A large field-of-view, single-cell-resolution two- and three-photon microscope for deep and wide imaging
Abstract In vivo imaging of large-scale neuronal activity plays a pivotal role in unraveling the function of the brain's circuitry. Multiphoton microscopy, a powerful tool for deep-tissue imaging, has received sustained interest in advancing its speed, field of view and imaging depth. However,...
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| Format: | Article |
| Language: | English |
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SpringerOpen
2024-11-01
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| Series: | eLight |
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| Online Access: | https://doi.org/10.1186/s43593-024-00076-4 |
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| author | Aaron T. Mok Tianyu Wang Shitong Zhao Kristine E. Kolkman Danni Wu Dimitre G. Ouzounov Changwoo Seo Chunyan Wu Joseph R. Fetcho Chris Xu |
| author_facet | Aaron T. Mok Tianyu Wang Shitong Zhao Kristine E. Kolkman Danni Wu Dimitre G. Ouzounov Changwoo Seo Chunyan Wu Joseph R. Fetcho Chris Xu |
| author_sort | Aaron T. Mok |
| collection | DOAJ |
| description | Abstract In vivo imaging of large-scale neuronal activity plays a pivotal role in unraveling the function of the brain's circuitry. Multiphoton microscopy, a powerful tool for deep-tissue imaging, has received sustained interest in advancing its speed, field of view and imaging depth. However, to avoid thermal damage in scattering biological tissue, field of view decreases exponentially as imaging depth increases. We present a suite of innovations to optimize three-photon microscopy for large field-of-view imaging at depths unreachable by two-photon microscopy. These techniques enable us to image neuronal activities of transgenic animals expressing protein calcium sensors in a ~ 3.5-mm diameter field-of-view with single-cell resolution in the deepest cortical layer of mouse brains. We further demonstrate simultaneous large field-of-view two-photon and three-photon imaging, subcortical imaging in the mouse brain, and whole-brain imaging in adult zebrafish. The demonstrated techniques can be integrated into typical multiphoton microscopes to enlarge field of view for system-level neural circuit research. |
| format | Article |
| id | doaj-art-d13c4a3b9eea4ebd9a9100ad8a630afd |
| institution | Kabale University |
| issn | 2097-1710 2662-8643 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | SpringerOpen |
| record_format | Article |
| series | eLight |
| spelling | doaj-art-d13c4a3b9eea4ebd9a9100ad8a630afd2024-12-15T12:07:08ZengSpringerOpeneLight2097-17102662-86432024-11-014111410.1186/s43593-024-00076-4A large field-of-view, single-cell-resolution two- and three-photon microscope for deep and wide imagingAaron T. Mok0Tianyu Wang1Shitong Zhao2Kristine E. Kolkman3Danni Wu4Dimitre G. Ouzounov5Changwoo Seo6Chunyan Wu7Joseph R. Fetcho8Chris Xu9School of Applied and Engineering Physics, Cornell UniversityDepartment of Electrical and Computer Engineering, Boston UniversitySchool of Applied and Engineering Physics, Cornell UniversityDepartment of Neurobiology and Behavior, Cornell UniversityHarvard T.H. Chan School of Public Health, Harvard UniversitySchool of Applied and Engineering Physics, Cornell UniversityDepartment of Molecular & Cellular Biology, Harvard UniversityMcGovern Institute for Brain Research at MITDepartment of Neurobiology and Behavior, Cornell UniversitySchool of Applied and Engineering Physics, Cornell UniversityAbstract In vivo imaging of large-scale neuronal activity plays a pivotal role in unraveling the function of the brain's circuitry. Multiphoton microscopy, a powerful tool for deep-tissue imaging, has received sustained interest in advancing its speed, field of view and imaging depth. However, to avoid thermal damage in scattering biological tissue, field of view decreases exponentially as imaging depth increases. We present a suite of innovations to optimize three-photon microscopy for large field-of-view imaging at depths unreachable by two-photon microscopy. These techniques enable us to image neuronal activities of transgenic animals expressing protein calcium sensors in a ~ 3.5-mm diameter field-of-view with single-cell resolution in the deepest cortical layer of mouse brains. We further demonstrate simultaneous large field-of-view two-photon and three-photon imaging, subcortical imaging in the mouse brain, and whole-brain imaging in adult zebrafish. The demonstrated techniques can be integrated into typical multiphoton microscopes to enlarge field of view for system-level neural circuit research.https://doi.org/10.1186/s43593-024-00076-4Three-photon microscopyTwo-photon microscopyLarge field of viewBrain imagingDEEPscope |
| spellingShingle | Aaron T. Mok Tianyu Wang Shitong Zhao Kristine E. Kolkman Danni Wu Dimitre G. Ouzounov Changwoo Seo Chunyan Wu Joseph R. Fetcho Chris Xu A large field-of-view, single-cell-resolution two- and three-photon microscope for deep and wide imaging eLight Three-photon microscopy Two-photon microscopy Large field of view Brain imaging DEEPscope |
| title | A large field-of-view, single-cell-resolution two- and three-photon microscope for deep and wide imaging |
| title_full | A large field-of-view, single-cell-resolution two- and three-photon microscope for deep and wide imaging |
| title_fullStr | A large field-of-view, single-cell-resolution two- and three-photon microscope for deep and wide imaging |
| title_full_unstemmed | A large field-of-view, single-cell-resolution two- and three-photon microscope for deep and wide imaging |
| title_short | A large field-of-view, single-cell-resolution two- and three-photon microscope for deep and wide imaging |
| title_sort | large field of view single cell resolution two and three photon microscope for deep and wide imaging |
| topic | Three-photon microscopy Two-photon microscopy Large field of view Brain imaging DEEPscope |
| url | https://doi.org/10.1186/s43593-024-00076-4 |
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