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Preparation of Samples for Electron Microscopy

Preparation of Samples for Electron Microscopy

To be visualized by an electron microscope, either transmission or scanning, biological samples need to be fixed (stabilized) so the electron beam does not destroy them and dried thoroughly (desiccated/dehydrated) so the vacuum does not affect them. Fixation needs to be done as quickly as possible because the sample properties will start changing as soon as it is removed from its natural environment. For example, in a tissue sample, the oxygen levels begin decreasing, causing an altered...

Overview of Electron Microscopy

Overview of Electron Microscopy

The wavelengths of visible light ultimately limit the maximum theoretical resolution of images created by light microscopes. Most light microscopes can only magnify 1000X, and a few can magnify up to 1500X. Electrons, like electromagnetic radiation, can behave like waves, but with wavelengths of 0.005 nm, they produce significantly greater resolution up to 0.05 nm as compared to 500 nm for visible light. An electron microscope (EM) can create a sharp image that is magnified up to 2,000,000X.

Scanning Electron Microscopy

Scanning Electron Microscopy

A scanning electron microscope (SEM) is used to study the surface features of a sample by using an electron beam that scans the sample surface in a two-dimensional manner. Typically, areas between ~1 centimeter to 5 micrometers in width can be imaged. SEM can be used to image bacteria, viruses, tissues as well as larger samples like insects. Conventional SEM gives a magnification ranging from 20X to 30,000X and spatial resolution of 50 to 100 nanometers.
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Transmission Electron Microscopy

Transmission Electron Microscopy

In 1931, physicist Ernst Ruska—building on the idea that magnetic fields can direct an electron beam just as lenses can direct a beam of light in an optical microscope—developed the first prototype of the electron microscope. This development led to the development of the field of electron microscopy. In the transmission electron microscope (TEM), electrons are produced by a hot tungsten element and accelerated by a potential difference in an electron gun, which gives them up to 400...

Immunogold Electron Microscopy

Immunogold Electron Microscopy

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Cryo-electron Microscopy

Cryo-electron Microscopy

Conventional electron microscopy (EM) involves dehydration, fixation, and staining of biological samples, which distorts the native state of biological molecules and results in several artifacts. Also, the high-energy electron beam damages the sample and makes it difficult to obtain high-resolution images. These issues can be addressed using cryo-EM, which uses frozen samples and gentler electron beams. The technique was developed by Jacques Dubochet, Joachim Frank, and Richard Henderson, for...

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Related Experiment Video

Updated: Feb 13, 2026

Optimizing Sample Preparation for Cryogenic Electron Microscopy

Optimizing Sample Preparation for Cryogenic Electron Microscopy

Published on: April 11, 2025

High speed/low dose analytical electron microscopy with dynamic sampling.

Karl A Hujsak¹, Eric W Roth², William Kellogg¹

¹Department of Materials Science & Engineering, Northwestern University, Evanston, Illinois, 60208-3108, United States.

Micron (Oxford, England : 1993)

|March 19, 2018

Summary

This summary is machine-generated.

A new Multi-Objective Autonomous Dynamic Sampling (MOADS) method significantly speeds up electron energy loss spectroscopy (EELS) and energy dispersive X-ray spectrometry (EDS) mapping. This technique reduces imaging time by over 10x, enabling detailed analysis of large or beam-sensitive samples.

Keywords:

Dose reduction Dynamic sampling Electron energy loss spectroscopy Energy dispersive X-Ray spectroscopy Machine learning Scanning transmission electron microscopy

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Area of Science:

Materials Science and Engineering
Analytical Chemistry
Microscopy and Imaging

Background:

Electron microscopy advancements enhance signal-to-noise ratios, enabling lower electron exposures for imaging materials and biological structures.
Analytical techniques like electron energy loss spectroscopy (EELS) and energy dispersive X-ray spectrometry (EDS) provide nanoscale elemental and bonding information.
Current spatially resolved spectrum imaging with EELS/EDS is time-consuming, requiring extensive data acquisition.

Purpose of the Study:

To introduce and demonstrate the Multi-Objective Autonomous Dynamic Sampling (MOADS) method for accelerating spectrum mapping in EELS and EDS.
To enable faster, more efficient acquisition of elemental and bonding information at high spatial resolutions.
To overcome limitations in current spectrum imaging techniques, particularly for large areas or beam-sensitive materials.

Main Methods:

Development of the Multi-Objective Autonomous Dynamic Sampling (MOADS) algorithm.
Integration of MOADS as a software add-on for commercial Scanning Transmission Electron Microscopes (STEMs) with Gatan Digital Micrograph interface.
Real-time construction of initial spectrum image guesses to predict informative measurement points.

Main Results:

MOADS accelerates spectrum mapping by over an order of magnitude compared to conventional methods.
Intelligent, on-the-fly selection of data points significantly reduces acquisition time and/or electron dose.
Demonstrated efficacy on prototypical analytical specimens and dose-sensitive materials.

Conclusions:

MOADS offers a substantial improvement in the speed and efficiency of EELS and EDS spectrum imaging.
This dynamic sampling approach facilitates the analysis of large-area maps and beam-sensitive materials previously considered infeasible.
Supervised dynamic sampling methods like MOADS are expected to expand the capabilities of analytical electron microscopy.