• 진공이야기
• /
• 제4권4호
• /
• pp.18-22
• /
• 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
• /
• 제38권2호
• /
• pp.73-79
• /
• 2008

현재 생물학 분야에서 세포 구조를 3차로 구현하기 위해 전자 현미경을 이용하는 연구는 매우 빠르게 발전하고 있다. 최근에는 시료의 구조를 변형시키는 화학 고정법 대신 시료를 빠른 시간 내에 동결고정시킴으로써 구조 변형 없이 세포 3차 구조 구현이 가능해졌다. 이러한 기술의 도입으로 단편이미지로만 세포 구조를 이해해왔던 지금까지와 달리 Cryo-ET를 통해 인위적인 구조 변형 없는 3차 구조 구현이 가능해졌고, 이 기법은 약물전달, 나노 공학 등 여러 학문 분야에 활용됨으로써 학문 발전에 이바지 할 것이다.

• Molecules and Cells
• /
• 제43권3호
• /
• pp.298-303
• /
• 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
• /
• 제47권3호
• /
• pp.171-175
• /
• 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
• /
• 한국현미경학회 2007년도 제38차 춘계학술대회 초록집
• /
• pp.29-31
• /
• 2007

• Applied Microscopy
• /
• 제52권
• /
• pp.9.1-9.5
• /
• 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
• /
• 제46권2호
• /
• pp.76-82
• /
• 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
• /
• 제39권1호
• /
• pp.65-72
• /
• 2009

Cryo-SEM은 수분을 함유하거나 액상 시료를 동결된 상태로 관찰하는 방법으로 rapid cooling, fracturing, sectioning, etching, coating과 같은 다소 복잡한 여러 과정의 전처리(Cryo-methods) 로 구성된다. 각 과정에서는 분석 목적이나 시료 특성(예: 시료크기, 물 함량 등)을 고려하여 최적의 조건을 설정하여야 한다. 본 연구에서는 cryo-SEM을 이용하여 남세균 (cyanobacteria, Synechocystis sp. PCC 6803)의 표면 및 내부 구조 관찰을 위한 etching time과 시료 처리에 관하여 살펴보았고 cryo-methods로 처리한 후 cryo-SEM으로 관찰한 이미지를 화학 고정한 일반 SEM(CSEM) 결과와 비교하였다. 관찰 결과, 화학 고정한 CSEM에서는 Synechocystis sp. PCC 6803 표면에 존재하는 pili가 잘 관찰되지 않았으며 파단된 내부 구조의 이미지를 얻을 수 없었으나 cryo-methods/cryo-SEM에서는 세포 표면의 pili 및 membrane의 형태를 관찰할 수 있었다. 또한 수화 상태에 따라 구조변형이 일어나는 biofilm의 구조도 관찰할 수 있었다. 이러한 결과로부터 cryo-methods/cryo-SEM은 수분을 함유하는 의생물 시료의 형태 구조를 분석하는 유용한 방법으로 사료된다.

• Applied Microscopy
• /
• 제38권3호
• /
• pp.275-278
• /
• 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.

• 대한화장품학회지
• /
• 제30권1호
• /
• pp.53-58
• /
• 2004

본 연구에서는 에멀젼의 미세구조 분석을 위한 동결처리 전자현미경 분석의 응용에 대해 소개하였다. 동결처리 전자현미경 분극법은 에멀젼의 미세구조 분석 및 해석에 유용하며, 이러한 미세구조 해석을 통해 에멀젼의 유동 특성 발현에 대한 메카니즘의 설명이 가능할 것으로 사료된다.