• Title/Summary/Keyword: Single molecular chemistry

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A Linear Beacon System Featuring an Internal Deoxyguanine Quencher Allows Highly Selective Detection of Single Base Mismatches

  • Lee, Young-Ae;Hwang, Gil-Tae
    • Bulletin of the Korean Chemical Society
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    • v.31 no.7
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    • pp.2011-2014
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    • 2010
  • The fluorescence intensity of a single-stranded oligonucleotide containing a fluorene-labeled deoxyuridine $(U^{Fl})$ unit increases by only 1.5-fold upon formation of its perfectly matched duplex. To increase the fluorescence signal during hybridization, we positioned a quencher strand containing a deoxyguanine (dG) nucleobase, functioning as an internal quencher, opposite to the $U^{Fl}$ unit to reduce the intrinsic fluorescence upon hybridization with a probe. From an investigation of the optimal length of the quencher strand and the effect of the neighboring base sequence, we found that a short strand (five-nucleotide) containing all natural nucleotides and dG as an internal quencher was effective at reducing the intrinsic fluorescence of a linear beacon; it also exhibited high total discrimination factors for the formation of perfectly matched and single base-mismatched duplexes. Such assays that function based on clear changes in fluorescence in response to single-base nucleotide mutations would be useful tools for accelerating diagnoses related to various diseases.

Single C-Reactive Protein Molecule Detection on a Gold-Nanopatterned Chip Based on Total Internal Reflection Fluorescence

  • Heo, Yunmi;Lee, Seungah;Lee, Sang-Won;Kang, Seong Ho
    • Bulletin of the Korean Chemical Society
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    • v.34 no.9
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    • pp.2725-2730
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    • 2013
  • Single C-reactive protein (CRP) molecules, which are non-specific acute phase markers and products of the innate immune system, were quantitatively detected on a gold-nanopatterned biochip using evanescent field-enhanced fluorescence imaging. The $4{\times}5$ gold-nanopatterned biochip (spot diameter of 500 nm) was fabricated by electron beam nanolithography. Unlabeled CRP molecules in human serum were identified with single-molecule sandwich immunoassay by detecting secondary fluorescence generated by total internal reflection fluorescence (TIRF) microscopy. With decreased standard CRP concentrations, relative fluorescence intensities reduced in the range of 33.3 zM-800 pM. To enhance fluorescence intensities in TIRF images, the distance between biochip surface and CRP molecules was optimally adjusted by considering the quenching effect of gold and the evanescent field intensity. As a result, TIRF only detected one single-CRP molecule on the biochip the first time.

Preparation of Anatase TiO2 Thin Films with (OiPr)2Ti(CH3COCHCONEt2)2 Precursor by MOCVD

  • Bae, Byoung-Jae;Lee, Kwang-Yeol;Seo, Won-Seok;Miah, Md. Arzu;Kim, Keun-Chong;Park, Joon T.
    • Bulletin of the Korean Chemical Society
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    • v.25 no.11
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    • pp.1661-1666
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    • 2004
  • The reaction of titanium tetraisopropoxide with 2 equiv of N,N-diethyl acetoacetamide affords Ti($O^iPr)_2(CH_3COCHCONEt_2)_2$ (1) as colorless crystals in 80% yield. Compound 1 is characterized by spectroscopic (Mass and $^1H/^{13}C$ NMR) and microanalytical data. Molecular structure of 1 has been determined by a single crystal X-ray diffraction study, which reveals that it is a monomeric, cis-diisopropoxide and contains a six coordinate Ti(IV) atom with a cis($CONEt_2$), trans($COCH_3$) configuration (1a) in a distorted octahedral environment. Variable-temperature $^1H$ NMR spectra of 1 indicate that it exists as an equilibrium mixture of cis, trans (1a) and cis, cis (1b) isomers in a 0.57 : 0.43 ratio at -20$^{\circ}C$ in toluene-$d_8$ solution. Thermal properties of 1 as a MOCVD precursor for titanium dioxide films have been evaluated by thermal gravimetric analysis and vapor pressure measurement. Thin films of pure anatase titanium dioxide (after annealing above 500$^{\circ}C$ under oxygen) have been grown on Si(100) with precursor 1 in the substrate temperature range of 350- 500$^{\circ}$ using a bubbler-based MOCVD method.

Biocompatible Formation of Silica/Titania Nanocomposite Shells on Living Chlorella Cells

  • Go, Eun-Hye;Yun, Yeon-Jeong;Jin, Seung-Uk;Hwang, Ji-Min;Lee, Gyu-Nam;Yang, Seong-Ho;Choe, In-Seong
    • Proceedings of the Korean Vacuum Society Conference
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    • 2012.02a
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    • pp.553-553
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    • 2012
  • The artificial shells of hard inorganic nanocomposites on individual cells would protect the cells physically and chemically, and control cell division. These emerging properties could be combined with cell-surface functionalizations for applications to cell-based sensors and assays as well as for fundamental studies on single-cell biology. In this work, individual Chlorella cells were encapsulated within a silica/titania nanocomposite shell in a biocompatible fashion that utilized a designed peptide, RKKRKKRKKRKKDDDDDDDD, as a catalytic template for formation of both $SiO_2$ and $TiO_2$ on the cell surface. The cell viability was maintained, and the division of the encapsulated Chlorella cells was controlled. The cell viability was enhanced compared with the $TiO_2$-shell formation. In addition, the incorporation of $TiO_2$ to the shell made it possible to anchor the ligands of interest to the shell via catechol chemistry. All in all, the combination of biological $SiO_2$ and abiolgical $TiO_2$ for the shell formation gave more tunability of the artificial shells compared with the $SiO_2$ or $TiO_2$ shells only.

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A New Analytical Method to Determine the Purity of Synthetic Fluorophores using Single Molecule Detection Technique

  • Song, Nam-Yoong;Kim, Hyong-Ha;Park, Tae-Sook;Yoon, Min-Joong
    • Journal of Photoscience
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    • v.12 no.2
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    • pp.87-93
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    • 2005
  • A new assay technique to distinguish between pure compounds and the isomeric mixtures has been suggested using single molecule (SM) fluorescence detection technique. Since the number of emission spots in a fluorophorespread film prepared from a genuine dye solution was determined by experimental condition, the deviation of spot numbers from the expected values could be considered to be an indication of lower purity of the sample solution. The lower limit of sample concentration for this assay was determined to be $5{\times}10^{-10}$ M to show uniform number of expected spots within 10% uncertainties in our experimental condition. An individual fluorescence intensity distribution for a mixture of isomers having doubly different emissivities was simulated by adding distributions obtained from Cy3 and nile red (NR) independently. The result indicated that the mixture could be identified from the pure compounds through the difference in the number of Gaussian functions to fit the distribution. This new assay technique can be applied to the purity test for synthetic biofluorophores which are usually prepared in small quantities not enough for classical ensemble assays.

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Studies of Molecular Orientation for Ferrielectric Liquid Crystal by Phase Transitions

  • Kim, S.W.;Choi, H.;Song, J.H.;KIm, J.H.;Kumar, S.;Choi, J.W.;Kim, Y.B.;Shin, S.T.
    • 한국정보디스플레이학회:학술대회논문집
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    • 2000.01a
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    • pp.61-62
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    • 2000
  • We have studied the molecular orientation by the phase transitions of the chiral smectic liquid crystals, 4-(1-Trifluoromethyl-6-ethoxy-hexyloxycarbonlphenyl)-4-nonyloxybiphenyl-4-carbo-xylate (R-TFMEOHPNBC) to seek the original solution of the zig-zag defect using two different experimental techniques; optical system and x-ray scattering. The phase sequence is gamma ferroelectric $(SmC{\gamma}\;^*)$ ${\rightarrow}$ smectic A (SmA) ${\rightarrow}$ isotropic (I). Existence of two layer spacing at chiral smectic phase gives a possibility of the molecular orientation in two different tilt angles, ${\theta}\;_1$ and ${\theta}\;_2$, which are separated each other to the layer normal at a given temperature. The gamma ferroelectric-like phase is, first, discovered in the single compound.

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Photobleaching Nonchemically Amplified Photoresists

  • Kim, Jin-Baek;Ganesan, Ramakrishnan;Kim, Kyoung-Seon;Choi, Jae-Hak
    • Proceedings of the Polymer Society of Korea Conference
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    • 2006.10a
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    • pp.176-176
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    • 2006
  • We designed and synthesized new type of nonchemically amplified polymeric and molecular resists and studied their lithographic performance. The polymeric resists [poly(DOBEMA-co-GBLMA)] show very high degree of photobleaching at 248 nm and moderate to good degree of photobleaching at 193 nm. The molecular resist (CDEOPE-POSS) shows high degree of photobleaching at 248 nm and modetate degree of photobleaching at 193 nm. CDEOPE-POSS possesses very high oxygen reactive ion etching resistance, which makes it suitable to be used as a bilayer resist. The lithographic studies of these resist materials suggest the feasibility of these materials to be used as single and bilayer resists.

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Molecular Dynamic Study of A Polymeric Solution (II). Solvent Effect

  • Oh In-Joon;Lee Young-Seek;Ree Tai-Kyue
    • Bulletin of the Korean Chemical Society
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    • v.4 no.2
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    • pp.87-91
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    • 1983
  • Molecular dynamic method has been applied to a single polymer chain immersed in a solvent. The interactions for the pairs, of two solvent molecules (SS), of a chain element and a solvent molecules (CS), and of two non-neighbor chain elements (CC) are given by the Lennard-Jones potential, and the interaction between two bonded chain elements is given by a harmonic potential. We changed the CS interaction parameter ${\varepsilon}_{CS}$ to 0.5, 1.0 and 2.0 times of the SS interaction ${\varepsilon}_{SS}$. We calculated the pair correlation functions for the SS, CS, and CC pairs, end-to-end distance and radius of gyration with the varying ${\varepsilon}_{CS}$ parameters. The results showed that a phase separation occurs between the polymer and solvent in the 0.5 system where ${\varepsilon}_{CS}$ = 0.5 {\varepsilon}_{SS}$. The autocorrelation functions for end-to-end distance and radius of gyration were also calculated.

Slipchip Device Development in Molecular Diagnostics

  • Qingtian Yin;Huiwen Bai;Ruijie Li;Youngung Seok
    • Korean Journal of Materials Research
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    • v.34 no.2
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    • pp.63-71
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    • 2024
  • Slipchip offers advantages such as high-throughout, low cost, and simple operation, and therefore, it is one of the technologies with the greatest potential for high-throughput, single-cell, and single-molecule analyses. Slipchip devices have achieved remarkable advances over the past decades, with its simplified molecular diagnostics gaining particular attention, especially during the COVID-19 pandemic and in various infectious diseases scenarios. Medical testing based on nucleic acid amplification in the Slipchip has become a promising alternative simple and rapid diagnostic tool in field situations. Herein, we present a comprehensive review of Slipchip device advances in molecular diagnostics, highlighting its use in digital recombinase polymerase amplification (RPA), loop-mediated isothermal amplification (LAMP), and polymerase chain reaction (PCR). Slipchip technology allows users to conduct reliable droplet transfers with high-throughput potential for single-cell and molecule analyses. This review explores the device's versatility in miniaturized and rapid molecular diagnostics. A complete Slipchip device can be operated without special equipment or skilled handling, and provides high-throughput results in minimum settings. This review focuses on recent developments and Slipchip device challenges that need to be addressed for further advancements in microfluidics technology.

In Situ Single Cell Monitoring by Isocyanide-Functionalized Ag and Au Nanoprobe-Based Raman Spectroscopy

  • Lee, So-Yeong;Jang, Soo-Hwa;Cho, Myung-Haing;Kim, Young-Min;Cho, Keun-Chang;Ryu, Pan Dong;Gong, Myoung-Seon;Joo, Sang-Woo
    • Journal of Microbiology and Biotechnology
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    • v.19 no.9
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    • pp.904-910
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    • 2009
  • The development of effective cellular imaging requires a specific labeling method for targeting, tracking, and monitoring cellular/molecular events in the living organism. For this purpose, we studied the cellular uptake of isocyanide-functionalized silver and gold nanoparticles by surface-enhanced Raman scattering (SERS). Inside a single mammalian cell, we could monitor the intracellular behavior of such nanoparticles by measuring the SERS spectra. The NC stretching band appeared clearly at ${\sim}2,100cm^{-1}$ in the well-isolated spectral region from many organic constituents between 300 and 1,700 or 2,800 and $3,600cm^{-1}$. The SERS marker band at ${\sim}2,100cm^{-1}$ could be used to judge the location of the isocyanide-functionalized nanoparticles inside the cell without much spectral interference from other cellular constituents. Our results demonstrate that isocyanide-modified silver or gold nanoparticle-based SERS may have high potential for monitoring and imaging the biological processes at the single cell level.