• Title, Summary, Keyword: electron microscopy

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Electron-Microscope Contributions to Autophagy Research and the Nobel Prize in Physiology or Medicine 2016

  • Rhyu, Im Joo
    • Applied Microscopy
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    • v.47 no.1
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    • pp.1-2
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    • 2017
  • Professor Yoshinori Ohsumi received the 2016 Nobel Prize in Physiology or Medicine for his contribution to autophagy research, which was first studied using electron microscopy. To celebrate and commemorate this historical moment, I describe the role of electron microscopy in autophagy research and suggest a role for next-generation electron microscopy in this research field.

Cryo-Transmission Electron Microscopy in Korean Society of Microscopy

  • Han, Sung Sik
    • Applied Microscopy
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    • v.47 no.4
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    • pp.215-217
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    • 2017
  • Dr. Jacques Dubochet, Dr. Joachim Frank, and Dr. Richard Henderson received the 2017 Nobel Prize for Chemistry for their efforts to develop effective ways to obtain high-resolution three-dimensional images of biomolecules using cryo-electron microscopy. Congratulations to the Nobel Prize in the field of electron microscopy, I will explain the scientific contributions of the three winners and introduce the role of cryo-electron microscopy (including cryo technology) in biology.

The Effects of Electron Beam Exposure Time on Transmission Electron Microscopy Imaging of Negatively Stained Biological Samples

  • Kim, Kyumin;Chung, Jeong Min;Lee, Sangmin;Jung, Hyun Suk
    • Applied Microscopy
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    • v.45 no.3
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    • pp.150-154
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    • 2015
  • Negative staining electron microscopy facilitates the visualization of small bio-materials such as proteins; thus, many electron microscopists have used this conventional method to visualize the morphologies and structures of biological materials. To achieve sufficient contrast of the materials, a number of imaging parameters must be considered. Here, we examined the effects of one of the fundamental imaging parameters, electron beam exposure time, on electron densities generated using transmission electron microscopy. A single site of a negatively stained biological sample was illuminated with the electron beam for different times (1, 2, or 4 seconds) and sets of micrographs were collected. Computational image processing demonstrated that longer exposure times provide better electron densities at the molecular level. This report describes technical procedures for testing parameters that allow enhanced evaluations of the densities of electron microscopy images.

Molecular Structure of Muscle Filaments Determined by Electron Microscopy

  • Craig, Roger
    • Applied Microscopy
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    • v.47 no.4
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    • pp.226-232
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    • 2017
  • Electron microscopy and X-ray diffraction have together played a key role in our understanding of the molecular structure and mechanism of contraction of muscle. This review highlights the role of electron microscopy, from early insights into thick and thin filament structure by negative staining, to studies of single myosin molecule structure, and finally to recent high-resolution structures by cryo-electron microscopy. Muscle filaments are designed for movement. Their labile structures thus present challenges to obtaining near-atomic detail, which are also discussed.

Nano-Resolution Connectomics Using Large-Volume Electron Microscopy

  • Kim, Gyu Hyun;Gim, Ja Won;Lee, Kea Joo
    • Applied Microscopy
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    • v.46 no.4
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    • pp.171-175
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    • 2016
  • A distinctive neuronal network in the brain is believed to make us unique individuals. Electron microscopy is a valuable tool for examining ultrastructural characteristics of neurons, synapses, and subcellular organelles. A recent technological breakthrough in volume electron microscopy allows large-scale circuit reconstruction of the nervous system with unprecedented detail. Serial-section electron microscopy-previously the domain of specialists-became automated with the advent of innovative systems such as the focused ion beam and serial block-face scanning electron microscopes and the automated tape-collecting ultramicrotome. Further advances in microscopic design and instrumentation are also available, which allow the reconstruction of unprecedentedly large volumes of brain tissue at high speed. The recent introduction of correlative light and electron microscopy will help to identify specific neural circuits associated with behavioral characteristics and revolutionize our understanding of how the brain works.

Three-Dimensional Automated Crystal Orientation and Phase Mapping Analysis of Epitaxially Grown Thin Film Interfaces by Using Transmission Electron Microscopy

  • Kim, Chang-Yeon;Lee, Ji-Hyun;Yoo, Seung Jo;Lee, Seok-Hoon;Kim, Jin-Gyu
    • Applied Microscopy
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    • v.45 no.3
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    • pp.183-188
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    • 2015
  • Due to the miniaturization of semiconductor devices, their crystal structure on the nanoscale must be analyzed. However, scanning electron microscope-electron backscatter diffraction (EBSD) has a limitation of resolution in nanoscale and high-resolution electron microscopy (HREM) can be used to analyze restrictive local structural information. In this study, three-dimensional (3D) automated crystal orientation and phase mapping using transmission electron microscopy (TEM) (3D TEM-EBSD) was used to identify the crystal structure relationship between an epitaxially grown CdS interfacial layer and a $Cu(In_xGa_{x-1})Se_2$ (CIGS) solar cell layer. The 3D TEM-EBSD technique clearly defined the crystal orientation and phase of the epitaxially grown layers, making it useful for establishing the growth mechanism of functional nano-materials.

Reconstruction of Neural Circuits Using Serial Block-Face Scanning Electron Microscopy

  • Kim, Gyu Hyun;Lee, Sang-Hoon;Lee, Kea Joo
    • Applied Microscopy
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    • v.46 no.2
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    • pp.100-104
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    • 2016
  • Electron microscopy is currently the only available technique with a spatial resolution sufficient to identify fine neuronal processes and synaptic structures in densely packed neuropil. For large-scale volume reconstruction of neuronal connectivity, serial block-face scanning electron microscopy allows us to acquire thousands of serial images in an automated fashion and reconstruct neural circuits faster by reducing the alignment task. Here we introduce the whole reconstruction procedure of synaptic network in the rat hippocampal CA1 area and discuss technical issues to be resolved for improving image quality and segmentation. Compared to the serial section transmission electron microscopy, serial block-face scanning electron microscopy produced much reliable three-dimensional data sets and accelerated reconstruction by reducing the need of alignment and distortion adjustment. This approach will generate invaluable information on organizational features of our connectomes as well as diverse neurological disorders caused by synaptic impairments.

Structural Analysis of Exosomes Using Different Types of Electron Microscopy

  • Choi, Hyosun;Mun, Ji Young
    • Applied Microscopy
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    • v.47 no.3
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    • pp.171-175
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    • 2017
  • Negative staining has been traditionally used for exosome imaging; however, the technique is limited to surface topology only and can cause staining artifacts. Therefore, to analyze the internal structure of exosomes, we employed a method of block preparation, thin sectioning, and electron tomography. In addition, an automatic serial sectioning technique with 15-nm thickness through focused ion beam was employed to observe the three-dimensional structure of exosomes of various sizes. Cryo-transmission electron microscopy revealed the near-to-native structure of exosomes.

HVEM Application to Electron Crystallography: Structure Refinement of $SmZn_{0.67}Sb_2$

  • Kim, Jin-Gyu;Kim, Young-Min;Kim, Ji-Soo;Kim, Youn-Joong
    • Applied Microscopy
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    • v.36 no.spc1
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    • pp.1-7
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    • 2006
  • The three-dimensional structure of an inorganic crystal, $SmZn_{0.67}Sb_2$ (space group $P4/nmm,\;a=4.30(3){\AA}\;and\;c= 10.27(1){\AA}$), was refined by electron crystallography utilizing high voltage electron microscopy (HVEM). Effects of instrumental resolution, image quality, beam damage and specimen tilting on the structure refinement have been evaluated. The instrumental resolution and image quality were the most important factors on the final results in the structure refinement, while the beam damage and specimen tilting effects could be experimentally minimized or controlled. The average phase errors $({\Phi}_{res})$ for the [001], [100] and [110] HVEM images of $SmZn_{0.67}Sb_2$ were $10.1^{\circ},\;9.6^{\circ}\;and\;6.8^{\circ}$, respectively. The atomic coordinates of $SmZn_{0.67}Sb_2$ were consistent within $0.0013{\AA}{\sim}0.0088{\AA}$, compared to the X-ray crystallography data for the same sample.