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加工设备的组合来测量生物分子，以提供高通量，单分子检测能力。该仪器将在连续流动过程中对单个生物分子进行分类和检测，与现有技术相比，它在灵敏度、准确度和分辨率方面都有显著提高。该提案旨在开发一种通用的免洗涤传感策略，该策略可以使用多通道共聚焦激光诱导荧光（LIF）和荧光相关光谱（FCS）在单分子水平上识别液体中的特定核酸和蛋白质，采用量子点作为荧光标签。作为该项目的一部分，将设计和制造功能性微流体芯片，以实现快速有效的分子分选。微芯片将具有用于多个同时测量的微通道阵列。这将允许在极低的样品体积和浓度下进行分析，并进行多路测量以提供高通量。所提出的仪器将是有用的，在许多生物分析领域，包括基因表达谱，基因分型，早期疾病诊断，药物筛选，以及分子结合的研究。该项目的影响将从研究扩展到各级教育和培训，并在进一步发展后可能扩大到商业应用。

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.