Fig. 1. Cervical lymphatic mapping and neck dissection model.
a Illustrative photographs depicting anatomic lymphatic mapping following injection of 5% Evans Blue dye into oral cavity (OC) subsites. b Top: Illustrative IVIS image to depict anatomic lymphatic mapping following injection of anti-LYVE-ef660 antibody into the tongue (1s exposure with the Cy5.5 channel on the IVIS 2000). Bottom: Representative image of a clearing-enhanced 3D (Ce3D) en bloc resected tongue-neck specimen stained with anti-LYVE-ef660 (1:100), imaged with the Leica SP8 confocal microscope. c Representative images demonstrating functional mapping following injection of SIINFEKL peptide/CpG adjuvant. Depicted are lymphatic basins, overlayed with heatmaps, to indicate the % CD11c + H-2kb SIINFEKL + cells identified by flow cytometry. d Putative tumor-draining lymph nodes (tdLNs) and non-draining lymph nodes (ndLNs) were harvested from 4MOSC1 tumor-bearing animals on day 10. Top: Representative H&E-stained tdLN and ndLN shown with (bottom) scoring for overall surface area and the number of reactive follicles (n = 10 independent samples/group). e Left: Representative tdLN with the focus of metastatic disease shown, stained by H&E and with anti-pan-CK antibody. Right: Quantification of the incidence of metastatic disease in tdLN and ndLN, shown in a contingency plot (n = 25/group, Fisher’s exact test). f Illustrative photographs demonstrating the key procedural steps of the murine neck dissection. (1) dissection to reflect the submandibular gland from the superficial lymphatic basin, (2) superficial lymphatic basin liberated from underlying tissues, (3) completed dissection of the deep lymphatic basin with the jugular venous plexus in situ along the floor of the neck, (4) closure. g Representative axial CT images of the neck obtained from an untreated 4MOSC1-tongue tumor-bearing animal or following neck dissection (arrowhead = cervical lymph node). h Cartoon image to diagram murine cervical lymphatic basins in the context of adjacent, critical head, and neck anatomy. The differences between experimental groups were analyzed using independent, two-sided Student t tests (d) or fisher’s exact test (e). All data represent averages ± SEM, except where indicated. ****P < 0.0001. ns not statistically significant. Source data are provided as a Source Data file.