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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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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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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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Miniaturized Sample Preparation for Transmission Electron Microscopy
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Contamination mitigation strategies for scanning transmission electron microscopy.

D R G Mitchell1

  • 1Electron Microscopy Centre, Australian Institute for Innovative Materials, Innovation Campus, University of Wollongong, North Wollongong, NSW 2500, Australia.

Micron (Oxford, England : 1993)
|April 18, 2015
PubMed
Summary

Specimen cleanliness is crucial for high-magnification scanning transmission electron microscopy (STEM). Beam showering and plasma cleaning are effective methods for preparing contamination-free specimens for atomic-scale imaging.

Keywords:
Hydrocarbon contaminationScanning transmission electron microscopySpecimen cleaning

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Area of Science:

  • Materials Science
  • Analytical Chemistry
  • Physics

Background:

  • Modern aberration-corrected scanning transmission electron microscopy (STEM) achieves atomic resolution.
  • High electron fluxes in STEM can lead to significant specimen contamination.
  • Specimen cleanliness is critical for accurate high-magnification STEM imaging and microanalysis.

Purpose of the Study:

  • To quantitatively assess the effectiveness of various specimen cleaning methods for high-magnification STEM.
  • To identify optimal cleaning strategies for different specimen types and sensitivities.

Main Methods:

  • Quantitative measurement of contamination rates.
  • Application and evaluation of baking, cooling, plasma cleaning, beam showering, and UV/ozone exposure.
  • Testing methods on various specimen types.

Main Results:

  • Beam showering is rapid, convenient, and highly effective across a broad range of specimens.
  • Oxidative plasma cleaning is also effective, particularly for specimens on carbon supports.
  • Cooling is a suitable method for electron beam-sensitive materials.

Conclusions:

  • A combination of preliminary cleaning (baking or plasma) followed by beam showering yields contamination-free specimens.
  • Effective specimen cleaning is essential for reliable atomic-scale imaging and analysis in high-flux STEM.