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Zhang, Peng,Park, Guenyoung,Kang, Seong Ho Korean Chemical Society 2014 Bulletin of the Korean Chemical Society Vol.35 No.4
A multi-channel microchip electrophoresis (MCME) method with parallel laser-induced fluorescence (LIF) detection was developed for rapid screening of H1N1 virus. The hemagglutinin (HA) and nucleocapsid protein (NP) gene of H1N1 virus were amplified using polymerase chain reaction (PCR). The amplified PCR products of the H1N1 virus DNA (HA, 116 bp and NP, 195 bp) were simultaneously detected within 25 s in three parallel channels using an expanded laser beam and a charge-coupled device camera. The parallel separations were demonstrated using a sieving gel matrix of 0.3% poly(ethylene oxide) ($M_r$ = 8,000,000) in $1{\times}$ TBE buffer (pH 8.4) with a programmed step electric field strength (PSEFS). The method was ~20 times faster than conventional slab gel electrophoresis, without any loss of resolving power or reproducibility. The proposed MCME/PSEFS assay technique provides a simple and accurate method for fast high-throughput screening of infectious virus DNA molecules under 400 bp.
Peng Zhang,Guenyoung Park,Seong Ho Kang 대한화학회 2014 Bulletin of the Korean Chemical Society Vol.35 No.4
A multi-channel microchip electrophoresis (MCME) method with parallel laser-induced fluorescence (LIF) detection was developed for rapid screening of H1N1 virus. The hemagglutinin (HA) and nucleocapsid protein (NP) gene of H1N1 virus were amplified using polymerase chain reaction (PCR). The amplified PCR products of the H1N1 virus DNA (HA, 116 bp and NP, 195 bp) were simultaneously detected within 25 s in three parallel channels using an expanded laser beam and a charge-coupled device camera. The parallel separations were demonstrated using a sieving gel matrix of 0.3% poly(ethylene oxide) (Mr = 8,000,000) in 1× TBE buffer (pH 8.4) with a programmed step electric field strength (PSEFS). The method was ~20 times faster than conventional slab gel electrophoresis, without any loss of resolving power or reproducibility. The proposed MCME/PSEFS assay technique provides a simple and accurate method for fast high-throughput screening of infectious virus DNA molecules under 400 bp.
Lee, Seungah,Park, Guenyoung,Chakkarapani, Suresh Kumar,Kang, Seong Ho Elsevier 2015 Biosensors & bioelectronics Vol.63 No.-
<P><B>Abstract</B></P> <P>Novel, fluorescence-free detection of biomolecules on nanobiochips was investigated based on plasmonic nanometal scattering in the evanescent field layer (EFL) using total internal reflection scattering (TIRS) microscopy. The plasmonic scattering of nanometals bonded to biomolecules was observed at different wavelengths by an electromagnetic field in the EFL. The changes in the scattering of nanometals on the gold-nanopatterned chip in response to the immunoreaction between silver nanoparticles and antibodies allowed fluorescence-free detection of biomolecules on the nanobiochips. Under optimized conditions, the TIRS immunoassay chip detected different amounts of immobilized antigen, i.e., human cardiac troponin I. The sandwich immuno-reaction was quantitatively analyzed in the dynamic range of 720zM–167fM. The limit of detection (<I>S</I>/<I>N</I>=4) was 600zM, which was ~140 times lower than limits obtained by previous total internal reflection fluorescence and dark field methods. These results demonstrate the possibility for a fluorescence-free biochip nanoimmunoassay based on the scattering of nanometals in the EFL.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Fluorescence-free detection of biomolecules on nanobiochips by total internal reflection scattering microscopy. </LI> <LI> Observation of the scattering shape and intensity of native nanometals by controlling optical components. </LI> <LI> Highly sensitive fluorescent-free detection of cTnI with a detection limit of 600zM. </LI> </UL> </P>