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Related Concept Videos

Overview of Electron Microscopy01:25

Overview of Electron Microscopy

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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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Super-resolution Fluorescence Microscopy01:37

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Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been...
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Overview of Microscopy Techniques01:22

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The early pioneers of microscopy opened a window into the invisible world of microorganisms. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes that leveraged nonvisible light, such as fluorescence microscopy that uses an ultraviolet light source and electron microscopy that uses short-wavelength electron beams. These advances significantly improved magnification, image resolution, and contrast. By comparison, the...
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Electron Microscope Tomography and Single-particle Reconstruction01:07

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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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Transmission Electron Microscopy01:15

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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...
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Preparation of Samples for Electron Microscopy01:20

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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...
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Related Experiment Video

Updated: Apr 11, 2026

Ultrastructural Localization of Endogenous LC3 by On-Section Correlative Light-Electron Microscopy
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Correlated light and electron microscopy: ultrastructure lights up!

Pascal de Boer1, Jacob P Hoogenboom2, Ben N G Giepmans1

  • 1Department of Cell Biology, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands.

Nature Methods
|May 29, 2015
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Summary

Correlated light and electron microscopy (CLEM) combines techniques to reveal cellular context and ultrastructure. This approach enhances the study of molecular dynamics and rare biological events.

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Area of Science:

  • Cell Biology
  • Microscopy Techniques

Background:

  • Microscopy has been crucial for observing living systems since the 17th century.
  • Fluorescence microscopy and electron microscopy (EM) offer distinct views of cellular components and ultrastructure.
  • Correlated light and electron microscopy (CLEM) integrates these techniques for comprehensive analysis.

Purpose of the Study:

  • To explore the advancements and applications of CLEM in biological research.
  • To highlight how CLEM overcomes limitations of individual microscopy methods.
  • To discuss the potential for broad implementation of CLEM in modern biology.

Main Methods:

  • Utilizing fluorescence microscopy with dyes and probes for molecular localization.
  • Employing electron microscopy (EM) for detailed cellular ultrastructure.
  • Integrating light and electron microscopy through CLEM protocols.

Main Results:

  • CLEM enables the study of protein dynamics in cellular context using fluorescent proteins (FPs).
  • Rare cellular events can be identified using light microscopy before detailed EM examination.
  • Advancements in probes, integrated microscopes, 3D EM, and super-resolution microscopy are improving CLEM.

Conclusions:

  • CLEM provides a powerful approach to bridge molecular localization and ultrastructural details.
  • Ongoing technological developments are making CLEM more accessible and versatile for biological studies.
  • CLEM is poised for widespread adoption, significantly advancing biological research.