• Title/Summary/Keyword: flow cytometer

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Cellular Immune Responses of the Manila Clam, Ruditapes philippinarum, Exposed to Cadmium Chloride (카드뮴에 노출된 바지락 (Ruditapes philippinarum) 의 세포성 면역 반응)

  • Park, Kyung-Il;Choi, Min-Soon;Park, Sung-Woo;Park, Kwan-Ha;Choi, Sang-Hoon;Yoon, Jong-Man
    • The Korean Journal of Malacology
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    • v.25 no.2
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    • pp.135-143
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    • 2009
  • The Manila clam, Ruditapes philippinarum, has been considered as a sentinel species due to dominant distribution along the coast of Korea and well developed regulatory system. In order to develop and understand immune responses of the Manila clams, clams were exposed to $50\;{\mu}g/L$ of cadmium chloride (Cd) for 8 days and monitored the cellular immune parameters of the hemocytes including blast cell composition, DNA damage, necrosis, apoptosis and hemocyte mortality using a flow cytometer. The results showed that all immune parameters analyzed in the present study increased remarkably compared to the controls and the increases were statistically significant. Apoptosis rate was higher than necrosis rate in the clams exposed to Cd suggesting that apoptosis was preferably induced by the concentration of Cd used in the present study. Our study indicates that the measurement of cellular immune responses of the Manila clam using flow cytometer will be a useful technique for assessment of heavy metal contamination in marine environment.

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FLOW CYTOMETRIC ANALYSIS OF LYMPHOCYTE AND CYCLING CELL DISTRIBUTION IN PERIAPICAL LESIONS (유세포 분석기를 이용한 치근단 병소의 임파구 조성 및 CYCLING CELL 분포에 관한 연구)

  • Oh, Tae-Seok;Lim, Sung-Sam
    • Restorative Dentistry and Endodontics
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    • v.18 no.2
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    • pp.317-340
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    • 1993
  • This study was designed 1) to compare the distributions of periapical inflammatory cells and 2) to identify lymphocytes and compare the lymphocyte distribution with T lymphocyte subpopulation and then 3) to examine the distribution of cycling cell in human dental periapical lesions. From each of the twenty-five human dental periapical lesions observed one small portion was fixed, embeded in paraffin, sectioned serially and stained with HE. The periapical inflammatory cells were counted to obtain the relative concentration of lymphocyte, plasma cell, macrophage and neutrophil. The large part of each lesion was analysed using Flow cytometer and monoclonal antibodies to obtain the relative concentration of T lymphocyte, B lymphocyte, T'helper cell and T suppressor/cytotoxic cell. In addition to that, seven human dental periapical lesions were examined with DNA analysis to observe the distribution of cycling cell. Following results were obtained: 1. 24 cases of the 32 periapical lesions examined were diagnosed as periapical granuloma and the remaining 8 cases as periapical cyst. Lymphocytes comprised 42.1% of total inflammatory cells in periapical granuloma and 41.8% in periapical cyst. Corresponding percentages for macrophages were 33.8% and 30.3%; for plasma cells, 15.9% and 19.0%; for neutrophils, 8.2% and 8.8%. 2. All of the periapical lesions examined had T lymphocyte, B lymphocyte, T helper cell, T suppressor/cytotoxic cell. And in all cases, T lymphocytes were observed predominantly more than B lymphocytes. 3. In 2 cases of the control group only T lymphocytes were found, and in the remaining 2 cases T lymphocytes were observed predominantly. 4. T helper cells were observed predominantly more than T suppressor/cytotoxic cells in all cases of perapical granulomas. 5. T suppressor/cytotoxic cells were observed predominantly more than T helper cells in 4 cases of periapical cysts (total 5 cases were examined) and only in one case T helper cells were more than T suppressor/cytotoxic cells. 6. In control group, T helper cells were predominant in 2 cases and T helper cells were equivalent to T suppressor/cytotoxic cells in one case. In remaining one case T suppressor/cytotoxic cells were predominant. 7. As the result of DNA analysis, the average proliferating indices of the various groups examined were measured as follows: in the control group 5.45%, in periapical granuloma 6.64%, in periapical cyst 10.1%. The highest index was observed in periapical cyst.

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Selection of Unnecessary Urine Culture Specimens Using Sysmex UF-5000 Urine Flow Cytometer (Sysmex UF-5000 소변 유세포분석기를 이용한 요배양 불필요 검체의 선별)

  • Song, Duyeal;Lee, Hyun-Ji;Jo, Su Yeon;Lee, Sun Min;Chang, Chulhun L.
    • Annals of Clinical Microbiology
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    • v.21 no.4
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    • pp.75-79
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    • 2018
  • Background: Urine culture is one of the most frequently requested tests in microbiology. Automated urine analyzers yield much infection-related information. The Sysmex UF-5000 analyzer (Sysmex, Japan) is a new flow cytometry urine analyzer capable of quantifying urinary particles, including bacteria, WBCs, and yeast-like cells (YLCs) and can provide a Gram stainability flag. In this work, we evaluated how many unnecessary urine cultures could be screened out using the UF-5000. Methods: We compared the culture results of 126 urine samples among 453 requested urine cultures (from sources other than the Urology and Nephrology departments) with urinalysis results. Urine cultures were considered positive if bacterial or YLC growth was ${\geq}10^4CFUs/mL$. Results: We used urinalysis cut-off values of $50/{\mu}L$ and $100/{\mu}L$ for bacteria and YLC, respectively. Forty eight of the 126 (38.1%, or 10.6% of 453 requested) cultures were below these cut-off values and did not contain any culture-positive samples. Conclusion: Bacteria and YLC counts generated using the UF-5000 analyzer could be used to screen out negative cultures and reduce urine culture volume by ~10% without sacrificing detection of positive cultures.

The effect of Astragali Radix and Armeniacae Semen on the apoptosis of immunocytes and nitric oxide production from peritoneal macrophages (황기와 행인(杏仁)이 면역세포(免疫細胞)의 Apoptosis 및 Nitric Oxide에 미치는 효과(效果))

  • Jeong Hyun-Woo;Moon Han-Ju
    • Herbal Formula Science
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    • v.6 no.1
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    • pp.175-186
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    • 1998
  • The purpose of this research was to investigate effects of Astragali Radix(AR) and Armeniacae Semen(AS) on T-lymphocytes and peritoneal macrophages in mice. The proliferation of thymocytes and splenocytes were teated using macroplate-reader. The apoptosis and sub-population of T-lymphocytes were tested using a flow cytometer. Nitric oxide production was tested using a Griess reagents. The result were obtained as follow; 1. AR increased the proliferation of thymocytes and splenocytes. 2. AS decreased the proliferation of thymocytes and splenocytes. 3. AR and AS decreased No production fron peritoneal macrophages 4. AR and AS were accelerate T-lymphocytes apoptosis. 5. AR and AS increased $T_C$ cells population, but decreased $T_H$ cells population of T-lymphocyte.

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Enhancement of Emulsion-mediated Gene Expression by Using Chitosan as a Pre-Condensing Agent

  • Chun, Soo-Kyung;Choi, Sung-Hee;Kim, Adele;Ahn, Woong-Shick;Kim, Chong-Kook
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.426.3-427
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    • 2002
  • The aim of this study was to enhance the transfection efficiency of emulsion-mediated gene expression by using chitosan, Conventional DNA/emulsion complexes and precondensed DNA/emulsion complexes were prepared by adding either naked or precondensed plasmids to cationic emulsion. The zeta potential. TEM, and size of transfection complexes were measured. In vitro transfection efficiency for boty complexes was also studied by several methods: flow cytometer, expression analysis by confocal microscope, RT-PCR, and in addition. (omitted)

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Influence for Carcinoma Cell and Lymphatic Cell of Acetyl Arsonate (아세틸 아르소네이트의 면역세포와 암세포에 미치는 영향: L1210, Sarcoma 180, MOLT-4 등 사람과 생쥐에 대한 작용비교)

  • Jeong, Yong-Ja;Seong, Yeong-Gi
    • YAKHAK HOEJI
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    • v.40 no.5
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    • pp.599-607
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    • 1996
  • Acetylarsonate was prepared for testing antitumor and immunological effects. It showed cytotoxicity directly on Sarcoma 180. L1210 and MOLT-4 by MTT assay. It did not seemed to trigger the mitosis of human lymphocytes in culture, but that showed the cytotoxicity with higher dose. The rosette formation and spleen weight of mouse which acetylarsonate was administered to for 2 weeks were increased. Furthermore, peripheral helper T- and cytotoxic/suppressor T-lymphocytes were increased in acetylarsonate-injected-mice significantly when it was estimated with simultaneous 2 color analysis using anti Lyt2-FITC and L3T4-PE monoclonal antibody by Flow cytometer.

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유식세포분석분리기의 원리 및 면역생물학에서의 역할

  • 정헌택
    • The Microorganisms and Industry
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    • v.17 no.1
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    • pp.38-50
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    • 1991
  • Flow cytometer(유식세포분석분리기:FCM)는 부유되어 있는 세포의 여러 특성을 측정할 수 있도록 고안되었기 때문에 동물의 체내에 있는 세포중 자연적으로 부유되어 있는 혈구세포를 분석하기가 가장 용이하다. 면역계에서 가장 중추적인 역할을 담당하며 면역반응에 특이성과 기억능력 등을 부여하는 림프구와 탐색작용을 가장 왕성히 나타내는 과립구나 단구 등이 혈액내에 존재하기 때문에 FCM을 이용하여 개체의 방어기능을 맡고 있는 면역세포를 분석하는 일이 가장 먼저 활발히 이루어졌음은 당연하였다. 도한 1970년대 면역학이 큰 발전에 이룩한 단크론항체가 Miller와 Kohler에 의하여 개발되었기 때문에 면역세포의 분화항원에 대한 여러 종류의 단크론항체는 FCM의 이용으로 면역세포를 여러 아형으로 분석하고 분리가 가능하게 하였다. 마지막으로 FCM이 최근에 개발된 것에 맞추어 세포성 면역학이 30년 전부터 급속도로 연구되었으므로 면역학적 연구가 FCM의 이용으로 더욱 활성화되었음은 주지의 사실이다. 이런 시점에서 저자는 FCM의 원리와 면역생물학에서의 FCM의 역할에 대하여 간단히 소개하려고 한다.

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Variations in endopolyploidy level during the short period of the early growing stage in the roots and leaves of maize (Zea mays) seedlings

  • Ogawa, Atsushi;Taguchi, Nanako;Miyoshi, Kazumitsu
    • Plant Biotechnology Reports
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    • v.4 no.2
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    • pp.117-123
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    • 2010
  • We used a flow cytometer to investigate the variations in endopolyploidy (the frequencies of nuclei with DNA contents equivalent to 4C through 16C) during the short period of the early growing stage in vigorously growing young tissues of maize seedlings. We examined different portions of the root and leaves that had been growing for 7 (day 7) and 13 (day 13) days after germination. Endoreplication showed two opposing phenomena without aging. In one case, the endopolyploidy of the first leaf was higher on day 13 than on day 7. In the latter case, endopolyploidy decreased, as clearly revealed by a comparison of the endopolyploidy of the second leaves and the 160-170 mm portion of the seminal root on days 7 and 13. Endopolyploidy was also lower in the top of the leaf. In roots, endopolyploidy was increased by the exogenous application of abscisic acid for only 1 day. The levels of endopolyploidy increased without an increase in cell size in the roots. These results showed that endoreplication occurs in actively growing and young tissue and that the variation can be induced in the short period examined.

Effects of Triterpenoids from Luvunga scandens on Cytotoxic, Cell Cycle Arrest and Gene Expressions in MCF-7 Cells

  • Taher, Muhammad;Al-Zikri, Putri Nur Hidayah;Susanti, Deny;Arief Ichwan, Solachuddin Jauhari;Rezali, Mohamad Fazlin
    • Natural Product Sciences
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    • v.22 no.4
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    • pp.293-298
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    • 2016
  • Plant-derived triterpenoids commonly possesses biological properties such as anti-inflammatory, antimicrobial, anti-viral and anti-cancer. Luvunga scandens is one of the plant that produced triterpenoids. The aims of the study was to analyze cell cycle profile and to determine the expression of p53 unregulated modulator of apoptosis (PUMA), caspase-8 and caspase-9 genes at mRNA level in MCF-7 cell line treated with two triterpenoids, flindissol (1) and 3-oxotirucalla-7,24-dien-21-oic-acid (2) isolated from L. scandens. The compounds were tested for cell cycle analysis using flow cytometer and mRNA expression level using quantitative RT-PCR. The number of MCF-7 cells population which distributed in Sub G1 phase after treated with compound 1 and 2 were 7.7 and 9.3% respectively. The evaluation of the expression of genes showed that both compounds exhibited high level of expression of PUMA, caspase-8 and caspase-9 as normalized to ${\beta}-actin$ via activation of those genes. In summary, the isolated compounds of L. scandens plant showed promising anticancer properties in MCF-7 cell lines.

Quantitation of Antigen-Antibody Reaction Condition for Development of Fluorescence Image-based CD4 Rapid Test (형광 영상 기반 CD4 신속 검사법 개발을 위한 항원-항체 반응 조건 정량화)

  • Kim, Subin;Kim, Jung Kyung
    • Journal of the Korean Society of Visualization
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    • v.13 no.1
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    • pp.35-42
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    • 2015
  • CD4+ T-cell count determines the effectiveness for antiretroviral therapy (ART) in patients with human immunodeficiency virus (HIV). Although ART slows the progression of HIV to AIDS, rapid counting of CD4+ T lymphocytes with a drop of patient's blood sample is urgently needed to ensure timely ART treatment in rural areas. Recently point-of-care CD4 testing devices have been developed by using non-flow based imaging cytometer incorporated with a sample cartridge where CD4+ T cells are reacted with fluorescently tagged specific antibodies. Here we conducted an experimental study using a conventional fluorescence microscope-based imaging system to quantitate the interaction of CD4 antibodies with CD4+ T cells at different reaction conditions. We demonstrated that a fast and affordable point-of-care CD4 test is feasible with a far less amount of antibodies and a shorter incubation time compared with a conventional sample preparation protocol for flow cytometry. We also proposed a general method to evaluate and compare the detection limit across different CD4 counting platforms by using fluorescently labelled microbeads for intensity calibration.