IRCCS Regina Elena National Cancer Institute 1 article published in JoVE Immunology and Infection Microfluidic Co-Culture Models for Dissecting the Immune Response in in vitro Tumor Microenvironments Adele De Ninno1, Francesca Romana Bertani1, Annamaria Gerardino1, Giovanna Schiavoni2, Martina Musella3, Claudia Galassi3, Fabrizio Mattei2, Antonella Sistigu3,4, Luca Businaro1 1CNR Institute for Photonics and Nanotechnology, 2Dept. of Oncology and Molecular Medicine, Istituto Superiore di Sanità, 3Istituto di Patologia Generale, Università Cattolica del Sacro Cuore, 4Tumor Immunology and Immunotherapy Unit, IRCCS Regina Elena National Cancer Institute In the age of immunotherapy and single-cell genomic profiling, cancer biology requires novel in vitro and computational tools for investigating the tumor-immune interface in a proper spatiotemporal context. We describe protocols to exploit tumor-immune microfluidic co-cultures in 2D and 3D settings, compatible with dynamic, multiparametric monitoring of cellular functions.