ArticleProceedings of the National Academy of Sciences of the United States of America2025
Volumetric imaging of the 3D orientation of cellular structures with a polarized fluorescence light-sheet microscope.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
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6 citing papers in PubMed.
- Efficient spatio-angular reconstruction enables high-fidelity mapping of six-dimensional structures and dynamics with polarized fluorescence microscopy.Nature communications · 2026Article
- Fused deep learning enables 6D single-molecule localization in polarization-resolved microscopy.Methods and applications in fluorescence · 2026Article
- Dual-view microscopy of single-molecule dipole orientations.Biophysical reports · 2025Article
- Observing biological spatio-angular structures and dynamics with statistical image reconstruction and polarized fluorescence microscopy.bioRxiv : the preprint server for biology · 2025Article
- Single molecule orientation and localization microscopy.Nature photonics · 2025Article
- Painting rich six-dimensional pictures using polarized fluorescence microscopy.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
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18 authors.
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Abstract
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.
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