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、微球和微槽叠层结构的基础上，设计和制作用于生物化学、分析化学和医学的微诊断芯片的原型作为实验，进一步总结了从这些原型学到的知识，并广泛地将微诊断芯片的设计准则作为一个上级概念展示出来。结果概要如下：(1)开发了用各种材料制造微针、球体、沟槽和层叠结构的基本制造技术。该技术通过激光加工和金属箔(针)抛光形成大约几十微米的针，开发了控制基质亲水性以产生单细胞培养液滴(球)的技术，即通过激光焊接(层压)将不锈钢基质粘合在一起的技术。(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。我们开发了免疫分析芯片，以不锈钢箔为电极，从凹槽的侧面向样品试剂施加电场，使样品与抗体在直径为2微米的乳胶蜂上发生反应。开发了单细胞培养芯片，观察单个细胞与试剂的反应，通过在芯片上形成亲水性区域，在所需位置生成包含单个细胞的细胞培养液，并通过芯片上形成的微通道仅向特定细胞投药。(3)通过原型设计，展示了微观空间与宏观空间的连接重要、根据检测原理存在最佳空间大小等设计原则。

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.