Improving fabrication protocols and design strategies, investigating on how scaffold architecture affects cell morphology, metabolism and phenotypic expression, predicting mechanical behaviors, have increasingly become crucial goals in the development of engineered tissue scaffolds. In the present study, an image-based analysis approach that provides an automatic tool to fully characterize engineered tissues fiber network topology was developed. The following micro architectural features are detected: fiber angle distribution, fiber connectivity, fiber overlap spatial density, fiber diameter. In order to demonstrate the potential of this approach Electrospun poly(ester urethane)urea (ES-PEUU) scaffolds were studied. Electrospun scaffolds were chosen for their recognized capability to recapitulate native soft tissue extra cellular matrix (ECM) morphology.