High-throughput microscopy system for ultrasensitive and multiplexed nanoarray analysis

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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.