Three Dimensional, Passivated-Electrode, Insulator-Based Dielectrophoresis (3D-PiDEP)

研究目的：研制新型介质电泳（DEP）芯片，该芯片由三维绝缘结构和钝化电极组成，用于低成本、高通量、选择性地分离和富集水中不同病原体。知识优势：我们将结合先进的3D硅微加工技术、软光刻技术和DEP技术，开发基于钝化电极绝缘体的三维介质电泳（3D-£kDEP）芯片，用于病原体富集。3D-£kDEP芯片由pdms -玻璃平台上的多个三维微结构阵列区域组成，通过利用3D绝缘体结构产生强电场梯度，并使用钝化电极施加所需的交流电压来选择性隔离/捕获包括病原体在内的生物颗粒，减轻了当前基于电极的DEP和基于绝缘体的DEP机制的缺点。3D-£kDEP芯片可用于表征DEP特性，并选择性捕获耐甲氧西林金黄色葡萄球菌（MRSA）和大肠杆菌（SMS-3-5）作为模型革兰氏阳性和阴性细菌。3D-£kDEP设备将通过对定量聚合酶链反应（qPCR）获得的结果进行基准验证，定量聚合酶链反应（qPCR）通过定量已知的目标基因拷贝对感兴趣的病原体特异性起作用。更广泛的影响：这项研究导致了一种新的DEP方法的发展，该方法在设计和操作上具有很高的灵活性，同时为生物流体中的颗粒分离和富集提供了可扩展的吞吐量。这项研究还促进了高中、本科和研究生阶段的教学。这包括：1)为本科生提供REU的机会；2)招募研究生参加一个高度跨学科的研究项目；3)为参加VT大学预科计划和学院外展活动的高中生提供实验室参观和演讲；4)将项目成果纳入不同部门的pi教授的课程；5)pi联合研讨会。此外，这项研究的结果将在同行评议的期刊上广泛传播给工程和科学界，并在多学科会议上发表。

Research Objective: The aim of this research is to develop new dielectrophoretic (DEP) chips comprising three-dimensional insulating structures and passivated electrodes for low-cost, high-throughput, and selective isolation and enrichment of different pathogens in water. Intellectual Merits: We will combine advanced 3D silicon micromachining technologies, soft lithography, and DEP techniques to develop three-dimensional passivated-electrode insulator-based dielectrophoresis (3D-£kDEP) chips for pathogen enrichment. The 3D-£kDEP chip, which consists of several zones with arrays of three-dimensional microstructures on a PDMS-glass platform, alleviates the shortcomings of the current electrode-based DEP and insulator-based DEP mechanisms by utilizing 3D insulator structures for generating strong electric field gradients and using passivated electrodes to apply the required AC voltage for selective isolation/trapping of bioparticles including pathogens. The 3D-£kDEP chips can be used to characterize the DEP properties and selectively trap Methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli (SMS-3-5) as model Gram positive and negative bacteria. 3D-£kDEP devices will be validated by benchmarking the results obtained against quantitative polymerase chain reaction (qPCR) which works by quantifying a target gene copy known to be specific to the pathogen of interest. Broader Impacts: This research leads into the development of a new DEP method with high flexibility in design and operation while providing scalable throughput for separation and enrichment of particles in biological fluids. The research also promotes teaching and learning at the high school, undergraduate, and graduate levels. This includes: 1) REU opportunities for undergraduates, 2) recruiting of graduate students into a highly interdisciplinary research program, 3) lab tours and lecture presentation for high school students participating in VT Pre-College Initiative and College outreach activities, 4) incorporation of the project results in the courses taught by the PIs in different departments, and 5) joint seminars by the PIs. Additionally, the outcome of this research will be widely disseminated to the engineering and scientific communities in peer-reviewed journals and in presentation at multidisciplinary conferences.