High-throughput microscopy system for ultrasensitive and multiplexed nanoarray analysis

用于超灵敏和多重纳米阵列分析的高通量显微镜系统

• 批准号: RTI-2017-00834
• 负责人: Juncker, David
• 金额: $ 10.85万
• 依托单位: McGill University
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 财政年份: 2016
• 资助国家: 加拿大
• 起止时间: 2016-01-01 至 2017-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=616776
• 关键词: throughput; microscopy; system; ultrasensitive; multiplexed

The single most significant factor that determines the outcome of many diseases is disease progression and diagnosis. Unfortunately, diseases are typically detected when symptoms appear when the diseases are already at an advanced stage. A hallmark of diseases is the dysregulation of proteins in cells, which are released into the blood and thus can be used as biomarkers for disease diagnosis. Early diagnosis of disease might therefore be possible if protein indicators of disease can be detected. Whereas current blood tests can detect a protein present as little as one part in 10 billion of total blood proteins, for early disease diagnosis, technologies that can measure multiple proteins simultaneously and are a thousand to a million times more sensitive are needed. Ultrasensitive protein analysis technologies have emerged recently, but they are complex, and multiplex protein measurement is either not feasible, or only possible at the expense of sensitivity and reliability. We propose to develop new technologies for ultrasensitive, multiplex detection of protein biomarkers that circulate in blood, as well as the protein content of individual nanometer-sized vesicles secreted by cancer cells, known as exosomes, that could be used to identify different types of cancers. We will develop novel platforms made of digital nanodot arrays that comprise millions of nanoscale-dots, each can capture proteins/single exosome using a low-cost patterning method we developed. Captured proteins/single exosome will be detected at single-molecule sensitivity, and the signal counted digitally for protein measurement and profiling of protein content on individual exosomes. For more fundamental study of the biology of disease, we will leverage our expertise in building nanodot arrays to create an artificial landscape of extracellular matrix proteins with well-defined yet complex parameters, to track both cell igration and intracellular signaling. Our research involves collaboration with heath, physics and engineering researchers, and fall under the integral theme of nanoarray fabrication, characterization and ultrasensitive signal signal detection. This NSERC-RTI proposal requests an automated and high sensitivity microscopy system that is critical for high-throughput imaging of nanoarrays. The proposed microscopy system will be automated, highly sensitive, and capable of rapid and high-throughput imaging, shortening a whole-day session that involves laborious manual adjustments into an hour of automatic image acquisitions. This instrument accelerate research that brings together experts in ultra-sensitive protein analysis, cell and caner biologists, and biomedical engineers, creating a unique, state-of-the-art microscopy facility to be used by many interdisciplinary trainees.

决定许多疾病结局的最重要因素是疾病的进展和诊断。不幸的是，疾病通常在症状出现时才被发现，而此时疾病已经处于晚期。疾病的一个标志是细胞中蛋白质的失调，这些蛋白质被释放到血液中，因此可以用作疾病诊断的生物标志物。因此，如果能够检测到疾病的蛋白质指标，就有可能对疾病进行早期诊断。虽然目前的血液检测只能检测到100亿分之一的蛋白质，但对于早期疾病诊断，需要能够同时测量多种蛋白质的技术，并且灵敏度要高1000到100万倍。超灵敏蛋白质分析技术是近年来出现的，但它们很复杂，多重蛋白质测量要么不可行，要么只能以灵敏度和可靠性为代价。