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Related Concept Videos

Preparation of Samples for Electron Microscopy01:20

Preparation of Samples for Electron Microscopy

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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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Scanning Electron Microscopy01:07

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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...
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Overview of Electron Microscopy01:25

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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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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.
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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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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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Processing Embryo, Eggshell, and Fungal Culture for Scanning Electron Microscopy
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Scanning Electron Microscopy Sample Preparation and Imaging.

Jenny Ngoc Tran Nguyen1, Amanda M Harbison2

  • 1Northern Virginia Community College, Manassas Campus, 6901 Sudley Road, Manassas, VA, 20109, USA. jennyngoctrannguyen@gmail.com.

Methods in Molecular Biology (Clifton, N.J.)
|May 15, 2017
PubMed
Summary
This summary is machine-generated.

Scanning electron microscopy (SEM) provides high-detail topographical and compositional images using electron beams. This technique enables 3D imaging of various specimens, including wet tissues, and aids molecular profiling.

Keywords:
ConductiveCoolstageDesiccatedElectronsImagingMagnificationNonconductiveScanning electron microscopy (SEM)Sputter coater

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Area of Science:

  • Materials Science
  • Biology
  • Microscopy

Background:

  • Scanning electron microscopes (SEM) offer high magnifications (20-130,000×) for detailed compositional and topographical imaging.
  • SEM utilizes a focused electron beam to generate images from scattered electrons, detected by charged detectors.
  • Variable pressure chambers allow for 3D imaging of both dry specimens and hydrated cells/tissues.

Purpose of the Study:

  • To detail sample preparation and imaging techniques for scanning electron microscopy.
  • To demonstrate SEM's utility in ultrastructural and intracellular analysis.
  • To supplement light microscopy for molecular profiling across diverse organisms.

Main Methods:

  • Sample preparation for desiccated, conductive specimens.
  • Coating desiccated, nonconductive samples with an electron conductive film (e.g., gold sputter coater).
  • Utilizing variable pressure chambers for imaging hydrated samples.

Main Results:

  • High-resolution, 3D topographical and compositional imaging is achievable with SEM.
  • Successful preparation and imaging protocols for various sample types were demonstrated.
  • SEM provides valuable ultrastructural data complementing other microscopy techniques.

Conclusions:

  • Scanning electron microscopy is a versatile tool for detailed biological and material analysis.
  • Proper sample preparation is crucial for obtaining high-quality SEM images.
  • SEM significantly enhances molecular profiling capabilities when combined with light microscopy.