A Multi-functional Optical Impedance Microscope for Live Cell Imaging

描述(由申请人提供)：项目摘要阻抗谱(EIS)是一种敏感的无标记技术，已被证明是广泛的活细胞研究的强大工具。测量整个表面上活细胞的局部阻抗是非常必要的，但目前的EIS技术到目前为止还不可能。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。拟议的新显微镜基于与传统EIS完全不同的原理。它不是电学测量阻抗，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，它引入了新的激发能力，包括：1)传感器芯片可以容易地制造和准备用于细胞连接；2)整个传感器芯片或选定的感兴趣区域可以被分析以进行详细的研究，这是重要的，因为它能够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)可以同时获得常规的表面等离子体共振图像，它提供了关于细胞/衬底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建造，以便如果需要的话，可以对同一样品获得原位相衬和荧光显微镜图像。该项目包括以下四项任务：1)建立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黏附行为之间的关系；4)测试和评估光学阻抗显微镜，用于进一步研究细胞的伤口愈合、毒理学和运动性。 公共卫生相关性(由申请人提供)：该项目旨在开发一种新的无标签显微镜，可以光学捕获活细胞的亚微米分辨率阻抗图像。此外，还可以同时获得常规的表面等离子体共振、光学和荧光显微图像。该项目的成功将为细胞动力学研究提供一种具有广泛应用前景的新工具。

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.