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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...

Electron Microscope Tomography and Single-particle Reconstruction

Electron Microscope Tomography and Single-particle Reconstruction

Transmission electron microscopy (TEM) can be used to determine the 3D structure of biological samples with the help of techniques such as electron microscope tomography and single-particle reconstruction. While single-particle reconstruction can examine macromolecules and macromolecular complexes in vitro conditions only, tomography permits the study of cell components or small cells in vivo.
Electron Tomography
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Atomic Force Microscopy

Atomic Force Microscopy

Atomic force microscopy (AFM) is a type of scanning probe microscopy that can analyze topographic details of various specimens like ceramics, glass, polymers, and biological samples. AFM offers over 1000 times more resolution than the optical imaging system. Images generated from AFM are three-dimensional surface profiles, offering an advantage over the flat, two-dimensional images from other imaging techniques.
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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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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.

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Related Experiment Video

Updated: May 22, 2026

User-friendly, High-throughput, and Fully Automated Data Acquisition Software for Single-particle Cryo-electron Microscopy

User-friendly, High-throughput, and Fully Automated Data Acquisition Software for Single-particle Cryo-electron Microscopy

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Paraxial charge compensator for electron cryomicroscopy.

John A Berriman¹, Peter B Rosenthal

¹Division of Physical Biochemistry, MRC National Institute for Medical Research, The Ridgeway, Mill Hill, London, NW7 1AA, United Kingdom.

Ultramicroscopy

|May 9, 2012

Summary

A new multi-hole aperture for transmission electron microscopy (TEM) allows simultaneous imaging and irradiation of separate specimen areas. This technique aids in mitigating charging effects for radiation-sensitive biological samples in cryomicroscopy.

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Last Updated: May 22, 2026

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Expression and Purification of the Human Lipid-sensitive Cation Channel TRPC3 for Structural Determination by Single-particle Cryo-electron Microscopy

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Single Particle Cryo-Electron Microscopy: From Sample to Structure

Single Particle Cryo-Electron Microscopy: From Sample to Structure

Published on: May 29, 2021

Area of Science:

Electron microscopy
Materials science
Biophysics

Background:

Charging effects in transmission electron microscopy (TEM) can degrade image quality and damage radiation-sensitive specimens.
Simultaneous irradiation and imaging of spatially separated regions is challenging with conventional TEM apertures.
Understanding charge mitigation mechanisms is crucial for high-resolution cryomicroscopy of biological samples.

Purpose of the Study:

To introduce a novel multi-hole condenser aperture for TEM.
To investigate its utility in simultaneously imaging and irradiating distinct specimen areas.
To explore charge compensation and neutralization strategies for biological specimens.

Main Methods:

Development and implementation of a multi-hole condenser aperture in a TEM.
Utilizing off-axis electron beams for simultaneous irradiation of specimen and support.
Employing paraxial charge compensation (PCC) to control specimen charging.
Observing charging effects on images of vitreous ice and frozen-hydrated specimens.

Main Results:

The multi-hole aperture enables simultaneous imaging and irradiation of spatially separated regions.
Irradiation of adjacent carbon support partially compensates for charging effects on the ice.
A through-space charge neutralization mechanism was observed for frozen-hydrated specimens.
PCC effectively controlled charge build-up, allowing for the study of charging effects.

Conclusions:

The multi-hole aperture is a valuable tool for investigating charging mechanisms in TEM.
This method offers a new approach for charge mitigation in cryomicroscopy of biological specimens.
The findings contribute to improving imaging conditions for radiation-sensitive materials.