Development of Bioprobe Integrated with Microneedle for Cellular Function Analysis

开发与微针集成的生物探针用于细胞功能分析

• 批准号: 18300150
• 负责人: SHIBATA Takayuki
• 金额: $ 10.5万
• 依托单位: Toyohashi University of Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18300150/
• 关键词: Microneedle; Atomic force microscope(AFM); Cellular function analysis; MEMS; BioMEMS; Micromachining; Cell surgery; Nanobiotechnology

In order to realize cellular function analysis on a chip-base system, we have developed a newly designed AFM probe in which a conventional sharp tip is superseded by a hollow silicon dioxide(SiO_2) microneedle connecting the root to a fluidic microchannel embedded into a silicon(Si) cantilever beam structure. The probe will be capable not only of performing AFM measurements but also of introducing desired biomolecules(nucleic acids, proteins, etc.) into living cells and extracting biomolecules expressed in the cells.The micromachining technique of a hollow SiO_2 microneedle with a sharpened tip has been developed on the basis of a combination of an anisotropic Deep reactive ion etching(DRIE) process for producing thorough holes into a Si substrate as a needle mold followed by wet oxidation and a selective etching process for leaving a SiO_2 microneedle structure, resulting that microneedles with a tip diameter of less than 0.1 μm(5.5 μm in outer diameter and 20 μm in length) was succes … More sfully fabricated. A circular-shaped microchannel(typically 6 μm in diameter) was also successfully fabricated into a Si cantilever structure. In additon, a novel electrostatic field-assisted direct bonding technique was developed.In order to investigate the mechanical stability of a microneedle array, penetration tests were performed with a gelatin as an artificial cell. The results indicated that the microneedles were expected to be sufficiently stiff to penetrate living cells without fracture. Insertion performance into gelatin and the possibility of liquid delivery with fabricated microneedles were also investigated. The displacement required for needle insertion increased linearly with an increase in surface area at the needle tip. It was also found that increasing mechanical oscillation frequency during insertion decreased remarkably the displacement probably due to an increase in viscous resistance. Moreover, the fluid ejection tests revealed the possibility of liquid delivery by employing fabricated microneedles with an inner diameter as small as 3.5 μm. It was also founded that the flow rate increased in proportion to the fourth power of the inner diameter of the needles that is subject to the Hagen-Poiseuille's equation. Less

为了在芯片系统上实现细胞功能分析，我们开发了一种新设计的AFM探针，该探针采用空心二氧化硅（SiO_2）微针代替传统的尖头，将根连接到嵌入硅（Si）悬臂梁结构中的流体微通道。该探针不仅能够进行AFM测量，而且能够将所需的生物分子（核酸、蛋白质等）引入活细胞并提取细胞中表达的生物分子。采用各向异性深度反应离子蚀刻（DRIE）工艺在Si衬底上形成小孔，然后湿氧化，再采用选择性蚀刻工艺留下SiO_2微针结构，在此基础上发展了尖端锋利的空心SiO_2微针的微加工技术。结果表明，制备出了针尖直径小于0.1 μm（外径5.5 μm，长20 μm）的微针。圆形微通道（直径通常为6 μm）也成功地制成了硅悬臂结构。此外，还开发了一种新的静电场辅助直接键合技术。为了研究微针阵列的机械稳定性，用明胶作为人工细胞进行了渗透试验。结果表明，微针有望有足够的硬度穿透活细胞而不断裂。还研究了微针在明胶中的插入性能以及用制备的微针输送液体的可能性。针插入所需的位移随针尖表面积的增加而线性增加。增加插入时的机械振荡频率会显著降低位移，这可能是由于粘性阻力的增加。此外，流体喷射试验还揭示了利用内径小至3.5 μm的自制微针输送液体的可能性。还发现，流速的增加与针内径的四次方成正比，这符合ha根-泊泽维尔方程。少

项目成果

Development of MEMS devices for cell surgery and cellular function analysis on a chip-based system (in Japanese)

在基于芯片的系统上开发用于细胞手术和细胞功能分析的 MEMS 设备（日语）

• 发表时间: 2007
• 期刊: The Papers of Technical Meeting on Linear Drives, IEE Japan LD-07-49
• 作者: N. Kato;T. Shibata;T. Sakai;S. Yamanaka;T. Kawashima
• 通讯作者: T. Kawashima

オンチップ細胞サージェリーシステムのためのマイクロニードルアレイの開発

片上细胞手术系统微针阵列的研制

• 发表时间: 2006
• 期刊: 表面技術協会第114回講演大会論文集
• 作者: 中西 彬;柴田隆行;川島貴弘;堀内 宰;峯田 貴;牧野英司
• 通讯作者: 牧野英司

細胞機能解析のためのマイクロニードル搭載型バイオプローブの開発(第1報)-先鋭化ニードルおよびマイクロチャネルの作製-

用于细胞功能分析的配备微针的生物探针的开发（第一份报告） - 锋利的针和微通道的制造 -

• 发表时间: 2007
• 期刊: 精密工学会春季大会講演論文集
• 作者: 加藤統久;柴田隆行;川島貴弘;堀内 宰;峯田 貴;牧野英司
• 通讯作者: 牧野英司

マイクロニードル搭載型バイオプローブ, およびマイクロニードル搭載型バイオプローブの作製方法

配备有微针的生物探针以及制造配备有微针的生物探针的方法

• 发表时间: 2006

Insertion performance of hollow microneedle array for cell surgery

细胞手术中空心微针阵列的插入性能

• 发表时间: 2008
• 期刊: Proc. Asia-Pacific Conference on Transducers and Micro-Nano Technology 2008(APCOT2008) (印刷中)
• 作者: T. Kawashima;T. Shibata;T. Sakai;N. Kato;M. Nomura;T. Mineta;E. Makino
• 通讯作者: E. Makino