Isolation of adoptively transferred CD8⁺ T cells in mouse tumor tissues for lipid peroxidation detection

The lipid peroxidation level of tumor-infiltrating CD8⁺ T cells is crucial for its activity and longevity. Here, we describe a protocol for effective and epitope-preserving dissociation of mouse tumors and subsequent leukocyte purification and lipid peroxidation staining of adoptively transferred CD8⁺ T cells. We use BODIPY 581/591 C11 to monitor the cellular lipid peroxidation level and detect its fluorescent change by flow cytometry, followed by analysis in FlowJo. This protocol is adaptable to intrinsic CD8⁺ T cells in tumors as well. For complete details on the use and execution of this protocol, please refer to Xiao et al. (2022)¹ and Ma et al. (2021).².