Fig. 7: Chorioallantoic membrane (CAM) assay experimental design.

Breeding of the fertilized eggs with automatic rotation started from embryonic development day zero (d 0). Eggs were opened and the CAM was prepared at day 6. On day 7, 1 × 106 cells of the respective head and neck squamous cell carcinoma (HNSCC) cell line were treated with 0, 105, 210, and 315 MPa high hydrostatic pressure (HHP) at 20 °C for 10 min. Following 7 days of incubation and tumor growth, euthanasia and harvesting of the CAM were performed. Near-infrared (NIR) fluorescence was measured, and the CAM tissue was prepared for histology.