Localized Surface Plasmon Resonance (LSPR) Biosensing Microarray for Multiplex, Real-time Single-Cell Immunophenotyping

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1263889 - KurabayashiThe immune system plays a critical role in protecting a living organism against invasions of viruses,bacteria, and parasitic worms and in distinguishing diseased tissue from healthy tissue. Leukocytes -white blood cells - are important blood constituents that support the immune system and haveseveral cellular subsets determined by their morphology and response to an antigen exposure. Comprehensive characterization of the immune cell functions is critical to diagnose, stratify, and monitor infections, malignancies, autoimmune disorders, and injuries or to study the efficacy of new drugs. In particular, quantitative analysis of the cytokine secretion profiles of individual cells is required for precise determination of the subtle variations in the cellular function and immune status across their sub-groups. However, there are some shortcomings in existing methods that need to be overcome for realizing rapid, efficient, dynamic analysis of leukocyte cytokine-secretion profiles. This research aims to establish a biosensing platform that enables high-throughput, multiplexed, dynamic measurement of cytokines secreted by target immune cells isolated from whole blood at thesingle-cell level. This single-cell functional assay efficiently permits on-chip isolation and stimulation of single cells and detection of cell-secreted proteins. Our approach will employ localized surface plasmon resonance (LSPR) imaging within microchamber arrays, each capturing a single cell. As a result of miniaturization of the whole system, both the sample volume and the total assay time canbe reduced. The microarray form of the microfluidic platform coupled with LSPR biosensing will allow for massively parallel label-free, real-time quantification and mapping of proteins secreted individual cells with a simple optics setup. The analysis performed for a large number of single cells will allow for immunophenotypical screening of leukocyte subsets while detecting subtle differences and time variations of their immune functions. If the isolation process only yields extraction of identical cell subsets, the large number of measurements per sample will lead to high statistical accuracy (i.e., reduced false positives). NontechnicalThis project will develop novel means to analyze immune cells present in blood. Specifically, the proposal will develop devices to capture individual immune cells and then monitor release of certain proteins from the captured cells. The levels and the types of proteins produced by the immune cells reflect on the state of health or disease of an individual. Therefore proposed devices will have significant diagnostic implications.