Subcellular three-dimensional imaging deep through multicellular thick samples by structured illumination microscopy and adaptive optics

Nat Commun. 2021 May 25;12(1):3148. doi: 10.1038/s41467-021-23449-6.

Abstract

Structured Illumination Microscopy enables live imaging with sub-diffraction resolution. Unfortunately, optical aberrations can lead to loss of resolution and artifacts in Structured Illumination Microscopy rendering the technique unusable in samples thicker than a single cell. Here we report on the combination of Adaptive Optics and Structured Illumination Microscopy enabling imaging with 150 nm lateral and 570 nm axial resolution at a depth of 80 µm through Caenorhabditis elegans. We demonstrate that Adaptive Optics improves the three-dimensional resolution, especially along the axial direction, and reduces artifacts, successfully realizing 3D-Structured Illumination Microscopy in a variety of biological samples.

Publication types

  • Evaluation Study
  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Artifacts
  • Ascomycota
  • Caenorhabditis elegans
  • Cell Line
  • Imaging, Three-Dimensional / instrumentation
  • Imaging, Three-Dimensional / methods*
  • Intravital Microscopy / instrumentation
  • Intravital Microscopy / methods*
  • Lighting / instrumentation*
  • Mice
  • Microscopy, Confocal / instrumentation
  • Microscopy, Confocal / methods
  • Microscopy, Fluorescence / instrumentation
  • Microscopy, Fluorescence / methods
  • Oryza / microbiology
  • Reproducibility of Results

Supplementary concepts

  • Pyricularia oryzae