Quantification of Biomolecules Based on Flow Cytometric Single-Molecule Detection and Sorting on a Microchip

该奖项支持开发一种新型仪器，用于使用显微镜和微制造设备的组合来测量生物分子，以提供高通量的单分子检测能力。该仪器将在连续流动中对单个生物分子进行分类和检测，与现有技术相比，在灵敏度、准确性和分辨率方面有了显著提高。该提案旨在开发一种通用的、免洗涤的传感策略，利用多通道共聚焦激光诱导荧光（LIF）和荧光相关光谱（FCS），利用量子点作为荧光标签，在单分子水平上识别液体中的特定核酸和蛋白质。作为该项目的一部分，将设计和制造功能微流控芯片，以实现快速有效的分子分选。该微芯片将有一个微通道阵列，用于多个并发测量。这将允许以极低的样品量和浓度进行分析，并进行多路测量以提供高通量。该仪器将用于许多生物分析领域，包括基因表达谱、基因分型、早期疾病诊断、药物筛选以及分子结合研究。该项目的影响将从研究扩展到几个层次的教育和培训，并可能在进一步发展后进行商业应用。

This award supports the development of a novel instrument for measurement of biomolecules using a combination of microscopy and a microfabricated device to provide high throughput, single-molecule detection capability. The instrument, which will sort and detect individual biomolecules during continuous flow, represents a significant enhancement in sensitivity, accuracy, and resolution in comparison to existing technology. The proposal aims to develop a generic, washing-free, sensing strategy that can identify specific nucleic acids and proteins in liquids at the single-molecule level using multi-channel confocal laser induced fluorescence (LIF) and fluorescence correlation spectroscopy (FCS), employing quantum dots as fluorescent tags. As part of the project, a functional microfluidic chip will be designed and fabricated to enable rapid and efficient molecular sorting. The microchip will have an array of microchannels for multiple, concurrent measurements. This will allow analysis with extremely low sample volume and concentration, and multiplexing of measurements to provide high-throughput. The proposed instrument will be useful in many bioanalytical areas including gene expression profiling, genotyping, early disease diagnostics, pharmaceutical screening, as well as the study of molecular binding. The impact of the project will extend beyond research to education and training at several levels and, potentially, commercial applications following further development.