On-Chip Multiplexed Adhesion Frequency Assay for Measuring Receptor-Ligand Interactions on Cells

项目摘要 细胞上的受体-配体相互作用介导细胞-细胞和细胞-环境通讯， 许多生物过程。粘附频率测定具有独特的测量细胞粘附的能力。 受体-配体相互作用在单细胞水平。测量可以提供重要的 关于生物过程质量的信息和用于选择有效的治疗药物的信息 细胞目前的粘附频率测定法使用微量移液器抽吸细胞进行两种相互作用 然后进行测量和小区转移。因此，它体积庞大，劳动密集型，手动- 可操作性和低吞吐量。为了最大限度地发挥粘附频率测定法在 生物医学研究和临床应用，有必要开发高通量 微型装置 鉴于发展高通量微型生物技术的巨大潜力和挑战， 粘附频率测定，我提出了一个变革性的技术路线-芯片上多路复用 粘附频率测定-这将寻求协同光学操纵和微流体。我 将带来来自不同领域的新想法，包括光学操纵，光学成像， 微流体，细胞生物学和设备物理学的开发，验证和应用， 建议的化验。具体来说，我将（1）解决多路复用测量和细胞分选 分析实施中的挑战，（2）表征，验证和优化预 包装的测定，和（3）将测定组装和包装到紧凑的装置中，并将其应用于 筛选用于疾病治疗的细胞。以前所未有的能力测量受体- 配体相互作用和细胞分选，拟议的测定将改变当前的细胞范式 分析并将量化细胞功能的最重要参数之一纳入 常规化验。一旦完全开发，芯片上的多路复用粘附频率测定将填补 在受体-配体的生物医学研究和临床应用中有许多未满足的需求 细胞间的相互作用。

Project Summary Receptor-ligand interactions on cells mediate cell-cell and cell-environment communications in many biological processes. Adhesion frequency assay has unique capability of measuring the receptor-ligand interactions at the single-cell level. The measurement can provide important information on the quality of biological processes and for the selection of potent therapeutic cells. Current adhesion frequency assay uses micropipettes to aspirate cells for both interaction measurement and cell transfer afterwards. As a result, it is bulky, labor-intensive, manual- operative, and low-throughput. In order to maximize the potential of adhesion frequency assay in biomedical research and clinical applications, it is necessary to develop high-throughput miniature device. In view of the tremendous potential and challenges in developing high-throughput miniature adhesion frequency assay, I propose a transformative technical route – on-chip multiplexed adhesion frequency assay – that will seek to synergize optical manipulation and microfluidics. I will bring in new ideas from different areas, including optical manipulation, optical imaging, microfluidics, cellular biology, and device physics for development, validation and applications of the proposed assay. Specifically, I will (1) address multiplexed-measurement and cell-sorting challenges in the implementation of the assay, (2) characterize, validate and optimize the pre- packaged assay, and (3) assemble and package the assay into a compact device and apply it to screen cells for disease therapy. With the unprecedented capabilities of measuring the receptor- ligand interactions and sorting cells, the proposed assay will change current paradigm of cell profiling and incorporate one of the most important parameters for quantifying cell functions into routine assays. Once fully developed, the on-chip multiplexed adhesion frequency assay will fill many unmet needs in both biomedical research and clinical applications of the receptor-ligand interactions on cells.