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Related Concept Videos

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

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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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Transmission Electron Microscopy01:15

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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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Immunogold Electron Microscopy01:20

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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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Cryo-electron Microscopy01:28

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Conventional electron microscopy (EM) involves dehydration, fixation, and staining of biological samples, which distorts the native state of biological molecules and results in several artifacts. Also, the high-energy electron beam damages the sample and makes it difficult to obtain high-resolution images. These issues can be addressed using cryo-EM, which uses frozen samples and gentler electron beams. The technique was developed by Jacques Dubochet, Joachim Frank, and Richard Henderson, for...
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Preparation of Samples for Electron Microscopy01:20

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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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Author Spotlight: Retinal Neuroscience Studies with Volume Electron Microscopy
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Progress and remaining challenges in high-throughput volume electron microscopy.

Jörgen Kornfeld1, Winfried Denk1

  • 1Max Planck Institute of Neurobiology, Am Klopferspitz 18, 82152 Planegg, Germany.

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Summary
This summary is machine-generated.

Automatic extraction of neural circuits from electron microscopy data is advancing rapidly. This progress brings the goal of reconstructing entire mammal brains closer within the next decade.

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Area of Science:

  • Neuroscience
  • Computational Biology
  • Microscopy

Background:

  • Reconstructing neural circuits is crucial for understanding brain function.
  • Previous methods for neural circuit extraction were limited in scale and accuracy.

Purpose of the Study:

  • To assess recent advancements in automatic neural circuit extraction from volume electron microscopy data.
  • To evaluate the feasibility of reconstructing entire mammalian brains in the near future.

Main Methods:

  • Utilizing advanced convolutional neural networks for data analysis.
  • Employing improved volume electron microscopy (VEM) datasets with enhanced quality and spatial extent.
  • Implementing novel sample preparation techniques like hot-knife partitioning and ion milling.
  • Leveraging multi-beam scanning electron microscopy to accelerate data acquisition.

Main Results:

  • Significant progress in the effectiveness of automatic neural circuit extraction.
  • Substantial improvements in the quality and spatial coverage of VEM datasets.
  • Conceptual advances in sample preparation and data acquisition methodologies.

Conclusions:

  • Reconstruction of entire mammalian brains is an achievable goal within the next decade.
  • Continued advancements in data analysis and imaging technologies are key drivers.
  • Methodological innovations are crucial for overcoming current bottlenecks in neuroscience research.