Abstract 3921: Precise spatial multiplexing for immune profiling in clinical FFPE tumor samples with ChipCytometry࣪

Abstract Highly multiplexed spatial biomarker analysis has the potential to inform the discovery of novel biomarkers and treatments with the potential to improve clinical outcomes. Clinical diagnoses and treatment decisions have traditionally been made with spatially resolved techniques like immunohistochemistry, that offer a wealth of information about cellular morphology and the spatial context of cells but are limited to a single plex. Newer technologies, such as single-cell RNA sequencing, identify cell subpopulations within tissue but do not capture their spatial distribution or intercellular interactions in situ. Here we present the analysis of clinical FFPE tumor samples using ChipCytometry, a novel precise spatial multiplexing technology that offers high-plex biomarker analysis, without compromising spatial context. ChipCytometry combines iterative immuno-fluorescent staining with high-dynamic range imaging to facilitate quantitative phenotyping with single-cell resolution. Standard FCS files are generated from multichannel OME-TIFF images, enabling identification of cellular phenotypes via flow cytometry-like hierarchical gating. In this study, we identify and quantify immune and epithelial cell phenotypes and subtypes in melanoma and non-small cell lung cancer FFPE samples. The results show precise expression levels for each marker in the assay in each individual cell in the sample, while maintaining spatial information about each cell. Spatial analysis of the samples reveals quantifiable heterogeneity of immune cell infiltration within the tumor samples, demonstrating the utility of the ChipCytometry platform for the in-depth immune profiling in clinical FFPE samples. -Bruker Confidential- Citation Format: Kevin Gamber, Arne Christians, Jennifer Brooks, Karen Kwarta, Crystal Winkeler. Precise spatial multiplexing for immune profiling in clinical FFPE tumor samples with ChipCytometry࣪ [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3921.