A city's economic growth and the inhabitants' wellbeing are highly affected by its topology and connecting networks, which, in turn, influence movement and flows in the city. Flow relates to how a city is developed, organized, managed, and built. The analysis of flow in cities is challenging but essential. In this study, the fields of urban design and animal science are combined, and a new approach for exploring the relationships between urban topology and physical flow is developed. Specifically, we establish an interdisciplinary methodology to evaluate mobility performance in various urban settings, utilizing experimental observations of the dynamic behavior of natural-biological agents, i.e., locusts, within physical city models. Our novel approach enriches the currently available toolbox by using living organisms as indicators for flow in physical city models. Our findings improve our understanding of the intricate flow interactions in urban settings.