An optimized protocol for generating fibrotic murine precision-cut lung slices

Precision-cut lung slices (PCLSs) offer significant advantages over both in vivo animal models and in vitro cell models, making them invaluable tools in respiratory research. Here, we present a protocol for generating fibrotic mouse PCLSs (mPCLSs), which offer the same benefits as human PCLSs but bypass issues of human tissue availability and related ethical concerns while also enhancing experimental homogeneity and reproducibility. This protocol details the steps for bleomycin (BLM)-induced pulmonary fibrosis, PCLS generation, visualization, and downstream analysis. For complete details on the use and execution of this protocol, please refer to Barbayianni et al.¹.