研究者業績
基本情報
研究分野
1経歴
8-
2017年4月 - 現在
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2007年10月 - 2017年3月
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2003年8月 - 2007年9月
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2003年1月 - 2003年7月
学歴
3-
1997年4月 - 2000年3月
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1995年4月 - 1997年3月
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1991年4月 - 1995年3月
委員歴
26-
2024年4月 - 現在
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2024年3月 - 現在
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2022年 - 現在
受賞
11-
2015年11月
論文
189-
Sensors and Materials 35(10) 4781-4781 2023年10月25日
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RSC Advances 13(31) 21118-21126 2023年7月12日Surface plasmon resonance is an optical phenomenon that can be applied for label-free, real-time sensing to directly measure biomolecular interactions and detect biomarkers in solutions. Previous studies using plasmonic nanohole arrays have monitored and detected various biomolecules owing to the propagating surface plasmon polaritons (SPPs). Extraordinary optical transmission (EOT) that occurs in the near-infrared (NIR) and infrared (IR) regions is usually used for detection. Although these plasmonic nanohole arrays improve the sensitivity and throughput for biomolecular detection, these arrays have the following disadvantages: (1) molecular diffusion in the solution (making the detection of biomolecules difficult), (2) the device fabrication's complexities, and (3) expensive equipments for detection in the NIR or IR regions. Therefore, there is a need to fabricate plasmonic nanohole arrays as biomolecular detection platforms using a simple and highly reproducible procedure based on other SPP modes in the visible region instead of the EOT in the NIR or IR regions while suppressing molecular diffusion in the solution. In this paper, we propose the combination of a polymer-based gold nanohole array (Au NHA) obtained through an easy process as a simple platform and dielectrophoresis (DEP) as a biomolecule manipulation method. This approach was experimentally demonstrated using SPP and LSPR modes (not EOT) in the visible region and simple, label-free, rapid, cost-effective trapping and enrichment of nanoparticles (trapping time: <50 s) and bovine serum albumin (trapping time: <1000 s) was realized. These results prove that the Au NHA-based DEP devices have great potential for real-time digital and Raman bioimaging, in addition to biomarker detection.
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Lab on a Chip 23(4) 692-701 2023年2月 査読有り最終著者責任著者Simultaneous electrorotation in microwells during chemical stimulation label-free monitoring effect of chemicals in single-cell manner.
MISC
129-
Proceedings of the Chemical Sensor Symposium 71 1-3 2022年9月
書籍等出版物
14講演・口頭発表等
168-
Hyper Bio Assembler for 3D Cellular Systems 2015年1月1日© Springer Japan 2015. Formation of line pattern with cells based on dielectrophoresis (DEP) was applied to simple and rapid distinction of cells with specific surface antigens from a cell population. Dispersed cells were accumulated to gap areas of the interdigitated band array (IDA) electrode modified with antibody within 5 s by negative DEP (n-DEP) and captured by immunoreactions. Unbounded cells without the specific antigen on the membrane were removed to form another pattern by switching the applied voltage of the band electrode. The time required for the assay was substantially short, 60 s for forcing and 60 s for the separation of unbounded cells. Furthermore, the present method does not require pretreatment such as target labeling or washing of unbound cells.
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2014 International Symposium on Micro-NanoMechatronics and Human Science, MHS 2014 2014年1月1日© 2014 IEEE. We present single-cell pairing of different types of cells with a rapid manipulation based on positive dielectrophoresis (p-DEP). The DEP device for the manipulation of cells consisted of a microfluidic channel with an upper indium tin oxide (ITO) electrode and a lower ITO electrode with 10,000 (100 × 100) microwells. The width (14 μm) and depth (25 μm) of the individual microwells restricted the size to two vertically aligned cells. Cells stained in blue and stained in green were continuously trapped in the microwells. Cells were paired within only 1 min and a pairing efficiency of 53% was achieved.
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World Automation Congress Proceedings 2014年1月1日© 2014 TSI Press. Rapid conversion of the line formation with the cells based on dielectrophoresis (DEP) was applied to simple and rapid distinction of cells with specific surface antigens from a cell population. The gap area of the interdigitated band array (IDA) electrode was modified with anti-CD33 antibody. Human promyelocytic leukemia cells (HL-60) which express CD33 surface antigen were accumulated on the surface in the gap area between both bands of the IDA electrode by negative DEP (n-DEP). Switching of the applied voltage of the band electrode brought about the removal of accumulated cells to form another pattern due to the formation of the different pattern of the electric field in the device. The modification with the antibody inhibits the removal of the cells with specific surface antigen due to the irreversible capture with immunoreactions during the first pattern formation. Therefore, we should discriminate the cells with specific antigen from the cell remained on the gap area. The time required for the assay was substantially short, 60 s for forcing and 60 s for the separation of unbounded cells. Furthermore, the present method does not require pretreatment such as target labeling or washing of unbound cells.
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18th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2014 2014年1月1日© 14CBMS. The rapid manipulation technique with positive dielectrophoresis (p-DEP) was applied to the formation of single-cell pairs in a microwells consisted of a microfluidic channel with an upper indium tin oxide (ITO) electrode and a lower microwell array electrode fabricated on an ITO substrate. Myeloma cells were trapped within 1 s in the microwells restricted the size to two vertically aligned cells by p-DEP. Different types of cells were paired within 1 min and a pairing efficiency of approximately 50% was achieved. The use of the present device allows large numbers of cell pairs (over 5,000 pairs) with the vertical alignment.
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電子情報通信学会技術研究報告 : 信学技報 2013年4月25日誘電泳動を用いると微粒子を数秒で迅速に操作することが可能である.我々は,この技術を迅速な免疫測定法へと応用展開している.抗体を固定化した基板と交互くし型マイクロアレイ電極を組み合わせて誘電泳動デバイスを作製し,電極に交流電圧を印加すると不均一電場が形成され微粒子に誘電泳動力が作用する.負の誘電泳動を用いて抗体固定化微粒子を抗体固定化基板上へ集積化させると,微粒子-基板間で免疫反応が進行し,わずか数分で微粒子は抗体固定化基板へ捕捉される.この捕捉された微粒子の数は,測定対象物質濃度に依存するため迅速な計測が可能となる.また,電圧印加停止後,未反応微粒子は基板から再分散するため洗浄工程が不要となる.デバイスに導入する初期微粒子濃度を制御することにより,検出濃度領域を制御できるため免疫測定の高感度化が期待できる.
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2013 International Symposium on Micro-NanoMechatronics and Human Science, MHS 2013 2013年1月1日We applied a rapid and simple fabrication method of the island pattern of particles and cells to discriminating cells with specific surface antigen. The Upper interdigitated microband array (IDA) electrode was mounted on the lower substrate with the same design to fabricate a microfluidic-channel device for dielectrophoretic manipulation. Grid formation of electrodes was fabricated by rotating the upper template IDA by 90° relative to the lower IDA. A suspension of particles modified with anti-CD33 antibody or/and HL60 cells was introduced into the channel. AC electric signal (typically 20 V peak-to-peak, 100 kHz) was then applied to the bands on the upper and lower IDA, resulting in the formation of island patterns at the intersections with low electric fields. The accumulated particles and cells were fixed to produce the complexes through the immunoreactions between the antibody immobilized on the particle and CD33 on the cell surface. The complexes were only produced by the corresponded pair of antigen-antibody. It is noted that the time required for single sensing is as short as 6 min in the presented procedure. Furthermore, the present method for a novel cell binding assay does not require pretreatment such as target labeling or washing of unbound cells. © 2013 IEEE.
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2012 International Symposium on Micro-NanoMechatronics and Human Science, MHS 2012 2012年12月1日The rapid and simple discrimination system of cells with specific surface antigen was developed based on the pattern switching of cellular array formed by the positive and negative dielectrophoresis (p-DEP and n-DEP). p-DEP was used to accumulate the cells and capture by the immunoreactions between the specific surface antigen and antibody immobilized on the surface, while n-DEP was to remove of unbound cells. The time required for the determination of the surface antigen is decreased to 60 s compared to that required by a cell binding assay using microtiter plates (30 min). Furthermore, the present method for a novel cell binding assay does not require pretreatment such as target labeling or washing of unbound cells. © 2012 IEEE.
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ECS Transactions 2012年12月1日Rapid accumulations of particles and living cells with negative dielectrophoresis (n-DEP) have been applied to develop the rapid and simple immunosensing method. Grid formation of electrodes was fabricated by rotating the upper template interdigitated microband array (IDA) electrode by 90° relative to the lower IDA. When AC electric signal was applied to the bands on the upper and lower IDA, island organization was rapidly formed at the intersections with low electric fields. The accumulated particles were fixed through the immunoreactions between the antibody immobilized on the particle surface and analytes in the solution. The presence of the specific antigens allowed the formation of fixed complexes of particles. It is noted that the time required for single sensing is as short as 5 min and separation steps are eliminated in the presented procedure. We demonstrated the rapid and simple immunosensing using the aggregation of particles accumulated with DEP. © 2012 The Electrochemical Society.
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電子情報通信学会技術研究報告 : 信学技報 2012年2月17日ターゲットとする表面抗原を発現した細胞を正の誘電泳動(p-DEP)により抗体固定化電極表面上に迅速に集積化し免疫反応により捕捉した.さらに,負の誘電泳動(n-DEP)を用いて,未反応の細胞を電極表面上から電極間ギャップへと移動させ表面抗原発現細胞を識別,分離した.ここでは,骨髄性白血病細胞(HL-60)に発現しているCD33抗原をモデルターゲットとして用い,抗CD33抗体固定化電極上に捕捉した.このp-DEPによる細胞の迅速な集積化とn-DEPによる未反応細胞の分離を用いると,アッセイに必要とする時間は60秒と極めて迅速であった.これは,通常の細胞結合性アッセイで必要とした時間(30分)と比較して明らかに優位性を有する.
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Proceedings of the 16th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2012 2012年1月1日In this paper, we describe a development of the rapid and simple discrimination system of cells with specific surface antigen by immobilization of cells accumulated by positive dielectrophoresis (p-DEP) via effective surface immunoreactions and removal of unbound cells by negative DEP (n-DEP). The time required for the determination of the surface antigen is decreased to 60 s compared to that required by a cell binding assay using microtiter plates (30 min). Furthermore, the present method for a novel cell binding assay does not require pretreatment such as target labeling or washing of unbound cells.
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Chemical sensors 2011年
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20th Anniversary MHS 2009 and Micro-Nano Global COE - 2009 International Symposium on Micro-NanoMechatronics and Human Science 2009年12月1日In the work, microfluidic device consisting of an interdigitated microarray (IDA) electrode was developed for a rapid, and separation-free immuno-sensors based on a manipulation technique of microparticles by dielectrophoresis (DEP). A poly-dimethylsiloxane (PDMS) substrate with microfluidic channel was placed on the IDA plate to allow to fabricating the device. On applying AC voltage to the IDA in a negative DEP (n-DEP) frequency region, goat anti-mouse immunoglobulin (anti-mouse IgG)-immobilized microparticles moved to the surface of PDMS substrate placed above the IDA by n-DEP force to accumulate at the designated areas of the PDMS surface, where anti-mouse IgG was precoated. When the fluorescence microparticles bearing anti-mouse IgG were suspended in an analyte (mouse IgG) solution, the microparticles trapped the analyte to form microparticle-conjugates. The conjugates were accumulated and captured at the designated areas of the PDMS surface via antibody-antigen-antibody (sandwich) reaction. The captured microparticles were detected selectively by fluorescence measurements at the focused, designated areas regardless of the presence of uncaptured microparticles in the suspended solution. Thus, the separation and washing-out steps, usually required for conventional immunoassay, are eliminated in the presented procedure. Since the formation of the sandwich structures was accelerated significantly by n-DEP, as short as 30 sec was enough to detect the immunoreaction at the surface. The fluorescence intensity of the captured microparticles at the designated area increased with the analyte in the range, 0.01 ∼ 10 ng/mL. The present procedure realizes a rapid, sensitive and separation-free immunoassay in a simple device. ©2009; IEEE.
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Proceedings of Conference, MicroTAS 2009 - The 13th International Conference on Miniaturized Systems for Chemistry and Life Sciences 2009年1月1日Endocrine disrupting chemicals that act like hormones is a potential hazard for human health. Therefore, it is necessary to develop a device that can detect easily hormone active chemicals. In this study, a microfludic device with some analytical chambers was fabricated for manipulating the yeast cells that were genetically engineered to respond to the presence of hormone active chemicals by synthesizing β-galactosidase (β-gal). Interdigitated array (IDA) electrodes with 40 electrode fingers were incorporated into the device for sensitive detection of the β-gal activity electrochemically, and hormone active chemicals were successfully detected by using the systems. © 2009 CBMS.
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ECS Transactions 2008年12月1日A coulometric signal accumulation system to detect a trace concentration of hydrogen peroxide (H2O2) has been applied to develop a novel electrochemical enzyme immunosensing system for insulin, an important hormone for clinical diagnosis of diabetes. Glucose oxidase (GOx)-labeled anti-insulin IgG antibody was trapped on the substrate immobilized with insulin by an indirect competitive immunoassay. H2O2 generated by the enzymatic reaction with the trapped conjugate was detected by the coulometric signal accumulation system. The H2O2 generated oxidized [Os(bpy)2Cl]+ to [Os(bpy)2Cl] 2+ in a horseradish peroxidase (HRP)-containing polymer layer on an electrode under the open-circuit condition, and then the [Os(bpy) 2Cl]2+ accumulated was reduced by applying a negative potential. The accumulation brought about a large current response and a low detection limit for H2O2 (0.5 nM). The enhanced sensitivity for H2O2 enabled the determination of a trace concentration of insulin with the immunoassay using the antibody modified with GOx, a H2O2-producing enzyme. © The Electrochemical Society.
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2008 International Symposium on Micro-NanoMechatronics and Human Science, MHS 2008 2008年12月1日Negative dielectrophoresis (n-DEP) have been used to manipulate microparticles with immunoreagents (antigens or antibodies) in a microfluidic channel, and applied to develop a rapid immunoassay system. A microfluidic device, with three-dimensional (3-D) microelectrodes fabricated on two substrates, was used to manipulate particle flow in the channel and to capture the particles in the caged area that was enclosed by the collector electrodes. Polystyrene microparticles (6 μm diameters) modified with anti-mouse immunoglobulin G (IgG) were manipulated and captured in the caged area by using n-DEP. A sandwich immunoassay was achieved by successively injecting a sample solution containing mouse antigen (IgG), and a solution containing FITC-labeled anti-mouse IgG antibody, into the channel. The fluorescence intensity from captured particles in the caged area increased with increasing concentrations (10 ng/ml to 10 μg/ml) of mouse IgG. The described system enables mouse IgG to be assayed in 40 min. This immunosensing system using the n-DEP technique is faster and simpler than conventional enzyme-linked immunosorbent assay (ELISA) using microtiter plates, and has the significant advantage that sensing requires simple and easy handling since unreacted immunomolecules are flushed from the signal detection area by the fluidic stream. The device can be reused by removing the microparticles. The automatic separation of free fractions from desired analytes and labeled antibodies can be achieved using a microfluidic device based on n-DEP. © 2008 IEEE.
担当経験のある科目(授業)
5-
2008年 - 現在化学実験 (兵庫県立大学)
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2020年10月 - 2023年2月バイオセンシングの化学 (兵庫県立大学)
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機器分析学 (兵庫県立大学)
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分析化学II (兵庫県立大学)
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分析化学I (兵庫県立大学)
所属学協会
10共同研究・競争的資金等の研究課題
64-
日本学術振興会 科学研究費助成事業 2024年4月 - 2027年3月
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日本学術振興会 科学研究費助成事業 基盤研究(B) 2023年4月 - 2026年3月
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日本学術振興会 科研費 2023年4月 - 2026年3月
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日本学術振興会 科学研究費助成事業 挑戦的研究(萌芽) 2022年6月 - 2024年3月
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日本学術振興会 科学研究費助成事業 基盤研究(B) 2020年4月 - 2023年3月