Structured illumination microscopy artefacts caused by illumination scattering

Philos Trans A Math Phys Eng Sci. 2021 Jun 14;379(2199):20200153. doi: 10.1098/rsta.2020.0153. Epub 2021 Apr 26.

Abstract

Despite its wide application in live-cell super-resolution (SR) imaging, structured illumination microscopy (SIM) suffers from aberrations caused by various sources. Although artefacts generated from inaccurate reconstruction parameter estimation and noise amplification can be minimized, aberrations due to the scattering of excitation light on samples have rarely been investigated. In this paper, by simulating multiple subcellular structure with the distinct refractive index from water, we study how different thicknesses of this subcellular structure scatter incident light on its optical path of SIM excitation. Because aberrant interference light aggravates with the increase in sample thickness, the reconstruction of the 2D-SIM SR image degraded with the change of focus along the axial axis. Therefore, this work may guide the future development of algorithms to suppress SIM artefacts caused by scattering in thick samples. This article is part of the Theo Murphy meeting issue 'Super-resolution structured illumination microscopy (part 1)'.

Keywords: aberration; illumination path; optical scattering; structured illumination microscopy.

MeSH terms

  • Animals
  • Artifacts
  • Biophysical Phenomena
  • Computer Simulation
  • Humans
  • Image Processing, Computer-Assisted / methods
  • Imaging, Three-Dimensional
  • Light
  • Microscopy, Fluorescence / instrumentation
  • Microscopy, Fluorescence / methods*
  • Microscopy, Fluorescence / statistics & numerical data
  • Optical Devices
  • Optical Phenomena
  • Phantoms, Imaging
  • Scattering, Radiation