Related Experiment Video

Updated: Jan 26, 2026

Micropatterning Transmission Electron Microscopy Grids to Direct Cell Positioning within Whole-Cell Cryo-Electron Tomography Workflows
09:53

Micropatterning Transmission Electron Microscopy Grids to Direct Cell Positioning within Whole-Cell Cryo-Electron Tomography Workflows

Published on: September 13, 2021

7.6K

Viral infection of cells in culture--approaches for electron microscopy

Paul Walther1, Li Wang, Sandra Liessem

  • 1Central Electron Microscopy Facility, Ulm University, Ulm D-89081, Germany.

Methods in Cell Biology
|September 28, 2010
PubMed

Insights

Investigating human cytomegalovirus (HCMV) infection pathways in human cells is crucial. Three electron microscopy techniques, including STEM tomography, reveal the complete viral cycle and membrane structure interactions.

Area of Science:

  • Virology
  • Microscopy
  • Cell Biology

Background:

  • Human cytomegalovirus (HCMV), a member of the Herpesviridae family, is a significant human pathogen.
  • Understanding the intricate steps of HCMV infection is essential for developing effective antiviral strategies.
  • Investigating viral entry, replication, and assembly within host cells requires advanced imaging techniques.

Purpose of the Study:

  • To present and discuss three electron microscopical approaches for studying HCMV infection.
  • To demonstrate the utility of these methods in visualizing the complete viral cycle in human macrophages and fibroblasts.
  • To highlight the application of these techniques for detailed analysis of viral membrane structures.

Main Methods:

  • Surface scanning electron microscopy (SEM) for visualizing cell surface interactions.
  • High-pressure freezing (HPF) combined with freeze-substitution (FS) for preserving cellular ultrastructure.
  • Electron tomography, specifically scanning transmission electron microscopy (STEM) tomography, for 3D imaging of viral particles and cellular structures.

Main Results:

  • The presented methods allow for comprehensive investigation and documentation of nearly all stages of the HCMV viral cycle.
  • These techniques are particularly effective for studying membrane-associated events during infection.
  • STEM tomography enables high-resolution three-dimensional imaging of viral particles (approx. 150 nm diameter) within semi-thin sections (approx. 500 nm).

Conclusions:

  • A combination of surface SEM, HPF-FS, and STEM tomography provides a powerful toolkit for studying HCMV infection.
  • These advanced electron microscopy approaches offer unprecedented insights into viral pathways and host-cell interactions.
  • The ability to perform 3D imaging is crucial for understanding the spatial organization of viral components and cellular responses.

More Related Videos

Correlative Light Electron Microscopy CLEM for Tracking and Imaging Viral Protein Associated Structures in Cryo-immobilized Cells09:18

Correlative Light Electron Microscopy CLEM for Tracking and Imaging Viral Protein Associated Structures in Cryo-immobilized Cells

Published on: September 7, 2018

81.8K
Using Click Chemistry to Measure the Effect of Viral Infection on Host-Cell RNA Synthesis09:35

Using Click Chemistry to Measure the Effect of Viral Infection on Host-Cell RNA Synthesis

Published on: August 9, 2013

12.2K

Related Concept Videos

Overview of Electron Microscopy01:25

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.
13.6K
Scanning Electron Microscopy01:07

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.
Fundamental Principles
Accelerated...
5.4K
Transmission Electron Microscopy01:15

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...
6.9K
Immunogold Electron Microscopy01:20

Immunogold Electron Microscopy

Immunoelectron microscopy utilizes immunogold labeling of endogenous proteins with specific antibodies to detect and localize these proteins in cells and tissues. The procedure provides insights into the distribution and quantification of protein under different stimulation conditions offering clues about their functions. Conjugating highly electron-dense gold particles with primary or secondary antibodies allow antigen detection on and within cells, with high resolution and specificity.
5.4K
Cryo-electron Microscopy01:28

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...
4.2K
Viral Recombination00:57

Viral Recombination

Cells are sometimes infected by more than one virus at once. When two viruses disassemble to expose their genomes for replication in the same cell, similar regions of their genomes can pair together and exchange sequences in a process called recombination. Alternatively, viruses with segmented genomes can swap segments in a process called reassortment.
25.0K