Develop a Dual Functional Nanoprobe for Integration of Fluorescence microscopy wi

描述（由申请人提供）：所提出的申请的目的是开发一种双功能探针，该探针可在单颗粒水平上检测到拉曼和荧光显微镜，以便将每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。有了这种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，不仅是高荧光和鲁棒性，但也可以报告其标记的配体的分子信息;（2）提供一种新的成像工具，整合了荧光显微镜和拉曼光谱的优势;（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质相互作用相关联。这些荧光和拉曼探针的应用并不局限于所提出的研究，它们也可以在生物成像中作为多路复用和多功能探针找到应用，以将荧光和拉曼图像与体外和体内水平的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中起关键作用，并为疾病诊断和新疗法开发提供基础。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，以便我们可以同时跟踪活细胞内单细胞水平的配体-受体复合物并对其进行化学成像。这项工作的成功将（1）提供一种新的成像探针，不仅是高荧光和鲁棒性，但也可以报告它标记的配体的分子信息;（2）提供一种新的成像工具，整合了荧光显微镜和拉曼光谱的优势;（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-受体相互作用相关联。

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.