A Multi-functional Optical Impedance Microscope for Live Cell Imaging

描述（由申请人提供）：项目概述电阻抗谱（EIS）是一种灵敏的无标签技术，已被证明是广泛活细胞研究的有力工具。测量整个表面上活细胞的局部阻抗是非常需要的，但目前的EIS技术还不可能实现。该项目旨在开发一种新的显微镜，可以捕获活细胞的高分辨率阻抗图像。提出的新显微镜的原理与传统的EIS完全不同。它不是电测量阻抗，而是以亚微米空间分辨率光学成像整个表面的局部阻抗。这简化了阻抗测量而不牺牲灵敏度，更重要的是，它引入了新的令人兴奋的功能，包括：1)传感器芯片可以很容易地制造和准备细胞附着；2)可以分析整个传感器芯片或选定感兴趣的区域进行详细研究，这一点很重要，因为它可以以最佳的灵敏度和空间分辨率跟踪单个细胞甚至单个细胞内的区域；3)可以同时获得常规表面等离子体共振图像，提供细胞/底物相互作用的详细信息；4)该仪器将在传统倒置光学显微镜的基础上构建，以便根据需要获得同一样品的原位相对比和荧光显微镜图像。项目包括以下四项任务：1)构建高分辨率光学阻抗显微镜系统；2)建立活细胞分析的数据采集、处理和分析算法；3)研究阻抗显微镜图像与细胞粘附行为的关系；4)测试和评估光学阻抗显微镜用于细胞的其他研究，包括伤口愈合，毒理学和运动性。

DESCRIPTION (provided by applicant): Project Summary Electrical impedance spectroscopy (EIS) is a sensitive label-free technique that has proved to be a powerful tool for a wide range of live cell studies. Measurement of the local impedance of live cells on an entire surface is highly desired, but so far has not been possible with current EIS technology. This project aims at the development of a new microscopy that can capture high-resolution impedance images of live cells. The proposed new microscopy is based on principles that are completely different from the conventional EIS. Instead of measuring impedance electrically, it images the local impedance of the entire surface optically with sub-micron spatial resolution. This simplifies the impedance measurement without sacrificing sensitivity and, more importantly, it introduces new exciting capabilities including: 1) sensor chips can be easily fabricated and prepared for cell attachment; 2) the entire sensor chip or selected region of interest can be analyzed for detailed studies, which is important because it enables the tracking of individual cells or even region within single cells with the best sensitivity and spatial resolution; 3) conventional surface plasmon resonance images can be obtained simultaneously, which provide detailed information on cell/substrate interaction; and 4) the instrument will be built based on a conventional inverted optical microscope, so that in-situ phase contrast and fluorescence microscopy images can be obtained for the same sample if desired. The project includes the following four tasks: 1) build a high-resolution optical impedance microscope system; 2) establish data acquisition, processing, and analysis algorithms for live cell analysis; 3) study the relationships between impedance microscopy images and cell adhesion behavior; and 4) test and evaluate the optical impedance microscope for additional studies of cells including wound healing, toxicology and motility. PUBLIC HEALTH RELEVANCE (provided by applicant): This project aims at the development of a new label-free microscopy that can capture sub-micron resolution impedance images of live cells optically. In addition, conventional surface plasmon resonance, optical and fluorescence microscopy images can be obtained simultaneously. The success of this project will provide a new tool that has a broad range of applications on cell dynamic studies.