Millimetric scale two-photon Bessel-Gauss beam light sheet microscopy with three-axis isotropic resolution using an axicon lens

Cléophace Akitegetse; Thomas Charland; Mireille Quémener; Charles Gora; Véronique Rioux; Michel Piché; Yves De Koninck; Martin Lévesque; Daniel C. Côté

doi:10.1117/1.NPh.10.3.035002

23 June 2023 Millimetric scale two-photon Bessel-Gauss beam light sheet microscopy with three-axis isotropic resolution using an axicon lens

Cléophace Akitegetse, Thomas Charland, Mireille Quémener, Charles Gora, Véronique Rioux, Michel Piché, Yves De Koninck, Martin Lévesque, Daniel C. Côté

Author Affiliations +

Neurophotonics, Vol. 10, Issue 3, 035002 (June 2023). https://doi.org/10.1117/1.NPh.10.3.035002

Abstract

Significance

Typical light sheet microscopes suffer from artifacts related to the geometry of the light sheet. One main inconvenience is the non-uniform thickness of the light sheet obtained with a Gaussian laser beam.

Aim

We developed a two-photon light sheet microscope that takes advantage of a thin and long Bessel-Gauss beam illumination to increase the sheet extent without compromising the resolution.

Approach

We use an axicon lens placed directly at the output of an amplified femtosecond laser to produce a long Bessel-Gauss beam on the sample. We studied the dopaminergic system and its projections in a whole cleared mouse brain.

Results

Our light sheet microscope allows an isotropic resolution of 2.4 μm in all three axes of the scanned volume while keeping a millimetric-sized field of view, and a fast acquisition rate of up to 34 mm² / s. With slight modifications to the optical setup, the sheet extent can be increased to 6 mm.

Conclusion

The proposed system’s sheet extent and resolution surpass currently available systems, enabling the fast imaging of large specimens.

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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Cléophace Akitegetse, Thomas Charland, Mireille Quémener, Charles Gora, Véronique Rioux, Michel Piché, Yves De Koninck, Martin Lévesque, and Daniel C. Côté "Millimetric scale two-photon Bessel-Gauss beam light sheet microscopy with three-axis isotropic resolution using an axicon lens," Neurophotonics 10(3), 035002 (23 June 2023). https://doi.org/10.1117/1.NPh.10.3.035002

Received: 17 March 2023; Accepted: 26 May 2023; Published: 23 June 2023

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