Fiber-Coupled Rotating Two-Photon Light-Sheet Microscopy for Physiological Vestibular Imaging in Larval Zebrafish
- 1Laboratoire Jean Perrin, IBPS, Sorbonne Université
Abstract
Physiological vestibular stimulation requires macroscopic rotation of the animal, making simultaneous high-resolution neural imaging technically demanding. Here we present a fiber-coupled rotating two-photon light-sheet microscope for physiological pitch-axis vestibular imaging in larval zebrafish. Two-photon excitation is attractive in this context because near-infrared excitation avoids visible-light contamination of the visual environment. Femtosecond pulses were delivered to the rotating microscope through a broadband hollow-core negative-curvature fiber previously characterized in static light-sheet configurations. The rotating instrument maintained the axial mechanical stability required for cellular-resolution imaging, with submicron mean axial displacement over the characterized angular range. Fiber performance under rotation was also stable: bend-induced transmission changes were small in the relevant curvature regime, and circular input polarization suppressed rotation-dependent polarization artefacts to below 1\% in the sample plane. The dominant rotation-sensitive optical limitation was not the fiber but perturbation of the thermal lens in the sample chamber. Reducing fluid motion along the excitation path suppressed this artefact and restored stable imaging during dynamic rotation. With this system, we recorded brain-wide vestibular responses during sinusoidal pitch stimulation and resolved anatomically organized cellular response patterns across the brain. These results establish rotating two-photon light-sheet microscopy as a practical solution for vestibular imaging in experiments that require physiological stimulation together with darkness or controlled visual conditions.
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BibTeX
@misc{trentesaux2026fibercoupled,
title = {{Fiber-Coupled Rotating Two-Photon Light-Sheet Microscopy for Physiological Vestibular Imaging in Larval Zebrafish}},
author = {H. Trentesaux and G. Migault and T. Panier and G. Debregeas and V. Bormuth},
howpublished = {bioRxiv},
year = {2026},
month = oct,
doi = {10.64898/2026.10.10.756414},
url = {https://www.biorxiv.org/content/10.64898/2026.10.10.756414v1},
note = {CC BY-NC-ND},
}
RIS
TY - UNPB
TI - Fiber-Coupled Rotating Two-Photon Light-Sheet Microscopy for Physiological Vestibular Imaging in Larval Zebrafish
AU - H. Trentesaux
AU - G. Migault
AU - T. Panier
AU - G. Debregeas
AU - V. Bormuth
PY - 2026
DA - 2026/10/10
PB - bioRxiv
DO - 10.64898/2026.10.10.756414
UR - https://www.biorxiv.org/content/10.64898/2026.10.10.756414v1
AB - Physiological vestibular stimulation requires macroscopic rotation of the animal, making simultaneous high-resolution neural imaging technically demanding. Here we present a fiber-coupled rotating two-photon light-sheet microscope for physiological pitch-axis vestibular imaging in larval zebrafish. Two-photon excitation is attractive in this context because near-infrared excitation avoids visible-light contamination of the visual environment. Femtosecond pulses were delivered to the rotating microscope through a broadband hollow-core negative-curvature fiber previously characterized in static light-sheet configurations. The rotating instrument maintained the axial mechanical stability required for cellular-resolution imaging, with submicron mean axial displacement over the characterized angular range. Fiber performance under rotation was also stable: bend-induced transmission changes were small in the relevant curvature regime, and circular input polarization suppressed rotation-dependent polarization artefacts to below 1\% in the sample plane. The dominant rotation-sensitive optical limitation was not the fiber but perturbation of the thermal lens in the sample chamber. Reducing fluid motion along the excitation path suppressed this artefact and restored stable imaging during dynamic rotation. With this system, we recorded brain-wide vestibular responses during sinusoidal pitch stimulation and resolved anatomically organized cellular response patterns across the brain. These results establish rotating two-photon light-sheet microscopy as a practical solution for vestibular imaging in experiments that require physiological stimulation together with darkness or controlled visual conditions.
ER -