Multi-photon fluorescence microscopy with adaptive optics

Tianyi Zheng; Adrian R. Liversage; Kayvan F. Tehrani; Jarrod A. Call; Peter A. Kner; Luke J. Mortensen

doi:10.1117/12.3003445

12 March 2024 Multi-photon fluorescence microscopy with adaptive optics

Tianyi Zheng, Adrian R. Liversage, Kayvan F. Tehrani, Jarrod A. Call, Peter A. Kner, Luke J. Mortensen

Proceedings Volume 12851, Adaptive Optics and Wavefront Control for Biological Systems X; 1285106 (2024) https://doi.org/10.1117/12.3003445
Event: SPIE BiOS, 2024, San Francisco, California, United States

Abstract

Mitochondria are extremely important organelles in the regulation of bone marrow and brain activity. However, live imaging of these subcellular features with high resolution in scattering tissues like brain or bone has proven challenging. In this study, we create a next-generation two-photon fluorescence microscope that leverages low-order wavefront correction by Shack-Hartmann wavefront sensor based on different metrics to achieve fast imaging of subcellular organelles of highly scattering living mice. Metrics include maximum intensity, minimum full width at half maximum (FWHM), and maximum energy of the point spread function (PSF), enabling accuracy and robustness of sensorless correction of the system. Using AO increases the fluorescence intensity and FWHM of the PSF and achieves fast imaging of subcellular organelles with 400nm resolution through 85 μm of highly scattering tissue. This study demonstrates a promising tool for imaging mitochondria and other organelles in optically distorting biological environments, which could facilitate the study of a variety of diseases connected to mitochondrial morphology and activity in a range of biological tissues.

Conference Presentation

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

Citation Download Citation

Tianyi Zheng, Adrian R. Liversage, Kayvan F. Tehrani, Jarrod A. Call, Peter A. Kner, and Luke J. Mortensen "Multi-photon fluorescence microscopy with adaptive optics", Proc. SPIE 12851, Adaptive Optics and Wavefront Control for Biological Systems X, 1285106 (12 March 2024); https://doi.org/10.1117/12.3003445

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