• Title/Summary/Keyword: label-free detection

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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.

Nanoplasmonics: Enabling Platform for Integrated Photonics and Sensing

  • Yeo, Jong-Souk
    • Proceedings of the Korean Vacuum Society Conference
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    • 2015.08a
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    • pp.75-75
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    • 2015
  • Strong interactions between electromagnetic radiation and electrons at metallic interfaces or in metallic nanostructures lead to resonant oscillations called surface plasmon resonance with fascinating properties: light confinement in subwavelength dimensions and enhancement of optical near fields, just to name a few [1,2]. By utilizing the properties enabled by geometry dependent localization of surface plasmons, metal photonics or plasmonics offers a promise of enabling novel photonic components and systems for integrated photonics or sensing applications [3-5]. The versatility of the nanoplasmonic platform is described in this talk on three folds: our findings on an enhanced ultracompact photodetector based on nanoridge plasmonics for photonic integrated circuit applications [3], a colorimetric sensing of miRNA based on a nanoplasmonic core-satellite assembly for label-free and on-chip sensing applications [4], and a controlled fabrication of plasmonic nanostructures on a flexible substrate based on a transfer printing process for ultra-sensitive and noise free flexible bio-sensing applications [5]. For integrated photonics, nanoplasmonics offers interesting opportunities providing the material and dimensional compatibility with ultra-small silicon electronics and the integrative functionality using hybrid photonic and electronic nanostructures. For sensing applications, remarkable changes in scattering colors stemming from a plasmonic coupling effect of gold nanoplasmonic particles have been utilized to demonstrate a detection of microRNAs at the femtomolar level with selectivity. As top-down or bottom-up fabrication of such nanoscale structures is limited to more conventional substrates, we have approached the controlled fabrication of highly ordered nanostructures using a transfer printing of pre-functionalized nanodisks on flexible substrates for more enabling applications of nanoplasmonics.

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Nanoplasmonic Spectroscopic Imaging and Molecular Probes

  • Choe, Yeon-Ho
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.02a
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    • pp.85-85
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    • 2013
  • Label-free, sensitive and selective detection methods with high spatial resolution are critically required for future applications in chemical sensor, biological sensor, and nanospectroscopic imaging. Here I describe the development of Plasmon Resonance Energy Transfer (PRET)-based molecular imaging in living cells as the first demonstration of intracellular imaging with PRET-based nanospectroscopy. In-vivo PRET imaging relied on the overlap between plasmon resonance frequency of gold nanoplasmonic probe (GNP) and absorption peak frequencies of conjugated molecules, which leads to create 'quantized quenching dips' in Rayleigh scattering spectrum of GNP. The position of these dips exactly matched with the absorption peaks of target molecules. As another innovative application of PRET, I present a highly selective and sensitive detection of metal ions by creating conjugated metal-ligand complexes on a single GNP. In addition to conferring high spatial resolution due to the small size of the metal ion probes (50 nm in diameter), this method is 100 to 1,000 folds more sensitive than organic reporter-based methods. Moreover, this technique achieves high selectivity due to the selective formation of Cu2+complexes and selective resonant quenching of GNP by the conjugated complexes. Since many metal ion ligand complexes generate new absorption peak due to the d-d transition in the metal ligand complex when a specific metal ion is inserted into the complex, we can match with the scattering frequency of nanoplasmonic metal ligand systems and the new absorption peak.

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Amperometric Immunosensor for Myeloperoxidase in Human Serum Based on a Multi-wall Carbon Nanotubes-Ionic Liquid-Cerium Dioxide Film-modified Electrode

  • Lu, Lingsong;Liu, Bei;Liu, Chenggui;Xie, Guoming
    • Bulletin of the Korean Chemical Society
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    • v.31 no.11
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    • pp.3259-3264
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    • 2010
  • A label-free amperometric immunosensor has been proposed for the detection of myeloperoxidase (MPO) in human serum. To fabricate such an immunosensor, a composite film consisting of N,N-dimethylformamide (DMF), multiwall carbon nanotubes (MWCNTs) and 1-ethyl-3-methyl imidazolium tetrafluoroborate ($EMIMBF_4$) suspension was initially formed on a glassy carbon electrode (GCE). Then cerium dioxide ($CeO_2$) dispersed by chitosan was coated on the GCE. After that, MPO antibodies (anti-MPO) were attached onto the nano$CeO_2$ surface. With a noncompetitive immunoassay format, the antibody-antigen complex formed between the immobilized anti-MPO and MPO in sample solution. The immunosensor was characterized by cyclic voltammetry, transmission electron microscopy (TEM) and scanning electron microscopy (SEM). The factors influencing the performance of the immunosensor were studied in detail. Under optimal conditions, the current change before and after the immunoreaction was proportional to MPO concentration in the range of 5 to $300\;ng\;mL^{-1}$ with a detection limit of $0.2\;ng\;mL^{-1}$.

Detection of Avidin Based on Rugate-structured Porous Silicon Interferometer

  • Koh, Young-Dae;Kim, Sung-Jin;Park, Jae-Hyun;Park, Cheol-Young;Cho, Sung-Dong;Woo, Hee-Gweon;Ko, Young-Chun;Sohn, Hong-Lae
    • Bulletin of the Korean Chemical Society
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    • v.28 no.11
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    • pp.2083-2088
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    • 2007
  • Biosensor based on rugate PSi interferometer for the detection of avidin has been described. Rugate PSi fabricated by applying a computer-generated pseudo-sinusoidal current waveform has been prepared for the application as a label-free biosensor based on porous silicon interferometer. The fabrication, optical characterization, and surface derivatization of a rugate PSi has been described. The method to fabricate biotinderivatized rugate PSi has been investigated. The surface and cross sectional morphology of rugate PSi are obtained with SEM. FT-IR spectroscopy is used to characterize the oxidation and functionalization reaction of rugate PSi sample. Binding of the avidin into the biotin-derivatized rugate PSi induces a change in refractive index. A red-shift of reflectivity by 18 nm in the reflectivity spectrum is observed, when the biotin-modified rugate PSi was exposed to a flow of avidin.

Immunoassay of haptoglobin and transferrin with proteinG-containing QCM sensor chip and unpurified antiserum (Protein G를 포함하는 수정미소저울 센서 칩과 정제되지 않은 항혈청을 이용한 헵토글로빈과 트랜스페린의 면역분석)

  • Ha, In-Young;Choi, Suk-Jung
    • Journal of Sensor Science and Technology
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    • v.17 no.5
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    • pp.380-386
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    • 2008
  • Quartz crystal microbalance immunosensor has a capacity to perform a label-free and real time detection of a trace amount of analyte through the specific interaction between antibody and antigen. However, immobilization of antibody molecules on the sensor surface is a troublesome procedure for researchers who are not experienced in chemistry. Protein G has a specific affinity to antibody and would serve as a capturing agent for antibody when immobilized on the sensor surface. In this work, we prepared a protein G sensor chip by immobilizing protein G on the surface of quartz crystal microbalance and examined its capability to detect human haptoglobin or human transferrin with unpurified corresponding antiserum. Specific and dose dependent response was observed when the protein G chip was used for detection of antigens after saturated with antiserum. We also verified several advantageous aspects of the protein G chip such as improved flexibility and sensitivity.

Reduced Graphene Oxide Field Effect Transistor for Detection of H+ Ions and Their Bio-sensing Application

  • Sohn, Il-Yung;Kim, Duck-Jin;Yoon, Ok-Ja;Tien, N.T.;Trung, T.Q.;Lee, N.E.
    • Proceedings of the Korean Vacuum Society Conference
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    • 2012.02a
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    • pp.195-195
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    • 2012
  • Recently, graphene based solution-gated field-effect transistors (SGFETs) have been received a great attention in biochemical sensing applications. Graphene and reduced graphene oxide (RGO) possess various advantages such as high sensitivity, low detection limit, label-free electrical detection, and ease of fabrication due to their 2D nature and large sensing area compared to 1D nanomaterials- based nanobiosensors. Therefore, graphene or RGO -based SGFET is a good potential candidate for sensitive detection of protons (H+ ions) which can be applied as the transducer in various enzymatic or cell-based biosensing applications. However, reports on detection of H+ ions using graphene or RGO based SGFETs have been still limited. According to recent reports, clean graphene grown by CVD or exfoliation is electrochemically insensitive to changes of H+ concentration in solution because its surface does not have terminal functional groups that can sense the chemical potential change induced by varying surface charges of H+ on CVD graphene surface. In this work, we used RGO -SGFETs having oxygen-containing functional groups such as hydroxyl (OH) groups that effectively interact with H+ ions for expectation of increasing pH sensitivity. Additionally, we also investigate RGO based SGFETs for bio-sensing applications. Hydroloytic enzymes were introduced for sensing of biomolecular interaction on the surface of RGO -SGFET in which enzyme and substrate are acetylcholinesterase (AchE) and acetylcholine (Ach), respectively. The increase in H+ generated through enzymatic reaction of hydrolysis of Ach by AchE immobilized on RGO channel in SGFET could be monitored by the change in the drain-source current (Ids).

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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.

Fabrication of Label-Free Biochips Based on Localized Surface Plasmon Resonance (LSPR) and Its Application to Biosensors (국소 표면 플라즈몬 공명 (LSPR) 기반 비표지 바이오칩 제작 및 바이오센서로의 응용)

  • Kim, Do-Kyun;Park, Tae-Jung;Lee, Sang-Yup
    • KSBB Journal
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    • v.24 no.1
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    • pp.1-8
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    • 2009
  • In the past decade, we have observed rapid advances in the development of biochips in many fields including medical and environmental monitoring. Biochip experiments involve immobilizing a ligand on a solid substrate surface, and monitoring its interaction with an analyte in a sample solution. Metal nanoparticles can display extinction bands on their surfaces. These charge density oscillations are simply known as the localized surface plasmon resonance (LSPR). The high sensitivity of LSPR has been utilized to design biochips for the label-free detection of biomolecular interactions with various ligands. LSPR-based optical biochips and biosensors are easy to fabricate, and the apparatus cost for the evaluation of optical characteristics is lower than that for the conventional surface plasmon resonance apparatus. Furthermore, the operation procedure has become more convenient as it does not require labeling procedure. In this paper, we review the recent advances in LSPR research and also describe the LSPR-based optical biosensor constructed with a core-shell dielectric nanoparticle biochip for its application to label-free biomolecular detections such as antigen-antibody interaction.

Emission Detection of Mercuric Ions in Aqueous Media Based-on Dehybridization of DNA Duplexes

  • Oh, Byul-Nim;Wu, Qiong;Cha, Mi-Sun;Kang, Hee-Kyung;Kim, Jin-Ah;Kim, Ka-Young;Rajkumar, Eswaran;Kim, Jin-Heung
    • Bulletin of the Korean Chemical Society
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    • v.32 no.9
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    • pp.3223-3228
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    • 2011
  • To quantify the presence of mercuric ions in aqueous solution, double-stranded DNA (dsDNA) of poly(dT) was employed using a light switch compound, $Ru(phen)_2(dppz)^{2+}$ (1) which is reported to intercalate into dsDNA of a right-handed B-form. Addition of mercuric ions induced the dehybridization of poly(dT)${\cdot}$poly(dA) duplexes to form a hairpin structure of poly(dT) at room temperature and the metal-to-ligand charge transfer emission derived from the intercalation of 1 was reduced due to the dehybridization of dsDNA. As the concentration of $Hg^{2+}$ was increased, the emission of 1 progressively decreased. This label-free emission method had a detection limit of 0.2 nM. Other metal ions, such as $K^+$, $Ag^+$, $Ca^{2+}$, $Mg^{2+}$, $Zn^{2+}$, $Mn^{2+}$, $Co^{2+}$, $Ni^{2+}$, $Cu^{2+}$, $Cd^{2+}$, $Cr^{3+}$, $Fe^{3+}$, had no significant effect on reducing emission. This emission method can differentiate matched and mismatched poly(dT) sequences based on the emission intensity of dsDNA.