Analysis of image reconstruction artifacts in structured illumination microscopy

Abstract

Structured Illumination Microscopy (SIM) is a super-resolution technique which enables to enhance the resolution of optical microscopes beyond the diffraction limit. The final super-resolution image quality strongly depends on the performance of SIM image reconstruction. Standard SIM methods require precise knowledge of the illumination pattern and assume the sample to be stationary during the acquisition of illumination patterned images. In the case of imaging live cells, the movements of the cell result in the occurrence of image reconstruction artifacts. To reduce this kind of artifacts the short acquisition time is needed. However, short exposure time causes low signal-to-noise ratio (SNR). Moreover, a drift of the specimen may distort the illumination pattern properties in each image. This issue together with the low SNR makes the estimation of reconstruction parameters a challenging task. Inaccurate assessment of spatial frequency, phase shift or orientation of the illumination pattern leads to incorrect separation and shift of spectral components in Fourier space. This results in unwanted image reconstruction artifacts and hampers the resolution enhancement in practice. In this paper, we analyze possible artifacts in super-resolution images reconstructed using super-resolution SIM technique (SR-SIM). An overview of typical image reconstruction artifact types is presented. Distinguishing image artifacts from newly resolved sample features is essential for future SIM applications in cell biology.