• Title, Summary, Keyword: flow cytometer

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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.

Phytoplankton in the Waters of the Ieodo Ocean Research Station Determined by Microscopy, Flow Cytometry, HPLC Pigment Data and Remote Sensing (현미경, Flow Cytometer, HPLC 색소자료 및 원격탐사를 이용한 이어도 관측기지 주변수의 식물플랑크톤 연구)

  • Noh, Jae-Hoon;Yoo, Sin-Jae;Lee, Jung-Ah;Kim, Hyun-Chul;Lee, Jae-Hak
    • Ocean and Polar Research
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    • v.27 no.4
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    • pp.397-417
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    • 2005
  • Phytoplankton community structure and distribution pattern in the surface water around the Ieodo Ocean Research Station were investigated during seven cruises carried out from July, 2003 to October, 2004. Samples were analyzed using various tools including a microscope, flow cytometer, and HPLC. Satellite images were used to analyze spatio-temporal phytoplankton biomass distribution. SeaWiFS chlorophyll a (chl a) images showed that spring blooms occurred in April-May near the Ieodo Station, and these waters were under the influence of Changjiang Dilute Water during July-October. Also, during the July-October period, HPLC pigments data showed increasing zeaxanthin concentrations, a marker pigment of cyanobacteria whereas increasing concentrations of various other pigments such as fucoxanthin, peridinin, prasinoxanthia alloxanthin, 19'-hexanoyloxyfucoxanthin and chlorophyll b were noted during spring blooms. Such pigment marker data were consistent with picoplankton data analyzed by flow cytometer and nano-microplankton analyzed by microscope. The pigment-CHEMTAX method was used to drive the phytoplankton group apportioned chi a. Diatoms, chlorophytes, dinoflagellates, and cryptophytes comprised 25.8, 20.7, 15.9, and 14.1%, respectively, of the total chl a in May. Average cyanobacteria concentrations in July-October contributed 25.4% of the total concentration. This was the highest percent contribution and was followed by chlorophytes, diatoms, and prymnesiophytes. This study discusses results from various methods, similarities and differences in the results among those methods, and the application range of the results from different analytical methods. Also, the study reveals a detailed phytolpankton community structure in the waters around the Ieodo Station, and suggests future monitoring considerations in relation to cell morphology, ecology and diversity factors according to taxonomic groups.

Flow Cytometric Analysis of Bovine Granulosa Cells : Changes of Cell Cycle During Follicular Maturation (Flow Cytometer를 이용한 소 과립막세포의 분석 : 난포성숙에 따른 세포주기의 변화)

  • 김해정;김동훈;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.17 no.4
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    • pp.279-285
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    • 1994
  • The objective of the present study was to characterize the cell cycles of granulosa cell populations during follicular maturation in cattle by using flow cytometer. Granulosa cells were isolated from bovine preovulatory antral follicles of F1(>10mm), F2(5~20mm), F3(3~4mm) and F4(1~2mm) diameter and fixed and stained with fluorochromes that selectively bine to DNA. Flow cytometer equipped with a laser excitation system was used to analyze the intensity of fluorescence from stained cells. Forward angle light-scatter(FSC) and 90$^{\circ}$light-scatter(SSC) signals were adopted to measure the size and the granularity of granulosa cells. As a results of FSC/SSC analysis, granulosa cell populations(G1 phase of cell cycle) from each follicle were relatively regular in size and granularity, regardless of follicular size. However, their distribution in granularity was greater than that in size. Most of granulosa cell populations collected from each follicle were distributed in G0/G1, S and G2/M phases. As the follicles approached to ovulation the percentage of cells in the proliferative phases of cell cycle (S and G2/M) decreased significantly, but there was a concomitant increase in the percentage of granulosa cells in G1 phase. Therefore, these data indicate the proportion of main populations to cell cycle of granulosa cells may be changed from proliferative phase to G1 phase during follicular maturation in cattle.

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A Flow Cytometrical Analysis of the Antitumor and Immunostimulatory Effects of LCT-CT, a Cold-water Extract Prepared from Rice Grasshopper Oxya japonica japonica Thunberg (벼메뚜기(Oxya japonica japonica Thunberg) 물 추출물 LCT-CT의 항암면역 활성에 관한 유세포 분석학적 연구)

  • Chung, Kyeong-Soo;Kim, Bit Na
    • YAKHAK HOEJI
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    • v.58 no.3
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    • pp.151-157
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    • 2014
  • Water extracts of rice grasshopper (Oxya japonica japonica Thurnberg) were prepared and their antitumor and immunostimulatory activities were investigated using a flow cytometer. When LCT-CT was ip injected into ICR mice at the dose of 33.3 mg/kg before and after the implantation of $4{\times}10^5$ cells/mouse of sarcoma 180 tumor cells, it inhibited the growth of the tumor cells by 96.6%, showed lymphoblstogenic activities on the splenic lymphocytes and increased the expression of CD25 molecules on the splenic T lymphocytes. When co-cultured with the splenic lymphocytes of a BALB/c mouse, LCT-CT showed strong immunostimulatory activities at the concentration of $25{\sim}100{\mu}g/ml$ by significantly increasing lymphoblasts ratio and CD25 expression.

Analysis of Complex Cell Cycle Occurring in the Rodent Testis by Laser Scanning Cytometer

  • 박미령;주학진;천영신;이미숙;김진회
    • Proceedings of the KSAR Conference
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    • pp.37-37
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    • 2001
  • 포유동물의 정자형성과정 (spermatogenesis) 은 유사분열과 감수분열이 동시에 일어나는 매우 복잡하지만, 효율적으로 생식세포를 증식, 분화시키는 시스템이다 정상적인 spermatogenesis가 일어나는 testis에서는 haploid (IN), diploid (2N), 그리고 tetraploid(4N)과 같은 핵형을 갖는 세포들이 일정한 비율로 존재한다. DNA flow cytometry(DNA FCM) 는 세포의 핵형(ploidy)을 신속·정확하게 측정하여, 1N, 2N 그리고 4N에 대한 비율을 예측할 수 있어서, 생식세포를 포함한 다양한 유형의 세포주기를 분석하는데 적용되어져 왔다. 세포주기 분석법 중 이와 같은 FCM이외에, flow cytometer와 static image cytometer를 결합시켜 새롭게 고안된 laser scanning cytometer (LSC)가 있다. 그리고, 이제까지 LSC를 사용한 spermatogenesis에 관한 연구에 대해서는 보고된 바가 없다. 본 실험은 설치류에 있어 각기 다른 발달단계에 있는 정상적인 정소세포를 분리하여 PI (propidium iodide) 로 DNA를 염색한 후, DNA함량을 LSC로 분석하였다. 이것을 FCM에 의한 정소세포의 DNA분석과 비교·검토하였으며, 이 방법을 정상적인 spermatogenesis 가 일어나지 않는 동물시스템에 적용시켰다. 생식세포를 소멸시키기 위해 항암제인 busulfan과 비타민 A를 결핍시켜 이것이 세포주기의 어떤 시점에서 어떻게 작용하여, 생식세포를 소멸시키는지 알아보았다. 위의 실험·분석결과로부터 LSC를 사용한 DNA함량과 핵형의 결정은 FCM과 동일한 수준의 정확성을 제시하였다. busulfan 또는 비타민 A의 결손은 살아있는 세포의 80% 이상이 2N의 핵형에 해당하는 G0/G1 기에 있는 것으로 나타났다. 그리고 1N:2N 및 4N:2N의 핵형비율의 변화를 가져왔다. 이러한 자극은 생식세포주기제어에 관여하며, 생식세포가 증폭하고 분화로 들어가는 단계를 차단, G0/G1 기에서 정체(arrest)되는 것으로 시사된다.

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A novel method for cell counting of Microcystis colonies in water resources using a digital imaging flow cytometer and microscope

  • Park, Jungsu;Kim, Yongje;Kim, Minjae;Lee, Woo Hyoung
    • Environmental Engineering Research
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    • v.24 no.3
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    • pp.397-403
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    • 2019
  • Microcystis sp. is one of the most common harmful cyanobacteria that release toxic substances. Counting algal cells is often used for effective control of harmful algal blooms. However, Microcystis sp. is commonly observed as a colony, so counting individual cells is challenging, as it requires significant time and labor. It is urgent to develop an accurate, simple, and rapid method for counting algal cells for regulatory purposes, estimating the status of blooms, and practicing proper management of water resources. The flow cytometer and microscope (FlowCAM), which is a dynamic imaging particle analyzer, can provide a promising alternative for rapid and simple cell counting. However, there is no accurate method for counting individual cells within a Microcystis colony. Furthermore, cell counting based on two-dimensional images may yield inaccurate results and underestimate the number of algal cells in a colony. In this study, a three-dimensional cell counting approach using a novel model algorithm was developed for counting individual cells in a Microcystis colony using a FlowCAM. The developed model algorithm showed satisfactory performance for Microcystis sp. cell counting in water samples collected from two rivers, and can be used for algal management in fresh water systems.

Formation of Reactive Oxygen Species and Cr(V) Entities in Chromium(VI) Exposed A549 Cells (크롬 6가 투여 후 A549 세포에서의 Reactive Oxygen Species와 크롬 5가의 발생)

  • 박형숙
    • Environmental health and toxicology
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    • v.11 no.1_2
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    • pp.49-57
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    • 1996
  • The production of reactive oxygen species on addition of hexavalent chromium (potassium dichromate, $K_2Cr_2O_7$ ) to lung cells in culture was studied using flow cytometer analysis. A Coulter Epics Profile flow cytometer was used to detect the formation of reactive oxygen species after $K_2Cr_2O_7$ was added to A549 cells grown to confluence. The cells were loaded with the dye, 2',7'-dichlorofluorescein diacetate, after which cellular esterases removed the acetate groups and the dye was trapped intracellularly. Reactive oxygen species oxidized the dye, with resultant fluorescence. Increased doses of Cr(VI) caused increasing fluorescence (10-fold higher than background at 200 gM). Addition of Cr(III) compounds, as the picolinate or chloride, caused no increased fluorescence. Electron paramagnetic resonance (EPR) spectroscopic studies indicated that three (as yet unidentified) spectral "signals" of the free radical type were formed on addition of 20, 50, 100 and 200 gM Cr(VI) to the A549 cells in suspension. Two other EPR 'signals" with the characteristics of Cr(V) entities were seen at field values lower than the standard free radical value. radical value.

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Study on natural killer cell activity and its characteristics during hepatocarcinogenesis in rats (랫드의 간암 발생과정에서 분리한 자연살해세포의 활성측정 및 특성연구)

  • Jeong, Ja-young;Lee, Kuk-kyung;Kil, Jwang-sup;Lee, Yong-soon
    • Korean Journal of Veterinary Research
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    • v.39 no.1
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    • pp.169-176
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    • 1999
  • The purposes of this study were to set up the method of the natural killer(NK) cell activity assay using the flow cytometer and to examine the characteristics and distribution of the NK cell during rat hepatocarcinogenesis. Forty five male 6 week-old specific pathogen free(SPF) Sprague-Dawley rats were randomly divided into three groups. Group I was the non-treated control and given normal diet and water. Group II was treated with diethylnitrosamine(DEN, 200mg/kg, i.p.) and partial hepatectomy. Group III was treated with DEN, partial hepatectomy and 0.05% phenobarbital sodium in water from 3 to 16 weeks. All animals were examined the morphology of the large granular lymphocyte(LGL), the LGL percent of the total lymphocytes and the LGL conjugation rate with YAC-1 cell in peripheral blood, spleen and liver. Moreover, activity of the LGL isolated from peripheral blood lymphocytes was determined using the flow cytometer. As results, LGL were observed in the peripheral blood, spleen and liver. LGL were observed the relatively faintly staining basophilic cytoplasm with granules, and eccentric, often kidney-shaped nuclei in Giemsa stain. Its size was $11{\sim}13{\mu}m$. LGL percentage of the isolated lymphocytes in peripheral blood, spleen and liver were 1.8~2.3%, 1.3~1.4% and 0.87~0.99%, respectively. LGL conjugation rate with YAC-1 cell was shown to be peripheral blood(9.3~10.3 %) > spleen(7.7~8.7%) > liver(5.6~7.0%). The activity of the LGL isolated from peripheral blood lymphocytes in Group I, II and III was 33.7%, 30.5% and 35.4%, respectively. However, all values were not significantly between groups.

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Cytogenetic Study of Diploid and Triploid Marine Medaka, Oryzias dancena (해산송사리, Oryzias dancena 유도 3배체의 세포유전학적 연구)

  • Park, In-Seok;Gil, Hyun Woo;Lee, Tae Ho;Nam, Yoon Kwon;Ko, Min Gyun;Kim, Dong Soo
    • Korean Journal of Ichthyology
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    • v.28 no.4
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    • pp.215-222
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    • 2016
  • Triploidy was induced in the marine medaka, Oryzias dancena by cold shock treatment ($0^{\circ}C$) of fertilized eggs for 30, 45, or 60 min, applied two minutes after fertilization. The triploid genotype was induced by each of the thermal shock regimes tested. The best result was obtained when the eggs were treated for 45 min, which induced triploidy in all the resulting fish. Triploidy was confirmed using chromosomal and flow cytometer analyses, and erythrocyte measurements. The surface areas and volumes of the erythrocytes of triploid fish were significantly larger than those of diploid fish, and their chromosome number (3N=72) was 1.5 times greater that for the diploids (2N=48). Based on a flow cytometer analysis, the triploid fish had approximately 1.5 times the cellular DNA content (2.40 pg/cell) of the diploid specimens (1.61 pg/cell). Data from this study provide the basis for the development of unique models for studying reproductive confinement in transgenic fish.

Flow Visualization in Microchannel Using Confocal Scanning Microscope (공초점 주사현미경을 통한 미세 유로에서의 유동 가시화)

  • Chang Jun Keun;Park Sung-Jin;Kim Jung Kyung;Han Dong Chul
    • Journal of the Korean Society of Visualization
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    • v.1 no.1
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    • pp.28-33
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    • 2003
  • This paper presents the visualization method in which 3-dimensional(3D) microchannel flow can be detected using a confocal scanning microscope. By soft-lithography, we fabricated various Bio-MEMS(Micro Electro-Mechanical System) devices such as a disposable microchip for a flow cytometer and a micro-mixer, which have 3D structures. Injecting aqueous fluorescent solution in the microfluidic devices, we measured the flow in a steady state by the confocal scanning microscope. At first, we explain the principle of the confocal scanning microscope. And then we show the results from 3D visualization of microscopic flow structures using the confocal scanning microscope.

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