• Title/Summary/Keyword: Fluorescence polarization

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SCREENING OF BENZODIAZEPINES IN URINE BY THE IMMUNOASSAY AND QUANTITATION BY GC-NPD METHOD

  • Park, Jongsei;Park, Jeongeum;Park, Myung-Ja
    • Toxicological Research
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    • v.7 no.1
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    • pp.21-27
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    • 1991
  • We developed a simple method to determine benzodiazepines in biological samples using electron capature detectors and nitrogen-phosphorous detectors. The extraction of 13 benezodiazepines in urine at pH 9.5 with toluene and its analysis in GC/NPD showed the peaks in 9-16 min. In this retention time range, the biological backaground was fairly low and the drugs could be identified in low concentrations. The benzodiazepines in urine samples were screened by the fluorescence polarization immunoassay and positive samples were confirmed by the GC/NPD method.

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Effect of Genistein on Antioxidative Defence System and Membrane Fluidity in Chick Skeletal Muscle Cells

  • Jiang, Z.Y.;Jiang, S.Q.;Lin, Y.C.;Ma, X.Y.;Xi, P.B.;Cao, T.;Wang, X.Q.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.8
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    • pp.1220-1225
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    • 2008
  • This study was conducted to investigate the protective effect of genistein on the antioxidative defence system and membrane fluidity in chick skeletal muscle cells after supplementation with 0, 20, 40, and $80{\mu}mol/L$ genistein in $50{\mu}mol/L$ $FeSO_4/H_2O_2$ treated cells for 24 h. Genistein supplementation recovered the decreased activity of total superoxide dismutase induced by $FeSO_4/H_2O_2$, significantly increased glutathione peroxidase activity (p<0.05) and decreased malondialdehyde production (p<0.05). The treatment of 80 mol/L genistein in $FeSO_4/H_2O_2$ treated cells decreased the secretion of creatine kinase (p<0.05). Fluorescence polarization values and microviscosities observed with $FeSO_4/H_2O_2$ treated cells were significantly higher than those observed with no $FeSO_4/H_2O_2$ treated cells. The addition of $80{\mu}mol/L$ genistein improved the increased fluorescence polarization value (p<0.05) caused by $FeSO_4/H_2O_2$ treatment. The microviscosity value was significantly decreased by adding genistein (p<0.05). In conclusion, genistein protected skeletal muscle cells from oxidative damage by improving antioxidative status and membrane fluidity.

Optimization of SNP Genotyping Assay with Fluorescence Polarization Detection

  • Cai Chun Mei;Van Kyujung;Kim Moon Young;Lee Suk-Ha
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.50 no.5
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    • pp.361-367
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    • 2005
  • Single nucleotide polymorphisms (SNPs) are valuable DNA markers due to their abundance and potential for use in automated high-throughput genotyping. Numerous SNP genotyping assays have been developed. In this report, one of effective and high throughput SNP genotyping assays, which was named the template-directed dye-terminator incorporation with fluorescence polarization detection (FP-TDI) was described. Although the most of this assay succeed, the objective of this work was to deter­mine the reasons for the failures, find ways to improve the assay and reduce the running cost. Ninety $F_2$-derived soybean, Glycine max (L.) Merr., RILs from a cross between 'Pureunkong' and 'Jinpumkong 2' were genotyped at four SNPs. FP measurement was done on $Victot^3$ microplate reader (perkinelmer Inc., Boston, MA, USA). Increasing the number of thermal cycles in the single-base extension step increased the separation of the FP values between the products corresponding to different genotypes. But in some assays, excess of heterozygous genotypes was observed with increase of PCR cycles. We discovered that the excess heterozygous was due to misincorporation of one of the dye­terminators during the primer extension reaction. After pyrophosphatase incubation and thermal cycle control, misincoporation can be effectively prevented. Using long amplicons instead of short amplicons for SNP genotyping and decreasing the amount of dye terminator and Acyclopol Taq polymerase to 1/2 or 1/3 decreased the cost of the assay. With these minor adjustments, the FP-TDI assay can be used more accurately and cost-effectively.

Membrane-Ordering Effects of Barbiturates on Pure Phospholipid Model Membranes

  • Knag, Jung-Sook;Chung, Young-Za;Cho, Goon-Jae;Byun, Won-Tan;Yun, Il
    • Archives of Pharmacal Research
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    • v.15 no.3
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    • pp.196-203
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    • 1992
  • Intramolecular excimer formation of 1, 3-di(1-pyrenyl)propane (Py-3-Py) and fluorescence polarization of 1, 6-diphenyl-1, 3, 5-hexatriene (DPH) were used to investigate the effects of barbiturates on the fluidity of model membranes of phosphatidycholine (SPMVPC), phosphatidylserine (SPMVPS), and phosphatidylinositol (SPMVPI) fractions of synaptosomal plasma membrane vesicles (SPMV) isolated from bovine cerebral cortex. In a dose-dependent manner, barbiturates decreased the excimer to monomer fluorescence intensity ratio (I'/I) of Py-3-Py and increased the anisotropy(r), rotational relaxation time (P), limiting anisotropy $(r_infty)$, and order parameter (S) of DPH in SPMVPC, SPMVPS and SPMVPI. This indicates that barbiturates decreased both the lateral and rotational diffusion of the probes in SPMVPC, SPMVPS and SPMVPI. The relative potencies of barbiturates in ordering the membranes were in the order: pentobarbital > hexobarbital > amobarbital > phenobarbital. This order correlates well with the anesthetic potencies of barbiturates and the potencies for enhancement of $\gamma$-aminobutyric acid-stimulated chloride uptake. Thus, it is strongly suggested that a close relationship might exist between the membrane ordering effects of barbiturates and the chloride fluxes across SPMV.

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Fabrication of Two-dimensional Photonic Crystal by Roll-to-Roll Nanoreplication (롤투롤 나노 복제 공정을 이용한 이차원 광결정 소자의 제작)

  • Kim, Young-Kyu;Byeon, Euihyeon;Jang, Ho-Young;Kim, Seok-Min
    • Journal of the Korean Society of Manufacturing Process Engineers
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    • v.12 no.5
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    • pp.16-22
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    • 2013
  • A two-dimensional photonic crystal structure was investigated using a roll-to-roll nanoreplication and physical vapor deposition processes for the inexpensive enhanced fluorescence substrate which is not sensitive to the polarization directions of excitation light source. An 8 inch silicon master having nano dot array with a diameter of 200 nm, a height of 100 nm and a pitch of 400 nm was prepared by KrF laser scanning lithography and reactive ion etching processes. A flexible polymer mold was fabricated by flat type UV replication process and a deposition of 10 nm nickel layer as an anti-adhesion layer. A roll mold was prepared by warping the flexible polymer mold on an aluminum roll base and a roll-to-roll UV replication process was carried out using the roll mold. After the deposition of ~ 100 nm $TiO_2$ layer on the replicated nano dot array, a 2 dimensional photonic crystal structure was realized with a resonance wavelength of 635 nm for both p- and s-polarized light sources.

Implementation of paper-based Ion concentration polarization phenomenon and Bio-sensor of commercialization concept (페이퍼기반 이온 농도 분극 현상 구현 및 상용화 컨셉의 바이오 센서)

  • Han, Sung Il;Kwak, Rhokyun;Lee, Jeong Hoon
    • Proceedings of the KIEE Conference
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    • 2015.07a
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    • pp.1219-1220
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    • 2015
  • 마이크로, 나노유체 (micro-, nanofluidics)을 이용한 종이 기반 분석 소자 (paper-based analytical devices, micro-PADs)에 대한 관심이 증가하고 있다. 종이 기반의 분석 소자는 초저가의 비용과 간단한 공정 방법으로 인하여 상용화 컨셉의 바이오센서로 각광받고 있다. 하지만, 종이 기반의 분석 소자는 낮은 검출 한계 (limit of detection, LOD)와 민감도 (sensitivity)의 제한이 있다. 그로 인해 우리는 이온 선택적 투과층 (ion permselective membrane, i.e. Nafion)을 종이 기반의 분석 소자와 결합하여 이온 농도 분극 (ion concentration polarization, ICP) 현상을 구현하여 낮은 검출한계와 민감도를 개선할 수 있었다. 접착력이 있는 테이프 표면에 이온 선택적 투과층을 패터닝 (patterning)하여 종이 기반 분석 소자와 결합하여 매우 간단하게 소자를 제작할 수 있었다. 따라서 종이 기반의 채널 양단에 직류 전압을 인가했을 때 발생하는 ICP 현상으로 인하여 형광 물질 (fluorescence dye)이 농축(preconcentration)되는 것을 확인할 수 있었다. 구체적으로, 초기 농도가 1.55 nM인 형광 물질을 이용하여 200 V의 외부전압을 인가했을 때, 500 초 이내에 1000 배 이상의 농축비를 얻을 수 있었다. 따라서, 외부 전압을 상용화된 건전지 출력값으로 낮출 수 있다면 다양한 종이 기반 분석 소자와 간단한 결합 방법을 통해 상용화 컨셉의 바이오센서로도 구현이 가능할 것이다.

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Epitaxial growth of oxide films using miscut substrates (Miscut된 기판을 이용할 산화물 박막의 에피 성장)

  • Bu Sang Don
    • Journal of the Korean Vacuum Society
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    • v.13 no.4
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    • pp.145-149
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    • 2004
  • We have grown piezoelectric oxide films by RF magnetron sputtering using miscut substrates. Films were Brown on(001) $SrTiO_3$ substrates with miscut angles from 0 to 8 degrees toward the (100) direction. Films on high miscut substrates (>$4^{\circ}$) showed almost the pure perovskite phase in x-ray diffraction and were nearly stoichiometric. In contrast, films on exact (001) $SrTiO_3$ contained a high volume fraction of pyrochlore phases. A film on an $8^{\circ}$ miscut substrate exhibits a polarization hysteresis loop with a remnent polarization of 20$\mu$C/$\textrm{cm}^2$ at room temperature.

Development of an Alignment Method for Retarders in isoSTED Microscopy

  • Ilkyu Park;Dong-Ryoung Lee
    • Current Optics and Photonics
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    • v.8 no.4
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    • pp.421-426
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    • 2024
  • The use of stimulated emission depletion (STED) microscopy has significantly improved resolution beyond the limits imposed by diffraction; Furthermore, STED microscopy adopts a 4Pi-geometry to achieve an isotropic improvement in resolution. In isoSTED microscopy, a polarizing beam splitter and retarders are used in a 4Pi cavity to split beams of identical power, generating constructive and destructive interference for lateral and axial resolution improvements, respectively. The precise alignment of the retarders is crucial for optimizing the performance of isoSTED microscopy, because this orientation affects the quality of the depletion focus, necessitating zero intensity at the center. Incomplete destructive interference can lead to unwanted fluorescence inhibition, resulting in degraded resolution and contrast. However, measuring the intensity and polarization state in each optical path of the 4Pi cavity is complex and requires additional devices such as a power meter. Here, we propose a simple and accurate alignment method for the 4Pi cavity in isoSTED microscopy. Our approach demonstrates the equal allocation of power between upper and lower beam paths and achieves complete destructive interference using a polarizing beam displacer and a single CCD camera positioned outside the 4Pi cavity.

Spectroscopic Studies of trans-1,2-Bispyrazylethylene (트란스-1,2-비스피라질에틸렌에 대한 분광학적 연구)

  • Sang Chul Shim;Han-Young Kang
    • Journal of the Korean Chemical Society
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    • v.22 no.4
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    • pp.239-244
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    • 1978
  • Spectroscopic studies of trans-1,2-bispyrazylethylene (BPE), one of the stilbene analogues, were carried out. In normal UV spectra, a distinct $n{\rightarrow}{\pi}^*$ absorption band is missing because of a strong, nearly isoenergetic ${\pi}{\rightarrow}{\pi}^*$ absorption band. The second derivative and low temperature $(77^{\circ}K)$ UV absorption spectra were taken and $n{\rightarrow}{\pi}^*$ absorption band was identified by these methods. The transition energies of ${\pi}{\rightarrow}{\pi}^*$ transitions were calculated by Pariser-Parr-Pople (PPP)-SCF-CI MO method. The calculated values showed good agreement with the observed spectral data. Luminescence studies were also carried out at low temperature. From the fluorescence spectra, fluorescence polarization studies, and PPP-SCF-CI MO calculation, the fluorescent state was determined to be a singlet $({\pi},\;{\pi}^*)$ state. This conclusion is in good agreement with the results obtained from alkaline salt effects on the fluorescence of this compound.

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Blood Glucose Measurement Principles of Non-invasive Blood Glucose Meter: Focused on the Detection Methods of Blood Glucose (무채혈 혈당 측정기의 혈당 측정 원리: 혈당 검출방법 중심으로)

  • Ahn, Wonsik;Kim, Jin-Tae
    • Journal of Biomedical Engineering Research
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    • v.33 no.3
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    • pp.114-127
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    • 2012
  • Recent technical advancement allows noninvasive measurement of blood glucose. In this literature, we reviewed various noninvasive techniques for measuring glucose concentration. Optical or electrical methods have been investigated. Optical techniques include near-infrared spectroscopy, Raman spectroscopy, optical coherence technique, polarization, fluorescence, occlusion spectroscopy, and photoacoustic spectroscopy. Electrical methods include reverse iontophoresis, impedance spectroscopy, and electromagnetic sensing. Ultrasound, detection from breath, or fluid harvesting technique can be used to measure blood glucose level. Combination of various methods is also promising. Although there are many interesting and promising technologies and devices, there need further researches until a commercially available non-invasive glucometer is popular.