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Three-Photon Adaptive Optics for Mouse Brain Imaging
Three-photon microscopy (3PM) was shown to allow deeper imaging than two-photon microscopy (2PM) in scattering biological tissues, such as the mouse brain, since the longer excitation wavelength reduces tissue scattering and the higher-order non-linear excitation suppresses out-of-focus background f...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2022
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9185169/ https://www.ncbi.nlm.nih.gov/pubmed/35692424 http://dx.doi.org/10.3389/fnins.2022.880859 |
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author | Sinefeld, David Xia, Fei Wang, Mengran Wang, Tianyu Wu, Chunyan Yang, Xusan Paudel, Hari P. Ouzounov, Dimitre G. Bifano, Thomas G. Xu, Chris |
author_facet | Sinefeld, David Xia, Fei Wang, Mengran Wang, Tianyu Wu, Chunyan Yang, Xusan Paudel, Hari P. Ouzounov, Dimitre G. Bifano, Thomas G. Xu, Chris |
author_sort | Sinefeld, David |
collection | PubMed |
description | Three-photon microscopy (3PM) was shown to allow deeper imaging than two-photon microscopy (2PM) in scattering biological tissues, such as the mouse brain, since the longer excitation wavelength reduces tissue scattering and the higher-order non-linear excitation suppresses out-of-focus background fluorescence. Imaging depth and resolution can further be improved by aberration correction using adaptive optics (AO) techniques where a spatial light modulator (SLM) is used to correct wavefront aberrations. Here, we present and analyze a 3PM AO system for in vivo mouse brain imaging. We use a femtosecond source at 1300 nm to generate three-photon (3P) fluorescence in yellow fluorescent protein (YFP) labeled mouse brain and a microelectromechanical (MEMS) SLM to apply different Zernike phase patterns. The 3P fluorescence signal is used as feedback to calculate the amount of phase correction without direct phase measurement. We show signal improvement in the cortex and the hippocampus at greater than 1 mm depth and demonstrate close to diffraction-limited imaging in the cortical layers of the brain, including imaging of dendritic spines. In addition, we characterize the effective volume for AO correction within brain tissues, and discuss the limitations of AO correction in 3PM of mouse brain. |
format | Online Article Text |
id | pubmed-9185169 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-91851692022-06-11 Three-Photon Adaptive Optics for Mouse Brain Imaging Sinefeld, David Xia, Fei Wang, Mengran Wang, Tianyu Wu, Chunyan Yang, Xusan Paudel, Hari P. Ouzounov, Dimitre G. Bifano, Thomas G. Xu, Chris Front Neurosci Neuroscience Three-photon microscopy (3PM) was shown to allow deeper imaging than two-photon microscopy (2PM) in scattering biological tissues, such as the mouse brain, since the longer excitation wavelength reduces tissue scattering and the higher-order non-linear excitation suppresses out-of-focus background fluorescence. Imaging depth and resolution can further be improved by aberration correction using adaptive optics (AO) techniques where a spatial light modulator (SLM) is used to correct wavefront aberrations. Here, we present and analyze a 3PM AO system for in vivo mouse brain imaging. We use a femtosecond source at 1300 nm to generate three-photon (3P) fluorescence in yellow fluorescent protein (YFP) labeled mouse brain and a microelectromechanical (MEMS) SLM to apply different Zernike phase patterns. The 3P fluorescence signal is used as feedback to calculate the amount of phase correction without direct phase measurement. We show signal improvement in the cortex and the hippocampus at greater than 1 mm depth and demonstrate close to diffraction-limited imaging in the cortical layers of the brain, including imaging of dendritic spines. In addition, we characterize the effective volume for AO correction within brain tissues, and discuss the limitations of AO correction in 3PM of mouse brain. Frontiers Media S.A. 2022-05-24 /pmc/articles/PMC9185169/ /pubmed/35692424 http://dx.doi.org/10.3389/fnins.2022.880859 Text en Copyright © 2022 Sinefeld, Xia, Wang, Wang, Wu, Yang, Paudel, Ouzounov, Bifano and Xu. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Neuroscience Sinefeld, David Xia, Fei Wang, Mengran Wang, Tianyu Wu, Chunyan Yang, Xusan Paudel, Hari P. Ouzounov, Dimitre G. Bifano, Thomas G. Xu, Chris Three-Photon Adaptive Optics for Mouse Brain Imaging |
title | Three-Photon Adaptive Optics for Mouse Brain Imaging |
title_full | Three-Photon Adaptive Optics for Mouse Brain Imaging |
title_fullStr | Three-Photon Adaptive Optics for Mouse Brain Imaging |
title_full_unstemmed | Three-Photon Adaptive Optics for Mouse Brain Imaging |
title_short | Three-Photon Adaptive Optics for Mouse Brain Imaging |
title_sort | three-photon adaptive optics for mouse brain imaging |
topic | Neuroscience |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9185169/ https://www.ncbi.nlm.nih.gov/pubmed/35692424 http://dx.doi.org/10.3389/fnins.2022.880859 |
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