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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.
Preparation of Samples for Electron Microscopy01:20

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...
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.
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...
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...

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Related Experiment Video

Updated: Jun 13, 2026

Scanning Electron Microscopy (SEM) Protocols for Problematic Plant, Oomycete, and Fungal Samples
10:57

Scanning Electron Microscopy (SEM) Protocols for Problematic Plant, Oomycete, and Fungal Samples

Published on: February 3, 2017

Environmental scanning electron microscopy (ESEM)--a versatile tool in studying plants.

Edith Stabentheiner1, Armin Zankel, Peter Pölt

  • 1Institute of Plant Sciences, University of Graz, Schubertstrasse 51, 8010, Graz, Austria. edith.stabentheiner@uni-graz.at

Protoplasma
|May 7, 2010
PubMed
Summary

Environmental scanning electron microscopy (ESEM) allows studying hydrated plant samples and dynamic processes without coating. This versatile tool offers two modes for diverse applications in plant science research.

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Area of Science:

  • Plant Science
  • Microscopy
  • Materials Science

Background:

  • Environmental scanning electron microscopy (ESEM) facilitates the study of hydrated and uncoated biological samples.
  • Water vapor in the ESEM chamber plays a crucial role in secondary electron detection and charge neutralization.
  • ESEM offers two distinct modes, expanding its applicability in scientific research.

Purpose of the Study:

  • To provide a guide for plant scientists on utilizing ESEM for diverse applications.
  • To showcase the capabilities of ESEM in observing both static and dynamic biological processes.
  • To present characteristic plant samples and their suitability for ESEM analysis.

Main Methods:

  • Utilized two ESEM modes: environmental/wet mode (sample cooling to 5°C, 4-6 Torr vapor pressure) to prevent dehydration, and low vacuum mode (≤1 Torr) for material contrast imaging with backscattered electron detectors.
  • Investigated various plant samples including leaf surfaces, trichomes, epicuticular waxes, inorganic layers, callus cells, and stigmatic tissue.
  • Demonstrated in situ observation of dynamic processes such as anther opening, leaf tensile testing, and hydration/dehydration experiments by manipulating vapor pressure.

Main Results:

  • ESEM successfully imaged hydrated and uncoated plant samples, including dehydration-sensitive tissues like callus cells and stigmatic tissue.
  • Dynamic processes, such as anther dehiscence and mechanical stress responses in leaves, were observed in situ.
  • Automated block-face imaging and serial sectioning via in situ ultramicrotomy were successfully applied.
  • ESEM demonstrated its versatility in plant science, highlighting both its strengths and limitations.

Conclusions:

  • Environmental scanning electron microscopy is a powerful and versatile tool for plant science research.
  • ESEM enables in situ observation of dynamic processes and the analysis of hydrated, uncoated samples.
  • The presented applications and sample types serve as a valuable resource for researchers exploring ESEM techniques.