Volumetric imaging of the 3D orientation of cellular structures with a polarized fluorescence light-sheet microscope.
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| Format: | Artículo científico |
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Proceedings of the National Academy of Sciences of the United States of America
2025
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| author | Chandler, Talon Guo, Min Su, Yijun Chen, Jiji Wu, Yicong Liu, Junyu Agashe, Atharva Fischer, Robert S Mehta, Shalin B Kumar, Abhishek Baskin, Tobias I Jaumouillé, Valentin Liu, Huafeng Swaminathan, Vinay Nain, Amrinder S Oldenbourg, Rudolf La Riviere, Patrick J Shroff, Hari |
| author_facet | Chandler, Talon Guo, Min Su, Yijun Chen, Jiji Wu, Yicong Liu, Junyu Agashe, Atharva Fischer, Robert S Mehta, Shalin B Kumar, Abhishek Baskin, Tobias I Jaumouillé, Valentin Liu, Huafeng Swaminathan, Vinay Nain, Amrinder S Oldenbourg, Rudolf La Riviere, Patrick J Shroff, Hari Chandler, Talon Guo, Min Su, Yijun Chen, Jiji Wu, Yicong Liu, Junyu Agashe, Atharva Fischer, Robert S Mehta, Shalin B Kumar, Abhishek Baskin, Tobias I Jaumouillé, Valentin Liu, Huafeng Swaminathan, Vinay Nain, Amrinder S Oldenbourg, Rudolf La Riviere, Patrick J Shroff, Hari |
| collection | PubMed - marine biology |
| contents | Volumetric imaging of the 3D orientation of cellular structures with a polarized fluorescence light-sheet microscope. Chandler, Talon Guo, Min Su, Yijun Chen, Jiji Wu, Yicong Liu, Junyu Agashe, Atharva Fischer, Robert S Mehta, Shalin B Kumar, Abhishek Baskin, Tobias I Jaumouillé, Valentin Liu, Huafeng Swaminathan, Vinay Nain, Amrinder S Oldenbourg, Rudolf La Riviere, Patrick J Shroff, Hari Imaging, Three-Dimensional Microscopy, Fluorescence Animals Humans Mice Microscopy, Polarization Polarized fluorescence microscopy is a valuable tool for measuring molecular orientations in biological samples, but techniques for recovering three-dimensional orientations and positions of fluorescent ensembles are limited. We report a polarized dual-view light-sheet system for determining the diffraction-limited three-dimensional distribution of the orientations and positions of ensembles of fluorescent dipoles that label biological structures. We share a set of visualization, histogram, and profiling tools for interpreting these positions and orientations. We model the distributions based on the polarization-dependent efficiency of excitation and detection of emitted fluorescence, using coarse-grained representations we call orientation distribution functions (ODFs). We apply ODFs to create physics-informed models of image formation with spatio-angular point-spread and transfer functions. We use theory and experiment to conclude that light-sheet tilting is a necessary part of our design for recovering all three-dimensional orientations. We use our system to extend known two-dimensional results to three dimensions in FM1-43-labeled giant unilamellar vesicles, fast-scarlet-labeled cellulose in xylem cells, and phalloidin-labeled actin in U2OS cells. Additionally, we observe phalloidin-labeled actin in mouse fibroblasts grown on grids of labeled nanowires and identify correlations between local actin alignment and global cell-scale orientation, indicating cellular coordination across length scales. |
| format | Artículo científico |
| id | pubmed_39982748 |
| institution | PubMed |
| language | en |
| publishDate | 2025 |
| publisher | Proceedings of the National Academy of Sciences of the United States of America |
| record_format | pubmed |
| spellingShingle | Volumetric imaging of the 3D orientation of cellular structures with a polarized fluorescence light-sheet microscope. Chandler, Talon Guo, Min Su, Yijun Chen, Jiji Wu, Yicong Liu, Junyu Agashe, Atharva Fischer, Robert S Mehta, Shalin B Kumar, Abhishek Baskin, Tobias I Jaumouillé, Valentin Liu, Huafeng Swaminathan, Vinay Nain, Amrinder S Oldenbourg, Rudolf La Riviere, Patrick J Shroff, Hari Imaging, Three-Dimensional Microscopy, Fluorescence Animals Humans Mice Microscopy, Polarization Volumetric imaging of the 3D orientation of cellular structures with a polarized fluorescence light-sheet microscope. Chandler, Talon Guo, Min Su, Yijun Chen, Jiji Wu, Yicong Liu, Junyu Agashe, Atharva Fischer, Robert S Mehta, Shalin B Kumar, Abhishek Baskin, Tobias I Jaumouillé, Valentin Liu, Huafeng Swaminathan, Vinay Nain, Amrinder S Oldenbourg, Rudolf La Riviere, Patrick J Shroff, Hari Imaging, Three-Dimensional Microscopy, Fluorescence Animals Humans Mice Microscopy, Polarization Polarized fluorescence microscopy is a valuable tool for measuring molecular orientations in biological samples, but techniques for recovering three-dimensional orientations and positions of fluorescent ensembles are limited. We report a polarized dual-view light-sheet system for determining the diffraction-limited three-dimensional distribution of the orientations and positions of ensembles of fluorescent dipoles that label biological structures. We share a set of visualization, histogram, and profiling tools for interpreting these positions and orientations. We model the distributions based on the polarization-dependent efficiency of excitation and detection of emitted fluorescence, using coarse-grained representations we call orientation distribution functions (ODFs). We apply ODFs to create physics-informed models of image formation with spatio-angular point-spread and transfer functions. We use theory and experiment to conclude that light-sheet tilting is a necessary part of our design for recovering all three-dimensional orientations. We use our system to extend known two-dimensional results to three dimensions in FM1-43-labeled giant unilamellar vesicles, fast-scarlet-labeled cellulose in xylem cells, and phalloidin-labeled actin in U2OS cells. Additionally, we observe phalloidin-labeled actin in mouse fibroblasts grown on grids of labeled nanowires and identify correlations between local actin alignment and global cell-scale orientation, indicating cellular coordination across length scales. |
| title | Volumetric imaging of the 3D orientation of cellular structures with a polarized fluorescence light-sheet microscope. |
| topic | Imaging, Three-Dimensional Microscopy, Fluorescence Animals Humans Mice Microscopy, Polarization |
| url | https://pubmed.ncbi.nlm.nih.gov/39982748/ |