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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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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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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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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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Targeted Studies Using Serial Block Face and Focused Ion Beam Scan Electron Microscopy
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Serial Block Face-Scanning Electron Microscopy as a Burgeoning Technology.

Andrea G Marshall1, Kit Neikirk1, Dominique C Stephens1

  • 1Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN, 37232, USA.

Advanced Biology
|May 28, 2023
PubMed
Summary
This summary is machine-generated.

Serial block face scanning electron microscopy (SBF-SEM) offers advanced 3D ultrastructural imaging for large volumes. This review covers SBF-SEM

Keywords:
3D EMCLEMSBF-SEMmachine learning

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Area of Science:

  • Neuroscience
  • Microscopy
  • Biotechnology

Background:

  • Serial block face scanning electron microscopy (SBF-SEM) is a powerful technique for 3D ultrastructural imaging.
  • Developed in 2004, SBF-SEM allows for high-resolution visualization of neuronal networks across large volumes.

Purpose of the Study:

  • To provide an overview of the advantages and challenges of SBF-SEM.
  • To review current and potential applications of SBF-SEM in biochemistry and clinical settings.
  • To consider the role of AI-based segmentation in SBF-SEM workflows.

Main Methods:

  • Review of existing literature on SBF-SEM.
  • Discussion of SBF-SEM's technical capabilities and limitations.
  • Exploration of complementary technologies like AI segmentation.

Main Results:

  • SBF-SEM provides superior x- and y-axis ranges for volumetric imaging compared to other EM techniques.
  • The technique enables nanometer-resolution 3D reconstruction of complex biological structures.
  • AI-based segmentation presents a promising avenue for optimizing SBF-SEM data analysis.

Conclusions:

  • SBF-SEM is a valuable tool for large-volume ultrastructural analysis, particularly in neuroscience.
  • Future applications in biochemistry and clinical diagnostics are anticipated.
  • Integration with AI tools can enhance the efficiency and scope of SBF-SEM.