• Title/Summary/Keyword: Fluorophore

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Research Method of Fatty Acids Transfer between Phospholipid Model Membranes (인지질 모델막에서의 지방산 이동에 관한 연구 방법)

  • 임병순;김혜경;김을상
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.4
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    • pp.743-750
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    • 1997
  • Direct measurement of the kinetics of free fatty acid transfer between phospholipid model membrane is technically limited by the rapid nature of the transfer process. Separation of membrane-bound fatty acid by centrifugation has shown that although the equilibrium distribution of free fatty acid is determined by this method, fatty acid transfer occurs too rapidly for accurate kinetic measurements. Recently fluorescence resonance energy transfer(FRET) assay has been developed to examine transfer of fatty acids between membranes. Donor membranes which has fluorescent fatty acid, anthroyloxy fatty acid(AOFA), is mixed with acceptor membranes which has non-interchangeable fluorescent quencher, nitrobenzo-xadiazol(NBD), using stopped flow apparatus. As the fluorescent fatty acids transfer from donor membrane to acceptor membrane, fluorescence intensity would be decreased and the rate and degree of fatty acid transfer can be analyzed. Fatty acid transfer between micelles is more complicated because of bile salt. Therefore in experiments with micelles, fluorescence self quenching assay is used. At high concentrations, a fluorophore tends to quench its own fluorescence causing a reduction in fluorescence intensity. Donor micelles contained self quenching concentrations of fluorophore and acceptor micelles had no fluorophore. Upon mixing of donor and acceptor micelles, the rate of transfer of the fluorophore from the donor to the acceptor was measured by monitoring the release in self quenching when its concentration in donor decreased over time.

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Spectrofluometry of sulfonamides using fluorescamine (Fluorescamine을 이용한 sulfonamide의 형광 분석법)

  • Lim, Jae-young;Han, Su-nam;Lee, Mun-han;Park, Jong-myung
    • Korean Journal of Veterinary Research
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    • v.30 no.1
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    • pp.41-49
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    • 1990
  • These experiments were carried out to develop a novel, simple, and rapid method to determine urinary sulfonamides using fluorescamine by spectrofluorometry. To get optimal conditions for the sulfonamide-fluorescamine reaction, sulfonamides such as sulfamethazine, sulfamerazine, sulfadimethoxine and sulfamonomethoxine, dissolved in buffers with various pH ranges were reacted with various concentrations of fluorescamine. and then, the fluorescence intensity and stability of the fluorophore were measured. To eliminate the interfering substances in urine, the fluorophore in buffers and urine with a definite pH range was extracted with some organic solvents. After then the fluorescence intensity was measured in organic and aquous phases. The results obtained were summarized as follows: 1. The maximal fluorescence of sulfonamides was presented in acidic state, pH 4.5~5.0, at 30 minutes after reaction. 2. The optimal concentration ratio of sulfamethazine and ffuorescamine was more than 1 : 40 in mole. 3. In pH 4.0, the intensity was maximal but was time-dependent, whereas in pH 8.0, the intensity was time-independent. 4. Sulfamethazine-fluorescamine conjugate could be dissolved in some of organic solvents in acidic state such as chloroform, n-butanol, and ethylacetate. 5. Sulfamethazine-flnorescamine conjugate in swine urine coule be extracted with ethylacetate in acidic state, pH 4.0~5.0.

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Luminescence Intensity Change Using N-Carbamoylglycine, N-Salicylideneaniline and Metal ions (N-Carbamoylglycine 및 N-Salicylideneaniline과 Metal ions들에 의한 발광 세기의 변화)

  • Kim, Ji Ung;Kim, Yeong Hae
    • Journal of the Korean Chemical Society
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    • v.46 no.6
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    • pp.502-508
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    • 2002
  • We have used PET chemosensors in the determination of N-carbamoylglycine. When N-carbam-oylglycine reacts with complex already made by the fluorophore and metal ion, the luminescence intensity can be changed and this phenomenon can be utilized in quantification. We used three metal ions, $Zn^{2+}$, $Ni^{2+}$, $Cu^{2+}$ and in order to investigate selectivity an acetic acid was used. $Ni^{2+}$ ion showed change in the eT mechanism by the anions. $Cu^{2+}$ ion showed the ability to distinguish N-carbamoylglycine from an acetic acid and it is noteworthy that $Zn^{2+}$ ion can change luminescence sensitively according to concentration.

Rhodamine derivative synthesis: dual-detectable chemosensor

  • Son, Young-A;Kim, Hyung-Joo;Park, June-Min;Lee, A-Reum;Lee, Jun-Hee;Lee, Do-Hyun
    • Proceedings of the Korean Society of Dyers and Finishers Conference
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    • 2012.03a
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    • pp.29-29
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    • 2012
  • Rhodamine chromophore/fluorophore have been attracted to many researchers due to its excellent photophysical properties. In this study, we have designed and synthesized a strong emissive fluorescent dye chemosensor for toxic elements. A rhodamine-based sensor was prepared by incorporation the rhodamine fluorophore and several functional host groups with high affinity to hazardous metal and anion. This sensor shows a high selectivity and an excellent sensitivity and is a dual-responsive colorimetric and fluorescent metal/anion-specific sensor. In addition, the 1:1 binding mode was proposed based on Job's plot method. Finally, computational calculation was simulated and calculated to approach for HOMO/LUMO of this dye chemosensor.

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Super-Resolution Optical Fluctuation Imaging Using Speckle Illumination

  • Kim, Min-Kwan;Park, Chung-Hyun;Park, YongKeun;Cho, Yong-Hoon
    • Proceedings of the Korean Vacuum Society Conference
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    • 2014.02a
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    • pp.403.1-403.1
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    • 2014
  • In conventional far-field microscopy, two objects separated closer than approximately half of an emission wavelength cannot be resolved, because of the fundamental limitation known as Abbe's diffraction limit. During the last decade, several super-resolution methods have been developed to overcome the diffraction limit in optical imaging. Among them, super-resolution optical fluctuation imaging (SOFI) developed by Dertinger et al [1], employs the statistical analysis of temporal fluorescence fluctuations induced by blinking phenomena in fluorophores. SOFI is a simple and versatile method for super-resolution imaging. However, due to the uncontrollable blinking of fluorophores, there are some limitations to using SOFI for several applications, including the limitations of available blinking fluorophores for SOFI, a requirement of using a high-speed camera, and a low signal-to-noise ratio. To solve these limitations, we present a new approach combining SOFI with speckle pattern illumination to create illumination-induced optical fluctuation instead of blinking fluctuation of fluorophore.. This technique effectively overcome the limitations of the conventional SOFI since illumination-induced optical fluctuation is possible to control unlike blinking phenomena of fluorophore. And we present the sub-diffraction resolution image using SOFI with speckle illumination.

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Physicochemical Properties of Recominant Hepatitis B Surface Antigen Expressed in Mammalian Cell(C127)

  • Lee, Young-Soo;Kim, Byong-Kak;Choi, Eung-Chil
    • Archives of Pharmacal Research
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    • v.21 no.5
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    • pp.521-526
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    • 1998
  • The physicochmical properties of recombinant hepatitis B surface antigen (r-HBsAg), which was expressed in C127 mammalian cell were studied. Using roller bottle culture in DMEM supplemented with fetal bovine serum, 10-15 mg/L of r-HBsAg was produced with about 31% of purification yield. The purity of r-HBsAg by HPLC was 99.8% and electron microscopic examination showed homogeneous spherical particle with 22 nm in diameter, a morphological characteristic of HBsAg. The density of r-HBsAg by CsCI density gradient method was 1.19g/ml and the isoelectric point by Mono $P^{TM}$ HR 5/20 column was 4.6. The analysis of subunit protein pattern using SDS-PAGE followed by scanning densitometry gave 81.3% of S protein and 18.7% of pre-S protein. fluorophore-assisted-carbohydrate-electrophoresis analysis showed the relative amount of carbohydrate to protein was 1.7% and it smajr component was N-acetyl glucosamine, which was about 39% of total carbohydrate. The relative amount of lipid to protein determined by vanillin phosphoric acid method was 32.5% and its major component was phospholipid, which was about 70% of total lipid. The physicochemical properties of C127 mammalian cell-derved r-HBsAg are similar to those of p-HBsAg, suggesting that the r-HBsAg can be used in developing a new preventive vaccine against hepatitis B.

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