Birefringent Fourier filtering for single molecule Coordinate and Height super-resolution Imaging with Dithering and Orientation

Abstract

Super-resolution imaging based on single molecule localization allows accessing nanometric-scale information in biological samples with high precision. However, complete measurements including molecule orientation are still challenging. Orientation is intrinsically coupled to position in microscopy imaging, and molecular wobbling during the image integration time can bias orientation measurements. Providing 3D molecular orientation and orientational fluctuations can offer new ways to assess the degree of alignment of protein structures, which cannot be monitored by pure localization. Here we demonstrate that by adding polarization control to phase control in the Fourier plane of the imaging path, all parameters can be determined unambiguously from single molecules: 3D spatial position, 3D orientation and wobbling or dithering angle. The method, applied to fluorescent labels attached to single actin filaments, provides precisions within tens of nanometers in position and few degrees in direction.

References in corpus (5)

• Fundamental Limits on Measuring the Rotational Constraint of Single Molecules using Fluorescence Microscopy
• Spatio-angular fluorescence microscopy I. Basic theory
• Spatio-angular fluorescence microscopy II. Paraxial 4$f$ imaging
• Dense Super-Resolution Imaging of Molecular Orientation via Joint Sparse Basis Deconvolution and Spatial Pooling
• Optimal birefringence distributions for star test polarimetry

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• Single-molecule orientation localization microscopy I: fundamental limits
• Quantum limits for precisely estimating the orientation and wobble of dipole emitters
• Periodic skyrmionic textures via conformal cartographic projections
• Using fluorescent beads to emulate single flurophores
• Vectorial phase retrieval in super-resolution polarization microscopy
• Imaging the dipole scattering of an optically levitated dielectric nanoparticle
• Resolving the Orientations of and Angular Separation between a Pair of Dipole Emitters
• Structured position-momentum entangled two-photon fields
• Dipole-Spread Function Engineering for 6D Super-Resolution Microscopy
• Interference of the scattered vector light fields from two optically levitated nanoparticles