• Title/Summary/Keyword: Microchip electrophoresis

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MEMS Fabrication of Microchannel with Poly-Si Layer for Application to Microchip Electrophoresis (마이크로 칩 전기영동에 응용하기 위한 다결정 실리콘 층이 형성된 마이크로 채널의 MEMS 가공 제작)

  • Kim, Tae-Ha;Kim, Da-Young;Chun, Myung-Suk;Lee, Sang-Soon
    • Korean Chemical Engineering Research
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    • v.44 no.5
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    • pp.513-519
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    • 2006
  • We developed two kinds of the microchip for application to electrophoresis based on both glass and quartz employing the MEMS fabrications. The poly-Si layer deposited onto the bonding interface apart from channel regions can play a role as the optical slit cutting off the stray light in order to concentrate the UV ray, from which it is possible to improve the signal-to-noise (S/N) ratio of the detection on a chip. In the glass chip, the deposited poly-Si layer had an important function of the etch mask and provided the bonding surface properly enabling the anodic bonding. The glass wafer including more impurities than quartz one results in the higher surface roughness of the channel wall, which affects subsequently on the microflow behavior of the sample solutions. In order to solve this problem, we prepared here the mixed etchant consisting HF and $NH_4F$ solutions, by which the surface roughness was reduced. Both the shape and the dimension of each channel were observed, and the electroosmotic flow velocities were measured as 0.5 mm/s for quartz and 0.36 mm/s for glass channel by implementing the microchip electrophoresis. Applying the optical slit with poly-Si layer provides that the S/N ratio of the peak is increased as ca. 2 times for quartz chip and ca. 3 times for glass chip. The maximum UV absorbance is also enhanced with ca. 1.6 and 1.7 times, respectively.

Ultra-fast Detection and Differentiation of Mycoplasma haemofelis and Candidatus M. Haemominutum in Korean Feral Cats by Microchip Electrophoresis with Programmed Field Strength Gradients

  • Kumar, Kailasa S.;Lee, Hee-Gu;Yoo, Dong-Jin;Kang, Seong-Ho
    • Bulletin of the Korean Chemical Society
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    • v.29 no.1
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    • pp.153-158
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    • 2008
  • A microchip-based capillary gel electrophoresis (MCGE) technique was developed for the ultra-fast detection and differentiation of Candidatus Mycoplasma haemominutum (Candidatus M. haemominutum, California strain) and Mycoplasma haemofelis (M. haemofelis, Ohio strain) in Korean feral cats through the application of programmed field strength gradients (PFSG) in a conventional glass double-T microchip. The effects of the poly (ethyleneoxide) (PEO) concentration and electric field strength on the separation of DNA fragments were investigated. The PCR-amplified products of Candidatus M. haemominutum (202-bp) and M. haemofelis (273-bp) were analyzed by MCGE within 75 s under a constant applied electric field of 117.6 V/cm and a sieving matrix of 0.3% PEO (Mr 8 000 000). When the PFSG was applied, MCGE analysis generated results 6.8-times faster without any loss of resolution or reproducibility. The MCGE-PFSG technique was also applied to eleven samples selected randomly from 33 positive samples. The samples were detected and differentiated within 11 s. The analysis time of the MCGE-PFSG technique was approximately 980-times faster than that using conventional slab gel electrophoresis.

Multi-step Reactions on Microchip Platform Using Nitrocellulose Membrane Reactor

  • Park, Sung-Soo;Joo, Hwang-Soo;Cho, Seung-Il;Kim, Min-Su;Kim, Yong-Kweon;Kim, Byung-Gee
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.8 no.4
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    • pp.257-262
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    • 2003
  • A straightforward and effective method is presented for immobilizing enzymes on a microchip platform without chemically modifying a micro-channel or technically microfabricating a column reactor and fluid channel network. The proposed method consists of three steps: the reconstitution of a nitrocellulose (NC) membrane on a plane substrate without a channel network, enzyme immobilization on the NC membrane, and the assembly of another substrate with a fabricated channel network. As a result, enzymes can be stably and efficiently immobilized on a microchip. To evaluate the proposed method, two kinds of enzymatic reaction are applied: a sequential two-step reaction by one enzyme, alkaline phosphatase, and a coupled reaction by two enzymes, glucose oxidase and peroxidase, for a glucose assay.

Diagnosis of the ORF Virus Using a Mixture of Sieving Gel Matrixes in Microchip Gel Electrophoresis (마이크로칩젤 전기영동에서 충진젤 혼합물을 이용한 ORF 바이러스의 진단)

  • Kim, Yun-Jeong;Chae, Joon-Seok;Kang, Seong-Ho
    • Journal of the Korean Chemical Society
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    • v.48 no.5
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    • pp.483-490
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    • 2004
  • We have developed a novel polymerase chain reaction (PCR)-microchip gel electrophoresis (MGE) method based on the sieving gel mixture of commercially available poly(vinylpyrrolidone) (PVP) and hydroxy ethyl cellulose (HEC) for the rapid detection and diagnosis of the orf virus (ORFV) from Korean indigenous goat. After amplification of 594-bp DNA fragment from the B2L gene of ORF virus, the amplicon was analyzed by the MGE separation. The glass microfluidic chip (64 mm total length (36 mm effective length)${\times}$90 ${\mu}$m width${\times}$20 ${\mu}$m depth) allowed the fast detection and diagnosis of ORFV in the mixture of 1.0% PVP ($M_r$ 360,000) and 1.0% HEC ($M_r$250,000) as a sieving matrix with better resolution and reproducibility of DNA fragments. Under the electric field of 277.8 V/cm, the 594-bp DNA was analyzed within 4 min. Compared to traditional slab gel electrophoresis, the PCR-MGE method was twenty times faster and an effective clinical method for the quantitative analysis of ORFV.

Disposable Microchip-Based Electrochemical Detector Using Polydimethylsiloxane Channel and Indium Tin Oxide Electrode (Polydimethylsiloxane 채널과 indium tin oxide 전극을 이용한 일회용 전기화학적 검출 시스템)

  • Yi In-Je;Kang Chi-Jung;Kim Yong-Sang;Kim Ju-Ho
    • The Transactions of the Korean Institute of Electrical Engineers C
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    • v.54 no.5
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    • pp.227-231
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    • 2005
  • We have developed a microsystem with a capillary electrophoresis (CE) and an electrochemical detector (ECD). The microfabricated CE-ECD systems are adequate for a disposable type and the characteristics are optimized for an application to the electrochemical detection. The system was realized with polydimethylsiloxane (PDMS)-glass chip and indium tin oxide electrode. The injection and separation channels (80 um wide$\ast$40 um deep) were produced by moulding a PDMS against a microfabricated master with relatively simple and inexpensive methods. A CE-ECD systems were fabricated on the same substrate with the same fabrication procedure. The surface of PDMS layer and ITO-coated glass layer was treated with UV-Ozone to improve bonding strength and to enhance the effect of electroosmotic flow. For comparing the performance of the ITO electrodes with the gold electrodes, gold electrode microchip was fabricated with the same dimension. The running buffer was prepared by 10 mM 2-(N-morpholino)ethanesulfonic acid (MES) titrated to PH 6.5 using 0.1 N NaOH. We measured olectropherograms for the testing analytes consisted of catechol and dopamine with the different concentrations of 1 mM and 0.1 mM, respectively. The measured current peaks of dopamine and catechol are proportional to their concentrations. For comparing the performance of the ITO electrodes with the gold electrodes, electropherograms was measured for CE-ECD device with gold electrodes under the same conditions. Except for the base current level, the performances including sensitivity, stability, and resolution of CE-ECD microchip with ITO electrode are almost the same compared with gold electrode CE-ECD device. The disposable CE/ECD system showed similar results with the previously reported expensive system in the limit of detection and peak skew. When we are using disposable microchips, it is possible to avoid polishing electrode and reconditioning.

Fabrication of electro phoresis microchips and effects of channel surface properties (마이크로 전기영동 소자의 제작과 유로 면 특성에 따른 전기삼투 및 전기영동 효과)

  • Kim, Min-Su;Cho, Seung-Il;Lee, Kook-Nyung;Kim, Yong-Kweon
    • Proceedings of the KIEE Conference
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    • 2003.10a
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    • pp.286-289
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    • 2003
  • We investigated the influence of the properties of substrate material on the separation efficiency in microchip electrophoresis. We fabricated the various microchips and studied separation efficiency in microchannels composed of a single material such as quartz, glass, polydimethylsiloxane (PDMS), and polymethylmetha crylate (PMMA), as well as hybrid micro channels composed of different materials. New fabrication process for glass chip was suggested and some treatment is added to improve fabrication process in other chip. Separation efficiency was compared by measuring migration times and bandwidths of EOF and analytes in each microchip. The efficiency is the function of migration time, which is affected by the electroosmotic flow (EOF), and bandwidth of an analyte. EOF is highly dependent upon the characteristics of a microchannel wall surface. Migration time was more reproducible in silica chips than that of PDMS chip and more band broadening was observed in the microchip composed of hybrid material due to non-uniformity of surface charge density at the walls of the channel.

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Fabrication and Characterization of Micro parts by Mechanical Micro Machining: Precision and Cost Estimation (기계식 마이크로 머시닝을 이용한 마이크로 형상의 특성과 비용 평가)

  • Kang, Hyuk-Jin;Choi, Woon-Yong;Ahn, Sung-Hoon
    • Journal of the Korean Society for Precision Engineering
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    • v.24 no.1 s.190
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    • pp.47-56
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    • 2007
  • Recently, demands on mechanical micro machining technology have been increased in manufacturing of micro-scale precision shapes and parts. The main purpose of this research is to verify the accuracy and cost efficiency of the mechanical micro machining. In order to measure the precision and feasibility of mechanical micro machining, various micro features were machined. Aluminum molds were machined by a 3-axis micro stage in order to fabricate microchips with $200{\mu}m$ wide channel for capillary electrophoresis, then the same geometry of microchip was made by injection molding. To evaluate the cost efficiency of various micro manufacturing processes, cost estimation for mechanical micro machining was conducted, and actual costs of microchips fabricated by mechanical micro machining, injection molding, and MEMS (Micro electro mechanical system) were compared.

An Electrochemical Detector Using Prussian Blue Electrodeposited Indium Tin Oxide Electrode (Prussian blue가 전착된 indium tin oxide 전극을 이용한 전기화학적 검출기)

  • Yi, In-Je;Kim, Ju-Ho;Kang, Chi-Jung;Kim, Yong-Sang
    • The Transactions of the Korean Institute of Electrical Engineers C
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    • v.54 no.10
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    • pp.449-452
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    • 2005
  • We fabricated an electrochemical detector (ECD) to catalyze redox reaction efficiently by electrodepositing Prussian blue (PB) on the indium tin oxide (ITO) electrode. Capillary electrophoresis (CE) and amperometric method were used. We investigated the PB surface properties by topography from atomic force microscopy (AFM). Also PB film thickness calibration with respect to deposition time and voltage was used to get better PB surFace. The PB thin film of dense and smooth surface could catalyze redox reaction efficiently. Comparing with CE-ECD microchip using bare-lTO electrode, proposed CE-ECD microchip using PB deposited electrode has shown better sensitivity by determining the detected peak current from the electropherograms while the concentration of tested analyzes was maintained the same. It is verified that detection limit can be lowered for 0.01 mM of dopamine and catechol respectively.

Amperometric detection of DNA using capillary electrophoresis on microchip (모세관 전기영동 마이크로칩을 이용한 디옥시리보핵산(DNA)의 전류법 검출)

  • Joo, Gi-Sung;Ha, Kon;Jha, Sandeep K.;Lee, Hyun-Ho;Yoon, Tae-Sik;Kang, C.J.;Kim, Yong-Sang
    • Proceedings of the KIEE Conference
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    • 2008.07a
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    • pp.1460-1461
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    • 2008
  • 마이크로칩 형태에서의 모세관 전기영동과 전류법을 이용하여 디옥시리보핵산(DNA) 단편들의 분리 검출하는 실험을 하였다. 마이크로 채널이 형성된 PDMS(polydimethylsiloxane)와 금 전극이 형성된 유리 기판을 접합하여 마이크로칩을 제작하였다. 20V/cm의 전계를 인가하여 100bp-1.5kbp 길이의 DNA 단편을 모세관 전기영동 하였을 때 250초내에 분리 검출되는 것을 확인하였다.

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