117/12.2671030

1 September 2023 Random-access two-photon holographic optogenetic stimulation combined with brain-wide functional light-sheet imaging in larval zebrafish

Antoine Hubert, Mattéo Dommanget-Kott, Sébastien Wolf, Thomas Panier, Georges Debrégeas, Volker Bormuth

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Proceedings Volume 12630, Advances in Microscopic Imaging IV; 1263007 (2023) https://doi.org/10.1117/12.2671030
Event: European Conferences on Biomedical Optics, 2023, Munich, Germany

Abstract

We describe a 3D holographic two-photon optogenetics setup based on acousto-optic deflectors combined with a custom-made compact light-sheet microscope setup. The system allows performing 3D-targeted perturbations of neuronal activity while simultaneously imaging the whole brain neuronal response over large field-of-view. This random-access holographic light patterning method offers high-speed sequential activation of dense neuronal assemblies spread over large areas while maintaining constant lateral and axial resolution. Here, we report on a systematic characterization of the stimulated neurons response and perform functional imaging on zebrafish (Danio Rerio) larvae while stimulating single neurons.

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Antoine Hubert, Mattéo Dommanget-Kott, Sébastien Wolf, Thomas Panier, Georges Debrégeas, and Volker Bormuth "Random-access two-photon holographic optogenetic stimulation combined with brain-wide functional light-sheet imaging in larval zebrafish", Proc. SPIE 12630, Advances in Microscopic Imaging IV, 1263007 (1 September 2023); https://doi.org/10.1117/12.2671030

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KEYWORDS

Holography

Neurons

Optogenetics

Optical lithography

Acoustooptics

Brain

Calcium

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