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Background: Understanding breast cancer progression and the relationships among biomarkers in the tumor and tumor microenvironment could go beyond established prognostic markers of breast cancer. Poor response and recurrence may indeed be a consequence of the highly heterogeneous spatial distribution of biomarkers within cancers, and/or the synergistic and antagonistic relationship between co-localized biomarkers. Recently, mechanical and physical microenvironmental signatures have emerged as relevant in determining cancer aggressiveness and invasiveness. This suggests that physical structures in the tumor tissue may drive favorable immune-tumor-stroma cell patterns. However, current assessment of tumor biopsies is limited in the ability to quantify and spatially resolve several different subtypes of cells and biomarkers within tumors. Methods: We developed a 60-marker imaging mass …