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; 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analysis on a chip-base system, we have developed a newly designed AFM probe in which a conventional sharp tip is superseded by a hollow silicon dioxide(SiO_2) microneedle connecting the root to a fluidic microchannel embedded into a silicon(Si) cantilever beam structure.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

为了实现芯片基碱系统上的细胞功能分析，我们开发了一种新设计的AFM探针，其中传统的锋利尖端被一个将根连接到嵌入硅（SI）danilever束结构中的流体微通道的微孔二氧化硅（SIO_2）微孔系取代。该探测器不仅能够进行AFM测量，还可以在细胞中引入所需的生物分子（核酸，蛋白质等），并提取在细胞中表达的生物分子。基于空心的Sio_2 microneedle的微加工技术，具有尖锐的尖端的基础，该过程是由AniS and and and reative far Intie fariie fariie fariie fariie fariie fariie fropic et a ansis and ansis andis and ansis of a ansis of and andis andis andis andiS andis andis andis andis andis of andis andis andiS andis andis andiS arties for for（将孔进入Si底物作为针头霉菌，然后进行湿氧化和留下SIO_2微针结构的选择性蚀刻过程，从而导致尖端直径小于0.1μm（外部直径为5.5μm，长度为20μm，长度为20μm）的微针成功制作了……更加成功地制作了。还将圆形微通道（通常直径为6μm）成功地制造成Si悬臂结构。在Additon中，开发了一种新型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，用明胶作为人造细胞进行穿透性测试。结果表明，预计微针具有足够的僵硬，可以穿透活细胞而不会裂缝。还研究了在明胶中的插入性能，并研究了用制造的微针的液体输送的可能性。针头插入所需的位移随着针头尖端的表面积增加而线性增加。还发现，插入过程中机械振荡频率的增加显着降低了位移可能是由于粘性阻力的增加所致。此外，流体射血测试通过采用内径高达3.5μm的造成的微针，揭示了液体递送的可能性。还建立的是，流速与受Hagen-Poiseuille方程的针头内径的第四功率成比例增加。较少的