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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.
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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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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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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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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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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.
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Related Experiment Video

Updated: Jun 20, 2025

3D Depth Profile Reconstruction of Segregated Impurities Using Secondary Ion Mass Spectrometry
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Shot noise-mitigated secondary electron imaging with ion count-aided microscopy.

Akshay Agarwal1, Leila Kasaei2, Xinglin He1

  • 1Department of Electrical and Computer Engineering, Boston University.

Arxiv
|July 23, 2024
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Ion count-aided microscopy (ICAM) offers a quantitative approach to nanoscale imaging by reducing noise and improving image quality. This technique enables lower radiation doses, making it ideal for imaging delicate samples.

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Area of Science:

  • Nanoscience
  • Microscopy
  • Materials Science

Background:

  • Nanoscale imaging relies on detecting secondary electrons from charged particle beams.
  • Measurement noise limits image quality and necessitates high particle doses, harming sensitive samples.
  • Current methods improve image quality but don't address fundamental noise sources or provide quantitative scaling.

Purpose of the Study:

  • Introduce a quantitative imaging technique, ion count-aided microscopy (ICAM).
  • Mitigate noise sources and enable dose reduction in nanoscale imaging.
  • Establish a physically meaningful scale for improved image interpretability.

Main Methods:

  • Developed ICAM, a statistically principled method for estimating secondary electron yield.
  • Implemented a change in data collection to reduce source shot noise.
  • Applied ICAM to helium ion microscopy for dose reduction and performance validation.

Main Results:

  • ICAM substantially reduces source shot noise.
  • Demonstrated a 3× dose reduction in helium ion microscopy.
  • Empirical results closely matched theoretical performance predictions.

Conclusions:

  • ICAM provides a quantitative imaging solution for nanoscale applications.
  • The technique facilitates imaging of dose-sensitive and fragile samples.
  • ICAM may enhance the attractiveness of heavy particle microscopy.