ArticleProceedings of the National Academy of Sciences of the United States of America2016
Dissection of molecular assembly dynamics by tracking orientation and position of single molecules in live cells.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 52 papers.
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Who cites it
52 citing papers in PubMed.
- SPIFFI enables single-shot super-resolution and multidimensional imaging.Nature methods · 2026Article
- Efficient spatio-angular reconstruction enables high-fidelity mapping of six-dimensional structures and dynamics with polarized fluorescence microscopy.Nature communications · 2026Article
- N-cadherin orientational order decreases with mechanical load at cardiomyocyte adherens junctions.bioRxiv : the preprint server for biology · 2026Article
- SurFlex microscopy: Measuring flexibility of surface-tethered biomolecules.Proceedings of the National Academy of Sciences of the United States of America · 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
- High contrast fluorescence polarization microscopy through double tagged photoswitchable fluorescent proteins.Npj imaging · 2025Article
- A Bayesian Model to Count the Number of Two-State Emitters in a Diffraction Limited Spot.Nano letters · 2025Article
- Volumetric imaging of the 3D orientation of cellular structures with a polarized fluorescence light-sheet microscope.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- Single-molecule orientation-localization microscopy: Applications and approaches.Quarterly reviews of biophysics · 2024Review
- Illuminating cellular architecture and dynamics with fluorescence polarization microscopy.Journal of cell science · 2024Review
- Overview of the super-resolution and advanced microscopy imaging session at the 21st IUPAB and the 62nd Biophysics Society of Japan joint congress.Biophysical reviews · 2024Article
- Permittivity tensor imaging: modular label-free imaging of 3D dry mass and 3D orientation at high resolution.Nature methods · 2024Article
- Three-dimensional spatio-angular fluorescence microscopy with a polarized dual-view inverted selective-plane illumination microscope (pol-diSPIM).bioRxiv : the preprint server for biology · 2024Article
- In Situ Direct Monitoring of the Morphological Transformation of Single Au Nanostars Induced by Iodide through Dual-Laser Dark-Field Microscopy: Unexpected Mechanism and Sensing Applications.Nanomaterials (Basel, Switzerland) · 2022Article
- Correlative imaging of the spatio-angular dynamics of biological systems with multimodal instant polarization microscope.Biomedical optics express · 2022Article
- 4polar-STORM polarized super-resolution imaging of actin filament organization in cells.Nature communications · 2022Article
- Optical tweezer and TIRF microscopy for single molecule manipulation of RNA/DNA nanostructures including their rubbery property and single molecule counting.Biophysics reports · 2021Article
- Single-Shot Quantitative Polarization Imaging of Complex Birefringent Structure Dynamics.ACS photonics · 2021Article
- A gain series method for accurate EMCCD calibration.Scientific reports · 2021Article
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Authors and funding
8 authors.
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Abstract
Regulation of order, such as orientation and conformation, drives the function of most molecular assemblies in living cells but remains difficult to measure accurately through space and time. We built an instantaneous fluorescence polarization microscope, which simultaneously images position and orientation of fluorophores in living cells with single-molecule sensitivity and a time resolution of 100 ms. We developed image acquisition and analysis methods to track single particles that interact with higher-order assemblies of molecules. We tracked the fluctuations in position and orientation of molecules from the level of an ensemble of fluorophores down to single fluorophores. We tested our system in vitro using fluorescently labeled DNA and F-actin, in which the ensemble orientation of polarized fluorescence is known. We then tracked the orientation of sparsely labeled F-actin network at the leading edge of migrating human keratinocytes, revealing the anisotropic distribution of actin filaments relative to the local retrograde flow of the F-actin network. Additionally, we analyzed the position and orientation of septin-GFP molecules incorporated in septin bundles in growing hyphae of a filamentous fungus. Our data indicate that septin-GFP molecules undergo positional fluctuations within ∼350 nm of the binding site and angular fluctuations within ∼30° of the central orientation of the bundle. By reporting position and orientation of molecules while they form dynamic higher-order structures, our approach can provide insights into how micrometer-scale ordered assemblies emerge from nanoscale molecules in living cells.
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