微流控芯片活体单细胞胞内组分释放过程实时动态监测

Nondestructive detection for living cell was achieved through the activation of stimulant, by which the components were released, being important for the study of living cell process. In this project, real-time dynamic detection of conponent release of living single cell based on microfluidic chip would be developed, on which cell manipulation, stimulated release, separation, enzyme-linked immunoassay and detection were integrated. Histamine and tryptase release was induced by introduction of proteinase activated receptor-2 agonists. The simultaneous detection of histamine and tryptase release from tonsilmast cells was achieved by enzyme-linked immunoassay.

用刺激剂对完整细胞进行刺激，使细胞释放出胞内组分，从而实现无损检测，对单个活体细胞生命过程的研究具有重要意义。本项目将细胞操纵、受激释放、分离、酶联免疫反应、检测集成在芯片上，构建集成化的微流控芯片电化学酶联免疫分析单细胞受激释放在线快速检测技术平台。通过导入刺激剂蛋白酶激活受体-2激动剂，引起胞吐释放类胰蛋白酶和组胺。用电化学酶联免疫分析法检测，实现单个肥大细胞受激释放类胰蛋白酶和组胺的同时检测。

合理设计、加工集成碳纤维微工作电极和对电极的PDMS/玻璃复合芯片，将细胞操纵、受激释放、分离、酶联免疫反应、检测集成在芯片上，构建集成化的微流控芯片电化学酶联免疫分析单细胞受激释放在线快速检测技术平台。用低噪声碳纤维微电极实时监测单个肥大细胞组胺的量子释放，经过芯片电泳分离，用电化学酶联免疫分析法检测类胰蛋白酶，实现单个肥大细胞受激释放组分类胰蛋白酶和组胺的同时检测。进一步利用纳米探针结合循环放大技术，实验肿瘤细胞的检测及诊疗一体化研究。