Collaborative Research: Plasmofluidic Nanoantenna-Superlens Biosensor for Single-Cell Functional Immunophenotyping

准确、实时地分析不同抗病血细胞的功能，对于监测人体的免疫状态，以及评估疾病阶段和药物疗效都非常有用。本研究的目的是开发一种新的方法，使单细胞功能免疫分析从人血滴。这样的平台最终将为临床医生提供一种新的测量方法，用于定量表征抗病血细胞。拟议的教育计划将对不同教育水平（K-12，本科和研究生），性别和种族的学生产生广泛的影响。准确、实时地分析不同免疫细胞亚群的功能，对于监测患者的免疫状态，以及评估疾病分期和药物疗效是必要的。特别地，来自个体免疫细胞的细胞因子分泌的定量和动态分析可用于确定患者中的细胞功能免疫表型。然而，免疫细胞的异质性使得这种定量表征具有挑战性，特别是在处理人血液样品时。本研究的目的是开发一种新的集成等离子体流体纳米天线-超透镜生物传感平台，该平台能够从微升人体血液中进行单细胞，多子集，多重功能免疫分析。该方法结合了一个有效的芯片上的单免疫细胞分离技术和一个平行的细胞因子分泌测定的基础上的无标记的等离子体纳米天线超透镜生物传感器。拟议的研究将通过追求以下目标来实现。 1)开发等离子体纳米天线-超透镜生物传感平台，用于快速和灵敏的芯片上细胞因子检测; 2）开发微流体单细胞分离技术，以实现从全血中发现的异质混合物中纯化白细胞亚群;和3）展示集成等离子体流体纳米天线-超透镜条形码微阵列生物传感器，用于单一免疫细胞的多重功能免疫表型分析。使用所提出的平台对大量个体免疫细胞进行的分析将建立一种新的方法，该方法通过检测白细胞亚群的免疫功能的差异和时间依赖性变化来进行白细胞亚群的免疫表型筛选。这样的平台最终将为临床医生提供一种新的工具，用于定量表征患者的免疫表型，因此可以进行精确的疾病诊断、监测和风险分层。拟议的教育计划将对不同教育水平（K-12，本科和研究生），性别和种族的学生产生广泛的影响。提案中开发的技术将被用作教育和外联活动的有效工具。将研究机会扩展到K-12学生和本科生将使他们接触到有益的科学发现，并鼓励他们追求未来的研究和工程职业。

Accurate and real-time analysis of the functions of different disease-fighting blood cells is very useful for monitoring the immune status of the human body, as well as evaluating the disease stage and drug efficacy. The objective of this research is to develop a new method that enables single-cell functional immune analysis from droplets of human blood. Such a platform would ultimately provide clinicians a new measurement for quantitatively characterizing the disease-fighting blood cells. The proposed educational plans will have broad impacts on students from various educational levels (K-12, undergraduate, and graduate), genders, and ethnicities. Accurate and real-time analysis of the functions of different immune cell subsets is needed for monitoring the immune status of patients, as well as for evaluating the disease stage and drug efficacy. In particular, quantitative and dynamic profiling of the cytokine secretions from individual immune cells is useful for determining the cellular functional immune phenotype in patients. However, the heterogeneity of the immune cells makes such quantitative characterization challenging especially when dealing with human blood samples. The objective of this research is to develop a new integrated plasmofluidic nanoantenna-superlens biosensing platform that enables single-cell, multi-subset, multiplex functional immune analysis from microliters of human blood. The approach combines an efficient on-chip single immune cell isolation technique and a parallel cell cytokine secretion assay based on the label-free plasmonic nanoantenna-superlens biosensor. The proposed research will be achieved by pursuing the following objectives. 1) development of plasmonic nanoantenna-superlens biosensing platform for rapid and sensitive on-chip cytokine detection; 2) development of microfluidic single cell isolation technique to achieve purification of leukocyte subsets from a heterogeneous mixture found in whole blood; and 3) demonstration of an integrated plasmofluidic nanoantenna-superlens barcode microarray biosensor for multiplex functional immunophenotyping of single immune cells. The analysis performed using the proposed platform for a large number of individual immune cells will establish a new approach that permits immunophenotypical screening of leukocyte subsets by detecting differences and time-dependent variations of their immune functions. Such a platform would ultimately provide clinicians a new tool for quantitatively characterizing the immunophenotypes of patients and would, therefore, allow precise disease diagnosis, monitoring, and risk stratification. The proposed educational plan will have broad impacts on students from different educational levels (K-12, undergraduate, and graduate), genders, and ethnicities. Technologies developed in the proposal will be used as effective vehicles for education and outreach activities. Extending research opportunities to K-12 students and undergraduates will expose them to the rewarding scientific discoveries and encourage them to pursue future studies and careers in engineering.