Spatial analysis of cellular arrangement using quantitative, single-cell imaging of protein multiplexing

• DOI: 10.1117/12.2610955
• Publication Date: 2022-04-04T19:55:17Z
• Authors: Alison Cheung; Dan Wang; Kela Liu; Sarah Hynes; Ben Wang; Simone Stone; Pamela Ohashi; Martin J. Yaffe
• Keywords: Multiplex; Immune checkpoint; Cancer Immunotherapy; Mass cytometry

Single cell phenotyping using molecular or protein multiplexing techniques is gaining momentum, especially in the characterization of cancer and tumor microenvironment. It has proven to be particularly useful studying extent heterogeneity cancer, profiling immune environment assess whether certain subsets could predictive treatment response. Using a sequential marker labelling system called Multiplex Immunofluorescence (MxIF, GE Research), we have developed quantitative image analysis computational tools for individual cells various types. The expressions T markers CD3, CD8, macrophage CD68, checkpoint proteins PD-1 PD-L1, together with proliferative (Ki67) cancer-specific PCK (pan-Cytokeratin) were studied on single 4um sections formalin-fixed, paraffinembedded (FFPE) ovarian tissue sections. We explored composition phenotype t-SNE quantified densities co-expression patterns binary counting. In addition types, their spatial localizations analyzed. Neighborhood was conducted co-occurrence matrices determine number times that particular type proximal one another. Cell-to-cell relationship assessed by quantifying Euclidean distances between These are being applied specimens from an immunotherapy clinical trial evaluate dynamic changes during course blockade therapy.