• Title/Summary/Keyword: spectrofluorometer

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Use of Ratiometric Probes with a Spectrofluorometer for Bacterial Viability Measurement

  • Cleach, Jerome;Watier, Denis;Le Fur, Bruno;Brauge, Thomas;Duflos, Guillaume;Grard, Thierry;Lencel, Philippe
    • Journal of Microbiology and Biotechnology
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    • v.28 no.11
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    • pp.1782-1790
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    • 2018
  • Assessment of microorganism viability is useful in many industrial fields. A large number of methods associated with the use of fluorescent probes have been developed, including fluorimetry, fluorescence microscopy, and cytometry. In this study, a microvolume spectrofluorometer was used to measure the membrane potential variations of Escherichia coli. In order to estimate the sensitivity of the device, the membrane potential of E. coli was artificially disrupted using an ionophore agent: carbonyl cyanide 3-chlorophenylhydrazone. The membrane potential was evaluated using two ratiometric methods: a Rhodamine 123/4',6-diamidino-2-phenylindole combination and a JC-10 ratiometric probe. These methods were used to study the impact of freezing on E. coli, and were compared with the conventional enumeration method. The results showed that it was beneficial to use this compact, easy-to-use, and inexpensive spectrofluorometer to assess the viability of bacterial cells via their membrane potential.

Fundamental Study on the Quantitative Analysis of Fluorescent Whitening Agent used for Papermaking (제지용 형광증백제의 정량분석에 대한 기초연구)

  • Lee, Ji-Young;Kim, Chul-Hwan;Lee, Hui-Jin;Gwak, Hye-Joeng
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.43 no.2
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    • pp.9-15
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    • 2011
  • Fluorescent whitening agent (FWA) is a widely used chemical in paper industry, but a systematic and scientific method on FWA analysis has not been established. We performed the basic researches on the fluorescence analysis of FWA. The fluorescence of FWA was investigated using a spectrofluorometer and a spectrophotometer. When FWA solution was analyzed using the spectrofluorometer, we found that the peak wavelength of the fluorescence emission was about 440 nm and that of the fluorescence excitation was about 370 nm irrespective of FWA types. Papers dyed with an internal FWA were prepared in a laboratory and the reflectance and the fluorescence index were measured using the spectrophotometer. It was confirmed that the optimum peak wavelength of the reflectance was 440 nm and the fluorescence index calculated from the CIE whiteness with and without UV light under a light source D65 was the best indicator to measure the fluorescence of FWAs exiting in papers.

Analysis of Two-Dimensional Fluorescence Spectra in Biotechnological Processes by Artificial Neural Networks I - Classification of Fluorescence Spectra using Self-Organizing Maps - (인공신경망에 의한 생물공정에서 2차원 형광스펙트럼의 분석 I - 자기조직화망에 의한 형광스펙트럼의 분류 -)

  • Lee Kum-Il;Yim Yong-Sik;Kim Chun-Kwang;Lee Seung-Hyun;Chung Sang-Wook;Rhee Jong Il
    • KSBB Journal
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    • v.20 no.4
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    • pp.291-298
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    • 2005
  • Two-dimensional (2D) spectrofluorometer is often used to monitor various fermentation processes. The change in fluorescence intensities resulting from various combinations of excitation and emission wavelengths is investigated by using a spectra subtraction technique. But it has a limited capacity to classify the entire fluorescence spectra gathered during fermentations and to extract some useful information from the data. This study shows that the self-organizing map (SOM) is a useful and interpretative method for classification of the entire gamut of fluorescence spectral data and selection of some combinations of excitation and emission wavelengths, which have useful fluorometric information. Some results such as normalized weights and variances indicate that the SOM network is capable of interpreting the fermentation processes of S. cerevisiae and recombinant E. coli monitored by a 2D spectrofluorometer.

The effect of octanol on the intracellular Ca2+ increase in submandibular acinar cells evoked by β-adrenergic activation (악하선 선포세포에서 β-아드레날린계 활성화 후 세포내 Ca2+ 농도 증가에 미치는 옥탄올의 효과)

  • Choi, Jeong-Iee
    • Journal of dental hygiene science
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    • v.2 no.1
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    • pp.47-51
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    • 2002
  • The concentration of intracellular $Ca^{2+}$ was measured by spectrofluorometer after rat submandibular acinar cells were loaded with fura-2/AM(fura-2). After isoproterenol and octanol were administered while letting submandibular gland acinar cell placed in a perfusion chamber flow through a standard solution, the changes of $Ca^{2+}$ concentration were measured. When they were administered separately, there showed little changes of intracellular $Ca^{2+}$ concentration. When they were administered at the same time, however the concentration of intracellar $Ca^{2+}$ was shown to increase. When forskolin, an adenylate cyclase activater, was administerd together with octanol the response looked similar to the response of isoproterenol. In case of the extracellular $Ca^{2+}$ was removed by omitting $Ca^{2+}$ in standard solution and treating EGTA, isoproterenol and forskolin does not affect to the concentration of intracellular $Ca^{2+}$. Therefore, it was certified that the increase of intracellular $Ca^{2+}$ was caused from outside the cell. In order to know that the $Ca^{2+}$ influx is related with capacitative entry pathway, godolinium, blocker of that pathway was treated. With the result of that experiment there was no complete control of the increase in the concentration of intracellular $Ca^{2+}$. However, speed and amount of $Ca^{2+}$ increase was comparatively diminished.

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NOTE - Determination of UVA Protection Factor with SPEX SkinSkan

  • Kawada, Akira;Kameyam, Hiroko;Sangen, Yoshiko;Aragane, Yoshinori;Tezuka, Tadashi;Hamagami, Ikuko
    • Journal of Photoscience
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    • v.10 no.2
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    • pp.215-216
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    • 2003
  • SPEX SkinSkan is a newly developed spectrofluorometer that can calculate transmission of UVA on the surface of human skin as a non-invasive method. We have investigated UVA protection factors of Japanese over-the-counter sunscreens using SPEX SkinSkan. This non-invasive method is useful to know UVA protection of sunscreens in vivo.

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Effect of Allergen Removed Rhus Verniciflua Extract on Inhibition of Tumor Metastasis (알러젠 제거 옻나무 추출물이 종양 전이 억제에 미치는 영향)

  • Park, Jae-Hyun;Moon, Goo
    • Journal of Korean Traditional Oncology
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    • v.15 no.1
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    • pp.47-61
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    • 2010
  • Objective : The aim of this present study is to evaluate the inhibitory effect of allergen removed Rhus verniciflua (ARV) on Matrix Metalloproteinase-9 (MMP-9), Matrix Metalloproteinase-2 (MMP-2) which is considered to have a clinically important role in tumor metastasis. Methods : The inhibitory effects of standardized extract of ARV on the MMP-2, MMP-9 were investigated by spectrofluorometer while the inhibitory effects on the active MMP-2, pro MMP-2, pro MMP-9 were investigated by zymography. Antimetastatic effect of standardized extract of ARV was investigated in vitro on human fibrosarcoma cell (HT1080)'s invasion through Matrigel. Results : The standardized extract of ARV showed inhibitory effects on the active MMP-2 (IC50, $1.01{\mu}g$/ml), active MMP-9 (IC50, $2.5{\mu}g$/ml) depending on concentrations which was determined by spectrofluorometer. The standardized extract of ARV showed inhibitory effects on the active MMP-2, pro MMP-2, pro MMP-9 depending on concentrations which was determined by zymography. However its inhibitory effect on pro MMP-9 was relatively weaker rather than active MMP-2, pro MMP-2. The standardized extract of ARV showed inhibitory effects in vitro on human fibrosarcoma cell (HT1080)'s invasion through Matrigel according to concentration. Conclusions : These results indicate that standardized extract of ARV has antimetastatic effect through inhibit again MMP-2, MMP-9. Also its inhibitory effect is more powerful on active MMP-2, pro MMP-2 than on active MMP-9, pro MMP-9. It is necessary to conduct further studies on other MMP families, TIMP, and each component of standardized extract of ARV.

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공액계 벤젠고리를 갖는 배위자에 의해 합성된 발광착체

  • Hwang, Hea-Eun;Kim, Dong-Ho;Bae, Jong-Bum;Ahn, Bong-Hee;Chung, Min-Chel
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2010.06a
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    • pp.151-151
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    • 2010
  • 본 연구에서는 배위자로 공액계 벤젠고리로 연결된 중심금속이 이리듐인 착물화합물과 합성하여 인광재료로서의 가능성을 실험하였다. 새로운 이리듐착물화합물의 화학적 구조를 알아보기 위해 $^1H$-NMR, $^{13}C$-NMR, UV-vis, spectrophotometer를 사용하였으며, 광 물리학적, 전기화학적 특성에 대한 측정은 spectrofluorometer, cyclic voltammetry를 통하여 측정하였다. 이리듐 단핵 및 이핵착체는 589~598nm의 영역에서 발광파장이 확인되었으며, DMSO[$5{\times}10^{-5}$]용액에서 양자효율이 단핵 착물화합물의 경우 0.1, 이핵착물화합물의 경우 0.13으로 나타났다. 이핵 이리듐착물화합물의 전기화학적 특성은 energy gap은 2.45eV, optical band gap은 2.58eV로 계산되었다.

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Biphasic Mechanical Responses of Rat Thoracic Aorta to Irradiation with $250{\sim}500\;nm$ Light (돼지 관상동맥 및 흰쥐 흉부대동맥에서 자외선 및 가시광선 조사시 파장에 따른 기계적 반응과 Cyclic GMP의 농도변화)

  • Kook, Hyun
    • The Korean Journal of Pharmacology
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    • v.31 no.3
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    • pp.285-290
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    • 1995
  • This study was undertaken to define the varying responses of vascular smooth muscle to different wavelengths of ultraviolet radiation and to relate them to the changes in cyclic GMP contents. The ring preparations of rat thoracic aorta with intact or removed endothelium were irradiated with the ultraviolet or visible light (UVR) of wavelengths in step of 10 nm between 250 and 500 nm from xenon lamp of a spectrofluorometer, and the changes in vascular tension were recorded. For cyclic GMP assay, the preparations, pretreated with phenylephrine as in the tension experinents, were frozen after irradiation and homogenated in trichloroacetic acid. The supernatant was extracted with ether and the cyclic GMP contents were measured with radioimmunoassay. In the endothelium-intact preparations, biphasic responses, vasoconstriction (UVR-contraction) followed by vasodilatation (UVR-dilatation), were observed. The maximal UVR-contraction was observed at 320 nm, while the maximal vasodilatation was elicited at 420 nm. In the endothelium-removed rings, however, only vasodilatation was observed, with the maximal vasodilatation taking place at 370 nm. The cyclic GMP contents were not affected by the Irradiation with 320 nm for 30 sec or 1 min in the endothelium-intact preparations, while it was significantly increased by 380 and 420 nm. In the endothelium-removed preparations, UVR of 370 nm markedly increased the cyclic GMP contents. The present study indicates that the increase in cyclic GMP is closely related to vasodilatation induced by UVR of 420 nm in the endothelium-intact or 370 nm in the denuded preparations, whereas it is not involved in the vasoconstriction induced by UVR of 320 nm in the intact rings, and the mechanism leading to UVR-contraction remains to be clarified. These observations suggest that nitric oxide-cyclic GMP system is closely related to the UVR-dilatation in rat aortic preparation, while it is not involved in the UVR contraction.

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Characteristics and thermal stability of SrAl2O4: Eu2+, Dy3+ long afterglow phosphors synthesized solid state reaction and polymerized complex method (고상반응법과 착체중합법으로 합성된 SrAl2O4: Eu2+, Dy3+ 축광성 형광체의 특성 및 열적 안정성 평가)

  • Kim, Tae-Ho;Hwang, Hae-Jin;Kim, Jin-Ho;Hwang, Kwang-Taek;Han, Kyu-Sung
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.26 no.5
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    • pp.193-200
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    • 2016
  • Characteristics of $SrAl_2O_4:Eu^{2+}$, $Dy^{3+}$ phosphorescent phosphors synthesized by solid state reaction and polymerized complex method were comparatively analyzed. In order to evaluate thermal stability of $SrAl_2O_4:Eu^{2+}$, $Dy^{3+}$ phosphorescent phosphors at high temperature, phosphorescent properties of $SrAl_2O_4:Eu^{2+}$, $Dy^{3+}$ were investigated with thermal treatment at $1250^{\circ}C$ under reducing atmosphere, which was the general heat treatment conditions for ceramic manufacturing process. The phosphorescent properties of thermally treated $SrAl_2O_4:Eu^{2+}$, $Dy^{3+}$ phosphors synthesized by solid state reaction and polymerized complex method were investigated. The crystal structure and crystallite size were observed through XRD analysis. Microstructure and particle size of thermally treated $SrAl_2O_4:Eu^{2+}$, $Dy^{3+}$ phosphors were analyzed by SEM and PSA. Photoluminescence and afterglow characteristics of thermally treated $SrAl_2O_4:Eu^{2+}$, $Dy^{3+}$ phosphorescent phosphors were measured by spectrofluorometer.