The structure of the microvascular network within the murine panniculus carnosus is highly anisotropic and adapted to the functional requirements of the tissue. To assess the rate of wound closure and functional characteristics of the regenerating vascular plexus, dorsal skinfold chambers (dsc) were implanted in adult mice enabling the use of intravital microscopy. As a result of heat-induced injury to the tissue, a non-perfused area defined as wound was created. Three days after wounding, vasodilatation of the surviving vessel segments was noted even in more distal areas of the plexus, with a peak in vessel junction density at day 6 (indicative of the period of active sprouting), and partial recovery of functional vessel density by day 9. Having established a longitudinal model of wound healing-induced angiogenesis, we sought to test the influence of different parameters in the progression of blood vessel growth. TNP-470 was the first anti-angiogenic agent to undergo clinical trials in human tumours. The main complication of anti-angiogenic therapy for tumours is its unavoidable effect on physiological processes such as wound healing. TNP-470 covalently binds and inhibits the metalloprotease methyonine aminopeptidase metAP-2 (Sin et al., 1997), which is enriched in endothelial cells, effectively arresting the cell cycle. Injection of TNP-470 led to a significant delay in wound closure with both the extent of vessel sprouting and recovery of tissue perfusion inhibited compared to vehicle-injected mice. Thus, in this experimental model, increase in functional vessel density depends upon endothelial cell proliferation (Machado et al., 2011). Blind-ended vessels (BEVs) at the leading edge of an expanding vascular plexus were recognized as major regulators of the structure of the developing network (Guerreiro-Lucas et al., 2008). Plasma leakage into the surrounding extracellular matrix occurring from BEVs could play an important role in preparing hypoxic tissue for vascular invasion. As such, plasma flux across the vessel wall was quantitated through the use on in vivo Fluorescence Recovery After Photobleaching (FRAP). Determination of time elapsed from bleaching until recovery of fluorescence intensity to half of baseline average (t1/2) revealed that the permeability of newly formed vessel segments (both flowing and blind-ended) is greater than that of pre-existing vessels between days 5-7 after injury. Furthermore, TNP-470 reduces macromolecular flux across the vessel wall of BEVs at the leading edge of the advancing vascular plexus, while increased FITC-dextran leakage was observed from pre-existing vessels after treatment with TNP-470 (p<0.05), consistent with induction of transient vascular damage, (Machado & Mitchell, 2011). This evidence contradicts previous reports where quiescent endothelium is not affected by TNP-470 (Kruger & Figg, 2000). Using in vivo FRAP in the dsc, these studies demonstrate the relationship between temporal changes in microvascular macromolecular flux and the morphology of maturing vascular segments. The combination of techniques used to characterize the morphology and function of the newly-formed blood vessels in this experimental model provided spatially and temporally resolved data, which is amenable for use within mathematical modeling approaches. The predictive power of these data-driven models will enable us to test for unwanted side effects of anti-angiogenic drugs, such as the increased leakiness of pre-existing vasculature.