A direct wavefront sensing device resilient to scattering for use in adaptive optics two-photon microscopy of the mouse brain

Sophia Imperato; Fabrice Harms; Cynthia Veilly; Mathias Mercier; Laurent Bourdieu; Alexandra Fragola

doi:10.1117/12.2621838

30 May 2022 A direct wavefront sensing device resilient to scattering for use in adaptive optics two-photon microscopy of the mouse brain

Sophia Imperato, Fabrice Harms, Cynthia Veilly, Mathias Mercier, Laurent Bourdieu, Alexandra Fragola

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Proceedings Volume PC12144, Biomedical Spectroscopy, Microscopy, and Imaging II; PC121440X (2022) https://doi.org/10.1117/12.2621838
Event: SPIE Photonics Europe, 2022, Strasbourg, France

Abstract

Optical microscopy allows to perform structural and functional imaging within large volume of tissues with subcellular resolution. Non-linear microscopy allows the interrogation of neuronal activity in mammalian brains but remains limited because of scattering and optical aberrations. To overcome these issues, Adaptive Optics (AO) strategies have been implemented to retrieve the microscope imaging quality while addressing important imaging depths. A first AO strategy implemented in non-linear microscopy relies on a sensorless configuration, but is a time-consuming iterative process hardly compatible with photobleaching issues. A second approach is based on direct wavefront sensing using Shack-Hartmann wavefront sensors and has proved its efficiency on in vivo experiments. However, this method fails at large depths because of the strong scattering of the emitted fluorescence. A method for direct wavefront sensing more resilient to scattering of the fluorescence emission would therefore facilitate the use of AO in optical microscopy. This work proposes an alternative method of direct wavefront measurement, which relies on the cross-correlation of images of an extended source obtained through a microlens array. This extended-source Shack-Hartmann wavefront sensor (ESSH) requires to be coupled to an optical sectioning method. Its efficiency has been proven when coupled to Light Sheet Fluorescence Microscopy in the adult drosophila brain in weekly scattering conditions. Here, we show that it allows quantitative aberration measurements through highly scattering fixed brain slices, up to four times the scattering length of the tissue. We demonstrate that it is more resilient to scattering compared to the current centroid-based approach. Taking advantage of its geometry, this new wavefront sensor also provides scattering coefficient measurements of biological tissues. Finally, we present its implementation on a two-photon microscope within a closed–loop configuration for in depth neuroimaging in mouse brain and compare its performances in scattering media to the classical centroid approach.

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Sophia Imperato, Fabrice Harms, Cynthia Veilly, Mathias Mercier, Laurent Bourdieu, and Alexandra Fragola "A direct wavefront sensing device resilient to scattering for use in adaptive optics two-photon microscopy of the mouse brain", Proc. SPIE PC12144, Biomedical Spectroscopy, Microscopy, and Imaging II, PC121440X (30 May 2022); https://doi.org/10.1117/12.2621838

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KEYWORDS

Wavefront sensors

Scattering

Adaptive optics

Brain

Tissues

Microscopy

Scatter measurement

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