Preparation of Melanoma Tissue for Multiplex Immunofluorescence Histology Analysis of Immune Infiltration and Spatial...

Immunotherapies extend survival in melanoma patients by increasing the immune system's capacity to detect tumors and mediate tumor cell death. Understanding mechanisms of antitumor immunity requires evaluating changes in immune infiltrate composition, activation, and spatial distribution within the tumor microenvironment (TME). Despite this need, relatively few methods allow for detailed analysis of both the structural organization and immunological characteristics of tumor-infiltrating immune cells in preclinical cancer models. In this chapter, we present a multiplex immunofluorescence staining approach for identifying, characterizing, and quantifying tumor-infiltrating immune cells in murine melanoma, allowing evaluation of how therapies influence immune infiltration in preclinical models.