Optimized protocol for immunophenotyping of melanoma and tumor-bearing skin from mouse

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.