• Title/Summary/Keyword: 형광 현미경

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하이브리드 SEM 시스템

  • Kim, Yong-Ju
    • Proceedings of the Korean Vacuum Society Conference
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    • 2014.02a
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    • pp.109-110
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    • 2014
  • 주사전자현미경(Scanning Electron Microscopy: SEM)은 고체상태에서 미세조직과 형상을 관찰하는 데에 가장 다양하게 쓰이는 분석기기로서 최근에 판매되고 있는 고분해능 SEM은 수 나노미터의 분해능을 가지고 있다. 그리고 SEM의 초점심도가 크기 때문에 3차원적인 영상의 관찰이 용이해서 곡면 혹은 울퉁불퉁한 표면의 영상을 육안으로 관찰하는 것처럼 보여준다. 활용도도 매우 다양해서 금속파면, 광물과 화석, 반도체 소자와 회로망의 품질검사, 고분자 및 유기물, 생체시료 nnnnnnnnn와 유가공 제품 등 모든 산업영역에 걸쳐 있다(Fig. 1). 입사된 전자빔이 시료의 원자와 탄성, 비탄성 충돌을 할 때 2차 전자(secondary electron)외에 후방산란전자(back scattered electron), X선, 음극형광 등이 발생하게 되는 이것을 통하여 topography (시료의 표면 형상), morphology(시료의 구성입자의 형상), composition(시료의 구성원소), crystallography (시료의 원자배열상태)등의 정보를 얻을 수 있다. SEM은 2차 전자를 이용하여 시료의 표면형상을 측정하고 그 외에는 SEM을 플랫폼으로 하여 EDS (Energy Dispersive X-ray Spectroscopy), WDS (Wave Dispersive X-ray Spectroscope), EPMA (Electron Probe X-ray Micro Analyzer), FIB (Focus Ion Beam), EBIC (Electron Beam Induced Current), EBSD (Electron Backscatter Diffraction), PBMS (Particle Beam Mass Spectrometer) 등의 많은 분석장치들이 SEM에 부가적으로 장착되어 다양한 시료의 측정이 이루어진다. 이 중 결정구조, 조성분석을 쉽고 효과적으로 할 수 있게 하는 X선 분석장치인 EDS를 SEM에 일체화시킨 장비와 EDS 및 PBMS를 SEM에 장착하여 반도체 공정 중 발생하는 나노입자의 형상, 성분, 크기분포를 측정하는 PCDS(Particle Characteristic Diagnosis System)에 대해 소개하고자 한다. - EDS와 통합된 SEM 시스템 기본적으로 SEM과 EDS는 상호보완적인 기능을 통하여 매우 밀접하게 사용되고 있으나 제조사와 기술적 근간의 차이로 인해 전혀 다른 방식으로 운영되고 있다. 일반적으로 SEM과 EDS는 별개의 시스템으로 스캔회로와 이미지 프로세싱 회로가 개별적으로 구현되어 있지만 로렌츠힘에 의해 발생하는 전자빔의 왜곡을 보정을 위해 EDS 시스템은 SEM 시스템과 연동되어 운영될 수 밖에 없다. 따라서, 각각의 시스템에서는 필요하지만 전체 시스템에서 보면 중복된 기능을 가지는 전자회로들이 존재하게 되고 이로 인해 SEM과 EDS에서 보는 시료의 이미지의 차이로 인한 측정오차가 발생한다(Fig. 2). EDS와 통합된 SEM 시스템은 중복된 기능인 스캔을 담당하는 scanning generation circuit과 이미지 프로세싱을 담당하는 FPGA circuit 및 응용프로그램을 SEM의 회로와 프로그램을 사용하게 함으로 SEM과 EDS가 보는 시료의 이미지가 정확히 일치함으로 이미지 캘리브레이션이 필요없고 측정오차가 제거된 EDS 측정이 가능하다. - PCDS 공정 중 발생하는 입자는 반도체 생산 수율에 가장 큰 영향을 끼치는 원인으로 파악되고 있으며, 생산수율을 저하시키는 원인 중 70% 가량이 이와 관련된 것으로 알려져 있다. 현재 반도체 공정 중이나 반도체 공정 장비에서 발생하는 입자는 제어가 되고 있지 않은 실정이며 대부분의 반도체 공정은 저압환경에서 이루어지기에 이 때 발생하는 입자를 제어하기 위해서는 저압환경에서 측정할 수 있는 측정시스템이 필요하다. 최근 국내에서는 CVD (Chemical Vapor Deposition) 시스템 내 파이프내벽에서의 오염입자 침착은 심각한 문제점으로 인식되고 있다(Fig. 3). PCDS (Particle Characteristic Diagnosis System)는 오염입자의 형상을 측정할 수 있는 SEM, 오염입자의 성분을 측정할 수 있는 EDS, 저압환경에서 기체에 포함된 입자를 빔 형태로 집속, 가속, 포화상태에 이르게 대전시켜 오염입자의 크기분포를 측정할 수 있는 PBMS가 일체화 되어 반도체 공정 중 발생하는 나노입자 대해 실시간으로 대처와 조치가 가능하게 한다.

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Conservation Treatment of the Seated Stone Bodhisattva Discovered in Pyeongchang and Restoration of the Statue Using 3D Digital Technologies (평창 발견 석조보살좌상의 보존처리와 3차원 디지털기술을 활용한 복원)

  • Jo, Seongyeon;Kwon, Yoonmi;Choi, Bobae
    • Conservation Science in Museum
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    • v.20
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    • pp.77-92
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    • 2018
  • A stone seated bodhisattva (Sinsu5971) was discovered in Pyeongchang-gun, Gangwon-do in 1974 and was transferred to the Chuncheon National Museum upon its opening in 2002. The statue had damage to wide areas and was thus difficult to restore. This study utilized 3D scanning and 3D printing technologies to identify the overall form of the statue and the degree of damage, which allowed the restoration of lost portions that otherwise could not have been accurately restored to their original shape. Prior to the conservation treatment, the pigments used to decorate the surface were investigated using an optical microscope, and their main components were analyzed with a p-XRF (Potable X-ray Fluorescence Analyzer). The deteriorated lacquered surface was stabilized using animal glue and consolidated with stone strengthener (OH-100). The investigation found that the surface of the statue was made of zeolite that was lacquered and then gilded. As for pigments, white lead was used for the white color and red lead and cinnabar were used for red. The lost portions were redesigned by mirroring the remaining parts with 3D technologies. However, it was difficult to affix the 3D printing outputs to the statue without visible gaps since the damaged parts suffered flection. The portions of the outputs to be connected to the statue were thus modified and supplemented. It was also difficult to collect data on the properties of 3D printing materials due to the lack of previous in-depth study. These obstacles are subjects for further study.

A Study of Material and Production Technique of Scroll Painting Ring in Joseon (조선시대 족자 장황에 사용된 고리의 재료 및 제작기법 연구)

  • Jang, Yeonhee;Yun, Eunyoung;Kwon, Yoonmi;Kim, Sooyeon
    • Conservation Science in Museum
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    • v.16
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    • pp.56-81
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    • 2015
  • Ring in the scroll painting is one of the mounting elements which are fixed string and tassel to hang, these differ widely in shape and produce a variety of metals. Most traditional shapes of ring have been lost, because of remount, there are used Japanese style rings in present.Therefore, this study examines to shape, production technique and analyze component traditional style rings of 19 traditional scroll paintings in National Museum of Korea for restoration of original style. Ring has been recorded official names; Wonhwan(Ring), Gukhwadong(Chrysanthemum shaped ornaments), Baemok(Ring-shaped nail) in Uigwe. Result of an optical microscope (Leica, M205A), Wonhwan has two type of production technique; one is cutting and bending a metal rod and other is cast. Baemok is made to forging process after metal rod or plate by casting alloy. Baemok decoration is metal plate cutting shape, and then decorates it with pattern by using kicking line engraving, chasing and so on. Component analysis result from portable X-ray fluorescence found various metals, such as, brass, iron silver-cooper. Brass based on copper and zinc used rings of 17 scroll painting. Baemok of Yun Sidal portrait is used iron and plated with a tin-lead alloy. Yi Seogu portrait is silver-cooper alloy in whole ring.

Mineralogical Characteristics and Provenance of Cu-bearing Green Minerals Used as Traditional Pigments (전통 안료로 사용된 구리함유 녹색광물의 광물학적 특성과 산지추정)

  • Do, Jin-Young;Jung, Jongmee
    • Journal of the Mineralogical Society of Korea
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    • v.31 no.2
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    • pp.123-135
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    • 2018
  • This study tried to find the clue to Seokrok province by comparing Seokrok used in painting culture properties with Seokrok ore from domestic occurrence and imported Seokrok ore. To this end, chemical and mineralogical characteristics of painting cultural properties were identified with portable X-ray Florescence (p-XRF), micro X-ray diffraction (micro XRD) and SEM/EDS Analysis. To obtain Pb isotopic ratio, the Pb contained in Seokrok has been analyzed with Thermal Ionization Mass Spectrometer. Atacamite (or botallackite) and small quantity of brochantite were identified from Seokrok in Dancheong, and malachite was also identified from Buddhist painting besides those two ingredients. Without distinction of type, most Seokrok used in painting cultural properties is atacamite composed of Cu and Cl. From Pb isotope analysis, it was found that Seokrok in painting cultural properties was closer to that of north Korea, north China and Japan than south Korea as in regional division for East North Asia suggested by Mabuchi. The Pb isotopic ratio of domestic green mineral belongs to the distribution of Seokrok inside the painting cultural properties but imported malachite showed considerably difference. Considering the fact that atacamite, the main mineral of Seokrok in painting cultural properties is rarely produced from southern mine of the Korean Peninsula and the result of Pb isotope analysis.

Expression of EGFP in Bovine Embryos after Intracytoplasmic Sperm Injection using Spermatozoa Co-cultured with Exogenous DNA (소 난자에 있어서 외래유전자가 도입된 정자의 직접 주입에 의한 EGFP 의 발현)

  • Lee, . H. C.;S. J. Uhm;S. Y. Ann;H. J. Chung;Park, H. D.;Lee, H. T.;K. S. Chung
    • Korean Journal of Animal Reproduction
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    • v.24 no.4
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    • pp.439-449
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    • 2000
  • This study was to investigate the expression of transgene after co-injection of spermatozoon and EGFP gene into mature oocytes in cattle. From frozen semen, spermatozoa were treated by DTT with 0.03% Tween-20, freezing and thawing or 0.02% Triton X-100 to disrupt their plasma membranes. The sperm injected oocytes were co-cultured with bovine oviduct epithelial cells in CRlaa, and expression of EGFP in embryos were observbed under epifluorescent microscope. Two pronuclei were formed in oocytes injected with sperm treated by DTT (44.6%), DTT-Tween-20 (48.4%), DTT-freezing (44.4%) and DTT-Triton X-100 (42.9%). Cleavage and blastocyst formation rates of bovine oocytes which injected with sperm treated by DTT, DTT-Tween-20, DTT-freezing, and DTT-Triton X-100 were 49.1, 58.5, 43.9, and 48.4% and 10.2, 13.0, 6.8, and 6.5%, respectively. Although the most of embryos were showing mosaicism, embryos expressing EGFP gene were observed in all treated groups. Therefore, these results indicate that membrane disrupted sperm could interact with exogenous DNA, and that this procedure may be useful to introduce foreign gene into bovine oocytes.

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Mini-transposon을 사용한 단백질의 세포내 분포 결정

  • 최의열
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1993.04a
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    • pp.134-134
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    • 1993
  • Tn3 Transposon을 이용한 Shuttle Mutagenesis방법으로 효모의 Genome 상에 무작위적으로 $\beta$-gal 표지 유전자를 삽입하고 효모생활사의 각 세포시기 마다 특이하게 발현되는 유전자를 X-Gal plate상에서 찾아내고 이들 효모 유전자의 단백질이 세포내에 어떤 위치에 분포하는가를 간접 면역현미경법으로 추적해보았다. 먼저 효모의 genomic library를 38bp의 Tn3 Termial repeat를 가지고 있지 않은 pHSS6 Vector에 patial fill-in 방법으로 조성하였으며 최종적으로 20 Genome equivalent에 해당하는 18개 Pool의 genomic library를 만들었다. 이들 library를 조사하여 본 결과 모든 클론이 평균 3kb 크기의 insert를 가지고 있었으며 이는 99.99%의 효묘 genome을 대표하였다. 특정한 유전자의 발현을 알아보기 위해 먼저 mini-Tn3로 shuttle mutagenesis를 실시하고 vegetative growth동안 발현되는 유전자를 X-gal을 사용하여 골라내었다. 지금까지 16823개의 클론을 조사하였는데 이중 13%에 해당하는 2187개가 X-gal plate상에서 양성반응을 보여주었다. 양성반응을 보여주는 융합단백질의 세포내 분포틀 anti-$\beta$-galactosidase 항체를 사용하여 추적해보았다. 항체론 이용한 형광염색결과 약 70%의 세포가 background이상의 염색을 보여주었으며 이중 novel한 염색 pattern을 나타내는 클론도 다수 탐지되었다. 이상의 결과를 종합하면 Tn3틀 이용한 Shuttle Mutagenesis 방법으로 지금까지 전통적인 유전학적인 접근 방식으로 탐지되지 않았던 다수의 새로운 효모 유전자를 찾아낼 수 있을 것으로 사료된다.tamine제중 triprolidine이 $K_{M}$ /K$_{H}$ 비가 가장 높았고 diphenidol이 가장 낮았다. 이상의 결과로 보아 항 histamine제의 muscarinic receptor 차단작용은 이들 약물의 항 alleragy 효과에 필요한 작용이 아니며 본 실험에서 추정된 항 histamine제의 H$_1$-receptor와 muscarinic receptor에 대한 상대적 역가는 이들 약물의 선택과 평가에 중요한 지표가 될수 있을 것으로 생각된다.ing ischemic insults. The nature of the receptor is being explored by molecular genetic techniques, and we have recently cloned two of the major subunits; some of the data will be presented.LIFO, 우선 순위 방식등을 선택할 수 있도록 확장하였다. SIMPLE는 자료구조 및 프로그램이 공개되어 있으므로 프로그래머가 원하는 기능을 쉽게 추가할 수 있는 장점도 있다. 아울러 SMPLE에서 새로이 추가된 자료구조와 함수 및 설비제어 방식등을 활용하여 실제 중형급 시스템에 대한 시뮬레이션 구현과 시스템 분석의 예를 보인다._3$", chain segment, with the activation energy of carriers from the shallow trap with 0.4[eV], in he amorphous regions.의 증발산율은 우기의 기상자료를 이용하여 구한 결과 0.05 - 0.10 mm/hr 의 범위로서 이로 인한 강우손실량은 큰 의미가 없음을 알았다.재발이 나타난 3례의 환자를 제외한 9례 (75%)에서는 현재까지 재발소견을 보이지 않고 있다.

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Traveling wave reactor atomic layer epitaxy process and characterization of ZnS and Tb-doped ZnS films (Traveling Wave Reactor Atomic Layer Epitaxy를 이용한 ZnS와 ZnS : Tb박막의 성장과 박막 특성의 연구)

  • 윤선진;남기수
    • Journal of the Korean Vacuum Society
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    • v.7 no.1
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    • pp.51-58
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    • 1998
  • ZnS and TB-doped ZnS (ZnS:Tb) thin films were grown by traveling wave reactor atomic layer epitaxy (AKE) and characterized using materials and surface analysis techniques. $ZnCl_2$, $H_2$S,and tris(2,26,6-tetramethyl-3,5-heptandionato) terbium ($Tb(TMHD)_3$) were used as the precursors in the growth of ZnS:Tb films. The dependence of Cl content in ZnS films on growth temperature was investigated using Rutherford backscattering spectrometry. The Cl content decreased from approximately 9 at.% to 1 at. % as increasing the growth temperature from 400 to $500^{\circ}C$. The segregation of Cl in near surface region was also observed by depth profiling using Auger electron spectroscopy. Scanning electron microscopic studies showed that the ALE-grown ZnS and ZnS:Tb film during ALE process using $Tb(TMHD)_3$was also investigated. Approximately 1 at.% of O in ZnS:Tb(0.5 at.%) film which showed a good crystallinity of hexagonal 2H structure.

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Characterization and Isolation of Mutants Involved in Cell Cycle Progression and Regulation in Saccharomyces cerevisiae (Saccharomyces cerevisiae에서 세포주기의 진행과 조절에 관련된 변이주들의 분리 및 특성화)

  • 박정은;임선희;선우양일
    • Korean Journal of Microbiology
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    • v.37 no.1
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    • pp.28-36
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    • 2001
  • These studies were carried out to understand the mechanisms of genes which are related in cell cycle progression at G1/S phase. Mutants involved in cell cycle progression and regulation in Saccharomyces cerevisiae were isolated and characterized. To isolate new mutants, we screened the sensitivity to ciclopirox olamine (CPO) which inhibits the cell cycle traverse at or very near the G1/S phase boundary in HeLa cell and budding yeast. As results, we isolated 30 mutants and named cos(ciclopirox olamine sensitivity: cos27∼cos57) mutants. To determine the phenotype of mutants, we examined the sensitivity to methyl-methane sulfonate (MMS) and hydroxyurea (HU). Several mutants were sensitive to MMS and HU. According to these Phenotypes, cos mutants were grouped into four. Group I mutants are cos27, cos28, cos32, cos33, cos36, cos37, cos40, cos42, cos46, cos50, cos52 and cos53 which show MMS, HU sensitivities and might act at a checkpoint pathway during S phase. Group II mutants are cos43 and cos48 which show MMS sensitivities and might act at a checkpoint pathway during Gl or G2 phase. Group III mutants are cos35, cos47, cos54, cos55 and cos56 which show HU sensitivities and might act at a progress pathway during S phase. Finally, Group IV mutants are cos29, cos30, cos31, cos34, cos38, cos39, cos41, cos44, cos45, cos49, cos51 and cos57 which show only CPO sensitivities. Moreover, we examined the terminal phenotype of mutants under fluorescent microscope and then found one of S phase checkpoint related mutant(cos37). Furthermore, we constructed the heterozygote strain between mutant and wild type haploid strains to study their genetic analysis of cos mutants.

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Transformation of the Diatom Phaeodactylum tricornutum with its Endogenous (E)-4-Hydroxy-3-methylbut-2-enyl Diphosphate Reductase Gene (Phaeodactylum tricornutum의 (E)-4-Hydroxy-3-methylbut-2-enyl Diphosphate Reductase 유전자의 형질전환)

  • Shin, Bok-Kyu;Jung, Yu-Jin;Kim, Sang-Min;Pan, Cheol-Ho
    • Journal of Applied Biological Chemistry
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    • v.58 no.3
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    • pp.273-279
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    • 2015
  • Phaeodactylum tricornutum is a model diatom that its genomic information and biological tools are well established. In this study, a gene encoding (E)-4-hydroxy-3-methylbut-2-enyl diphosphate reductase (PtHDR), a terminal enzyme of the methylerythritol phosphate pathway regulating chlorophyll and carotenoid biosynthesis, was isolated from P. tricornutum. The isolated gene was cloned into pPha-T1 vector containing fcpA promoter to prepare pPha-T1-HDR plasmid. As a positive control, pPha-T1-eGFP plasmid was constructed with egfp gene. Stable nuclear transformation was carried out with these plasmids by particle bombardment method and zeocin resistant colonies of P. tricornutum were selected on f/2 agar plate. In result, transformation efficiency was evaluated according to the amount of plasmid DNA coated with gold particles. Integration of introduced plasmids was confirmed with genomic DNA of each transformant by polymerase chain reaction. The eGFP fluorescence was visible in the cytoplasm, indicating that eGFP was successively expressed in P. tricornutum system. The transcript level of exogenous Pthdr gene was evaluated with the obtained transformants. The results presented here demonstrated that introduction of Pthdr gene into P. tricornutum chromosome succeeded and expression of PtHDR was enhanced under the fcpA promoter.

Establishment and Maintenance of Embryonic Stem-like Cell Lines from In Vitro Produced Bovine Blastocysts (체외수정 유래 소 배반포로부터 유사 배아 줄기 세포의 확립 및 유지)

  • Lee, Yu-Yeon;Kim, Sun-Uk;Kim, Ji-Su;Song, Bong-Seok;Cho, Yoon-Jeong;Park, Jung-Sun;Yu, Dae-Yeul;Jin, Dong-Il;Lee, Kyung-Kwang;Koo, Deog-Bon
    • Reproductive and Developmental Biology
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    • v.31 no.3
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    • pp.215-220
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    • 2007
  • This study was conducted to examine the establishment of bovine ES-like cells having pluripotency. The hatched blastocysts derived from culture of in vitro fertilized embryos for 10 to 12 days dissociated mechanically into ICM-and trophectoderm-rich clumps using needle, and cultured onto mitotically-inactivated MEF feeder layer. The primary colonies originated from ICM cells were detached mechanically 7 days after seeding and subsequent subculture was conducted at intervals of every 5 to 7 days. Two ES -like cell lines were established and maintained over 40 passages. Self-renewal of the established lines was confirmed by examining the alkaline phosphatase activity, stem cell-specific marker profiles including SSEA isotopes, Oct-4 and STAT3. Moreover, the established cell lines could produce anchorage-independent embryoid bodies (EBs) with gradual decrease of Oct-4 transcript level in time-dependent manner.