• Applied Microscopy
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• 제48권1호
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• pp.1-5
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• 2018

Cryo-transmission electron microscopy (cryo-TEM) allows us to perform structural analysis of a analyses of large protein complexes, which are difficult to analyze using X-ray crystallography or nuclear magnetic resonance. The most common examples of proteins used are ribosomes and proteasomes. In this paper, we briefly describe the advantage of cryo-TEM and the process of two-dimensional classification by considering a human proteasome as an example.

• Applied Microscopy
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• 제52권
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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.

• 진공이야기
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• 제4권4호
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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.

• 대한화장품학회지
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• 제30권1호
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• pp.53-58
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• 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.

• Applied Microscopy
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• 제42권1호
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• pp.49-52
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• 2012

For TEM study of biological samples or polymers that are contained in organic structure, it is often required that the sample is prepared by using ultramicrotome and stained with proper agents to increase the contrast of organic structure. In this study, we investigated an efficient TEM sample preparation method for ductile heterogeneity material by using ultramicrotomy. Cryo-ultramicrotomy is a suitable method that is capable of rendering sample hardness for various ductile materials. However, it has several factors to consider, such as experimental cost, working time and finding the optimal staining conditions. To satisfy these considerations, we prepared TEM sample by using ultramicrotome without cryofunction, and secured the sample hardness by applying the staining process prior to ultrathin sectioning. The cross-linked polyethylene structure in the sample was stained with the 2% $RuO_4$ solution in a sealed test tube for 24 hours at $4^{\circ}C$. After the sample staining, ultrathin sections of sample were prepared using ultramicrotome. As a result, it was revealed that the difficulties associated with staining of ultrathin sections prepared by low-temperature conditions were improved. In addition, appropriate staining depth of sample could be selected for sectioning process. The quality of TEM sample obtained by using this method was better than that of cryo-ultramicroscopy. Finally, it is expected that our method could be effectively applied in TEM sample preparation for a variety of nano-bio convergence materials.

• Applied Microscopy
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• 제38권3호
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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.

• 대한화장품학회지
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• 제46권2호
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• pp.133-145
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• 2020

본 연구는 용해도가 낮은 수용성 활성물질인 바이오틴(biotin)의 안정화 및 용해도 증가를 목적으로 나노리포좀을 활용하였다. 이번 실험을 통해 바이오틴 나노리포좀의 안정성에 pH가 큰 영향을 준다는 사실을 확인할 수 있었으며, pH 상승이 바이오틴 활성에 튼 영향을 미치지 않음을 확인하였다. 또한 제타사이저(zetasizer)로 입자크기, 제타전위(zeta potential) 및 다분산지수(polydispersity index)를 측정하여 안정성을 평가하였다. 입자크기는 평균 100 ~ 250 nm, 제타전위 -80 ~ -30 mV로 나노리포좀 제조가 가능함을 확인하였다. 바이오틴 나노리포좀 내의 바이오틴 캡슐화율(capsulation efficiency)을 측정하기 위해 dialysis membrane method (DMM)를 이용하여 평가하였으며, 이를 통해 알지닌을 첨가시킨 바이오틴 나노리포좀이 일반 바이오틴 나노리포좀보다 캡슐화율이 5 배 높은 것으로 측정되었다. 바이오틴 나노리포좀의 경피흡수율을 측정하기 위해 in vitro franz diffusion cell method를 통해 확인하였으며, cryogenic transmission electron microscopy (cryo - TEM)을 통해 바이오틴 나노리포좀이 잘 형성되었는지 확인하였다. 본 논문을 통하여 모발건강과 밀접한 관계가 있는 것으로 소개된 바이오틴을 약물전달체(drug delivery carrier)인 나노리포좀에 캡슐화시켜 기존의 낮은 용해도 및 석출되는 문제를 보완한 바이오틴 나노리포좀을 만들 수 있음을 확인하였다.

• 대한화장품학회지
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• 제42권1호
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• pp.9-13
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• 2016

마이셀을 이용한 가용화 제형은 화장품 산업에서 스킨 로션, 토너, 미스트 등 다양한 제형으로 이용되고 있다. 마이셀은 입자 자체가 매우 작기 때문에 효능 물질의 담지체 역할보다는 향을 가용화시키는 정도로 활용되고 있다. 본 연구에서는 효능 물질인 ${\beta}$-sitosterol을 담지 할 수 있는 새로운 마이셀을 개발하기 위하여 투명한 외관을 갖는 swollen micelle을 고려하였다. 특히, 효능 성분과의 용해도 계수를 고려하여 swollen micelle을 제조함으로써 난용성 효능 성분이 마이셀 내부에 안정하게 존재할 수 있는 새로운 방법을 개발하였다. 이렇게 만들어진 swollen micelle의 안정도는 동적광산란장치(dynamic light scattering, DLS)를 이용하여 확인하였고, 투명한 입자의 외관과 모양은 육안 관찰 및 cryo-TEM을 통해 확인하였다. 또한, DSC를 이용한 열분석을 통해 난용성 효능 성분인 ${\beta}$-sitosterol이 swollen micelle 내에서 안정하게 존재함을 확인하였다. 본 연구를 통해 용해도 계수를 고려한 swollen micelle은 난용성 효능 성분의 새로운 담지체로서 이용할 수 있음을 확인하였다.

• 대한화장품학회지
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• 제49권2호
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• pp.127-139
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• 2023

본 연구에서는 의약품 원료로 잘 알려진 minoxidil과 화장품 원료인 diaminopyrimidine oxide (DAO) 활성성분으로 사용하여 니오좀(niosome)의 물성평가와 더불어 인공피부에 대한 경피투과율을 비교하였다. 나노입자의 니오좀을 제조하기 위해 고압유화(high pressure homogenization) 방법을 이용하였으며 제타사이저(zetasizer)로 물성평가를 진행하였다. 활성성분을 포함한 니오좀의 입자크기는 HLB에 따라 평균 99 ~ 123 nm, 제타전위(zeta potential)는 -60 ~ -81 mV의 범위로 측정되었다. DSC (differential scanning colorimetry)를 통해 결정성 성분인 minoxidil이 니오좀 내에 무결정 상태로 균일하게 용해되어 있음을 확인하였다. 경피투과량을 확인 및 비교하기 위해 in vitro Franz diffusion cell 방법으로 측정하였으며, 니오좀 제형이 대조군인겔 제형보다 minoxidil의 경우 3.4배, DAO의 경우 11.1배 높은 투과율을 보였다. 또한 minoxidil과 DAO 니오좀의 경피투과 비교 시 유사한 경향을 보였으며, 상대적으로 DAO의 투과량이 많았다. HLB 값을 달리한 니오좀 제형을 Cryo-TEM을 이용하여 형상을 관찰하였으며, 모두 소포체가 형성되었으며 SUV (small unilamella vesicle)와 LUV (large unilamella vesicle)의 중간 형태임을 확인하였다. 본 연구를 통하여 탈모에 효과적인 약물인 minoxidil과 화장품 원료인 DAO 성분을 니오좀 제형에 캡슐화시킴으로써 효과적으로 피부에의 전달을 기대할 수 있다.

• 한국재료학회:학술대회논문집
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• 한국재료학회 2012년도 춘계학술발표대회
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• pp.55.2-55.2
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• 2012

Intensive theoretical and experimental studies have been carried out at Korean Institute of Science and Technology (KIST) on controlling the bio absorbing rate of the Mg alloys with high mechanical strength through tailoring of electrochemical potential. Key technology for retarding the corrosion of the Mg alloys is to equalize the corrosion potentials of the constituent phases in the alloys, which prevented the formation of Galvanic circuit between the constituent phases resulting in remarkable reduction of corrosion rate. By thermodynamic consideration, the possible phases of a given alloy system were identified and their work functions, which are related to their corrosion potentials, were calculated by the first principle calculation. The designed alloys, of which the constituent phases have similar work function, were fabricated by clean melting and extrusion system. The newly developed Mg alloys named as KISTUI-MG showed much lower corrosion rate as well as higher strength than previously developed Mg alloys. Biocompatibility and feasibility of the Mg alloys as orthopedic implant materials were evaluated by in vitro cell viability test, in vitro degradation test of mechanical strength during bio-corrosion, in vivo implantation and continuous observation of the implant during in vivo absorbing procedures. Moreover, the cells attached on the Mg alloys was observed using cryo-FIB (focused ion beam) system without the distortion of cell morphology and its organ through the removal of drying steps essential for the preparation of normal SEM/TEM samples. Our Mg alloys showed excellent biocompatibility satisfying the regulations required for biomedical application without evident hydrogen evolution when it implanted into the muscle, inter spine disk, as well as condyle bone of rat and well contact interface with bone tissue when it was implanted into rat condyle.