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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 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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Sample Processing and Benchmarking for Multibeam Optical Scanning Transmission Electron Microscopy.

B H Peter Duinkerken1, Arent J Kievits2, Anouk H G Wolters1

  • 1Department of Biomedical Sciences, University of Groningen, University Medical Centre Groningen, Antonius Deusinglaan 1, Groningen, AV 9713, The Netherlands.

Microscopy and Microanalysis : the Official Journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada
|April 2, 2025
PubMed
Summary
This summary is machine-generated.

Multibeam optical scanning transmission electron microscopy (OSTEM) offers an order of magnitude speed increase over conventional methods. This high-throughput technique provides comparable image quality for biological ultrastructure analysis, enabling larger-scale studies.

Keywords:
FAST-EMelectron microscopymultibeam STEMoptical STEM

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Area of Science:

  • Cell Biology
  • Microscopy Techniques
  • Biotechnology

Background:

  • Electron microscopy (EM) is crucial for visualizing biological ultrastructure.
  • High-throughput EM is needed for analyzing larger biological samples and volumes.
  • Multibeam optical scanning transmission EM (OSTEM) promises increased imaging throughput.

Purpose of the Study:

  • To evaluate the compatibility of multibeam OSTEM with standard sample preparation.
  • To determine the impact of machine settings on multibeam OSTEM image quality.
  • To compare the speed and quality of multibeam OSTEM with conventional EM techniques.

Main Methods:

  • Utilized multibeam OSTEM with multiple beamlets and optical electron separation.
  • Employed standard high-contrast staining protocols for biological tissue samples.
  • Investigated optimal acceleration voltage (5 kV), section thickness, and pixel dwell time.

Main Results:

  • Multibeam OSTEM achieved an order of magnitude higher throughput than conventional EM.
  • Generated high-quality images comparable to standard transmission EM modalities.
  • Demonstrated flexibility in stain types, with optimal results using embedding methods.

Conclusions:

  • Multibeam OSTEM is a viable high-throughput EM technique for ultrastructure analysis.
  • Achieves comparable image quality to traditional methods while significantly increasing speed.
  • Enables analysis of biological ultrastructure across larger scales and volumes.