Development of a high-resolution scanning electrochemical microscope to characterize micro-bio devices

开发高分辨率扫描电化学显微镜来表征微生物设备

• 批准号: 15201030
• 负责人: MATSUE Tomokazu
• 金额: $ 32.45万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15201030/
• 关键词: Scanning Probe Microscopy; Micro-nano device; Biophysics; Research Instrument for Biology; Cellular array chip; ultramicroelectrode; near-field microscopy; 走査型電気化学顕微鏡; ナノプローブ; ディップ・ペン・ナノリソグラフィー; GFP; DNA; 距離制御; バイオチツプ; 走査型近接場光学顕微鏡; 大腸菌; バイ

We developed a high-resolution scanning electrochemical microscope (SECM) towards characterization of various micro-bio devices. Our SECM-based multifunctional measuring system provides near-field scanning optical microscopic (NSOM) information by featuring an optical fiber-electrode probe. The size of the electrode-, optical fiber-, or optical fiber-electrode- probe is less than 100nm, which allows us high-resolution images of protein arrays and cellular arrays. The feedback mechanism controlling the probe-sample distance is based on the shearing force, by which high-quality images of rough surfaces with the height differences larger than 10μm and fragile samples such as living cells under physiological conditions were captured.1, Nano-electrode probes were fabricated with the Ti/P sputtering and the subsequent parylene C-vapor deposition polymerization. The effective electrode radius estimated from the cyclic voltammogram for ferrocyanide has found to be less than 50nm. The optical f … More iber-electrode probes with the radii less than 100nm were made as the same manner. The optical aperture size was also less than 100 nm which was confirmed from the cross section of the NSOM image of the quantum-dot (QD) particles with tens nm-diameters.2, The feedback mechanism controlling the probe-sample distance was improved by vertically moving the probe with the width of 〜 100nm to reduce the damage applied to the samples. This feedback mode, defined as "standing approach (STA) mode" (Yamada et al., Anal. Chem., 2005, 77, 1785-179), has allowed to simultaneous electrochemical and topographic images of the axon and the cell body of the single PC-12 cell under physiological conditions for the first time. The STA-mode feedback imaging sometimes works better than the tip-sample regulation for the commercially available AFM. For example, relatively larger polystyrene beads (with 〜 10μm diameter) can be imaged with our STA-mode SECM, whereas not imaged with the conventional AFM instrument.3, Towards the progress in the construction of the micro-bio devices, various techniques to pattern baiomaterials were attempted. Especially, collagen-gel embedded culture of bacteria and mammalian cells with ultra-small volume (tens to several nL) has been successfully demonstrated on glass or silicon chip to microfabricate various types of cellular arrays. Micro-contact printing (μCP), microfluidics, dip-pen nanolithography (DPN), and negative dielectric phoresis (n-DEP) have been also applied to assemble protein and cellular arrays. Less

我们开发了一种高分辨率扫描电化学显微镜（SECM），用于表征各种微生物装置。我们基于secm的多功能测量系统通过光纤电极探头提供近场扫描光学显微镜（NSOM）信息。电极探针、光纤探针或光纤电极探针的尺寸小于100nm，这使我们能够获得蛋白质阵列和细胞阵列的高分辨率图像。控制探针-样品距离的反馈机制基于剪切力，通过剪切力捕获高差大于10μm的粗糙表面和生理条件下的脆弱样品（如活细胞）的高质量图像。1、采用Ti/P溅射和聚对二甲苯c -气相沉积法制备纳米电极探针。由循环伏安图估计的亚铁氰化物的有效电极半径小于50nm。采用相同的方法制备了更多半径小于100nm的光纤电极探针。量子点（QD）粒子的光学孔径也小于100 nm，这从直径为数十nm的量子点（QD）粒子的NSOM图像截面上得到证实。2、改进了控制探针-样品距离的反馈机制，通过垂直移动宽度为~ 100nm的探针，减少了对样品的损伤。这种反馈模式被定义为“站立接近（STA）模式”（Yamada et al., 2004）。化学。（2005, 77, 1785-179），首次实现了生理条件下单个PC-12细胞轴突和细胞体的同时电化学和地形图像。对于市售的AFM， sta模式反馈成像有时比尖端样品调节效果更好。例如，相对较大的聚苯乙烯珠（直径~ 10μm）可以用STA-mode SECM成像，而传统的AFM仪器无法成像。随着微生物器件建设的进展，人们尝试了各种材料的图案技术。特别是，胶原凝胶嵌入细菌和哺乳动物细胞的超小体积（几十到几个nL）培养已经成功地在玻璃或硅芯片上进行了微制造各种类型的细胞阵列。微接触印刷（μCP）、微流体、浸笔纳米光刻（DPN）和负介电电泳（n-DEP）也被用于组装蛋白质和细胞阵列。少

项目成果

Fabrication of Microbial Chip Using Collagen Gel Microstructure.

使用胶原凝胶微结构制造微生物芯片。

• 发表时间: 2003
• 期刊: Lab on a Chip 3
• 作者: T.Kaya

細胞の形質転換方法及び形質導入方法

细胞转化和转导方法

• 发表时间: 2005

Y.Torisawa: "Proliferation assay on a silicon chip applicable for tumors extirpated from mammalians"International Journal of Cancer. 109. 302-308 (2004)

Y.Torisawa：“适用于哺乳动物摘除肿瘤的硅芯片上的增殖测定”国际癌症杂志。

Dielectrophoretic micropatterning with microparticle monolayers covalently linked to glass surfaces

• DOI: 10.1021/la048111p
• 发表时间: 2004-12-07
• 期刊: LANGMUIR
• 影响因子: 3.9
• 作者: Suzuki, M;Yasukawa, T;Matsue, T
• 通讯作者: Matsue, T

Proliferation Assay on Chip Applicable for Tumors Extirpated from Mammalians

适用于从哺乳动物中摘除的肿瘤的芯片增殖测定

• 发表时间: 2004
• 期刊: Int. J. Cancer 109
• 作者: Y.Torisawa