Abstract In the present study, we developed a novel model to image angiogenesis of lymph node cancer metastasis. Transgenic nude mice carrying GFP under the control of the nestin promoter (ND-GFP mice), 6 to 8 weeks old, were used as hosts. ND-GFP mice express GFP in nascent blood vessels. B16F10-RFP cells were injected into the efferent lymph vessel of the inguinal lymph node of ND-GFP mice to establish experimental metastasis in the axillary lymph node. The axillary lymph nodes of the mice were imaged at days 0, 3, 7, 14, and 28 after injection of tumor cells. Three days after cancer-cell transplantation, nascent blood vessels were imaged by GFP fluorescence in the tumor-involved lymph nodes in the ND-GFP mice. Ten days after cancer-cell transplantation, ND-GFP expression demonstrated an angiogenesis network in the tumor-involved lymph node. Fourteen days after cancer-cell transplantation, ND-GFP expression in the lymph node tumor blood vessels began to be extinguished as the blood vessels matured. Twenty-eight days after cancer-cell transplantation, ND-GFP expression in blood vessels in the lymph node tumors was completely extinguished. This model is useful for direct visualization of nascent tumor angiogenesis in lymph nodes with metastatic cancer, a very critical site for early metastasis. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3437.