Establishment of the design guidelines of laminated microdiagnostics chip using micro needle, sphere, and groove structures

本研究的目的是在使用微针、微球和微槽的层叠结构的基础上，设计并制作用于生物化学、分析化学和医学的微诊断芯片的原型，作为实验，将从这些原型中进一步获得的知识进行归纳，并将微诊断芯片的设计方针作为上位概念广泛地展示。（1）开发了用各种材料制造微针、球体、凹槽和层压结构的基本制造技术。通过激光加工和抛光从金属箔（针）形成约几十微米的针的技术，控制基底的亲水性以产生单细胞培养液滴（球体）的技术，开发了使用激光焊接（层压）结合由不锈钢制成的基板的技术。（2）将这些技术应用于免疫分析芯片和单细胞培养芯片等微诊断芯片的原型制作中，研制了免疫分析芯片，该芯片以不锈钢箔为电极，在直径为2 μ m的乳胶粒上，通过对样品试剂施加电场，使样品与抗体发生反应。我们开发了单细胞培养芯片，通过在芯片上的亲水区域形成图案以在所需位置上产生含有单细胞的细胞培养溶液的液滴，并通过在芯片上形成的微通道仅向特定细胞施用试剂，来观察单细胞与试剂的反应。（3）此外，通过原型制作，示出了其中微观空间与宏观空间的连接是重要的、根据检测原理存在最佳空间尺寸等的设计准则。

The purpose of this research is designing and making the prototypes of microdiagnostics chip for biochemistry, analytical chemistry, and medicine as an experiment on the basis of the laminating structure using micro needle, sphere, and groove, generalizing the knowledge further acquired from those prototyping, and showing the design guideline of a microdiagnostics chip widely as a superordinate concept.The outline of the results is shown below.(1)Fundamental fabrication technique which produces micro needle, sphere, groove, and laminating structure with various materials was developed.The technique which forms an about dozens of micrometers needle by laser processing and polish from metal foil(needle), the technique which controls the hydrophilicity of a substrate to generate single cell culture solution drops(sphere), the technique which bonds the substrates made from stainless steel using laser welding(laminating), were developed.(2)These technique ware applied to the prototyping of microdiagnostics chips, such as an immunoassay chip and a single-cell culture chip.We developed the immunoassay chip to react sample with antibody on the latex beeds with a diameter of 2 micrometers by applying electric field to the sample reagent from sides of a groove using the wall made from stainless steel foil as the electrodes. And we developed the single cell culture chip to observe a reaction of a single cell with reagent, by patterning hydrophilic area on the chip to generate drop of cell culture solution containing a single cell on the desired position, and medicating only a specific cell with the reagent through the microchannel formed on the chip.(3)Moreover, the design guidelines in which connection of micro space with macro space is important, there is optimal space size according to a detection principle, and so on, were shown through the prototyping.