Protocol for the generation, viability assessment, and downstream processing of murine precision-cut skin slices

Precision-cut tissue slices (PCTS) are an ex vivo model of viable organ sections widely used for therapeutic screening, toxicity assessment, and infection modeling. Here, we present a protocol for the generation, viability assessment, and downstream processing of murine Precision-cut Skin Slices (PCSS). We describe steps for preparing and agarose embedding skin samples, generating PCSS, maintaining them in culture, detecting lactate dehydrogenase (LDH), and quantifying ATP. We then detail procedures for RNA/protein extraction, the quantification of PCSS homogenates, and histological and TUNEL analysis of PCSS.