• Title/Summary/Keyword: Fluorescent Dye

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Metronidazole Reduced Ammonia Toxicity in Human Hep G2 cell and Rat Hepatocytes (Hep G2 세포와 rat 간세포에서 Metronidazole에 의한 암모니아 독성 감소)

  • Kim, Bo-Ae;Kim, Hyun-Jung;Kim, You-Young
    • KSBB Journal
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    • v.23 no.5
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    • pp.381-386
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    • 2008
  • Lipophilic ammonia is toxic gas and can easily diffuse across cell membranes. Excess ammonia is implicated in the pathogenesis of several metabolic disorders including hepatic encephalopathy and may result in the death. The purpose of this study was to clarify the inhibition effect of metronidazole on liver cell damage due to ammonia in human Hep G2 cell and rat hepatocytes. The effects of metronidazole were studied in ammonium chloride treated human Hep G2 cell (75 mM) and rat hepatocyte (100 mM) following $0.1{\mu}M$ metronidazole treatment. In MTZ+AC group, cell viabilities increased prominently and LDH activities decreased over 25% than AC group. Furthermore, ammonia level according to ammonium chloride treatment reduced over 30% and lipid peroxidation as an index of cell membrane damage decreased more than twice. By comparison with control, catalase activity showed more than 30% reduction in AC group while less than 10% reduction in MTZ+AC group, respectively. In addition, MTZ+AC group showed the similar cell structure as control in cell morphology study by using light microscope, and represented fluorescent intensity decrement compared with AC group in fluorescent microscopic study with avidin-TRITC fluorescent dye. And cleaved PARP expression due to ammonia reduced twofold or more in MTZ+AC group. As the results suggest, metronidazole may protect the liver cell by inhibiting cell damages due to ammonia and be used for an effective antagonist of ammonia in hyperammonemia.

Simple Classification of Male Mouse Germ Cells using Hoechst 33258 Staining (Hoechst 33258 Staining을 이용한 웅성 생쥐 성세포의 간편 분류)

  • Kim, Kyoung Guk;Park, Young Sik
    • Journal of Embryo Transfer
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    • v.30 no.3
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    • pp.213-218
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    • 2015
  • In the study for a differentiation and development of spermatogonial cells, the researchers should commonly require a simple, fast and reasonable method that could evaluate the developmental stage of male germ cells without any damage and also relentlessly culture them so far as a cell stage aiming at experimental applications. For developing the efficient method to identify the stage of sperm cells, the morphological characteristics of sperm cells were investigated by staining the cells with blue fluorescent dye Hoechst 33258, and a criterion for male germ cell classification was elicited from results of the previous investigation, then the efficiency of the criterion was verified by applying it to assort the germ cells recovered from male mice in age from 6 to 35 days. As morphological characteristics, spermatogonia significantly differed from spermatocytes in size, appearance and fluorescent patches of nucleus, and spermatids could also be distinguished from spermatozoa by making a difference in the volume and shape of nucleus and the shape and fluorescence of tail. Aforesaid criterion was applicable for classifying in vitro cultured sperm cells by verifying its efficiency and propriety for assorting the stages of testicular germ cells. However, the fluorescent staining showed that germ cells in mouse testis should be dramatically differentiated and developed at 21 days and 35 days of age, which were known as times of sexual puberty and maturity in male mice, respectively. In conclusion, the results indicated that this simple criterion for sperm cell classification using fluorescence staining with Hoechst 33258 may be highly efficient and reasonable for spermatogenesis study.

Studies on Preconcentration and Electrophoretic Mobility of Fluorescent Dyes Depending on Flow Velocity and Concentration in the Electromembrane System (전기막 시스템에서 유속과 농도에 따른 형광염료의 농축 및 전기영동 이동도에 관한 연구)

  • Minsung, Kim;Bumjoo, Kim
    • Applied Chemistry for Engineering
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    • v.34 no.1
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    • pp.45-50
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    • 2023
  • Microfluidic preconcentration technologies, which collect or extract low-abundance analytes in a specific location, have been spotlighted in various applications such as medical and bio-engineering. Here, we conducted extensive studies on the variables to be considered when concentrating target samples based on electrophoresis in an electromembrane system utilizing an ion exchange membrane. Using negatively charged Alexa Fluor 488 and positively charged Rhodamine 6G as fluorescent dyes, we examined the effects of flow velocity of the main channel, channel electrolyte concentration, and applied voltage on sample preconcentration. As a result, it was found that the preconcentration of the target sample occurs much better when the flow velocity is slow and the concentration of the main channel containing the sample is high, given that the channel concentration ratio (main and buffer) is constant. In addition, based on the experimental data acquired in this study, the electrophoretic mobility values of Alexa Fluor 488 and Rhodamine 6G were experimentally calculated and compared.

A Study on Optical Properties of Nanocomposite Composed of Au Nanorods and Organic Dyes (금 나노막대와 유기 염료로 구성된 나노복합체의 광학특성 연구)

  • Kim, Ki-Se;Yoo, Seong Il;Sohn, Byeong-Hyeok
    • Clean Technology
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    • v.20 no.2
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    • pp.141-145
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    • 2014
  • In this study, we studied optical properties on the layer-by-layer (LbL) assemblies consisting of Au nanorods and organic dyes. For this purpose, poly (allylamine hydrochloride), PAH and poly (styrene sulfonate), PSS were selected as ionic polymers and rhodamine B isothiocyanate (RB) was utilized as an organic dye based on its spectral overlap with plasmon band of Au nanorods. In the view point of assembling methods, RB was covalently attached to PAH, then, LbL structure of Au [PSS/PAH]2/PSS/PAH-RB was prepared by sequential coating of PAH, PSS, PAH-RB on Au nanorods. Since the prepared LbL assembly exhibits both plasmonic and fluorescent properties, we studied the mutual nanorod-dye properties by dissolving Au nanorods.

FLUORESCENT LABELLING OF MC3T3 CELL LINE BY 5-(AND-6)-CARBOXY-2', 7'-DICHLOROFLUORESCEIN DIACETATE, SUCCINIMIDYL ESTER MIXED (MC3T3 preosteoblast cell line의 5-(and-6)-carboxy-2',7'-dichlorofluorescein diacetate, succinimidyl ester mixed에 의한 fluorescent labelling)

  • Kook, Min-Suk
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.31 no.6
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    • pp.461-467
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    • 2005
  • Background. 5-(and-6)-carboxy-2',7'-dichlorofluorescein diacetate, succinimidyl ester mixed (CFSE) is the fluorescent labelling agent of living cells and used to trace the cells in vivo after transplatnation of various cells. The CFSE labelled cells can maintain fluorescence for up to 7 days after labelling. The MC3T3-E1 cell line (MC3T3) has been used for many studies about osteoblast, which is well known as a mouse preosteoblast. So the CFSE would be used to trace the transplanted MC3T3. However there are few reports about CFSE labelling of MC3T3. This study is aimed to know about adequate concenturation and incubation time of CFSE to MC3T3. Materials and methods. The MC3T3 was incubated in a humidified atmosphere of 95% air with 5% $CO_2$ at $37^{\circ}C$ using ${\alpha}$-minimal essential medium (${alpha}$-MEM) containing10% FBS and gentamycin. Ten mM CFSE solution in dimethylsulphoxide (DMSO: 1%) was diluted with phosphate buffered saline (PBS) and final concentration of culture medium was, respectively, 5, 10, 15, 20, 25 and 30 ${{\mu}M$. Then the MC3T3 was incubated with CFSE in a humidified atmosphere of 95% air with 5% $CO_2$ at $37^{\circ}C$ for 5, 10, 15, 20, 25, 30, 35, 40 and 45 minutes in each concentration. The fluorescence of CFSE labelled cells was analysed with a inverted fluorescence microscope. The duration of cell labelling was also studied. Trypan blue dye exclusion test was done for cell viability. Results. For concentration between 5 and 10 ${\mu}M$, CFSE did not significantly label the MC3T3 in vitro. The destruction of MC3T3 was observed at the concentration of 20 ${\mu}M$. In the concentration of 15 ${\mu}M$, the best labelling was obtained at an incubation period between 15 and 30 minutes. The MC3T3 labelled with an incubation period of 15 minutes at 15 ${\mu}M$ was still fluorescent 7 days after CFSE labelling. The mean cell viability was 95.93%. Conclusion. These results suggests an incubation period of 15 minutes at 15 ${\mu}M$ of CFSE provides best labelling of MC3T3 in vitro.

Red OLEDs containing the dotted-line doped layer structure in its emitting region.

  • Lee, Chang-Min;Han, Jeong-Whan
    • 한국정보디스플레이학회:학술대회논문집
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    • 2004.08a
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    • pp.612-615
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    • 2004
  • We present an extremely high efficient red organic light-emitting diodes (OLEDs) using a fluorescent dye 4-(dicyanomethylene)-2-tert-butyl-6-(1,1,7,7-tetramethyljulolidyl-9-enyl)-4H-pyran (DCJTB) doped into an emitting region which consists of multiple pairs of a doped and an undoped layer. An emitting region of OLEDs composes of a tris-(8-hydroxyquinolinato) aluminum (Alq3) codoped with rubrene of 5% wt. or a mixture of Alq3 and rubrene (1:1). The luminance yield of the codoped device and the mixed device are 6.5 cd/A and 9.2 cd/A at 10 mA/$cm^2$, respectively. We have considerably improved the luminance yields of red OLEDs as much as ${\sim}$90% at 10 mA/$cm^2$ compared with that of the device doped with only DCJTB. We attribute it to both the emitting assist dopant (rubrene) and the dotted-line doping structure in an emitting region of OLED.

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Efficient orange-red OLED using a new DCM derivative as a doping molecule

  • Hwang, Do-Hoon;Lee, Jong-Don;Lee, Moon-Jae;Lee, Chang-Hee
    • 한국정보디스플레이학회:학술대회논문집
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    • 2004.08a
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    • pp.579-581
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    • 2004
  • A new DCM derivative containing the phenoxazine moiety (DCPXZ) has been synthesized for use as a red fluorescent dye molecule in organic light-emitting diodes (OLEDs). The photoluminescence and electroluminescence properties of DCPXZ were examined. The maximum photoluminescence of DCPXZ in chloroform solution ($10^{-5}$ mol) was observed at 616 nm. EL devices were fabricated with the structure ITO/PEDOT-PSS/Cu-PC(15nm)/${\alpha}$-NPD(45nm)/$Alq_3$:DCPXZ(30nm)/$Alq_3$(30nm)/LiF(0.5nm)/Al. The maximum EL emission for the 2.0% DCPXZ-doped device was at 608 nm with CIE coordinates (0.57, 0.42). The EL device exhibited a maximum brightness of 15,000 cd/$m^2$ at 19.4 V and a power efficiency of 1.04 lm/W at a luminance of 100 cd/$m^2$.

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고삼투압이 재조합 Erythropoietin의 생산과 당쇄구조에 미치는 효과

  • Jeong, Yeon-Tae;Kim, Jeong-Hoe
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.221-224
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    • 2001
  • Effect of hyperosmotic pressure on growth of recombinant Chinese hamster 。 vary cells and Erythropoietin (EPO) production was investigated. Cells were cultivated in batch modes at various osmolalities. When the osmolality increased from 314 to 463mOsm/Kg, specific EPO productivity (qp) was increased up to 1.6-fold but cell growth was inhibited. EPO has a complex oligosaccharide structure that plays an important role in biological activity in vivo. To investigate the influence of hypoerosmotic pressure on the glycosylation, structural analysis of oligosaccharide was calTied out. Recombinant human EPO was produced by CHO cells grown under various osmotic pressure and purified from culture supernatants by heparin-sepharose affinity column and immunoaffinity column. N-linked oligosaccharides were released enzymatically and isolated by paper chromatography. The isolated oligosaccharides were labeled with fluorescent dye, 2-aminobenzamide and analyzed with MonoQ anion exchange chromatography and GlycosepN amide chromatography for the assignment of GU (glucose unit) value. Glycan analysis by HPLC showed that neutral (asialo) oligosaccharide was increased slightly with an increase in osmolality. In portion of sialylated glycan, total relative amount of mono- and di-sialyated glycan was increased but that of tri- and tetra-sialylated glycan decreased as osmolality was increased.

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Surface Temperature Measurements in Microscale with Temperature Sensitive Fluorescence (온도 민감 형광을 이용한 마이크로 스케일 표면온도 측정)

  • Jung, Woon-Seop;Kim, Sung-Wook;Kim, Ho-Young;Yoo, Jung-Yul
    • Proceedings of the KSME Conference
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    • 2004.11a
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    • pp.1225-1230
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    • 2004
  • The effects of substrate material on the local heating performance of microheaters are studied by both numerical analysis and experiment. Transient conduction analysis shows that the substrate material with low thermal conductivity is critical to the local heating and fast response. A measurement technique for surface temperature field in microscale is newly proposed, which uses temperature sensitive fluorescent dye coated on the surface. The measured surface temperature fields on microheater arrays fabricated on different substrates are presented.

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Micro-LIF Measurement in a Micro-channel Using an Micro Laser Light Sheet (마이크로 레이저 평면빔을 이용한 마이크로채널 내에서의 Micro-LIF 측정)

  • Yoon, Sang-Youl;Kim, Jae-Min;Kim, Su-Hun;Kim, Kyung-Chun
    • Proceedings of the KSME Conference
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    • 2004.11a
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    • pp.1540-1545
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    • 2004
  • Measurement of concentration fields in a micro-channel is the crucial technology in the area of Lab-on-a-chip to be used for various bio-chemical applications. It is wel-known that the only possible way to measure the concentration field in the micro-channel is using micro-LIF(Laser Induced Fluorescence) method. However, an accurate concentration field at a given cross plane in a micro-channel has not been made so far due to the limit of light illumination. The present study demonstrates a novel method to provide an ultra thin laser sheet beam having 5 microns thickness by a micro focus laser line generator. Nile Blue A was used as fluorescent dye for LIF measurement. The laser sheet beam illuminates an exact plane of concentration measurement in the micro-channel to increase the signal to noise ratio and reduce the depth uncertainty considerably.

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