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X-ray Imaging01:24

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German physicist Wilhelm Röntgen (1845–1923) was experimenting with electrical current when he discovered that a mysterious and invisible "ray" would pass through his flesh but leave an outline of his bones on a screen coated with a metal compound. In 1895, Röntgen made the first durable record of the internal parts of a living human: an "X-ray" image (as it came to be called) of his wife’s hand. Scientists worldwide quickly began their own experiments with...
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The size of the unit cell and the arrangement of atoms in a crystal may be determined from measurements of the diffraction of X-rays by the crystal, termed X-ray crystallography.
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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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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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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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Focussed Ion Beam Milling and Scanning Electron Microscopy of Brain Tissue
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Imaging of post-mortem human brain tissue using electron and X-ray microscopy.

Amanda J Lewis1, Christel Genoud2, Mélissa Pont1

  • 1Center for Cellular Imaging and NanoAnalytics (C-CINA), Biozentrum, University of Basel, Switzerland.

Current Opinion in Structural Biology
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Summary

Electron microscopy of post-mortem human brain (PMHB) faces challenges but offers vital insights into neurodegenerative diseases. This review explores current electron microscopy techniques and their applications for studying PMHB.

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Area of Science:

  • Neuroscience
  • Pathology
  • Microscopy

Background:

  • Post-mortem human brain (PMHB) analysis is critical for understanding neurodegenerative diseases.
  • Electron microscopy (EM) provides essential ultrastructural details of brain tissue.
  • PMHB imaging presents unique challenges due to uncontrollable variables.

Purpose of the Study:

  • To review the importance of EM imaging in studying PMHB.
  • To discuss the challenges and limitations associated with PMHB electron microscopy.
  • To cover recent advancements in EM techniques for PMHB analysis.

Main Methods:

  • Review of current literature on electron microscopy techniques applied to PMHB.
  • Discussion of established and emerging EM methods, including cryo-electron tomography (cryo-ET).
  • Exploration of correlative hybrid imaging techniques.

Main Results:

  • EM imaging of PMHB is crucial for advancing neurodegenerative disease research.
  • Significant challenges exist in obtaining high-quality EM data from PMHB.
  • Recent developments like cryo-ET and hybrid methods offer improved imaging capabilities.

Conclusions:

  • Despite challenges, EM analysis of PMHB is indispensable for understanding neurodegeneration.
  • Continued development of EM techniques is vital for overcoming PMHB imaging limitations.
  • Advanced EM methods enhance the potential for discovering disease mechanisms in PMHB.