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

本研究的目的是在微针、微球、微槽层合结构的基础上，设计制作生物化学、分析化学和医学微诊断芯片的原型作为实验，进一步推广这些原型所获得的知识，并将微诊断芯片的设计指南作为一个上级概念广泛展示。结果概要如下所示。(1)发展了用各种材料制造微针、微球、微槽、微层合结构的基本制造技术。发展了用激光加工和抛光金属箔形成几十微米左右的针的技术（针）、控制基板亲水性产生单细胞培养液滴（球）的技术（球）、用激光焊接将不锈钢基板粘合（层压）的技术。这些技术被应用于微诊断芯片的原型，如免疫测定芯片和单细胞培养芯片。我们研制了一种免疫分析芯片，利用不锈钢箔制成的壁作为电极，从凹槽两侧施加电场作用于样品试剂，使样品与抗体在直径为2微米的乳胶床上发生反应。我们研制了单细胞培养芯片，观察单细胞与试剂的反应，通过在芯片上绘出亲水性区域，使含有单细胞的细胞培养液滴在需要的位置上，通过芯片上形成的微通道，用试剂只给特定的细胞用药。(3)通过原型设计，揭示了微观空间与宏观空间之间的联系、根据检测原则存在最优空间尺寸等设计准则。

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.