Three-dimensional structured illumination microscopy using Lukosz bound apodization reduces pixel negativity at no resolution cost

Abstract

Abstract:The quality of the reconstructed image in structured illumination microscopy (SIM) depends on various aspects of the image filtering process. To optimize the trade-off between resolution and ringing artifacts, which lead to negative intensities, we extend Lukosz-bound filtering to 3D SIM and derive the parametrization of the 3D SIM cut-off. We compare the use of the Lukosz-bound as apodization filter to triangular apodization and find a tenfold reduction in the most negative pixel value with a minimal resolution loss. We test this algorithm on experimental SIM images of tubulin filaments and DAPI stained DNA structure in cancer cells and find a substantial reduction in the most negative pixel value and the percentage of pixels with a negative value. This means that there is no longer a need to clip the final image to avoid these negative pixel values.

Successful optimization of reconstruction parameters in structured illumination microscopy – a practical guide

“…An additional radial frequency weighting of the deconvolved spectrum allows further suppressing of high frequency spectral components leaving low frequency components unchanged. Different shapes for the apodization function were suggested such as and exponentially weighted distance transform (22) or a Luckosz bound, which should additionally ensure a non-negative point spread function (31,32).…”