Urban design is a complex and challenging process that encompasses many disciplines, such as urban planning, architecture, and landscape. On the urban design scale, it’s important to consider short paths compatible with pedestrian behaviors, especially in public spaces, in terms of designing healthier and more accessible urban areas. But conventional design processes, which are mostly based on the designer’s observations or intuitions, fail to satisfy user behaviors in usage. Studies reveal that P. polycephalum slime molds have the potential to guide good urban planning. However, it is noteworthy that there are not enough studies yet in the disciplines of architecture and urban design on the design of pedestrian ways in small-scale areas, and there is a need for research. In this direction, this study is the first step of a method that is aimed at being developed for the disciplines of architecture and urban design in the long process. The main hypothesis is that a design template can be created as a result of graphical analysis of living slime mold experiments on the map, especially in undesigned empty urban areas, for architects and planners, and it can provide creative suggestions to the designer. In this direction, in this study, a graphic design template proposal was created based on live cell experiments at a site in Izmir, Turkey, which the local government aims to design as a pedestrianized zone and open for competition. Slime molds are used directly as a computer rather than as inspiration for a computer, and a pedestrian-oriented design template proposal develops with the behavioral models of slime molds. Compared to the award-winning designs developed for the same area, it has been shown that even within the framework of simple graphical measurements, it gives better results and creates more optimal routes on main and side roads. It is envisaged that this study can play a pioneering role for studies involving more detailed mathematical models that can begin in the field of architecture and urban design.