• Title/Summary/Keyword: Label-Free

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Avidin Induced Silver Aggregation for SERS-based Bioassay

  • Sa, Youngjo;Chen, Lei;Jung, Young Mee
    • Bulletin of the Korean Chemical Society
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    • v.33 no.11
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    • pp.3681-3685
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    • 2012
  • We developed a simple and effective method for the SERS-based detection of protein-small molecule complexes and label-free proteins using avidin-induced silver aggregation. Upon excitation with light of the appropriate wavelength (633 and 532 nm), the aggregated silver nanoparticles generate a strong electric field that couples with the resonance of the molecules (atto610 and cytochrome c), increasing the characteristic signals of these molecules and resulting in sensitive detection. The detection limit of biotin with the proposed method is as low as 48 ng/mL. The most important aspect of this method is the induction of silver aggregation by a protein (avidin), which makes the silver more biocompatible. This technique is very useful for the detection of protein-small molecule complexes.

The Coordination of Pyridyl-N to Pentacyanoferrate for the Electrochemical Detecting Small Organic Molecules

  • Choi, Young-Bong;Jeon, Won-Yong;Kim, Hyug-Han
    • Bulletin of the Korean Chemical Society
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    • v.34 no.2
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    • pp.595-599
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    • 2013
  • The coordination of pyridyl-N to pentacyanoferrate for the detection of small organic antigens in solution is presented. The unique contribution of this paper is the direct conjugation of pyridyl-N in small organic antigens to pentacyanoferrate. Pentacyanoferrate is promising as an electrochemical label owing to its good electro-chemical properties, which can be utilized to generate an electrical signal in homogeneous electrochemical immunoassays. The facilely synthesized pyridyl-N to pentacyanoferrate was characterized by the electrochemical and spectroscopic methods. Hippuric acid (HA) has been detected competitively on the interaction of free HA and pentacyanoferrate-(4-aminomethylpyridine-hippuric acid) (Fe-HA) to its antibody, with the detection limit of 0.50 ${\mu}g\;mL^{-1}$. While pentacyanoferrate-based immunoassay is in its simplicity and infancy, the proposed immunoassay offers attractive opportunities for developing pyridyl-N-based the electrochemical detection of small organic antigens in the health care area.

Identification of Irradiated Foods by Using DNA, Immunochemical, and Biological Methods

  • Kim, Kyeung-Eun;Yang, Jae-Seung
    • Preventive Nutrition and Food Science
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    • v.4 no.4
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    • pp.276-282
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    • 1999
  • Ionizing radiation is considered to be an efficient technology to improve food safety and to extend food shelf-life in the food industry, and it has been used in food processing with a number of attributes. Food labeling should be established to enable the consumer to choose food freely, based on label information. A variety of methodologies to determine the physical, chemical, microbiological, and biological changes due to irradiation has been investigated in order to discriminate the irradiated and unirradiated food products for the consumer's free choice in food selection. However, no satisfactory method has been developed so far. In this review, various approaches based on DNA, immunochemical, and biological methods are addressed.

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Data analysis for quantitative proteomics research (프로테오믹스 연구를 위한 정량분석 데이터의 해석)

  • Kwon Kyung-Hoon
    • KOGO NEWS
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    • v.6 no.1
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    • pp.24-28
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    • 2006
  • 프로테오믹스는 생물체 안에 포함되어 있는 단백질을 통합적으로 연구한다. 단백질을 동정(Protein identification)하고, 단백질의 상태를 분석(Protein characterization)하며, 단백질의 양적 변화를 관찰(Protein quantitation)한다. 단백질에 대한 분석, 특히 질량분석기에 의해 초고속으로 대량의 단백질 데이터를 생산하는 프테테오믹스의 연구는 정량적인 단백질 발현양상분석의 정확도를 높이고 분석시간을 단축하기 위해 다양한 실험기법과 데이터 분석기법을 동원하고 있다. 1) 단백질의 양적 차이나 양적 변화의 관찰은 바이오마커를 발굴하고 생명현상의 메카니즘을 규명하여 그 결과를 신약개발에 활용하기 위한 기초 연구이다. 이 글에서는 프로테오믹스 연구의 초창기부터 사용되어온 2차원 전기영동법에 의해 생성되는 2D-gel image에서의 스팟(spot)분석법과 함께, 탄뎀 질량분석기를 사용하는 ICAT, SILAC 등의 동위 원소를 사용한 라벨링(labeling) 방법, 라벨링을 하지 않는 label-free 방법 등 프로테오믹스에서의 정량분석법에 대한 기본 개념을 살펴보고, 이들에서의 데이터 분석 기술의 적용에 대해 간략히 소개하였다.

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Development of New Biochip and Genome Detection Using an Non-labeling Target DNA (차세대형 바이오칩의 개발 및 비수식화 표적 DNA를 이용한 유전자 검출)

  • Choi, Yong-Sung;Park, Dae-Hee;Kwon, Young-Soo;Kawai, Tomoji
    • Proceedings of the KIEE Conference
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    • 2002.11a
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    • pp.51-53
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    • 2002
  • This research aims to develop a multiple channel electrochemical DNA chip using micro-fabrication technology. At first, we fabricated a high integrated type DNA chip array by lithography technology. Several probe DNAs consisting of thiol group at their 5-end were immobilized on the sold electrodes. Then target DNAs were hybridized by an electrical force. Redox peak of cyclic-voltammogram showed a difference between target DNA and mismatched DNA in the anodic peak current. Therefore, it is able to detect a various genes electrochemically after immobilization of a various probe DNA and hybridization of label-free DNA on the electrodes simultaneously. It suggested that this DNA chip could recognize the sequence specific genes.

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Three-dimensional Refractive-index Distributions of Individual Angiosperm Pollen Grains

  • Park, Chansuk;Lee, SangYun;Kim, Geon;Lee, SeungJun;Lee, Jaehoon;Heo, Taehyun;Park, Yoonjeong;Park, YongKeun
    • Current Optics and Photonics
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    • v.2 no.5
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    • pp.460-467
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    • 2018
  • Three-dimensional (3D) refractive-index (RI) imaging and quantitative analyses of angiosperm pollen grains are presented. Using optical diffraction tomography, the 3D RI structures of individual angiosperm pollen grains were measured without using labeling or other preparation techniques. Various physical quantities including volume, surface area, exine volume, and sphericity were determined from the measured RI tomograms of pollen grains. Exine skeletons, the distinct internal structures of angiosperm pollen grains, were identified and systematically analyzed.

Optimization of vertical SOI slot optical waveguide with confinement factor and sensitivity for integrated-optical biochemical sensors (구속계수와 감지도에 기반한 집적광학 바이오케미컬 센서에 적합한 수직 SOI 슬롯 광 도파로 최적화)

  • Jung, Hongsik
    • Journal of Sensor Science and Technology
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    • v.30 no.3
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    • pp.131-138
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    • 2021
  • The optimization of the specifications of vertical silicon on insulator (SOI) slot optical waveguides suitable for integrated-optical biochemical sensors was performed through computational analysis of the confinement factor of the guided mode distributed in the slot in addition to analytical examination of the TE mode. The optimized specifications were confirmed based on sensitivity in terms of the change in the refractive index of the biochemical analyte. When the slot width, rail width, and height were set to 120 nm, 200 nm, and 320 nm, respectively, the confinement factor was evaluated to be about 56% and the sensitivity was at least 0.9 [RIU/nm].

Nonparaxial Imaging Theory for Differential Phase Contrast Imaging

  • Jeongmin Kim
    • Current Optics and Photonics
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    • v.7 no.5
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    • pp.537-544
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    • 2023
  • Differential phase contrast (DPC) microscopy, a central quantitative phase imaging (QPI) technique in cell biology, facilitates label-free, real-time monitoring of intrinsic optical phase variations in biological samples. The existing DPC imaging theory, while important for QPI, is grounded in paraxial diffraction theory. However, this theory lacks accuracy when applied to high numerical aperture (NA) systems that are vital for high-resolution cellular studies. To tackle this limitation, we have, for the first time, formulated a nonparaxial DPC imaging equation with a transmission cross-coefficient (TCC) for high NA DPC microscopy. Our theoretical framework incorporates the apodization of the high NA objective lens, nonparaxial light propagation, and the angular distribution of source intensity or detector sensitivity. Thus, our TCC model deviates significantly from traditional paraxial TCCs, influenced by both NA and the angular variation of illumination or detection. Our nonparaxial imaging theory could enhance phase retrieval accuracy in QPI based on high NA DPC imaging.

Quantitative Alpha Fetoprotein Detection with a Piezoelectric Microcantilever Mass Sensor (압전 마이크로캔틸레버 질량센서를 이용한 정량적 알파태아단백 검출)

  • Lee, Sangk-Yu;Cho, Jong-Yun;Lee, Yeol-Ho;Jeon, Sang-Min;Cha, Hyung-Joon;Moon, Wonk-Yu
    • Journal of the Korean Society for Nondestructive Testing
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    • v.31 no.5
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    • pp.487-493
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    • 2011
  • Alpha fetoprotein(AFP), which is serological marker for hepatocellular carcinoma, was quantitatively measured by its normal concentration, 10 ng/ml, with a label-free piezoelectric microcantilever mass sensor. The principle of detection is based on changes in the resonant frequency of the piezoelectric microcantilever before and after target molecules are attached to it, and its resonant frequency is measured electrically using a conductance spectrum. The resonant frequency of the developed sensor is approximately 1.34 MHz and the mass sensitivity is approximately 175 Hz/pg. The sensor has high reliability as mass sensor by reducing the effect of surface stress on resonant frequency due to attached proteins. 'Dip and dry' technique was used to react the sensor with reagents for immobilizing AFP antibody on the sensor and detecting AFP antigen. The measured mass of the detected AFP antigen was 6.02 pg at the concentration of 10 ng/ml, and 10.67 pg at 50 ng/ml when the immunoreaction time was 10 min.

Real-Time Monitoring of Mitochondrial ATP Synthesis and Hydrolysis by Surface Infrared Spectroscopy

  • Yamaguchi, Ryo-Taro;Hirano-Iwata, Ayumi;Aonuma, Yuki;Yoshimura, Yuya;Shinohara, Yasuo;Kimura, Yasuo;Niwano, Michio
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.02a
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    • pp.108-109
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    • 2013
  • Mitochondria play key roles in the production of cell's energy. Their dominant function is the synthesis of adenosine 5'-triphosphate (ATP) from adenosine diphosphate (ADP) and phosphate (Pi) through the oxidative phosphorylation. Evaluation of drug-induced mitochondrial toxicity has become increasingly important since mitochondrial dysfunction has recently been implicated in numerous diseases including cancer and diabetes mellitus. Mitochondrial functions have been monitored via oxygen consumption, mitochondrial membrane potential, and more importantly via ATP synthesis since ATP synthesis is the most essential function of mitochondria. Various analytical methods have been employed to investigate ATP synthesis in mitochondria, including high performance liquid chromatography (HPLC), bioluminescence technique, and pH measurement. However, most of these methods are based on destructive analysis or indirect monitoring through the enzymatic reaction. Infrared absorption spectroscopy (IRAS) is one of the useful techniques for real-time, label-free, and direct monitoring of biological reactions [1,2]. However, the strong water absorption requires very short path length in the order of several micrometers. Transmission measurements with thin path length are not suitable for mitochondrial assays because solution handlings necessary for evaluating mitochondrial toxicity, such as rapid mixing of drugs and oxygen supply, are difficult in such a narrow space. On the other hand, IRAS in the multiple internal reflection (MIR) geometry provides an ideal optical configuration to combine solution handling and aqueous-phase measurement. We have recently reportedon a real-time monitoring of drug-induced necrotic and apoptotic cell death using MIR-IRAS [3,4]. Clear discrimination between viable and damaged cells has been demonstrated, showing a promise as a label-free and real-time detection for cell-based assays. In the present study, we have applied our MIR-IRAS system to mitochondria-based assays by monitoring ATP synthesis in isolated mitochondria from rat livers. Mitochondrial ATP synthesis and hydrolysis were in situ monitored with MIR-IRAS, while dissolved oxygen level and solution pH were simultaneously monitored with O2 and pH electrodes, respectively. It is demonstrated that ATP synthesis and hydrolysis can be monitored by the IR spectral changes in phosphate groups in adenine nucleotides and MIR-IRAS is useful for evaluating time-dependent drug effects of mitochondrial toxicants.

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