Electrochemical Imaging of in vitro Tight Junctions with Scanning Ion Conductance

使用扫描离子电导对体外紧密连接进行电化学成像

• 批准号: 7896927
• 负责人: Lane A. Baker
• 金额: $ 22.15万
• 依托单位: TRUSTEES OF INDIANA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-06-01 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7896927
• 关键词: Adherent Culture; Biological; Biological Models; Blood; Blood - brain barrier anatomy; Caco-2 Cells; Canis familiaris; Cell Line; Cell model; Cells; Colorectal Adenocarcinoma; Cornea; Cystic Fibrosis; Disease; Drug Delivery Systems; Electrical Resistance; Electron Microscopy; Endothelial Cells; Epithelial; Epithelial Cells; Exploratory/Developmental Grant for Diagnostic Cancer Imaging; Goals; Histocompatibility Testing; Human; Image; In Vitro; Individual; Ion Transport; Ions; Kidney; Knowledge; Location; Malignant Neoplasms; Measurement; Measures; Membrane; Methods; Microscopy; Modeling; Monitor; Optics; Outcome; Pathway interactions; Play; Renal tubule structure; Research; Resolution; Role; Scanning; Slice; Stimulus; System; Techniques; Testing; Tight Junctions; Tissues; Translating; Work; base; electrical measurement; in vitro Model; in vivo; innovation; interstitial; membrane model; nanometer; nanopore; nanoscale; public health relevance; response; scanning ion conductance microscopy; tool

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).

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）作为研究细胞接触、间隙和空间的动态工具。我们的长期目标是了解纳米尺度紧密连接的传输。该应用的目的是使用体外模型获得数十纳米水平的紧密连接传输的定量和定性测量。该应用的中心假设是扫描离子电导显微镜可用于以分辨率对通过各个紧密连接的传输进行成像。本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 通过紧密连接的合成模拟物进行运输成像的能力。第二个具体目标是建立 SICM，使用包含培养上皮细胞的体外模型定性和定量检查紧密连接在纳米尺度运输中的作用。所提出的工作具有创新性，因为它使用新平台来收集紧密连接传输的空间和时间信息。预期结果是创建一种可用于研究纳米尺度紧密连接传输的工具。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。 公共健康相关性：该应用描述了一种表征紧密连接的新方法，该连接调节跨上皮组织界面的运输。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）相关的疾病以及跨相对不可渗透的生物界面（例如血脑屏障）的药物输送的研究。