DNA-barcode-based Multiplex Immunofluorescence Imaging to Analyze FFPE Specimens from Genetically Reprogrammed Murine MelanomaSachin S. Surwase *1,2,3, Xin Ming M. Zhou *3,4, Kathryn M. Luly 1,2,3, Qingfeng Zhu 5,6,7, Niki Talebian 6, Robert A. Anders 5,6,7,8,9, Jordan J. Green 1,2,3,5,8,9,10,11,12, Stephany Y. Tzeng 1,2,3, Joel C. Sunshine 1,3,4,5,6,8,9
1Department of Biomedical Engineering, Johns Hopkins University, 2Translational Tissue Engineering Center, Johns Hopkins University, 3Johns Hopkins Translational ImmunoEngineering Center, Johns Hopkins University, 4Department of Dermatology, Johns Hopkins University, 5Department of Oncology, Johns Hopkins University, 6Department of Pathology, Johns Hopkins University, 7Convergence Institute, Sidney Kimmel Comprehensive Cancer Center Johns Hopkins University School of Medicine, 8Bloomberg Kimmel Institute for Cancer Immunotherapy, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, 9Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, 10Institute for Nanobiotechnology, Johns Hopkins University, 11Departments of Neurosurgery and Ophthalmology, Johns Hopkins University School of Medicine, 12Departments of Materials Science & Engineering and Chemical & Biomolecular Engineering, Johns Hopkins University
This protocol provides a step-by-step guide for designing a multiplex immunofluorescence antibody panel for DNA-barcode-based imaging of murine FFPE melanoma tissues. We also describe an image analysis pipeline using open-source tools for generating spatial proteomics insights into the murine melanoma tumor immune microenvironment.
