High resolution two-photon imaging in deep mouse brain through thinned-skull based on alpha-FSS adaptive optics

Zhentao She; Zhongya Qin; Jianan Qu

doi:10.1117/12.2692204

12 March 2024 High resolution two-photon imaging in deep mouse brain through thinned-skull based on alpha-FSS adaptive optics

Zhentao She, Zhongya Qin, Jianan Qu

Author Affiliations +

Proceedings Volume 12847, Multiphoton Microscopy in the Biomedical Sciences XXIV; 128470G (2024) https://doi.org/10.1117/12.2692204
Event: SPIE BiOS, 2024, San Francisco, California, United States

Abstract

Imaging the brain in its natural state with high spatial resolution is a challenging task for visualization techniques. Two-photon microscopy offers a better approach to image deep brain tissue due to its inherent sectioning ability compared to single-photon confocal microscopy. Despite this, the optical heterogeneity of the skull severely compromises imaging contrast and spatial resolution, limiting imaging depth to the superficial layer. Recently, a new adaptive optics method called alpha-FSS has been applied to three-photon microscopy, enabling high-resolution imaging in deep brain tissue through the intact skull and demonstrating strong ability to correct large aberrations and scattering. In this study, we combined alpha-FSS adaptive optics with two-photon microscopy and demonstrated that this method works well in two-photon imaging system as well, which is much more widely used than three-photon microscopy. Using this system, we achieved subcellular resolution transcranial imaging of layer 5 pyramidal neurons up to 650um below pia in living mice. We also demonstrated the ability to perform functional calcium imaging with high sensitivity, as well as high-precision laser-mediated microsurgery through thinned skull. In summary, we applied alpha-FSS to two photon system and proved its ability to achieve near non-invasive high-resolution imaging in deep brain tissue.

Conference Presentation

(2024) Published by SPIE. Downloading of the abstract is permitted for personal use only.

Citation Download Citation

Zhentao She, Zhongya Qin, and Jianan Qu "High resolution two-photon imaging in deep mouse brain through thinned-skull based on alpha-FSS adaptive optics", Proc. SPIE 12847, Multiphoton Microscopy in the Biomedical Sciences XXIV, 128470G (12 March 2024); https://doi.org/10.1117/12.2692204

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