Electrochemical Imaging of in vitro Tight Junctions with Scanning Ion Conductance

描述（由申请人提供）：本申请旨在开发扫描离子电导显微镜（SICM）作为研究细胞-细胞接触、间隙和空间的动态工具。我们的长期目标是在纳米尺度上了解通过紧密连接的运输。本申请的目的是获得定量和定性测量的运输通过紧密连接在几十纳米的水平，使用体外模型。该应用程序的中心假设是，扫描离子电导显微镜可用于图像传输通过个别紧密连接的分辨率。所提出的研究的基本原理是，使用SICM可以测量空间和时间响应，从而可以分析紧密连接。我们的研究有两个具体目标。第一个具体目标是建立SICM通过合成模拟紧密连接的运输成像能力。第二个具体的目的是建立SICM定性和定量研究的作用，紧密连接的运输在纳米尺度上使用体外模型包括培养的上皮细胞。所提出的工作是创新的，因为它使用了一个新的平台来收集空间和时间信息紧密连接传输。预期的结果是创建一个工具，可用于研究在纳米尺度上通过紧密连接的运输。在这些水平上监测运输的能力将为研究生物模型提供新的机会。 公共卫生相关性：本申请描述了一种新的方法来表征调节上皮组织界面转运的紧密连接。这项研究将开发一种工具，可以影响与紧密连接（例如癌症，囊性纤维化）相关的疾病的研究，并通过相对不可渗透的生物界面（例如血脑屏障）进行药物递送。

DESCRIPTION (provided by applicant): This application seeks to develop scanning ion conductance microscopy (SICM) as a dynamic tool for the study of cell-cell contacts, gaps and spaces. Our long-term goal is to understand transport through tight junctions at the nanometer scale. The objective of this application is to obtain quantitative and qualitative measurements of transport through tight junctions at the level of tens of nanometers using in vitro models. The central hypothesis of the application is that scanning ion conductance microscopy can be used to image transport through individual tight junctions at resolutions. The rationale for the proposed research is that using SICM both spatial and temporal response can be measured, allowing analysis of tight junctions. Our study has two specific aims. The first specific aim is to establish capabilities of SICM for imaging of transport through synthetic mimics of tight junctions. The second specific aim is to establish SICM for qualitatively and quantitatively examining the role of tight junctions in transport at the nanometer scale using in vitro models comprised cultured epithelial cells. The proposed work is innovative because it uses a new platform to collect both spatial and temporal information tight junction transport. The expected outcomes are to create a tool that can be used to study transport through tight junctions at the nanometer scale. The ability to monitor transport at these levels will provide new opportunities to study biological models. PUBLIC HEALTH RELEVANCE: This application describes a new way to characterize tight junctions that regulate transport across the interface of epithelial tissues. This study will develop a tool that can impact research in disease related to tight junctions (e.g. cancer, cystic fibrosis) and drug delivery across relatively impermeable biological interfaces (e.g. the blood brain barrier).