• 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.

• Journal of Life Science
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• v.19 no.5
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• pp.620-624
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• 2009

D-Ala2-Leu5-enkephalin (DADLE), a hibernation inducer, can induce hibernation-like state in vivo and in vitro. We treated U937 human leukemia cancer cells with DADLE and investigated its possible effect on transcription and proliferation. Treatment of U937 cells with DADLE resulted in growth inhibition and induction of apoptotic cell death on high-dose as measured by MTT assay and DNA flow cytometer analysis. Bcl-XL, c-IAP-2 and survivin genes especially showed decreases in mRNA levels. DADLE treatment also inhibited the levels of cyclooxygenase (COX)-2 mRNA without alteration of COX-1 expression. DNA flow cytometer analysis revealed that DADLE caused arrest of the cell cycle on low-dose, which was associated with a down-regulation of cyclin E at the transcriptional level. DADLE treatment induced a marked down-regulation of cyclin-dependent kinase (Cdk)-2, -4 and -6. In addition, treatment with DADLE decreased telomere associated genes such as, c-myc and TERT, and increased TEP-1 in U937 cells. These results suggest that DADLE can be an inhibition agent in the cell cycle of the human leukemia cancer U937 cell.

• Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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• 2019.05a
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• pp.278-280
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• 2019

Flow cytometer is one of the instrument that can measure various optical properties of a single cell or microparticle. These parameters including size, granularity, and fluorescence intensity are determined by the physical and optical interaction of the cells with excitation light source. However, users have some difficulties such as high cost, size of instrument, and limited fluorescence selectivity. In addition, abundant data is also unintentionally acquired even though user wants to have a single optical parameter. For these reasons, the use of flow cytometer is more challenging for researchers to apply their study. Therefore, the proposed study aims to develop a low-cost portable fluorescence acquisition system using a commercially available light-emitting diode and photodiode. It is designed by a 3D printer, and fluorescence selectivities are increased by changing of the light source / optical filter / detection sensor. Various number sets of fluorescently labeled cells were measured, and its feasibility was evaluated through the proposed system. As a result, acquried fluorescence intensities were proportional to the concentration of the cells and showed high linearity.

• International Journal of Oral Biology
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• v.30 no.2
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• pp.65-69
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• 2005

Periodontitis is a multi-microbial disease and the comparison of a series of periodontopathogenic and non-periodontopathogenic bacteria in terms of microbe-host interaction may provide clues to understand the microbial etiology of the disease better. When we deal with twenty different bacterial species in a study, the first technical issue is how to measure the accurate concentration and use the same number of bacterial cells. We measured bacterial concentration by enumerating bacteria stained with SYTOX green for constant time using a flow cytometer and compared the results with those obtained by plate counting. Concentrations calculated by two different methods were very close. Therefore, flow cytometric counting allowed the rapid analysis of live/dead bacteria, offering the advantage of turbidity measurement and that of colony counting together.

• Korean Journal of Veterinary Research
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• v.32 no.4
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• pp.649-661
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• 1992

This study was performed to compare DNA content by flow cytometer (FCM) and glutathione S-transferase placental form (GST-P) positive foci for searching objective and accurated properties of tumor. Sprague-Dawley rats aged six weeks were divided into three groups and group 1 and 2 of rats were given an intraperitoneal injection of diethylnitrosamine at 200mg/kg body weight and group 3 of rats were given saline. Three weeks after beginning of the experiment, all groups were performed partial hepatectomy. Group 1 of rats were begun to feed on diets containing 0.02% 2-acetylaminofluorene as a promoter for six weeks, group 2 and 3 of rats were begun to feed on basal diets. At 4, 6, and 8 weeks after initiation, all groups of rats were killed, livers were extracted for H & E stain, immunohistochemical stain, and DNA ploidy analysis. In quantitative analysis for GST-P positive lesion number and area by using Image Analyzer, group 1 and 2 represented significant difference in comparison with group 3. In ploidy distribution, diploid cells of group 1 and 2 were increased significantly in comparison with those of group 3 at 4, 6, and 8 weeks after initiation, respectively tetraploid cells were reduced. But S-phase cells were not changed significantly. It is concluded that ploidy change by FCM is useful as objective data for early detection in hepatocarcinogenesis. Therefore, methodology and study of DNA content are carried out for more objective and accurate ploidy analysis in liver tumor.

• Toxicological Research
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• v.14 no.4
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• pp.475-481
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• 1998

This study was performed to investigate the modifying effect of the general (GPA) and the fermented pilose antler (FPA) on experimental hepatocarcinogenesis and Natural Killer cell activity in rats. Specific pathogen free, 5-week male Sprague-Dawley rats were divided into four groups. To induce hepatocarcinogenesis, diethylnitrosamine (DEN, 200 mg/kg, i.p.) was used as a tumor initiator and was given in a single dose at experimental onset. All rats were given a partial hepatectomy (PH) at 3 weeks after experimental onset. Sodium phenobarbital (PB, 0.05% in diet), GPA (0.075% in diet) and FPA (0. 075% in diet) were given from 2 to 8 weeks. Group I of the initiation control group was only given DEN. As initiation-promotion group, Group II was given DEN and then PB. Group III and IV were given DEN-PB-GPA and DEN-PB-FPA, respectively. In hematological analysis, as compared with Group I. the number of white blood cells were significantly increased in the GPA (p<0.01) and the FPA treated group (p<0.05), respectively. Natural killer (NK) cell activity by flow cytometer (FCM) analysis was higher in group of treated with the GPA (35%) than that of the FPA (27.5%), but not significant. Result of the immunohistochemical staining of the glutathione S-transferase placental form (GST-p) indicated that the number of and area of the pre-neoplastic lesions was not significantly changed in Group III and IV compared Group II, respectively. In conclusion, the GPA and the FPA treatment significantly increased the number od WBC in peripheral blood, but the enhancing NK activity and the modifying effect on the experimental hepatocarcinogenesis were not observed.

• Ocean and Polar Research
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• v.26 no.2
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• pp.273-286
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• 2004

The effects of environmental forcing on autotrophic picoplankton distributional patterns were investigated for convergence ($5^{\circ}N$), divergence ($9^{\circ}N-10^{\circ}30'N$) and oligotrophic ($17^{\circ}N$) sites in the tropical eastern Pacific during 2001 and 2003 KODOS (Korea Deep Ocean Study) cruises. The distributions of picoplankton populations - Prochlorococcus, Synechococcus and picoeukaryotes algae - were determined by flow cytometric analyses. Latitudinal variations in abundance maxima, vertical profiles, integrated abundance (0-150 m), and estimated carbon biomass were contrasted for each site according to three hydrological conditions. Prochlorococcus showed consistently high abundance in the surface mixed layers of all sites at $1\;{\times}\;10^5{\sim}3\;{\times}\;10^5\;cells\;ml^{-1}$ and showed declining abundance below these layers. However, these decreasing rates were not particularly sharp showing considerably high abundance at $1\;{\times}\;10^4\;cells\;ml^{-1}$ or higher even at 100 m depth. Vertical profiles of Synechococcus and picoeukaryotes were generally parallel to each other in all sites. A clear abundance maximum was observed at divergence site at or slightly above the pycnocline depth. Higher abundance was observed at the surface mixed layer for convergence site but a sharp decrease was observed below the pycnocline. However, there was no significant abundance fluctuation with depth at more oligotrophic site ($17^{\circ}N$). Integrated cell abundance of Prochlorococcus was high in the oligotrophic site at $2.17\;{\times}\;10^{13}\;m^{-2}$, and low in the convergence site at $0.88\;{\times}\;10^{13}\;m^{-2}$. However, opposite pattern was observed for Synechococcus and picoeukaryotes where relatively high integrated cell abundance was shown in the convergence site. Estimated carbon biomass of Prochlorococcus contributed 30.4-80.3% of total autotrophic picoplankton carbon showing the highest contribution in the oligotrophic site and the lowest contribution in the convergence site. Synechococcus contribution of total autotrophic picoplantkon carbon biomass was lower than 5.8% for most of sites except the convergence site where Synechococcus contributed 23.2% of picoplankton carbon biomass. Carbon biomass of picoeukaryotes was 18.8-46.4% showing the highest carbon biomass at the convergence site. Overall, Prochlorococcus showed higher cell abundance and carbon biomass and exhibited different reaction to hydrological conditions when compare with the other two major autotrophic picoplankton groups.

• Korean Journal of Clinical Laboratory Science
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• v.49 no.4
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• pp.382-389
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• 2017

Currently, reticulocyte experimental calculation technology used in clinical laboratories are divided two types: manual and automated. Manual reticulocyte counting using a microscopy lacks accuracy due particularly to its low reproducibility, affecting the accuracy of manual reticulocyte count. Moreover, Automatic blood corpuscle analyzer flow cytometry is difficult to be used in underdeveloped countries and small scale laboratories due to relatively high cost. Therefore, this study tried to find a new method to complement these drawbacks. The aim of this study was to compare the stained reticulocytes count by spectrophotometer and also to analyze the statistics of spectrophotometer and flow cytometer. The same 8 EDTA samples were repeated 36 times to compare the agreement between spectrophotometer and flow cytometer. This study measured the specimen diluted 600 times at 700~780 nm by 10 differences. Wavelength between 710 to 730 by absorbance showed a positive correlation between standard data and test data (r=0.967, p<0.01), presenting a correlation between variables. Statistical analyses of regression for test and standard parametric data, the optimal dilution factor was 600 times. Therefore, this study tried to technical utilizes such as contributing economical for the reticulocyte absorbance apply from the auto spectrophotometer, a monitoring system for the reticulocyte relation anemia, etc. Therefore, more extensive studies, including an auto chemical analyzer application, will be needed.

• BMB Reports
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• v.29 no.2
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• pp.127-132
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• 1996

Germinal centers (GCs) are formed in peripheral lymphoid tissues in response to protein antigens. In order to see if immunoglobulin isotype switching takes place in GC B-cells, we isolated GC B-cells (PNA positive cells) from mouse popliteal lymph nodes by a flow cytometer after the staining of lymph node cells with PNA-FITC and anti-B220-PE, and determined the expression of ${\gamma}1$ germline transcript and ${\gamma}1$ mRNA by RT-PCR. ${\gamma}1$ germline transcript and ${\gamma}1$ mRNA were amplified specifically in cDNAs from hybridoma expressing IgG1 or splenocytes stimulated LPS plus IL-4. Germinal center B-cells formed in popliteal lymph nodes of mice immunized with chicken ovalbumin were isolated 7 days after immunization. We sorted GC B-cells five times. Immunoglobulin ${\gamma}1$ germline transcripts were expressed in germinal center B-cells in three out of five sorts whereas two out of five sorts did not express ${\gamma}1$ germline transcripts in GC B-cells. The contents of GC B-cells ranged from 5 to 7% of total lymph node cells in most flow cytometric analyses but those of two sorted cells which did not express ${\gamma}1$ germline transcripts were out of normal range. These results imply that isotype switching of immunoglobulins may take place in GCs.

• Korean Journal of Pharmacognosy
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• v.28 no.4
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• pp.286-292
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• 1997

We studied the screening condition for immune regulatory responsor. We focused on the T-lymphocytes leer this purpose. Mouse fetal thymic organ culture (FTOC) system and flow cytometric analysis were mainly used in this experiment. Even if FTOC is carried out in vivo condition, the pattern of thymic development in the condition of FTOC is similar to that of in vivo condition. In this regard, FTOC system might be very powerful tool to screen the immune regulator, especially concerning on T cells. To establish the optimum condition of FTOC to screen the Immune regulator, we focused on the optimum amount of dose and culture period. The cell number and surface antigens on T cells were also analysed by using hemacytometer and flow cytometer. To monitor the differentiation event, anti-CD3, anti-CD4 and anti-CD8 antibodies were used. Alkoxyglycerol and Phellodendri Cortex were used fur positive and negative control, respectively. Astragalus membranceus was used as test sample. From our analysis, we reached to conclusions that the best dose of extract is $50\;{\mu}g/ml$ of culture medium, the best culture period is for 9 days, and ethanol used as solvent has no toxicity to FTOC.