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Construction of biochemical analytical systems using the microchip technology

利用微芯片技术构建生化分析系统

基本信息

批准号：
14206014
负责人：
ABO Mitsuru
金额：
$ 33.11万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14206014/
关键词：
micro-nanodevice environmental analysis cell system on chip allergy sensor マイクロチップイムノアッセイ PDMS 肥満細胞ヒスタミンマクロファージ TNF-α 酵素活性 TLM ステロイドホルモン ELISA

项目摘要

Biochemical analytical systems using microchip technology were developed. In the field of environmental analyses, analytical systems for steroid hormones in marine gastropods and for nitrite and sulfite in river were constructed. Analytical systems for bioactive compounds such as cytokines were also developed. An immunoassay microchip for TNF-α from mouse macrophages and a bioassay microchip for histamine from rat peritoneal mast cells were constructed.First, we developed a simple method for micro-fabrication on a glass plate by photolithography using a high viscosity photoresist as an etching mask and then the fabricated glass was used for the template of a polydimethylsiloxane (PDMS) microchip. We also developed a novel fluorescence detection unit for microchip analyses.The analytical sensitivity for steroid hormones in marine gastropods, using the immunoassay microchip, was improved by one order of magnitude and the analysis time was reduced to one third, compared to the conventional ELISA method.In the allergic drug screening system for the detection of histamine released from mast cells, the number of the cells, required for the assay, was reduced to 1%, compared with conventional bulk systems and the real-time monitoring of the histamine release was achieved by the continuous flow of cell culture medium.Microbioassay system for an anti-cancer agent equipped with a microfluidic gradient mixer. The simultaneous gradient assay was conducted easily by simply flowing two fluids in microchannels. This system demonstrated that cell viability of a stomach cancer cell line changed according to the concentrations of an anti-cancer agent.

开发了使用微芯片技术的生化分析系统。在环境分析领域，建立了海洋腹足类中甾体激素和河流中亚硝酸盐和亚硫酸盐的分析系统。还开发了用于生物活性化合物如细胞因子的分析系统。首先，采用高粘度光刻胶作为掩膜，在玻璃板上进行微加工，然后将微加工后的玻璃作为聚二甲基硅氧烷（PDMS）微芯片的模板，构建了小鼠巨噬细胞TNF-α免疫检测微芯片和大鼠腹腔肥大细胞组胺生物检测微芯片。我们还开发了一种新型的用于微芯片分析的荧光检测单元，与传统的ELISA方法相比，该免疫芯片对海洋腹足类中类固醇激素的分析灵敏度提高了一个数量级，分析时间缩短到三分之一。与传统的本体系统相比，组胺释放量降低到1%，并通过细胞培养液的连续流动实现了组胺释放的实时监测。通过简单地使两种流体在微通道中流动，可以容易地进行同时梯度测定。该系统证明胃癌细胞系的细胞活力根据抗癌剂的浓度而改变。