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Updated: Jun 20, 2026
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Three-dimensional Characterization of Interorganelle Contact Sites in Hepatocytes using Serial Section Electron Microscopy
Published on: June 9, 2022
Three-dimensional imaging by optical sectioning in the aberration-corrected scanning transmission electron
G Behan1, E C Cosgriff, Angus I Kirkland
1Department of Materials, University of Oxford, Oxford OX1 3PH, UK.
Summary
Depth resolution in scanning transmission electron microscopy optical sectioning is limited by object size due to missing transfer function information. Three-dimensional imaging is possible for specific samples like nanoparticles with prior information.
Area of Science:
- Microscopy
- Optical Physics
- Materials Science
Background:
- Optical sectioning in scanning transmission electron microscopy (STEM) is crucial for 3D imaging.
- Depth resolution is a key parameter affecting the quality of optical sectioning.
- Lateral extent of objects can influence imaging fidelity.
Purpose of the Study:
- To measure and analyze the depth resolution in STEM optical sectioning for extended objects.
- To investigate the impact of object lateral extent on depth resolution.
- To explore methods for improving 3D information retrieval in challenging imaging scenarios.
Main Methods:
- Experimental optical sectioning of laterally extended objects using STEM.
- Analysis of depth resolution dependence on numerical aperture and object lateral extent.
- Evaluation of deconvolution methods and the utility of prior information.
Main Results:
- Depth resolution is confirmed to depend on objective lens numerical aperture.
- A novel finding is the dependence of depth resolution on object lateral extent, attributed to a missing cone of information.
- Deconvolution methods show limited utility, but 3D information is achievable for specific samples (e.g., supported nanoparticles) using prior information.
Conclusions:
- Object lateral extent significantly impacts STEM optical sectioning depth resolution.
- Advanced imaging strategies, including confocal geometry and prior information, are necessary for robust 3D reconstruction.
- Further research is needed on dynamical diffraction effects in crystals for improved optical sectioning.
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