Protocol to culture and image pancreatic ductal adenocarcinoma tumor slices to study T cell migration

Here, we describe a protocol for culture and live cell imaging of tumor slices. This approach studies carcinoma and immune cell dynamics in complex tumor microenvironments (TME) with nonlinear optical imaging platforms. Using a tumor-bearing mouse model of pancreatic ductal adenocarcinoma (PDA), we detail steps to isolate, activate, and label CD8(+) T lymphocytes and later introduce them to live murine PDA tumor slice explants. The techniques described in this protocol can improve our understanding of cell migration in complex microenvironments ex vivo. For complete details on the use and execution of this protocol, please refer to Tabdanov et al. (2021).(1)