Develop a Dual Functional Nanoprobe for Integration of Fluorescence microscopy wi

描述(申请人提供)：本申请的目标是开发一种可在拉曼显微镜和荧光显微镜下在单粒子水平上检测的双功能探针，以便将两种技术的优势结合到单一成像工具中，以应对生物成像中的挑战。有了这个新的成像探针和工具，我们计划解决一个长期的生物成像挑战，即在活细胞内的分子水平上实时成像配体和受体之间的相互作用。这项拟议工作的成功将(1)提供一种新的成像探针，它不仅高度荧光和强大，而且可以报告其标记的配体的分子信息；(2)提供一种结合荧光显微镜和拉曼光谱优势的新成像工具；(3)使我们能够在活细胞内的化学水平上将配体的细胞动力学与配体结合蛋白的相互作用联系起来。这些荧光和拉曼探针的应用不仅限于拟议的研究，它们还可以作为多路复合和多功能探针在生物成像中的应用，在体外和体内水平上将荧光和拉曼图像与电子显微镜图像相关联。 公共卫生相关性：配体-受体相互作用在调节细胞功能方面发挥着关键作用，并为疾病诊断和新疗法的开发提供了基础。然而，在分子水平上全面了解配体和受体之间的体内相互作用仍然具有很大的挑战性。我们的研究应用旨在使用双功能探针将荧光显微镜与拉曼光谱相结合，以便我们可以在活细胞内的单细胞水平上同时跟踪和化学成像配体-受体复合体。这项工作的成功将(1)提供一种新的成像探针，不仅具有高度的荧光和健壮，而且可以报告其标记的配体的分子信息；(2)提供一种结合荧光显微镜和拉曼光谱优点的新的成像工具；(3)使我们能够在活细胞内的化学水平上将配体的细胞动力学与配体-受体的相互作用联系起来。

DESCRIPTION (provided by applicant): The objective of the proposed application is to develop a dual functional probe which is detectable at single-particle level for both Raman and fluorescence microscopy, so that the strengths of each technique can be combined into a single imaging tool for tackling challenges in bioimaging. With this new imaging probe and tool, we plan to address a long-term bioimaging challenge that is real-time imaging of interactions between ligands and receptors at the molecular level inside live cells. Success of this proposed work will (1) provide a new imaging probe that is not only highly fluorescent and robust but can also report molecular information for ligands it labels; (2) offer a new imaging tool which integrates strengths of fluorescence microscopy and Raman spectroscopy; (3) enable us to correlate cellular dynamics of ligands with ligand-binding protein interactions at the chemical level inside live cells. The applications of these fluorescent and Raman probes are not limited to the proposed studies; they can also find applications in bioimaging as multiplexing and multifunctional probes to correlate fluorescence and Raman images with electron microscopy images at in vitro and in vivo level. PUBLIC HEALTH RELEVANCE: Ligand-receptor interactions play a pivotal role in regulating cellular functions and provide a foundation for disease diagnosis and new therapy development. However, comprehensive understanding of in vivo interactions between ligand and receptors at the molecular level remains highly challenging. Our research application aims to integrate fluorescence microscopy with Raman spectroscopy using a dual functional probe, so that we can simultaneously track and chemically image of ligand-receptor complexes at the single-cell level inside live cells. Success of this proposed work will (1) provide a new imaging probe that not only is highly fluorescent and robust but also can report molecular information of ligands it labels; (2) offer a new imaging tool which integrate strengths of fluorescence microscopy and Raman spectroscopy; (3) enable us to correlate cellular dynamics of ligands with ligand-receptor interactions at the chemical level inside live cells.