• Vacuum Magazine
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• v.4 no.4
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• pp.18-22
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• 2017

Transmission electron microscopy (TEM) is a versatile and powerful technique that enables direct visualization of biological samples of sizes ranging from whole cell to near-atomic resolution details of a protein molecule. Thanks to numerous technical breakthroughs and monumental discoveries, 3D electron microscopy (3DEM) has become an indispensable tool in the field of structural biology. In particular, development of cryo-electron microscopy(cryo-EM) and computational image processing played pivotal role for the determination of 3D structures of complex biological systems at sub-molecular resolution. Here, basis of TEM and 3DEM will be introduced, especially focusing on technical advancements and practical applications. Also, future prospective of constantly evolving 3DEM field will be discussed, with an anticipation of great biological discoveries that were once considered impossible.

• 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.

• Molecules and Cells
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• v.43 no.3
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• pp.298-303
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• 2020

Cryo-electron microscopy (cryo-EM) is now the first choice to determine the high-resolution structures of huge protein complexes. Grids with two-dimensional arrays of holes covered with a carbon film are typically used in cryo-EM. Although semi-automatic plungers are available, notable trial-and-error is still required to obtain a suitable grid specimen. Herein, we introduce a new method to obtain thin ice specimens using real-time measurement of the liquid amounts in cryo-EM grids. The grids for cryo-EM strongly diffracted laser light, and the diffraction intensity of each spot was measurable in real-time. The measured diffraction patterns represented the states of the liquid in the holes due to the curvature of the liquid around them. Using the diffraction patterns, the optimal time point for freezing the grids for cryo-EM was obtained in real-time. This development will help researchers rapidly determine high-resolution protein structures using the limited resource of cryo-EM instrument access.

• 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.

• 한국전자현미경학회:학술대회논문집
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• 2007.05a
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• pp.29-31
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• 2007

• Applied Microscopy
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• v.52
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• pp.9.1-9.5
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• 2022

The compact smooth muscle 10S myosin II is a type of a monomer with folded tail and the heads bending back to interact with each other. This inactivated form is associated with regulatory and enzymatic activities affecting myosin processivity with actin filaments as well as ATPase activity. Phosphorylation by RLC can however, shuttle myosin from the inhibited 10S state to an activated 6S state, dictating the equilibrium. Multiple studies contributed by TEM have provided insights in the structural understanding of the 10S form. However, it is only recently that the true potential of Cryo-EM in deciphering the intramolecular interactions of 10S myosin state has been realized. This has led to an influx of new revelations on the 10S inactivation, unfolding mechanism and association in various diseases. This study reviews the gradual development in the structural interpretation of 10S species from TEM to Cryo-EM era. Furthermore, we discuss the utility of Cryo-EM in future myosin 10S studies and its contribution to human health.

• Applied Microscopy
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• v.46 no.2
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• pp.76-82
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• 2016

Aloe vera has been used in the pharmaceutical, food and cosmetic industry for its therapeutic properties. However, there are not many current studies on the microstructure of A. vera compared to studies on the chemical constituents and health efficacy of A. vera. Therefore, we compared the morphology of an A. vera leaf using an optical microscope, a conventional scanning electron microscope (SEM) and a cryo-SEM. Especially, this study focused on observing the gel in the inner leaf of A. vera, which is challenging using standard imaging techniques. We found that cryo-SEM is most suitable method for the observation of highly hydrated biomaterials such as A. vera without removing moisture in samples. In addition, we found the optimal analytical conditions of cryo-SEM. The sublimation conditions of $-100^{\circ}C$ and 10 minutes possibly enable the surface of the inner leaf of A. vera to be observed in their "near life-like" state with retaining moisture. The experiment was repeated with A. arborescens and A. saponaria to confirm the feasibility of the conditions. The results of this study can be applied towards the basic research of aloe and further extend previous knowledge about the surface structures of the various succulent plants.

• Applied Microscopy
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• v.39 no.1
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• pp.65-72
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• 2009

Cryo-SEM which enables specimens to be observed in frozen form has been used to study liquid specimens in their native states. Cryo-methods, sample preparation for cryo-SEM, are quite complex and involve several discrete but vitally interconnected steps which are rapid cooling, fracturing, sectioning, etching and coating. It is important to select practical techniques and to optimize conditions of each steps considering analytic purpose and specimen characters, viz., sample dimension, water contents. In this study, etching methods and sample preparation before freezing had been studied for observation of cyanobacteria, Synechocystis sp. PCC 6803 using cryo-SEM and their cryo-SEM images were compared to Conventional SEM (CSEM) images treated by chemical fixation. We could observe the improved morphological images of the pili of the surface and membranes of Synechocystis sp. PCC 6803 and the three-dimensional architectures of their biofilm, which were difficult to observe using chemical fixation and CSEM. These results suggest that cryo-methods/cryo-SEM are useful techniques for morphological study of biological specimen.

• 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.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.30 no.1
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• pp.53-58
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• 2004

In this study, we describe the application of electron microscopes that incorporate freeze treatments or cryo systems to achieve the characterization of the microstuctures of emulsions. We confirmed that the preparations of freezing replica method and with cryo systems were useful to clarify the microstuctures of the emulsions. This methodology will be able to contribute to understanding the relation between microstuctures and rheology of emulsions.