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Three-dimensional Characterization of Interorganelle Contact Sites in Hepatocytes using Serial Section Electron Microscopy

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Electron tomography combining ESEM and STEM: a new 3D imaging technique.

P Jornsanoh¹, G Thollet, J Ferreira

¹Université de Lyon, INSA-Lyon, MATEIS, UMR CNRS 5510, F-69621 Villeurbanne Cedex, France.

Ultramicroscopy

|August 26, 2011

Summary

This study introduces a new electron tomography method combining Scanning Transmission Electron Microscopy (STEM) and Environmental Scanning Electron Microscopy (ESEM) for 3D material structure analysis. This technique offers a balance of resolution and large tomogram size, ideal for non-conductive samples.

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Three-dimensional Characterization of Interorganelle Contact Sites in Hepatocytes using Serial Section Electron Microscopy

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Preparation and Observation of Thick Biological Samples by Scanning Transmission Electron Tomography

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Published on: March 12, 2017

Area of Science:

Materials Science
Electron Microscopy
Nanotechnology

Background:

Characterizing the 3D structure of materials is crucial for understanding their properties.
Existing electron tomography techniques often face limitations in resolution, sample type, or tomogram size.
Environmental Scanning Electron Microscopy (ESEM) offers unique advantages for analyzing samples under various conditions.

Purpose of the Study:

To develop and apply a novel electron tomography technique by integrating Scanning Transmission Electron Microscopy (STEM) within an Environmental Scanning Electron Microscopy (ESEM) framework.
To enable the 3D structural characterization of materials with improved resolution and larger tomogram sizes.
To provide a method suitable for non-conductive samples and materials with low atomic numbers.

Main Methods:

Development of a dedicated stage for electron tomography within an ESEM chamber.
Utilizing a STEM configuration for high-resolution imaging.
Employing a wide range of tilt angles, unhindered by chamber space limitations.
Application to materials science samples to demonstrate performance.

Main Results:

Achieved a resolution of a few tens of nanometers.
Generated large tomogram sizes suitable for thick samples.
Demonstrated good contrast for low atomic number materials.
Successfully characterized the 3D structure of various materials science samples.

Conclusions:

The developed STEM-in-ESEM electron tomography technique offers a valuable approach for 3D material characterization.
The method effectively balances resolution and tomogram size, particularly for non-conductive and low-atomic-number materials.
The optimized stage design and wide tilt range enhance the applicability and performance of ESEM-based tomography.