Cell-mediated Cytotoxicity Assay: Total Cytotoxicity Test

1. Prepare samples and controls., 2. Dilute 10X Injection Buffer 1:10 by adding 45 mL diH20., 3. Reconstitute FAM-FLIVO with 50 µL DMSO., 4. Dilute FAM-FLIVO 1:12 with 550 µL 1X Injection Buffer., 5. Inject 100 µL intravenously., 6. Let FAM-FLIVO circulate 30-60 minutes., 7. View live tumor through a window chamber using a fluorescence microscope., 8. If not viewing directly, excise tissue., 9. If desired, label with additional stains, such as Hoechst 33342, or an antibody., 10. If desired, fix cells., 11. Analyze with a fluorescence microscope, flow cytometer, or a window chamber system. FAM-FLIVO excites at 492 nm and emits at 520 nm.