Angiogenesis is essential for tumor growth and a promising target for cancer therapy. Blood vessel monitoring is an
indispensable tool for evaluation and development of anti-angiogenic drugs. Here, we report a new noninvasive in vivo
imaging tool, named dynamic fluorescence imaging (DyFI), for the simultaneous measurement of multiple vascular
parameters, such as vascular density, perfusion rate, and permeability, using spatiotemporal profiles of indocyanine green.
Using DyFI in a tumor xenograft model, we quantitatively measured multiple vascular parameters in tumors and normal
tissues with high spatial resolution. The multimodality of this method allowed us to find a negative spatial correlation
between perfusion and permeability. Moreover, DyFI was effective for revealing the early effects of an anti-angiogenic
drug; these findings were validated using two-photon microscopy. We suggest that DyFI could be a useful tool for the
preclinical development of anti-angiogenic drugs.