Full three-dimensional imaging deep through multicellular thick samples with subcellular resolution by structured illumination microscopy and adaptive optics

Abstract

Structured Illumination Microscopy enables live imaging with resolutions of ~120 nm. 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 140 nm lateral and 585 nm axial resolution in tissue culture cells, C. elegans, and rice blast fungus. We demonstrate that AO improves resolution and reduces artifacts, making full 3D SIM possible in thicker samples.

To be more precise: the role of intracellular trafficking in development and pattern formation

“…The bottom plot represents the axial resolution of the techniques (μm). Single objective light sheet (SOLS) [110], 3D structured illumination microscopy (3D SIM) [111,112], spinning disk (SD) [113], widefield (WF) [114,115], lattice light sheet microscopy (LLSM) [116,117], dual inverted selective plane illumination microscopy (diSPIM) [118], axially swept light sheet microscopy (ASLM) [119], swept confocally-aligned planar excitation (SCAPE) [120], IsoView [121], 2-photon/ 3-photon Bessel light sheet [122], raster adaptive optics polyscope (RAO-polyscope) [123], 2-photon random access mesoscope (2P RAM) [124], mesolens-widefield [125], 2-photon planar Airy [126].…”

Structured illumination microscopy artefacts caused by illumination scattering

“…By combining AO with SIM imaging, Lin et al . presented an improved resolution (140 nm laterally and 585 nm axially) and reduced artefact imaging in 3D SR imaging [19]. Liu et al .…”

“…They demonstrated the feasibility of IsoSense for aberration correction in a deformable-mirror-based SIM [18]. By combining AO with SIM imaging, Lin et al presented an improved resolution (140 nm laterally and 585 nm axially) and reduced artefact imaging in 3D SR imaging [19]. Liu et al simulated the effects of the biased thickness of coverslip, the tilted coverslip, the mismatched refractive index (RI), and the misalignment of incident beams concerning the back focal plane of the objective [20].…”

Structured illumination microscopy artifacts caused by illumination scattering

“…They demonstrated the feasibility of IsoSense for aberration correction in a deformable-mirror-based SIM [18]. By combining AO with SIM imaging, Lin et al presented an improved resolution (140 nm laterally and 585 nm axially) and reduced artifact imaging in 3D SR imaging [19].…”

“…They demonstrated the feasibility of IsoSense for aberration correction in a deformable-mirror-based SIM [18]. By combining AO with SIM imaging, Lin et al presented an improved resolution (140 nm laterally and 585 nm axially) and reduced artifact imaging in 3D SR imaging [19]. Liu et al simulated the effects of the biased thickness of coverslip, the tilted coverslip, the mismatched refractive index (RI), and the misalignment of incident beams concerning the back focal plane of the objective[20].…”