Spatio-angular fluorescence microscopy II. Paraxial 4$f$ imaging

Abstract

We investigate the properties of a single-view fluorescence microscope in a 4$f$ geometry when imaging fluorescent dipoles without using the monopole or scalar approximations. We show that this imaging system has a spatio-angular band limit, and we exploit the band limit to perform efficient simulations. Notably, we show that information about the out-of-plane orientation of ensembles of in-focus fluorophores is recorded by paraxial fluorescence microscopes. Additionally, we show that the monopole approximation may cause biased estimates of fluorophore concentrations, but these biases are small when the sample contains either many randomly oriented fluorophores in each resolvable volume or unconstrained rotating fluorophores.

21 pages, 12 figures. Portions of this work previously appeared as arXiv:1812.07093, which was split during refereeing

References in corpus (4)

• Fundamental precision bounds for three-dimensional optical localization microscopy with Poisson statistics
• Measurement-based estimation of global pupil functions in 3D localization microscopy
• Fundamental Limits on Measuring the Rotational Constraint of Single Molecules using Fluorescence Microscopy
• Spatio-angular fluorescence microscopy I. Basic theory

Cited by in corpus (7)

• Birefringent Fourier filtering for single molecule Coordinate and Height super-resolution Imaging with Dithering and Orientation
• 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 III. Constrained angular diffusion, polarized excitation, and high-NA imaging
• Dipole-Spread Function Engineering for 6D Super-Resolution Microscopy