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Electromagnetics and Photonics
Washington University in St. Louis
Electrical & Systems Engineering Publications and Presentations
- Keyword
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- Diffusion (2)
- Fluorescence microscopy (2)
- Chemical solutions (1)
- Cholesterol condensing (1)
- Environment-sensitive fluorophores (1)
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- Fluorescence (1)
- Lateral diffusion (1)
- Molecules (1)
- Nanofibers (1)
- Nanoparticles (1)
- Peptides and proteins (1)
- Phase mask design (1)
- Phospholipid bilayer (1)
- Point spread function engineering (1)
- Polarization-sensitive imaging (1)
- Polymers (1)
- Rotational diffusion (1)
- Rotational dynamics (1)
- Single-molecule imaging (1)
- Single-molecule spectroscopy (1)
- Single-molecule tracking (1)
- Super-resolution microscopy (1)
- Thin films (1)
- Transient binding (1)
Articles 1 - 6 of 6
Full-Text Articles in Physics
6d Single-Fluorogen Orientation-Localization Microscopy For Elucidating The Architecture Of Beta-Sheet Assemblies And Biomolecular Condensates, Tingting Wu, Weiyan Zhou, Jai S. Rudra, Rohit V. Pappu, Matthew D. Lew
6d Single-Fluorogen Orientation-Localization Microscopy For Elucidating The Architecture Of Beta-Sheet Assemblies And Biomolecular Condensates, Tingting Wu, Weiyan Zhou, Jai S. Rudra, Rohit V. Pappu, Matthew D. Lew
Electrical & Systems Engineering Publications and Presentations
We develop six-dimensional single-molecule orientation-localization microscopy (SMOLM) to measure the 3D positions and 3D orientations simultaneously of single fluorophores. We show how careful optimization of phase and polarization modulation components can encode phase, polarization, and angular spectrum information from each fluorescence photon into a microscope’s dipole-spread function. We used the transient binding and blinking of Nile red (NR) to characterize the helical structure of fibrils formed by designed amphipathic peptides, KFE8L and KFE8D, and the pathological amyloid-beta peptide Aβ42. We also deployed merocyanine 540 to uncover the interfacial architectures of biomolecular condensates.
Six-Dimensional Single-Molecule Imaging With Isotropic Resolution Using A Multi-View Reflector Microscope, Oumeng Zhang, Zijian Guo, Yuanyuan He, Tingting Wu, Michael D. Vahey, Matthew D. Lew
Six-Dimensional Single-Molecule Imaging With Isotropic Resolution Using A Multi-View Reflector Microscope, Oumeng Zhang, Zijian Guo, Yuanyuan He, Tingting Wu, Michael D. Vahey, Matthew D. Lew
Electrical & Systems Engineering Publications and Presentations
Imaging of both the positions and orientations of single fluorophores, termed single-molecule orientation-localization microscopy, is a powerful tool for the study of biochemical processes. However, the limited photon budget associated with single-molecule fluorescence makes high-dimensional imaging with isotropic, nanoscale spatial resolution a formidable challenge. Here we realize a radially and azimuthally polarized multi-view reflector (raMVR) microscope for the imaging of the three-dimensional (3D) positions and 3D orientations of single molecules, with precisions of 10.9 nm and 2.0° over a 1.5-μm depth range. The raMVR microscope achieves 6D super-resolution imaging of Nile red molecules transiently bound to lipid-coated spheres, accurately resolving …
Resolving The Three-Dimensional Rotational And Translational Dynamics Of Single Molecules Using Radially And Azimuthally Polarized Fluorescence, Oumeng Zhang, Weiyan Zhou, Jin Lu, Tingting Wu, Matthew D. Lew
Resolving The Three-Dimensional Rotational And Translational Dynamics Of Single Molecules Using Radially And Azimuthally Polarized Fluorescence, Oumeng Zhang, Weiyan Zhou, Jin Lu, Tingting Wu, Matthew D. Lew
Electrical & Systems Engineering Publications and Presentations
We report a radially and azimuthally polarized (raPol) microscope for high detection and estimation performance in single-molecule orientation-localization microscopy (SMOLM). With 5000 photons detected from Nile red (NR) transiently bound within supported lipid bilayers (SLBs), raPol SMOLM achieves 2.9 nm localization precision, 1.5° orientation precision, and 0.17 sr precision in estimating rotational wobble. Within DPPC SLBs, SMOLM imaging reveals the existence of randomly oriented binding pockets that prevent NR from freely exploring all orientations. Treating the SLBs with cholesterol-loaded methyl-β-cyclodextrin (MβCD-chol) causes NR’s orientational diffusion to be dramatically reduced, but curiously NR’s median lateral displacements drastically increase from 20.8 to …
Single-Molecule Localization Microscopy Of 3d Orientation And Anisotropic Wobble Using A Polarized Vortex Point Spread Function, Tianben Ding, Matthew D. Lew
Single-Molecule Localization Microscopy Of 3d Orientation And Anisotropic Wobble Using A Polarized Vortex Point Spread Function, Tianben Ding, Matthew D. Lew
Electrical & Systems Engineering Publications and Presentations
Within condensed matter, single fluorophores are sensitive probes of their chemical environments, but it is difficult to use their limited photon budget to image precisely their positions, 3D orientations, and rotational diffusion simultaneously. We demonstrate the polarized vortex point spread function (PSF) for measuring these parameters, including characterizing the anisotropy of a molecule’s wobble, simultaneously from a single image. Even when imaging dim emitters (∼500 photons detected), the polarized vortex PSF can obtain 12 nm localization precision, 4°–8° orientation precision, and 26° wobble precision. We use the vortex PSF to measure the emission anisotropy of fluorescent beads, the wobble dynamics …
Imaging The Three-Dimensional Orientation And Rotational Mobility Of Fluorescent Emitters Using The Tri-Spot Point Spread Function, Oumeng Zhang, Jin Lu, Tianben Ding, Matthew D. Lew
Imaging The Three-Dimensional Orientation And Rotational Mobility Of Fluorescent Emitters Using The Tri-Spot Point Spread Function, Oumeng Zhang, Jin Lu, Tianben Ding, Matthew D. Lew
Electrical & Systems Engineering Publications and Presentations
Fluorescence photons emitted by single molecules contain rich information regarding their rotational motions, but adapting single-molecule localization microscopy (SMLM) to measure their orientations and rotational mobilities with high precision remains a challenge. Inspired by dipole radiation patterns, we design and implement a Tri-spot point spread function (PSF) that simultaneously measures the three-dimensional orientation and the rotational mobility of dipole-like emitters across a large field of view. We show that the orientation measurements done using the Tri-spot PSF are sufficiently accurate to correct the anisotropy-based localization bias, from 30 nm to 7 nm, in SMLM. We further characterize the emission anisotropy …
Measuring 3d Molecular Orientation And Rotational Mobility Using A Tri-Spot Point Spread Function, Oumeng Zhang, Tianben Ding, Jin Lu, Hesam Mazidi, Matthew D. Lew
Measuring 3d Molecular Orientation And Rotational Mobility Using A Tri-Spot Point Spread Function, Oumeng Zhang, Tianben Ding, Jin Lu, Hesam Mazidi, Matthew D. Lew
Electrical & Systems Engineering Publications and Presentations
We present a method to measure the molecular orientation and rotational mobility of single-molecule emitters by designing and implementing a Tri-spot point spread function. It can measure all degrees of freedom related to molecular orientation and rotational mobility. Its design is optimized by maximizing the theoretical limit of its measurement precision. We evaluate the precision and accuracy of the Tri-spot PSF by measuring the orientation and effective rotational mobility of single fluorescent molecules embedded in a polymer matrix.