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Related Concept Videos

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...
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.
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...
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.
Overview of Microscopy Techniques01:22

Overview of Microscopy Techniques

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...
Electron Microscope Tomography and Single-particle Reconstruction01:07

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
Electron tomography can be performed either in TEM or STEM (scanning transmission...

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Related Experiment Video

Updated: May 18, 2026

Large-scale Scanning Transmission Electron Microscopy (Nanotomy) of Healthy and Injured Zebrafish Brain
10:09

Large-scale Scanning Transmission Electron Microscopy (Nanotomy) of Healthy and Injured Zebrafish Brain

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Environmental scanning electron microscopy in cell biology.

J E McGregor1, L T L Staniewicz, S E Guthrie Neé Kirk

  • 1The School of Biological Sciences, University of Bristol, Bristol, UK. Juliette.McGregor@bristol.ac.uk

Methods in Molecular Biology (Clifton, N.J.)
|October 3, 2012
PubMed
Summary

Environmental scanning electron microscopy (ESEM) allows imaging of hydrated samples with minimal preparation. This technique offers higher resolution than light microscopy, with careful control needed to prevent sample damage.

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Area of Science:

  • Microscopy and Imaging Technologies
  • Materials Science
  • Biological Sciences

Background:

  • Conventional scanning electron microscopy (CSEM) requires extensive sample preparation, often introducing artifacts.
  • Optical microscopy offers lower resolution compared to electron microscopy techniques.
  • Environmental scanning electron microscopy (ESEM) enables imaging of hydrated and insulating samples under an electron beam.

Purpose of the Study:

  • To introduce the fundamental principles of ESEM imaging.
  • To present imaging protocols for various biological samples using ESEM.
  • To highlight the advantages and considerations for using ESEM in scientific research.

Main Methods:

  • Utilizing Environmental Scanning Electron Microscopy (ESEM) for sample analysis.
  • Implementing minimal sample preparation techniques suitable for ESEM.
  • Adjusting chamber humidity and beam energy to optimize imaging and prevent artifacts.
  • Employing secondary electron imaging for topographic analysis of plant and mammalian cells.
  • Using transmission imaging techniques for bacterial samples.

Main Results:

  • ESEM provides higher resolution than optical microscopy but lower than CSEM.
  • Minimal sample preparation in ESEM reduces artifact introduction.
  • Successful imaging of hydrated samples, including live cells in some cases.
  • Demonstrated protocols for imaging plant epidermis, mammalian cells, and bacteria.

Conclusions:

  • ESEM is a valuable technique for imaging hydrated and insulating samples with minimal preparation.
  • Careful control of environmental conditions and beam parameters is crucial for artifact-free imaging.
  • ESEM offers a practical alternative to CSEM for certain applications, particularly in biological sciences.