Fig. 8

Fluorescence multiplex immunohistochemistry (mIHC) of draining LNs harvested at different time points after i.m. immunization. a Representative seven-color multispectral image of a lymph node collected 4 h after i.m. injection of HA-OVA, scanned at ×20 (upper panel) and ×40 (lower panel) magnification. The white arrows in the ×40 image show examples of an OVA⁺ DC (1, CD11c), OVA⁺ lymphatic endothelial cell (2, LYVE-1), and OVA⁺ macrophage (3, F4/80). b Quantification of OVA⁺ cell density (cells/mm²) in LNs of mice subjected to i.m. injection with 10 µg of OVA alone, mixed with alum, or conjugated to HA. c Percentage of OVA⁺ cells among LYVE-1⁺, CD11c⁺, and F4/80⁺ cells. Cumulative data from three independent experiments are presented (n = 3 mice; 6 LNs/group). Multiple t-tests were performed (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; P > 0.05 if not indicated)