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Scanning Electron Microscopy

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Updated: May 9, 2026

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

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

Published on: March 12, 2017

Integration of a high-NA light microscope in a scanning electron microscope.

A C Zonnevylle¹, R F C Van Tol, N Liv

¹Department of Imaging Science and Technology, Faculty of Applied Sciences, Delft University of Technology, Lorentzweg, the Netherlands.

Journal of Microscopy

|July 30, 2013

Summary

We developed an integrated light-electron microscope for simultaneous high-resolution imaging. This novel correlative microscopy technique enables simultaneous optical and scanning electron microscopy on the same sample area.

Keywords:

Cathodoluminescence microscopy cellular imaging correlative microscopy fluorescence microscopy light microscopy scanning electron microscopy

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Area of Science:

Correlative microscopy
Nanotechnology
Materials Science

Background:

Simultaneous imaging with light and electron microscopy is challenging.
Existing methods often lack precise alignment and high resolution.

Purpose of the Study:

To present an integrated light-electron microscope for simultaneous high-resolution imaging.
To enable correlative light and electron microscopy on the same sample area.

Main Methods:

An inverted high-numerical aperture (NA) objective lens was integrated inside a scanning electron microscope (SEM).
The light and SEM objective lenses share a common axis and focal plane.
In situ epifluorescence microscopy with a NA of 1.4 was performed using vacuum-compatible immersion oil.

Main Results:

Simultaneous high-resolution optical and scanning electron microscopy was achieved.
Precise alignment of fields of view was demonstrated (sub-10 μm).
A 40-nm fluorescent nanoparticle showed a full width at half maximum (FWHM) of 380 nm, with uncompromised SEM performance.

Conclusions:

The integrated instrument offers new possibilities for correlative light and electron microscopy.
This technique is applicable to life sciences, physics, and chemistry.
The system provides flexibility in light microscope component mounting.