Quantitative Immune Profiling of Human Tumor Tissues with Multiplexed ChipCytometry

Abstract Cancer therapies that rely on manipulating or engineering immune cells have shown much promise in recent years for effectively combating a broad array of cancer types. In order to determine the effectiveness of such therapies, it is necessary to accurately profile the complement of immune cell types present in the tumor microenvironment. Methods to reliably obtain immune cell signatures require the combination of powerful resolution to discriminate cell boundaries, broad dynamic range to capture markers of varying expression levels, and accurate cell segmentation across a wide range of cell sizes and morphologies. Many methods for quantitative immune cell profiling have demonstrated shortcomings when it comes to the aforementioned attributes. To remedy these shortcomings, we present here ChipCytometry, an imaging technology for immune profiling of cells and sectioned tissues. ChipCytometry is a fluorescence-based imaging system that utilizes multiplexed immuno-fluorescence staining in combination with high-dynamic-range (HDR) imaging to facilitate quantitative phenotyping of individual cells within tissue samples. Employing this technology to profile metastatic and primary tumors, multiple biomarkers were stained in iterative multiplex assays to profile single cells with an AI-powered cell segmentation algorithm. The HDR imaging allows for both strong and weak fluorescent signals to be simultaneously obtained without loss of sensitivity. This immune profiling was completed on 6 different cryosectioned human tumor tissues (head & neck, liver, lung, breast, colon, and pancreas).