• Title/Summary/Keyword: 유세포분석기

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Enumeration of Weissella cibaria phage with cytometry, epifluorescence microscopy, and plaque assay (유세포분석기, 형광현미경, 용균반검사 분석을 이용한 Weissella cibaria 박테리오파지 정량분석 및 상관관계분석)

  • Park, Won Jeong;Lim, Ga-Yeon;Park, Jong-Hyun
    • Korean Journal of Food Science and Technology
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    • v.50 no.2
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    • pp.244-247
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    • 2018
  • Quantitative analysis for non-host infection bacteriophage was conducted for their enumeration. Flow cytometry and epifluorescence microscopy (EPM) were selected as counting methods. Correlation analysis was performed based on the plaque assay method on the existing host infection and consisted of Pearson correlation statistical analysis, regression analysis, and difference analysis. Analyses of 12 samples with flow cytometry and plaque assay methods showed that there was a correlation of 96.7% with Pearson correlation value r=0.967, $R^2$ 0.9352, and difference value of 1.063. Analyses of 12 samples with EPM and plaque assay methods showed that there was a correlation of 99.0% with Pearson correlation value r=0.990, $R^2$ 0.9811, and difference value of 1.605. Therefore, flow cytometry and epifluorescence microscopy would be effective for enumeration of Weissella cibaria bacteriophage with plaque assay.

In vivo Imaging Flow Cytometer (세포 이미징 기능을 겸비한 생체 유세포 분석기)

  • Lee, Ho
    • Journal of the Korean Society of Visualization
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    • v.5 no.1
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    • pp.9-11
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    • 2007
  • We introduce an in vivo imaging flow cytometer, which provides fluorescence images simultaneously with quantitative information on the cell population of interest in a live animal. As fluorescent cells pass through the slit of light focused across a blood vessel, the excited fluorescence is confocally detected. This cell signal triggers a strobe beam and a high sensitivity CCD camera that captures a snap-shot image of the cell as it moves down-stream from the slit. We demonstrate that the majority of signal peaks detected in the in vivo flow cytometer arise from individual cells. The instrument's capability to image circulating T cells and measure their speed in the blood vessel in real time in vivo is demonstrated. The cell signal irradiance variation, clustering percentage, and potential applications in biology and medicine are discussed.

Design of microcontroller-based cancer cell detection system (마이크로컨트롤러를 활용한 표적 암세포 검출기 개발)

  • Cho, Kyoungrae;Seo, Jeong-hyeok;Choe, Se-woon
    • Proceedings of the Korean Society of Computer Information Conference
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    • 2019.07a
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    • pp.395-396
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    • 2019
  • 표적 암세포 또는 마이크로미터 단위 이하의 미세 입자의 검출하기 위해 다양하게 사용되는 유세포분석기는 레이저와 같은 특정 파장의 광원 및 파장대별 선택이 가능한 필터, 고성능 광탐지기로 구성되어있다. 이 분석기는 표적물질의 광학적 산란특징에 기인하여 입상도와 크기, 농도 등의 물리적 정보제공이 가능하나, 고가이며 사용자의 요구에 따른 다양한 광원 및 필터의 선택이 제한적이라는 단점을 가지고 있다. 본 논문에서는 상용화된 마이크로컨트롤러, 발광다이오드, 광검출기와 필름형식의 광학필터를 사용하여 저가의 표적 암세포 검출기를 개발하고자 한다. 또한, 3D 프린터를 사용하여 사용자의 필요에 따른 설계변화와 이동이 용이하므로, 공간의 제약을 받지 않고 다양한 분야에 적용이 가능하다. 제안된 검출기의 광감지 수치는 세포의 농도에 따라 높은 선형성을 보였으며, 세포수량기를 사용한 결과 값과 비교하였을 때 무의미한 통계학적 차이를 보였다.

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A flowcytometric determination of DNA content in Pacific abalone, Haliotis discus hannai cell (유동세포분석에 의한 참전복(Haliotis discus hannai) 세포내 DNA 함량 분석 최적화)

  • Park, In-Seok
    • Korean Journal of Environmental Biology
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    • v.38 no.2
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    • pp.248-253
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    • 2020
  • The level at which analyses of DNA content might contribute more significantly to the genetic mechanisms of evolution lies in the events of speciation. The object of this study was to investigate the DNA content of abalone (Haliotis discus hannai) and determine the optimal tissue samples for measuring the DNA content of abalone by flowcytometry without fixation. The DNA content (pg/nucleus) of gill tissue (2.5±0.08), which was contaminated with protozoa, was significantly lower than that of muscle tissue (3.2±0.02), mantle tissue (3.2±0.02) (p<0.05), and a standard reference standard, while the DNA contents of muscle tissue and mantle tissue were higher than that of the standard reference. Considering the results of this study, DNA content analysis with flowcytometry is an acute and rapid method by which muscle tissue and mantle tissue are the most appropriate sample for measuring the DNA content of abalone without fixation.

SONICATED EXTRACT OF TREPONEMA DENTICOLA IMPAIRS THE LYMPHOCYTE PROLIFERATION (Treponema denticola sonicated extract에 의한 임파구 세포주기 차단 기전에 관한 연구)

  • Lee, Woo-Cheol;Bruce J. Shenker
    • Restorative Dentistry and Endodontics
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    • v.27 no.5
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    • pp.473-478
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    • 2002
  • 근관내 spirochetes의 존재유무가 명확하게 밝혀져 있지 않았으나 최근 PCR을 사용한 연구에서 Treponema denticola균주가 감염근관의 50% 이상의 경우에서 발견됨에 따라 이 세균이 치수 및 치근단 질환에 관여하는지에 대한 관심 이 높아졌다. 하지만 그 정확한 기전은 아직 밝혀져 있지 않다. 이와 관련하여 Shenker등이 T. denticola의 sonicated extract에서 순수분리된 단백질 (SIP)이 임파구 proliferation을 방해함을 보고한바 있다. 따라서 본 연구의 목적은 면역억제단백질 SIP이 어떤 기전에 의해서 임파구증식을 억제하는지를 밝히는 데 있다. 건강한 혈액 공여자로부터 추출해낸 T세포에 PHA (phytohemagglutinin)로 증식자극을 주게되는데 이 과정에서 SIP을 처리하거나 처리하지 않은 경우를 비교하여 세포주기 진행과정을 유세포분석기 (Becton-Dickinson FACS$^{tarplus}$) 를 통하여 평가하였다. 실험결과 세단계의 chromatography과정을 통해 순수정제된 SIP은 50kDa와 56kDa의 두가지 polypeptide로 구성되어 있고 0.25$\mu\textrm{g}$으로 처리된 T 임파구는 42.5%의 [$^3$H]thymidine incorporation 억제가 그리고, 0.5$\mu\textrm{g}$으로 처리한 경우는 75.1%의 억제가 일어나 dose-dependent한 양상이 나타났다. Propidium iodide와 유세포 분석기를 사용하여 세포주기를 분석한 결과 medium으로만 처리한 경우 97%이상의 임파구는 G$_0$/G$_1$ phase에 머물러 있었으나 PHA자극을 받은 경우 G$_0$/G$_1$ phase에서 58%, S phase에서 34.6%, G$_2$/M phase에서 7.4%로 분포되어 나타났다. SIP으로 전처리한 경우 세포 증식이 감소하여 0.25$\mu\textrm{g}$을 첨가한 경우 75.1%가 G$_0$/G$_1$ phase에 머물러 있었고 더 강한 농도의 0.5$\mu\textrm{g}$을 첨가한 경우는 87.7%가 G$_0$/G$_1$ phase에서 S phase로 진행되지 않고 머물러있었다. 따라서 SIP으로 전처리된 T 임파구는 그 증식이 G$_0$/G$_1$ phase에서 차단된 것으로 보인다. 이러한 면역억제현상이 in vitro 상태뿐 아니라 in vivo에서도 진행된다면 spirochete가 치수 및 치근단 질환의 병인론에 연관된 면역반응저하기전에 중요한 역할을 하는 것으로 추론할 수 있다.

Significant Attenuation of Aden-associate Virus Gene Expression by Catechol-conjugated Heparin Surface Coating (카테콜기가 도입된 헤파린의 표면고정화에 의한 아데노연관바이러스의 발현 억제에 관한 연구)

  • Do, Minjae;Lee, Slgirim;Jang, Jae-Hyung;Lee, Haeshin
    • Journal of Adhesion and Interface
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    • v.17 no.4
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    • pp.149-154
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    • 2016
  • In this study, natural polymer-based virus neutralizing agent was developed in an attempt to replace the conventional sterilization method for mammalian cell culture. A catechol conjugated heparin was synthesized by using EDC chemistry, and it show unique binding ability to virus which has heparin affinity (adenovirus, adeno-associated virus). To evaluate neutralization ability of catechol conjugated heparin, adeno-associated virus was used for test model, instead of using a pathogenic virus. The catechol conjugated heparin exhibited resistance to high concentration of salt and complete inactivation of adeno-associated virus. The result suggests that the catechol conjugated heparin, which is biocompatible and efficiency, may replace conventional sterilization method for mammalian cell culture.

Effect of silver nanoparticles on the performance of riverbank filtration: Column study (강변여과에서의 은나노입자의 영향 : 실험실규모 컬럼 실험)

  • Lee, Donghyun;No, Jin-Hyeong;Kim, Hyun-Chul;Choi, Jae-Won;Choi, Il-Hwan;Maeng, Sungkyu
    • Journal of Korean Society of Water and Wastewater
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    • v.29 no.1
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    • pp.77-88
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    • 2015
  • Soil column experiments were evaluated effects of silver nanoparticles (i.e., 0, 2.5, 5, and 10 mg/L) on the microbial viability which is strongly associated with the degradation of organic matter, pharmaceutically active compounds(PhACs) and biological oxidation of nitrogenous compounds during river bank filtration. The addition of silver nanoparticles resulted in almost no change in the aqueous matrix. However, the intact cell concentration decreased with addition of silver nanoparticles from 2.5 to 10 mg/L, which accounted for 76% to 82% reduction compared to that of control (silver nanoparticles free surface water). The decrease in adenosine triphosphate was more pronounced; thus, the number and active cells in aqueous phase were concurrently decreased with added silver nanoparticles. Based on the florescence excitation-emission matrix and liquid chromatograph - organic carbon detection analyses, it shows that the removal of protein-like substances was relatively higher than that of humic-like substances, and polysaccharide was substantially reduced. But the extent of those substances removed during soil passage was decreased with the increasing concentration of silver nanoparticles. The attenuation of ionic PhACs ranged from 55% to 80%, depending on the concentration of silver nanoparticles. The attenuation of neutral PhACs ranged between 72% and 77%, which was relatively lower than that observed for the ionic PhACs. The microbial viability was affected by silver nanoparticles, which also resulted in inhibition of nitrifiers.

FLOW CYTOMETRIC ANALYSIS OF LYMPHOCYTES IN NORMAL AND INFLAMED PULP (유세포분석기를 이용한 정상치수조직과 염증성 치수조직 내의 임파구 분포에 관한 연구)

  • Kim, Seon-Ah;Bae, Kwang-Shik;Im, Seong-Sam
    • Restorative Dentistry and Endodontics
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    • v.22 no.1
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    • pp.374-387
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    • 1997
  • The purpose of this study was to examine the distribution of lymphocyte populations in normal, reversibly inflamed and irreversibly inflamed human dental pulp tissues using flow cytometry. Flow cytometry, with specific antibody and fluorochrome reagent allows us to know cellular properties of hematolymphoid cells by measuring fluorescence of stained cells. Before extirpation of pulps in routine endodontic treatment, the clinical diagnosis were performed by symptom. The extirpated pulp tissues were divided into normal pulp group (N=5), reversible pulpit is group(N=10) and irreversible pulpitis group(N=7). The specimen was placed into RPMI 1640 medium, minced into small pieces, and then digested in medium with collagenase. The cell suspension was resuspended in PBS for monoclonal antibody staining of T lymhocytes(CD3+), B lymphocytes (CD19+), T helper cell (CD4+) and T supressor cell (CD8+). The percentages of cells were counted by FACStar(BD) flow cytometer. Following results were obtained; 1. In the most normal and inflamed pulps, the percentages of T lymphocyte, B lymphocytes, T helper cell and T suppressor/cytotoxic cell were less than 1 % in total counted pulpal cells. 2. The higher percentages of T, B, T helper and T suppressor cells were observed in irreversible pulpitis group as compared with the normal pulp and reversible pulpitis group but the differences between groups were not statistically significant (p>0.05). 3. The percentages of T helper cells (CD4 + cells) were greater than that of T suppressor/cytotoxic cells (CD8 + cells) in the inflamed pulps.

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Cytogenetic Analysis of the Triploid Pacific Abalone, Haliotis discus hannai (북방전복, Haliotis discus hannai 3배체의 세포유전학적 연구)

  • Jee, Young-Ju;Chang, Young-Jin
    • The Korean Journal of Malacology
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    • v.28 no.1
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    • pp.37-43
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    • 2012
  • In this study, we invesgated a cytogenetic analysis of the Pacific triploid abalone, Haliotis discus hannai induced by low temperature treatment. We got a lot of mitotic metaphase chromosome spreads from the triploid and diploid Pacific abalones' hatched larvae (trochophores). The chromosome number of diploid abalone was 2n = 36 and that of triploid abalone was 3n = 54, so the chromosome number of triploid abalone was 1.5 times higher than that of diploid abalone. We developed a modified flow cytometric method for Pacific abalone from the existing methods. We uesd 51 months aged triploid and diploid Pacific abalones' hemolymph to get the DNA contents by flow cytometry. The DNA content of diploid abalone was 1.743 pg/cell and the DNA content of triploid abalone was 1.49 times higher than that of diploid one. It proved that triploid abalone was consisted with two sets of maternal diploid and one set of paternal genome.