1. Optimized protocol for immunophenotyping of melanoma and tumor-bearing skin from mouse
- Author
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Sai Sakktee Krisna, Christophe Goncalves, Dany Plourde, Fan Huang, Jörg H. Fritz, Natascha Gagnon, Wilson H. Miller, and Sonia V. del Rincón
- Subjects
Science (General) ,Skin Neoplasms ,Cell ,ved/biology.organism_classification_rank.species ,Immunology ,Melanoma, Experimental ,Mice, Transgenic ,Biology ,General Biochemistry, Genetics and Molecular Biology ,Flow cytometry ,Immunophenotyping ,Q1-390 ,Mice ,Immune system ,Model Organisms ,medicine ,Protocol ,Animals ,Flow Cytometry/Mass Cytometry ,Model organism ,Cancer ,Skin ,Mice, Inbred BALB C ,General Immunology and Microbiology ,medicine.diagnostic_test ,ved/biology ,General Neuroscience ,Melanoma ,Innate lymphoid cell ,biochemical phenomena, metabolism, and nutrition ,medicine.disease ,Flow Cytometry ,Mice, Inbred C57BL ,medicine.anatomical_structure ,Cell isolation ,Cancer research ,bacteria - Abstract
Summary While isolating immune cells from spleens and lungs is routinely achieved using flow cytometry, it is challenging to isolate viable immune cells from skin. Here, we describe a step-by-step protocol for skin digestion using a murine melanoma model, which is amenable for detection of low abundant immune cell populations including group 2 innate lymphoid cells., Graphical abstract, Highlights • Optimized skin and melanoma digestion protocol for immune phenotyping • Detection of ILC2s and other immune cells isolated from skin and melanoma samples • Detailed gating strategies to identify ILC2s, T cells, and myeloid cell subsets, While isolating immune cells from spleens and lungs is routinely achieved using flow cytometry, it is challenging to isolate viable immune cells from skin. Here, we describe a step-by-step protocol for skin digestion using a murine melanoma model, which is amenable for detection of low abundant immune cell populations including group 2 innate lymphoid cells.
- Published
- 2021