Structured illumination microscopy with noise-controlled image reconstructions

Nat Methods. 2021 Jul;18(7):821-828. doi: 10.1038/s41592-021-01167-7. Epub 2021 Jun 14.

Abstract

Super-resolution structured illumination microscopy (SIM) has become a widely used method for biological imaging. Standard reconstruction algorithms, however, are prone to generate noise-specific artifacts that limit their applicability for lower signal-to-noise data. Here we present a physically realistic noise model that explains the structured noise artifact, which we then use to motivate new complementary reconstruction approaches. True-Wiener-filtered SIM optimizes contrast given the available signal-to-noise ratio, and flat-noise SIM fully overcomes the structured noise artifact while maintaining resolving power. Both methods eliminate ad hoc user-adjustable reconstruction parameters in favor of physical parameters, enhancing objectivity. The new reconstructions point to a trade-off between contrast and a natural noise appearance. This trade-off can be partly overcome by further notch filtering but at the expense of a decrease in signal-to-noise ratio. The benefits of the proposed approaches are demonstrated on focal adhesion and tubulin samples in two and three dimensions, and on nanofabricated fluorescent test patterns.

Publication types

  • Research Support, Non-U.S. Gov't
  • Video-Audio Media

MeSH terms

  • Algorithms
  • Animals
  • Cell Line
  • Green Fluorescent Proteins / genetics
  • Humans
  • Image Processing, Computer-Assisted / methods*
  • Imaging, Three-Dimensional / methods
  • Mice
  • Microscopy / methods*
  • Signal-To-Noise Ratio
  • Zyxin / analysis
  • Zyxin / genetics

Substances

  • ZYX protein, human
  • Zyxin
  • Green Fluorescent Proteins