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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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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.
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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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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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A Simple Approach for Thickness Measurements Using Electron Probe Microanalysis.

Mouad Essani1,2, Victor Krawiec1, Emmanuelle Brackx1

  • 1CEA, DEN, DMRC, Université de Montpellier, F-30207Marcoule, France.

Microscopy and Microanalysis : the Official Journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada
|February 15, 2021
PubMed
Summary

This study introduces a fast electron probe microanalysis (EPMA) method for measuring thin sample thickness without needing instrumental parameters. The technique uses transmitted electron energy, determined by substrate X-ray signals, to accurately assess thickness.

Keywords:
X-ray emissionelectron microscopyelectron probe microanalysismicroparticlesthickness measurementthin samples

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Area of Science:

  • Materials Science
  • Analytical Chemistry
  • Physics

Background:

  • Accurate thickness measurement is crucial for analyzing thin samples, especially those with complex geometries.
  • Traditional methods for thickness determination in electron probe microanalysis (EPMA) can be complex and require knowledge of instrumental parameters.
  • Non-bulk samples and particles often present challenges for precise thickness quantification.

Purpose of the Study:

  • To develop a simple, fast, and parameter-independent method for thickness measurements using EPMA.
  • To enable accurate thickness determination for samples thinner than the electron interaction volume.
  • To provide a versatile technique applicable to various sample shapes, including irregular morphologies.

Main Methods:

  • The method relies on measuring the distance electrons travel within the sample before transmission.
  • Samples are placed on a substrate that generates an X-ray signal upon interaction with transmitted electrons.
  • The characteristic X-ray intensity of the substrate is used to determine the energy of transmitted electrons, correlating to sample thickness.

Main Results:

  • The developed EPMA method accurately determined the thickness of spherical K411 glass and cylindrical U–Ce oxide particles (0.2–4 μm).
  • Measured thicknesses showed excellent agreement with actual values for the studied particles.
  • The technique demonstrated potential for determining local thickness in thin samples with irregular shapes.

Conclusions:

  • The novel EPMA method offers a straightforward and efficient approach for thickness measurements of thin samples.
  • It overcomes limitations of traditional methods by not requiring instrumental parameter knowledge.
  • The technique is valuable for analyzing non-bulk samples with complex geometries and irregular morphologies, enhancing EPMA applications.