基于电渗诱导压力流的流式细胞术微流控方法及其应用研究

Flow cytometry is a popular and significant analytical cell-biology technique which is used extensively to characterize the physical and chemical characteristics of cells. However, conventional flow cytometers are often bulky, expensive, complicated in operation and require high sample consumption, preventing the researchers from using it. Microfluidic technology has gained much scientific interest in the last decades and been applied and adopted across many areas, and also demonstrated that it is a potentially valuable tool for flow cytometric analysis. Although numerous efforts have been taken in the development of flow cytometry based on microfluidics, breakthrough progress has not been made yet. In this project, a novel electrokinetically-induced pressure-driven method for microfluidic system is proposed. Numerical and experimental analysis will be performed to investigate the method. By using the electrokinetically-induced pressure-driven method, a novel microfluidic chip integrated with multiple functions including cell driving, focusing, counting, sorting and detection will be developed towards the flow cytometric analysis. The detailed research methodologies are as below: (1) Numerical and experimental study of the fundamental of electrokinetically-induced pressure-driven flow; (2) Numerical and experimental investigation for the on-chip functional units design. 3) Develop a method for miniaturizing the detection system. The theoretical study of the electrokinetically-induced pressure-driven flow in the microfluidic chip in this proposal will enrich the theory of fluid mechanics in the microscale, overcome the technical difficulties in current microfluidic flow cytometry, hence providing a new perspective for the development of the microfluidic flow cytometry and further expanding its applications.

目前，流式细胞术已经成为当代最先进和最重要的细胞定量分析技术。但是，昂贵的价格、巨大的体积、复杂的操作以及超高的消耗却让普通科研人员望而却步。随着微流控技术的进步，很多科学小组将微流控的概念引入到流式细胞术当中，但相关努力缺乏突破性进展。为了克服这一困难，本课题提出了基于电渗诱导压力流的驱动新方法，并以此为基础设计和制作集成细胞驱动、聚焦、计数、鉴定、分选一体化的新型微流控芯片，力求发展一种流式细胞术微流控新方法。具体研究内容涵括三个方面：（1）基于电渗诱导压力流理论，探究微流控流式细胞术的各个功能单元的机制并进行理论分析、数值模拟和实验验证；（2）构建集成各个功能单元的微流控芯片并进行试验验证；（3）发展微型化的检测方法。本课题的研究突破了现有的流式细胞术微流控方法的瓶颈，丰富了微尺度下流体力学的理论，为研制便携式流式细胞仪提供了新的视角和思路，为流式细胞术的应用开拓了更广阔的空间。

本项目以微流控技术为核心，以微尺度下流体力学为核心要素，创新性地建立了基于电渗诱导压力流的理论模型，突破性解决了现有的流式细胞术微流控方法瓶颈问题，进而构建了微流控芯片，进一步将其应用于微颗粒/细胞驱动、聚焦、计数、分选和鉴定，显示了微流控芯片技术与生物医学的交叉融合。主要研究内容有：1）构建尺度-电场耦合作用下的流场模型来揭示细胞在微通道内的运输和聚焦机制；2）研究微流控芯片中对微颗粒/细胞进行分选的机制；3）研究检测系统微型化的方法；4）研究液态金属微电极与微流控芯片结构的一体化工艺方法；5）研究基于电渗诱导压力流的流式细胞术的应用和功能评价。. 自项目实施以来，发表11篇高质量研究论文（SCI收录10篇）；申请中国技术发明专利5项，授权3项；培养博士研究生2名，硕士研究生3名；受邀参加国际/国内会议8次，并与国内外多名领域专家开展学术交流，推动了我国微流控新技术以及应用的发展。