Development of Microfluidic Nano-Liter Platform for Single Cell Dynamic Monitorin

描述(申请人提供)：调节免疫功能的中心过程之一涉及T细胞与抗原提呈细胞(APC)建立细胞-细胞联系，例如 通过称为免疫突触(IS)的接触区作为树突状细胞(DC)。这种突触的重要性是相当大的，因为T细胞与淋巴结中的DC的相互作用是产生抗原特异性免疫反应的关键步骤之一，从而激活NAVE T细胞。尽管显微成像技术取得了很大进步，但在单个细胞水平上动态监测细胞和诱导分泌模式的技术发展滞后，以避免由于细胞群体中的不同反应而产生的模糊性。这项拟议工作的目标是开发一种微流控液滴方法，用于动态监测在免疫反应的这一关键事件中细胞间通信所发生的生化事件。特别是，当前的提案概述了一种新的方法的集成，以监测活细胞表面表达的变化以及DC和T细胞之间形成的实际免疫突触(IS)。除了细胞表面研究外，本文开发的微流控液滴方法还将使我们能够利用微流控反应液滴中包含的微球传感器收集IS形成过程中分泌分子的数据。因此，活细胞分泌和细胞表面变化将在一个不同的微环境中在单个细胞水平上同时进行监测，这是使用传统技术不可能实现的。

DESCRIPTION (provided by applicant): One of the central processes regulating immune function involves T cells establishing cell-cell contacts with antigen-presenting cells (APCs) such as dendritic cells (DCs) via a contact zone known as the immunological synapse (IS). The importance of this synapse is considerable since interaction of T cells with DCs in lymph nodes is one of the critical steps in the generation of antigen specific immune responses which activate the na¿ve T cells. Despite great technical advances in microscopic imaging, the development of techniques to dynamically monitor the cellular and the induced secretion patterns at the IS interaction on a single cell level to avoid ambiguities that arise due to heterogeneous responses in cell populations is lagging. It is the goal of the proposed work to develop a microfluidic droplt approach for dynamic monitoring of the biochemical events that transpire upon cell-cell communication in this key event of the immune response. In particular, the current proposal outlines the integration of a novel approach to monitor live cell surface expression changes as well as the actual immunological synapse (IS) formed between DCs and T cells. In addition to cell surface studies, the microfluidic droplet approach developed herein will also allow us to collect data on secreted molecules during IS formation utilizing microsphere sensors incorporated in the microfluidic reaction droplets. Thus, live cell secretion and cell surface changes will be monitored on a single cell level simultaneously in a distinct microenvironment, feat not possible using conventional techniques.