ArticleProceedings of the National Academy of Sciences of the United States of America2012
Simultaneous, accurate measurement of the 3D position and orientation of single molecules.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 87 papers.
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Who cites it
87 citing papers in PubMed.
- Optimal excitation polarization schemes to boost the precision of single-molecule orientation imaging.Biomedical optics express · 2026Article
- Fused deep learning enables 6D single-molecule localization in polarization-resolved microscopy.Methods and applications in fluorescence · 2026Article
- Emission dipole orientation reveals dynamic single-molecule interactions with 2D crystals at solvent interfaces.Nature communications · 2026Article
- 4polar3D single molecule imaging of 3D orientation in dense actin networks using ratiometric polarization splitting.Nature communications · 2026Article
- D‑Blur: A Deep Learning Approach for Mapping Subdiffraction Diffusion with Motion-Blurred Images.Chemical & biomedical imaging · 2025Article
- Dual-view microscopy of single-molecule dipole orientations.Biophysical reports · 2025Article
- Electrokinetic nanoparticle transport in an interconnected porous environment: Decoupling cavity escape and directional bias.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- Long axial range 3D single-particle tracking using birefringent substrates.Nature communications · 2025Article
- High contrast fluorescence polarization microscopy through double tagged photoswitchable fluorescent proteins.Npj imaging · 2025Article
- Single-molecule orientation-localization microscopy: Applications and approaches.Quarterly reviews of biophysics · 2024Review
- POLCAM: instant molecular orientation microscopy for the life sciences.Nature methods · 2024Article
- Tracking the extensive three-dimensional motion of single ions by an engineered point-spread function.Nature communications · 2024Article
- Reaching the precision limit with tensor-based wavefront shaping.Nature communications · 2024Article
- Multicolor two-photon light-patterning microscope exploiting the spatio-temporal properties of a fiber bundle.Biomedical optics express · 2024Article
- Combining deep learning approaches and point spread function engineering for simultaneous 3D position and 3D orientation measurements of fluorescent single molecules.Optics communications · 2023Article
- Metal-Induced Energy Transfer (MIET) for Live-Cell Imaging with Fluorescent Proteins.ACS nano · 2023Article
- Imaging and positioning through scattering media with double-helix point spread function engineering.Journal of biomedical optics · 2023Article
- Tuning axial and lateral localization precision in 3D super-resolution microscopy with variable astigmatism.Optics letters · 2022Article
- Deep learning in single-molecule imaging and analysis: recent advances and prospects.Chemical science · 2022Review
- Deep-SMOLM: deep learning resolves the 3D orientations and 2D positions of overlapping single molecules with optimal nanoscale resolution.Optics express · 2022Article
27 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
8 authors.
Funding
Abstract
Recently, single molecule-based superresolution fluorescence microscopy has surpassed the diffraction limit to improve resolution to the order of 20 nm or better. These methods typically use image fitting that assumes an isotropic emission pattern from the single emitters as well as control of the emitter concentration. However, anisotropic single-molecule emission patterns arise from the transition dipole when it is rotationally immobile, depending highly on the molecule's 3D orientation and z position. Failure to account for this fact can lead to significant lateral (x, y) mislocalizations (up to ∼50-200 nm). This systematic error can cause distortions in the reconstructed images, which can translate into degraded resolution. Using parameters uniquely inherent in the double-lobed nature of the Double-Helix Point Spread Function, we account for such mislocalizations and simultaneously measure 3D molecular orientation and 3D position. Mislocalizations during an axial scan of a single molecule manifest themselves as an apparent lateral shift in its position, which causes the standard deviation (SD) of its lateral position to appear larger than the SD expected from photon shot noise. By correcting each localization based on an estimated orientation, we are able to improve SDs in lateral localization from ∼2× worse than photon-limited precision (48 vs. 25 nm) to within 5 nm of photon-limited precision. Furthermore, by averaging many estimations of orientation over different depths, we are able to improve from a lateral SD of 116 (∼4× worse than the photon-limited precision; 28 nm) to 34 nm (within 6 nm of the photon limit).
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.