Polymer Dot Nanoparticles for Detection of Single Molecules in Live Cells

描述(申请人提供)：拟议研究的目标是开发一种名为“聚合物点”的新型荧光纳米颗粒，并测试该纳米颗粒用于使用传统荧光显微镜方法检测活细胞中单分子的可行性。聚合物点纳米粒子是一种很有前途的新型荧光纳米粒子技术，它基于荧光聚酰亚胺共轭聚合物，与传统的荧光染料和纳米粒子相比，在亮度上有很大的提高。该项目的具体目标包括优化和表征单分子检测的相关优值系数，开发和测试针对细胞内特定生物分子的生物偶联策略，以及展示在活细胞中检测单个纳米颗粒。将使用一系列表征技术，包括荧光光谱、单分子光谱、原子力显微镜、电泳和荧光显微镜。与传统的荧光染料相比，在亮度和光稳定性方面有望提高100到10,000倍，满足活细胞中基于荧光的单分子检测的要求。还有望展示简便、灵活的生物偶联方法，用于靶向感兴趣的特定生物分子，并检测活细胞中个别标记的生物分子。这种新型荧光纳米颗粒平台的发展将促进未来涉及监测活细胞内单分子运输和生化事件的研究，并可能为新型生物检测和生物传感器的开发提供基础。

DESCRIPTION (provided by applicant): The objective of the proposed research is to develop a novel class of fluorescent nanoparticles called "Polymer Dots" and test the feasibility of the nanoparticles for detection of single molecules in live cells using conventional fluorescence microscopy methods. Polymer dot nanoparticles are a promising new fluorescent nanoparticle technology based on fluorescent pi-conjugated polymers which provide substantial improvements in brightness as compared to conventional fluorescent dyes and nanoparticles. The specific aims of the project include optimizing and characterizing relevant figures of merit for single molecule detection, development and testing of bioconjugation strategies for targeting specific biomolecules within cells, and demonstration of single nanoparticle detection in living cells. A range of characterization techniques will be employed, including fluorescence spectroscopy, single molecule spectroscopy, atomic force microscopy, electrophoresis, and epifluorescence microscopy. Improvements in brightness and photostability of a factor of 100 to 10,000 as compared to conventional fluorescent dyes are expected, meeting the requirements for fluorescence-based single molecule detection in living cells. Demonstration of facile, flexible bioconjugation methods for targeting specific biomolecules of interest and detection of individual labeled biomolecules within living cells are also expected. The development of this novel fluorescent nanoparticle platform will facilitate future research involving monitoring transport and biochemical events of single molecules within living cells and could also provide the basis for the development of novel bioassays and biosensors.