ABSTRACT
Point spread function (PSF) engineering is used in single emitter localization to measure the emitter position in 3D and possibly other parameters such as the emission color or dipole orientation as well. Advanced PSF models such as spline fits to experimental PSFs or the vectorial PSF model can be used in the corresponding localization algorithms in order to model the intricate spot shape and deformations correctly. The complexity of the optical architecture and fit model makes PSF engineering approaches particularly sensitive to optical aberrations. Here, we present a calibration and alignment protocol for fluorescence microscopes equipped with a spatial light modulator (SLM) with the goal of establishing a wavefront error well below the diffraction limit for optimum application of complex engineered PSFs. We achieve high-precision wavefront control, to a level below 20 mλ wavefront aberration over a 30 minute time window after the calibration procedure, using a separate light path for calibrating the pixel-to-pixel variations of the SLM, and alignment of the SLM with respect to the optical axis and Fourier plane within 3 µm (x/y) and 100 µm (z) error. Aberrations are retrieved from a fit of the vectorial PSF model to a bead z-stack and compensated with a residual wavefront error comparable to the error of the SLM calibration step. This well-calibrated and corrected setup makes it possible to create complex '3D+λ' PSFs that fit very well to the vectorial PSF model. Proof-of-principle bead experiments show precisions below 10 nm in x, y, and λ, and below 20 nm in z over an axial range of 1 µm with 2000 signal photons and 12 background photons.
ABSTRACT
The effects of birefringence on the light distribution in the focal region of a high-NA optical system are investigated with use of the Debye approach to vector diffraction theory. The attention is limited to uniaxially birefringent media with symmetry axis along the optical axis of the imaging system. The radially (p) and tangentially (s) polarized fields in the exit pupil produce spots in the focal region that are defocused with respect to each other. For small birefringence values the relative defocus causes a distortion and broadening of the spot; for larger values the two spots separate completely. As a corollary to the theory it is shown that there is a tangential tornadolike flow of energy in the focal region when the polarization in the entrance pupil is elliptical.
ABSTRACT
This study covers a ten-year period of UCI/RAM total knee arthroplasty (Dow-Corning-Wright). Two hundred twenty-nine knees out of 516 done during this period were evaluated in an office setting. Additionally, 70 mail responses were obtained, and 52 patients were found to be deceased. This gives a 68% followup of all cases done during this period. Surgical technique, postoperative course, functional results, and complications are reviewed.
