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DNA-barcode-based Multiplex Immunofluorescence Imaging to Analyze FFPE Specimens from Genetically...

JoVE Journal
Cancer Research

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JoVE Journal Cancer Research

DNA-barcode-based Multiplex Immunofluorescence Imaging to Analyze FFPE Specimens from Genetically Reprogrammed Murine Melanoma

DNA-barcode-based Multiplex Immunofluorescence Imaging to Analyze FFPE Specimens from Genetically Reprogrammed Murine Melanoma

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1,181 Views

09:58 min

June 6, 2025

DOI: 10.3791/68124-v

Sachin S. Surwase*^1,2,3, Xin Ming M. Zhou*^3,4, Kathryn M. Luly^1,2,3, Qingfeng Zhu^5,6,7, Niki Talebian⁶, 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.

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