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We demonstrate single-molecule fluorescence imaging beyond the optical diffraction limit in 3 dimensions with a wide-field microscope that exhibits a double-helix point spread function (DH-PSF). The DH-PSF design features high and uniform Fisher information and has 2 dominant lobes in the image plane whose angular orientation rotates with the axial (z)(More)
An extension of Nomarski differential interference contrast microscopy enables isotropic linear phase imaging through the combination of phase shifting, two directions of shear, and Fourier space integration using a modified spiral phase transform. We apply this method to simulated and experimentally acquired images of partially absorptive test objects. A(More)
We demonstrate quantitative multicolor three-dimensional (3D) subdiffraction imaging of the structural arrangement of fluorescent protein fusions in living Caulobacter crescentus bacteria. Given single-molecule localization precisions of 20-40 nm, a flexible locally weighted image registration algorithm is critical to accurately combine the super-resolution(More)
When a single molecule is detected in a wide-field microscope, the image approximates the point spread function of the system. However, as the distribution of molecules becomes denser and their images begin to overlap, existing solutions to determine the number of molecules present and their precise three-dimensional locations can tolerate little to no(More)
Rotating point spread functions (PSFs) present invariant features that continuously rotate with defocus and are important in diverse applications such as computational imaging and atom/particle trapping. However, their transfer function efficiency is typically very low. We generate highly efficient rotating PSFs by tailoring the range of invariant rotation(More)
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(More)
We demonstrate three-dimensional tracking of fluorescent microparticles, with a computational optical system whose point spread function (PSF) has been engineered to have two twisting lobes along the optical axis, generating a three-dimensional (3D) double-helix (DH) PSF. An information theoretical comparison in photon limited systems shows that the DH-PSF(More)
We demonstrate the use of a 30-period dielectric stack structure as a highly dispersive device to spatially separate two beams with a 4-nm wavelength difference by more than their beam width. Unlike previous devices, our structure is simple to fabricate and relatively compact. We discuss possible applications of our device within wavelength-division(More)