Accurate estimation of the illumination pattern’s orientation and wavelength in sinusoidal structured illumination microscopy

Abstract

Structured illumination microscopy is able to improve the spatial resolution of wide-field fluorescence imaging by applying sinusoidal stripe pattern illumination to the sample. The corresponding computational image reconstruction requires precise knowledge of the pattern’s parameters, which are its phase (φ) and wave vector (<b>p</b>). Here, a computationally inexpensive method for estimation of <b>p</b> from the raw data is proposed and illustrated with simulations. The method estimates <b>p</b> through a selective discrete Fourier transform at tunable subpixel precision. This results in an accurate <b>p</b> estimation for all the illumination patterns and subsequently improves the superresolution image recovery by a factor of 10 around sharp edges as compared to an integer pixel approach. The technique as presented here is of major interest to the large variety of custom-build systems that are used. The feasibility of the presented method is proven in comparison with published data.