Noninvasive, longitudinal near‐infrared fluorescence (NIRF) imaging was used to detect and quantify lymphangiogenesis following a full‐dermis thickness incision in the presence and absence of locally administered vascular endothelial growth factor‐C (VEGF‐C), a well‐known regulator of lymphangiogenesis. Peripheral cytokines/chemokines were also measured in treated and sham‐injected animals. Lymphangiogenesis was detected via NIRF imaging by day 7–8 and confirmed by intravital microscopy, while angiogenesis was observed by day 2–3 postincision (PI). All lymph vessel parameters quantified were significantly greater on wounded vs. nonwounded sides of mice. Lymph vessel parameters appeared larger on wounded sides of VEGF‐C– relative to NaCl‐treated mice, although differences were not significant. Interleukin‐1α and interleukin‐22 were significantly elevated at day 7 PI relative to respective preincision levels in VEGF‐C‐treated mice, and decreased by day 21 PI to levels nearing those measured preincision. For the majority of cytokines/chemokines measured, mean responses were significantly greater in VEGF‐C– vs. NaCl‐treated animals. Local VEGF‐C administration may stimulate lymphangiogenesis during tissue repair and regeneration via mediating systemic cytokine/chemokine levels. NIRF imaging can be utilized to detect lymphangiogenesis during wound healing, and offers a promising platform to complement current methods for monitoring wound status and studying the effects of growth factors on healing.