Fluorescent images prior to the popliteal lymphadenectomy (A, G), at 27 days post-surgery (B, H), and 21 days after B16F10 cell inoculation (C, I).

<p>At 3 weeks post inoculation, intravital color images showed B16F10 metastasis to the ILN (D), IsLN (E), and LLN (F) in a mouse with lymphatic drainage to ILN and IsLN across the popliteal wound site as demonstrated by the presence of black melanin pigment in LNs. In a mouse with lymphatic drainage to the ILN (I), ILN metastasis (J) was observed, but not in the mouse IsLN which was not involved in the lymphatic drainage pathway (K). In addition, in-transit metastasis was detected proximal to the wound site (L). White arrow, ICG injection site. Double arrow, PLN. Open arrowhead, popliteal efferent collecting lymphatic vessel. Arrowhead, ILN. Open arrow, B16F10 metastasis. Black arrow, in-transit metastasis. Scale bar, 1 mm.