Dipole-Spread Function Engineering for 6D Super-Resolution Microscopy

Abstract

Fluorescent molecules are versatile nanoscale emitters that enable detailed observations of biophysical processes with nanoscale resolution. Because they are well-approximated as electric dipoles, imaging systems can be designed to visualize their 3D positions and 3D orientations, so-called dipole-spread function (DSF) engineering, for 6D super-resolution single-molecule orientation-localization microscopy (SMOLM). We review fundamental image-formation theory for fluorescent di-poles, as well as how phase and polarization modulation can be used to change the image of a dipole emitter produced by a microscope, called its DSF. We describe several methods for designing these modulations for optimum performance, as well as compare recently developed techniques, including the double-helix, tetrapod, crescent, and DeepSTORM3D learned point-spread functions (PSFs), in addition to the tri-spot, vortex, pixOL, raPol, CHIDO, and MVR DSFs. We also cover common imaging system designs and techniques for implementing engineered DSFs. Finally, we discuss recent biological applications of 6D SMOLM and future challenges for pushing the capabilities and utility of the technology.

arXiv commentThis is a preprint of the following chapter: Tingting Wu and Matthew D. Lew, Dipole-Spread Function Engineering for 6D Super-Resolution Microscopy, published in Coded optical imaging, edited by Jinyang Lang, 2023, Springer Nature reproduced with permission of Springer Nature

References in corpus (13)

• Dielectric Metasurfaces for Complete Control of Phase and Polarization with Subwavelength Spatial Resolution and High Transmission
• Deep-STORM: super-resolution single-molecule microscopy by deep learning
• DeepSTORM3D: dense three dimensional localization microscopy and point spread function design by deep learning
• Birefringent Fourier filtering for single molecule Coordinate and Height super-resolution Imaging with Dithering and Orientation
• Fundamental precision bounds for three-dimensional optical localization microscopy with Poisson statistics
• Spatio-angular fluorescence microscopy I. Basic theory
• Single-molecule orientation localization microscopy II: a performance comparison
• Single-molecule orientation localization microscopy I: fundamental limits
• Quantum limits for precisely estimating the orientation and wobble of dipole emitters
• Spatio-angular fluorescence microscopy II. Paraxial 4$f$ imaging
• Dense Super-Resolution Imaging of Molecular Orientation via Joint Sparse Basis Deconvolution and Spatial Pooling
• Using fluorescent beads to emulate single flurophores
• Towards optimal point spread function design for resolving closely spaced emitters in three dimensions