• Title/Summary/Keyword: Cryo-electron tomography

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Toward High-Resolution Cryo-Electron Microscopy: Technical Review on Microcrystal-Electron Diffraction

  • Lee, Sangmin;Chung, Jeong Min;Jung, Hyun Suk
    • Applied Microscopy
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    • v.47 no.4
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    • pp.223-225
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    • 2017
  • Cryo-electron microscopy (cryo-EM) is arguably the most powerful tool used in structural biology. It is an important analytical technique that is used for gaining insight into the functional and molecular mechanisms of biomolecules involved in several physiological processes. Cryo-EM can be separated into the following three groups according to the analytical purposes and the features of the biological samples: cryo-electron tomography (cryo-ET), cryo-single-particle reconstruction, and cryo-electron crystallography. Cryo-tomography is a unique EM technique that is used to study intact biomolecular complexes within their original environments; it can provide mechanistic insights that are challenging for other EM-methods. However, the resolution of reconstructed three-dimensional (3D) models generated by cryo-ET is relatively low, while single-particle reconstruction can reproduce biomolecular structures having near-atomic resolution without the need for crystallization unless the samples are large (>200 kDa) and highly symmetrical. Cryo-electron crystallography is subdivided into the following two categories according to the types of samples: one category that deals with two-dimensional (2D) crystalline arrays and the other category that uses 3D crystals. These two categories of electron-crystallographic techniques use different diffraction data obtained from still diffraction and continuous-rotation diffraction. In this paper, we review crystal-based cryo-EM techniques and focus on the recently developed 3D electron-crystallographic technique called microcrystal-electron diffraction.

Techniques for Cryo-electron Tomography in Biological Field (생물학분야에서 Cryo-electron Tomography 활용기법)

  • Mun, Ji-Young;Lee, Kyung-Eun;Han, Sung-Sik
    • Applied Microscopy
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    • v.38 no.2
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    • pp.73-79
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    • 2008
  • In Biology, Studies Using Electron Microscopy for making Cell Structure to 3D reconstruction very fast development. Recently, by using Cryo fixation, we can see cell 3D structure without structural change, instead of using chemical fixation which can change cell structure. Before using this technology, we could understand cell structures only in 2D images. But now, through cryo-ET, 3D reconstruction of cell structure without artificial structure changes can be possible and this technology will give us many advantages in Drug delivery and Nanothechnology.

Workflow of Cryo-Electron Microscopy and Status of Domestic Infrastructure

  • Choi, Ki Ju;Shin, Jae In;Lee, Sung Hun
    • Applied Microscopy
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    • v.48 no.1
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    • pp.6-10
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    • 2018
  • Cryo-electron microscopy (cryo-EM) allows the analysis of the near-native structures of samples such as proteins, viruses, and sub-cellular organelles at the sub-nano scale. With the recent development of analytical methods, this technique has achieved remarkable results. The importance of cryo-EM gained wide recognition due to last year's award of the Nobel Prize in Chemistry. To help promote the knowledge of this technique, this paper introduces the basic workflows of cryo-EM and domestic cryo-EM service institutes.

A Glance of Electron Tomography through 4th International Congress on Electron Tomography (International Congress on Electron Tomography에 대한 간략한 이해와 현황)

  • Rhyu, Im-Joo;Park, Seung-Nam
    • Applied Microscopy
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    • v.38 no.3
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    • pp.275-278
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    • 2008
  • Electron tomography (ET) is an electron microscopic technique for obtaining a 3-D image from any electron microscopy specimen and its application in biomedical science has been increased thanks to development of electron microscopy and related technologies during the last decade. There are few researches on ET in Korea during this period. Although the importance of ET has been recognized recently by many researchers, initial approach to electron tomographic research is not easy for beginners. The 4th International Congress on Electron Tomography (4 ICET) was held on Nov $5{\sim}8$, 2006, at San Diego. The program dealt instrumentation, reconstruction algorithm, visualization/quantitative analysis and electron tomographic presentation of biological specimen and nano particles. 1 have summarized oral presentations and analyzed the posters presented on the meeting. Cryo-electron microscopic system was popular system for ET and followed conventional transmission electron microscopic system. Cultured cell line and tissue were most popular specimens analyzed and microorganisms including bacteria and virus also constituted important specimens. This analysis provides a current state of art in ET research and a guide line for practical application of ET and further research strategies.

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.