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New Photostable Nanoprobes for Real-time Imaging of Single Live Cells

用于单个活细胞实时成像的新型光稳定纳米探针

基本信息

批准号：
9468747
负责人：
X. Nancy Xu
金额：
$ 8.96万
依托单位：
OLD DOMINION UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-08-01 至 2021-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9468747
关键词：
Address Binding Biochemical Biological Markers Biomedical Research Biosensor CD44 gene Cancer Biology Cell physiology Cells Color Complex Darkness Detection Disease Early Diagnosis Early treatment Educational Activities Event Fluorescence Fluorescence Microscopy Halogens Health Hour Image Imaging Device In Situ Individual Institution Lasers Ligands Light Malignant Neoplasms Malignant neoplasm of ovary Microscope Molecular Molecular Probes Nanoscopy Noise Optics Peroxisome Proliferator-Activated Receptors Photobleaching Population Proteins Research Activity Research Infrastructure Resolution Role Signal Pathway Signal Transduction Signal Transduction Pathway Source Students Time Time Study Tumor stage Work cancer stem cell cancer therapy cell type design educational atmosphere fluorescence imaging fluorophore imaging detector imaging modality imaging platform imaging probe instrument instrumentation liquid crystal live cell imaging molecular imaging nanoparticle nanoprobe neoplastic cell novel optical spectra outreach programs public health relevance quantitative imaging receptor single molecule tool

项目摘要

DESCRIPTION: New Photostable Nanoprobes for Real-time Imaging of Single Live Cells. Different types of cells express trace amounts of distinctive sets of receptors, enabling them serve as biomarkers for cell identification. Binding of a few ligand molecules with its receptors on single live cells can initiate dynamic cascades of cellular signaling pathways, alter cellular functions and cause diseases. Multiple types of ligand-receptor (L-R) interactions typically work together to regulate and alter the cellular functions. Such signaling pathways can take minutes, hours or days. Thus, it is crucial to simultaneously visualize different types of receptors on single live cells with single-molecule sensitivity in order to quantify various types of receptor molecules, to directly capture how L-R interactions work together to achieve given cellular functions, and to effectively detect rare subsets of single live cells. Currently, fluorescence microscopy using fluorescence imaging probes is the primary workhorse for live cell imaging. Unfortunately, fluorescence probes (fluorophor, fluorescence protein, QD) suffer intrinsic photobleaching, making them unable to continuously capture the dynamic events of single live cells over hours. Photobleaching also makes quantitative analysis over time difficult. Separation of different excitation and emission of various fluorophores leads to complex and expensive instrument and restricts their capacity of multiplexing study of multiple types of molecules simultaneously. All fluorescence imaging methods including conventional, confocal and super-resolution fluorescence microscopy suffer these same fundamental limitations. In this proposal, we aim to develop a novel molecular imaging platform including photostable multicolored and multifunctional single molecule nanoparticle optical biosensors (SMNOBS) and far-field photostable optical nanoscopy (PHOTON) for real-time study of functions of single live cells. To demonstrate the-proof-of-concept, we will use them to quantitatively image and molecular characterize roles and functions of multiple types of the receptors on rare subsets of ovarian cancer stem cells (oCSCs) in highly heterogeneous tumor cell populations and to study their differentiation mechanisms. These powerful new tools are expected to address a wide range of pressing biochemical and biomedical questions about molecular and real-time characterization of functions of single receptor molecules on single live cells and their related signaling pathways in real time. The proposed study will enable us to create an interdisciplinary educational environment for students; expose them to leading-edge biomedical research program; involve more students in biomedical research activities; enhance and strengthen institutional biomedical research program and infrastructure.