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Related Concept Videos

Overview of Microscopy Techniques01:22

Overview of Microscopy Techniques

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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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Overview of Electron Microscopy01:25

Overview of Electron Microscopy

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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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Scanning Electron Microscopy01:07

Scanning Electron Microscopy

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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.
Fundamental Principles
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Transmission Electron Microscopy01:15

Transmission Electron Microscopy

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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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Preparation of Samples for Electron Microscopy01:20

Preparation of Samples for Electron Microscopy

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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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Electron Microscope Tomography and Single-particle Reconstruction01:07

Electron Microscope Tomography and Single-particle Reconstruction

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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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Co-localizing Kelvin Probe Force Microscopy with Other Microscopies and Spectroscopies: Selected Applications in Corrosion Characterization of Alloys
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High-performance probes for light and electron microscopy.

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Researchers developed novel

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Area of Science:

  • Neuroscience
  • Molecular Biology
  • Biochemistry

Background:

  • Traditional epitope tags have limitations in resolving weakly expressed proteins.
  • Need for advanced imaging tools to visualize cellular structures and protein localization with higher precision.

Purpose of the Study:

  • To engineer a new class of fluorescent protein-based probes for enhanced molecular visualization.
  • To develop highly antigenic molecules for robust immunolabeling in diverse biological systems.

Main Methods:

  • Engineering of 'spaghetti monster' fluorescent proteins (smFPs) with multiple peptide epitopes.
  • Utilizing IgG antibody binding for immunolabeling.
  • Testing smFP probes in cultured neurons, mouse brains, and fly brains.
  • Application in advanced imaging techniques like super-resolution fluorescence imaging and electron microscopy.

Main Results:

  • smFPs demonstrated excellent distribution in neurons, including dendrites, spines, and axons.
  • Enabled sensitive localization of weakly expressed proteins.
  • Achieved robust, orthogonal multicolor visualization of proteins and cell populations.
  • Improved single-molecule tracking and RNA sequencing yield in living cells.

Conclusions:

  • smFP probes offer a powerful new tool for neuroscience research.
  • They significantly expand the capacity for simultaneous imaging channels.
  • Facilitate advanced experiments in connectomics, transcriptomics, and protein localization.