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Quantitative, Architectural Analysis of Immune Cell Subsets in Tumor-Draining Lymph Nodes from Breast Cancer Patients and Healthy Lymph Nodes 1 2 1 1 1 A. Francesca Setiadi , Nelson C. Ray , Holbrook E. Kohrt , Adam Kapelner , Valeria Carcamo-Cavazos , 1 1 3 4 2 Edina B. Levic , Sina Yadegarynia , Chris M. van der Loos , Erich J. Schwartz , Susan Holmes , Peter P. Lee1* 1 Department of Medicine, Stanford University School of Medicine, Stanford, California, United States of America, 2 Department of Statistics, Stanford University, Stanford, California, United States of America, 3 Department of Pathology, Academic Medical Center, Amsterdam, The Netherlands, 4 Department of Pathology, Stanford University School of Medicine, Stanford, California, United States of America Abstract Background: To date, pathological examination of specimens remains largely qualitative. Quantitative measures of tissue spatial features are generally not captured. To gain additional mechanistic and prognostic insights, a need for quantitative architectural analysis arises in studying immune cell-cancer interactions within the tumor microenvironment and tumor- draining lymph nodes (TDLNs). Methodology/Principal Findings: We present a novel, quantitative image analysis approach incorporating 1) multi-color tissue staining, 2) high-resolution, automated whole-section imaging, 3) custom image analysis software that identifies cell types and locations, and 4) spatial statistical analysis. As a proof of concept, we applied this approach to study the architectural patterns of T and B cells within tumor-draining lymp