• Title/Summary/Keyword: 미세조작기술

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Atomic Force Microscopy(AFM) based Single Cell Manipulation and High Efficient Gene Delivery Technology (원자간력 현미경을 이용한 단일세포 조작 및 고효율 유전자 도입기술)

  • Han, Sung-Woong;Nakamura, Chikashi;Miyake, Jun;Kim, Woo-Sik;Kim, Jong-Min;Chang, Sang-Mok
    • Korean Chemical Engineering Research
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    • v.47 no.5
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    • pp.538-545
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    • 2009
  • The principle and application of a scanning probe microscopy(SPM) are reviewed briefly, and a low-invasive single cell manipulation and a gene delivery technique using an etched atomic force microscopy(AFM) probe tip, which we call a nanoneedle, are explained in detail. The nanoneedle insertion into a cell can be judged by a sudden drop of force in a force-distance curve. The probabilities of nanoneedle insertion into cells were 80~90%, which were higher than those of typical microinjection capillaries. When the diameter of the nanoneedle was smaller than 400 nm, the nanoneedle insertion into a cell over 1 hour had almost no influence on the cell viability. A highly efficient gene delivery and a high ratio of expressed gene per delivered DNA compared the conventional major nonviral gene delivery methods could be achieved using the gene modified nanoneedle.

스마트 랩 개념과 도입 사례

  • Kim, Yeong-Su
    • Bulletin of the Korea Photovoltaic Society
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    • v.7 no.1
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    • pp.27-32
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    • 2021
  • 코로나-19 팬데믹으로 인류는 많은 어려움에 처하게 되었다. 그러나 바이러스 감염 예방을 위해 대량 시료에 대한 정확하고 신속한 분석이 필요함에 따라 실험실 자동화 기술의 상용화가 크게 앞당겨졌다. 스마트 랩이란 실험실 자동화를 위한 H/W인 실험실 자동화 시스템(LAS)와 실험 절차 및 데이터를 통합 관리하는 S/W인 실험실 데이터 관리 시스템 (LIMS)의 결합으로 이뤄진 통합 연구 시스템이라고 할 수 있다. 스마트 랩을 통해 사용자는 24시간 연속적인 장비 가동을 통한 분석 물량 확대는 물론 전처리와 같은 반복 작업 감소, 정확도 향상 등의 자동화로 인한 장점과 함께 시험 방법의 동일성 유지, 시료별 이력 추적, 조작 방지 등의 소프트웨어적인 장점을 동시에 얻을 수 있다. 스마트 랩은 지금도 발전하고 있으며 일부 대기업과 연구소에서 도입을 추진하고 있는 태동기라고 할 수 있다. 최근 우리나라가 선도하고 있는 페로브스카이트 태양전지와 같이 수분에 취약한 유무가 복합 재료의 in-line 시험이나 조성의 미세한 조정에 따라 변화하는 물성 변화를 추적 조사하는데 필요한 많은 측정과 데이터 처리 등 태양전지 분야에서도 앞으로 그 활약이 기대된다.

Magnetic Propwrties of High Quality $Nd_{12}Fe_{80}B_{6}(Nb, M)$ (M=Ti/Cu/Ga) Melt-Spun Ribbons (고특성 $Nd_{12}Fe_{80}B_{6}(Nb, M)$ (M=Ti/Cu/Ga)급속응고리본의 자기특성)

  • 김윤배;김창석;김동환;이갑호;김택기
    • Journal of the Korean Magnetics Society
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    • v.2 no.1
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    • pp.44-49
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    • 1992
  • Magnetic properties and microstructures of $Nd_{12}Fe_{80}B_{6}(Nb,\;M)$ (M=Ti/Cu/Ga) melt-spun ribbons prepared by single wheel technique have been studied. The results of microstructural study have shown that Ga is effective for the orientation of c-axis normal to the ribbon plane. The Ga-added melt-spun ribbon, $Nd_{12}Fe_{80}B_{6}(Nb,\;Ga)$, quenched at $V_{s}=17.9\;m/s$ was mostly composed of fine grains of about 30 nm in size with the textured free-side surface. The powder of this ribbon aligned in mag-netic field showd a high remanence of 0.87 T which was about 5 % higher than that of ribbon itself. It is believed that there is a possibility to fabricate a new type of HIREM melt-spun ribbon with highly textured free-side surface.

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A Study on Hydrophobic Surface Treatment for Microfluidic System Fabrication Based on SLA 3D Printing Method (SLA 3D 프린팅 방식 기반의 미세 유체 시스템 제작을 위한 소수성 표면 처리 연구)

  • Jae Uk Heo;Seo Jun Bae;Do Jin Im
    • Korean Chemical Engineering Research
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    • v.62 no.1
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    • pp.105-111
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    • 2024
  • The SLA (Stereolithography Apparatus) method is a type of 3D printing technique predicated on the transformation of liquid photocurable resin into a solid form through UV laser exposure, and its application is increasing in various fields. In this study, we conducted research to enhance the hydrophobicity and transparency of SLA 3D printing surfaces for microfluidic system production. The enhancement of surface hydrophobicity in SLA outputs was attainable through the application of hydrophobic coating methods, but the coating durability under different conditions varied depending on the type of hydrophobic coating. Additionally, to simultaneously achieve the required transparency and hydrophobic properties for the fabrication of microfluidic systems, we applied hydrophobic coatings to the proposed transparency enhancement method from prior research and compared the changes in contact angles. Teflon coating was proposed as a suitable hydrophobic coating method for the fabrication of microfluidic systems, given its excellent transparency and high coating durability in various environmental conditions, in comparison to titanium dioxide coating. Finally, we produced an Electrophoresis of Charged Droplet (ECD) chip, one of the digital microfluidics systems, using SLA 3D printing with the proposed Teflon coating method (Fluoropel 800). Droplet manipulation was successfully demonstrated with the fabricated chip, confirming the potential application of SLA 3D printing technology in the production of microfluidic systems.

Single Cell Oil-Recent Trends in Microbial Production and Utilization (미생물에 의한 단세포유지의 생산과 이용)

  • Kim, Yong-Ro;Yoon, Suk Hoo
    • Korean Journal of Food Science and Technology
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    • v.47 no.6
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    • pp.687-697
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    • 2015
  • With the shortage of edible fats and oils and depletion of fossil fuels in many countries, microbial lipids is emerging as one of the most promising sources of fats and oils in the global market. Oleaginous microorganisms, also called single cell oils (SCOs), can accumulate lipids more than 25% in the cell volume. Triacylglycerols are the major storage lipids. SCOs offer several advantages for lipid production as follows: SCOs have short life span which would shorten production time, cultivation conditions are not affected by climate and place; the production process is easy to control. There are a number of oleaginous yeasts, molds, and microalgae. Furthermore, the lipid productivity of SCOs can be enhanced through strain improvement and the optimization of cultivation conditions. The new strains developed using recent advanced biotechnical methods showed greatly improved oleaginicity. Further, hydrolysates of lignocellulosic materials can be used as carbon sources for economic production of SCO.

Studies on nuclear transplantation in mouse embryos. I. Functional differences between maternal and paternal genomes (생쥐 수정란의 핵이식에 관한 연구 I. 모성 및 부성 genome의 기능차이에 관한 연구)

  • Choe, Sang-yong;Park, Choon-saeng;Lee, Hyo-jong;Park, Hee-sung
    • Korean Journal of Veterinary Research
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    • v.30 no.2
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    • pp.123-127
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    • 1990
  • By nuclear transplantation technology twenty eight mice have been produced after transfer of heterozygous biparental eggs. Also heterozygous gynogenetic eggs with two female pronuclei and heterozygous androgenetic eggs with two male pronuclei have been obtained by injecting a male or female pronucleus with Sendai virus into the perivitelline space of enucleated haploid zygotes at pronuclear stage. The success rate of enucleation, karyoplast injection and fusion of both the pronuclei was 80.3, 83.4 and 81.8%, respectively. The overall pronuclei fusion rates by this technique were 56, 50 and 56% in biparental, gynogenetic and androgenetic eggs, respectively. The evidence was ascertained that the gynogenetic and androgenetic eggs were also able to develop in vitro up to blastocyst stage, even though their developmental potential was greatly diminished beyond 2-cell stage. The gynogenetic eggs were able to develop in vivo up to day 10 of pregnancy, while the androgenetic eggs failed to develop in vivo during the same period.

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Dielectrophoresis for Control of Particle Transport: Theory, Electrode Designs and Applications (입자 이동 제어를 위한 유전영동: 이론, 전극 구조 및 응용분야)

  • Lee, Minji;Kim, Ji-Hye;Koo, Hyung-Jun
    • Korean Chemical Engineering Research
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    • v.57 no.2
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    • pp.149-163
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    • 2019
  • Under non-uniform electric field, a directional force along the electric field gradient is applied to matter having permanent or induced dipoles. The transport of particles by the directional force is called dielectrophoresis (DEP). Since the strength and direction of the DEP force depend on parameters, such as permittivity and conductivity of particles and surrounding media, and frequency of the applied AC electric field, particle can be precisely manipulated by controlling the parameters. Moreover, unlike electrophoresis, DEP can be applied to any particles where dipole is effectively induced by electric field. Such a DEP technique has been used in various fields, ranging from microfluidic engineering to biosensor and microchip research. This paper first describes the fundamentals of DEP, and discusses representative microelectrode designs used for DEP study. Then, exemplary applications of DEP, such as separation, capture and self-assembly of particles, are introduced.

Improvement of Silkworm Egg Microinjection Using 3D Printing Technology (3D 프린팅 기술을 이용한 누에 알 미세주입 기술 개선)

  • Jeong, Chan Young;Lee, Chang Hoon;Seok, Young-Seek;Yong, Sang Yeop;Kim, Seong-Wan;Kim, Kee Young;Park, Jong Woo
    • Korean journal of applied entomology
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    • v.61 no.1
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    • pp.249-254
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    • 2022
  • Silkworms, which have for long been used as an insect resource for industrialization, have recently attracted attention as potential bio-factories for the production of novel biomaterials. In this regard, material production is typically achieved based on transformation technology, mediated via microinjection, in which a target gene is inserted into eggs containing an embryo. However, an essential step in the microinjection procedure is egg fixation, which can be a time-consuming and laborious task. Therefore, in this study, using the 3DCADian program, we adopted a 3D printing approach to model egg liners and glue drawers, which can contribute to facilitating egg alignment and fixation, thereby enhancing transformation efficiency by reducing time consumption and fatigue. After rendering using Fusion 360, the two supplementary tools were produced by printing with nylon resin (PA12) and Sinterit Lisa Pro. Subsequent analysis of the time required to fix eggs on glass slides using the two manufactured tools, revealed that the processing time was reduced by approximately 18.6% when the two tools were used compared with when these tools were not used. These innovations not only reduced fatigue but also contributed to more effective use of the microscope and manipulator for microinjection. Consequently, we believe that with additional research and refinement, the egg liner and glue drawer developed in this study could be used to enhance silkworm transformation efficiency and study similar transformation systems in other industrial insects.

The Production of Chimeric Mice by Embryonic Stem Cell Carrying Human Luteinizing Hormone Gene (사람 LH 호르몬유전자를 도입한 배아주세포에 의한 카이미라 생쥐 생산)

  • ;Tokunaga, T.;Imai, H.
    • Journal of Embryo Transfer
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    • v.9 no.3
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    • pp.243-247
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    • 1994
  • 최근 의약적으로 유용한 단백질을 대량 생산키 위한 실현 가능한 방법이 유전자변환 가축의 이용과 관련되어 발전되어 왔다. 이러한 유전자 변환동물은 이종의 단백질을 유즙속으로 분비시키는 생체반응기로서 이용되고 있다. 이러한 전략적 목적을 위해 현재 유전자 변환동물의 생산을 위한 이용에 있어 여러 가지 방법들이 보고되고 있다. 그러나 ES 세포의 사용이 이러한 방법들 사이에서 가장 실질적인 것으로 추정되고 있다. 본 실험에서는 유전자 구축을 위해 사람 황체 호르몬(human luteinizing hormone; hLH)의 전사를 유도하기 위해 각각 2.2 및 0.5 kb의 토끼 $\beta$-casein pronoter 단편을 이용하여 생쥐의 유선에 hLH를 발현시키도록 조절하고 발현이 thynidine kinase(TK) pronoter에 의해 좌우되는 neo 유전자를 selectable marker로서 plasnid속에 삽입하였다. 그 결과 생긴 구축 유전자는 각각 pCas 2.2와 pCas 0.5로 명명하였다. 구축된 유전자로 2$\times$107의 TT-2 ES세포를 170V, 550$\mu$F로 100$\mu$g의 선상 plasmid에 의해 electroporation 시켰다. 감염된 colony들은 250$\mu$g/$m\ell$ G418을 함유하는 ESM 배양액에서 선별 7일 이후에 회수하여 성공적으로 감염된 ES세포는 PCR 및 Southern blot에 의해 확인되었고 그들 중 나머지는 trypsin 처리 후 각각 미세조작과 공배양 기술을 사용하여 ICR 생쥐의 8세포기 수정란 속에 도입하였다. 결국 24시간 동안 37$^{\circ}C$, 5% $CO_2$에서 배양된 배반포를 chimera의 생산을 위해 위임신 유기된 G418 선발처리 이후 400 및 275개의 ES 세포 colony가 생존하였으며, 3개의 ES 세포으 colony 의 genome 속에 임의적으로 plamid가 삽입된 것을 Southern blot에 의해 확인되었다. 총 13 chimera 생쥐가 3 colony로부터 생산되었으나 germ-line chimera는 현재 조사중이다. chimera 생산빈도는 공배양 기술보다 주입방법에서 현저히 높았다.

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Simple Methods for Production of Chimeric Mouse by Coculture with TT2 Embryonic Stem Cells (TT2 Embryonic Stem Cell 을 이용한 Chimeric Mouse 생산에 있어서 간단한 공배양방법)

  • Cho, Y.Y.;Moon, S.J.;Kang, M.J.
    • Korean Journal of Animal Reproduction
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    • v.24 no.4
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    • pp.451-455
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    • 2000
  • Gene targeting are very useful tools for the research on the gene function in vivo, mass production of foreign materials and biomedical approach of therapeutic process. But this process is very complicated and necessary highly skilled technique, because it is very different from ES cell origin, genetic background of embryo, and experimental conditions. We investigated the productivity ability of chimeric mouse after aggregation with TT2 ES cells. Increse of ES cell density caused gradual decrease in embryo development in vitro and in th $\varepsilon$ production of chimeric mice in vivo. One million ES cell density for the aggregation was very efficient to produce high percentage chimeric mice in their coat color. These results suggested that appropriate cell density plays a key role in the development and production of chimeric mice by a 8-cell aggregation method.

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