Hydrophobic Barriers for Directing Physarum polycephalum Propulsion and Navigation

[Image: see text] Physarum polycephalum (P. polycephalum) is a unicellular protist with unique properties, such as learning and remembering in its cultured environment without a brain or central nervous system. The organism has been extensively used in morphology, taxis, and positive feedback dynamics studies. However, the lack of standardization of materials and substrate designs used in P. polycephalum studies has significantly limited conducting such studies, increasing the cost and time. In this study, we introduce a method to control the direction and migration of P. polycephalum by drawing hydrophobic lines and patterns. Our study succeeded in controlling the movement of P. polycephalum by setting a variety of hydrophobic designs such as complete barrier, single-slit barrier, taper barrier, dumbbell barrier, and one-side-opened rectangular barrier, suggesting the effectiveness of the hydrophobic barrier in regulating the propulsion and navigation of the organisms. Moreover, we demonstrated that utilizing such geometric constraints can reduce the experimental time required for toxicity testing based on P. polycephalum by more than 300%. Our techniques open new possibilities for studying the biophysical properties and behaviors of P. polycephalum, while also facilitating toxicity testing.