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Animal behavior is an essential element in behavioral neuroscience study. However, most behavior studies in small animals such as fruit flies ( Drosophila  melanogaster ) have been performed in a limited spatial chamber or by tethering the fly's body on a fixture, which restricts its natural behavior. In this paper, we developed the Transparent Omnidirectional Locomotion Compensator (TOLC) for a freely walking fruit fly without tethering, which enables its navigation in infinite space. The TOLC maintains a position of a fruit fly by compensating its motion using the transparent sphere. The TOLC is capable of maintaining the position error &lt; 1 mm for 90.3% of the time and the heading error &lt; 5° for 80.2% of the time. The inverted imaging system with a transparent sphere secures the space for an additional experimental apparatus. Because the proposed TOLC allows us to observe a freely walking fly without physical tethering, there is no potential injury during the experiment. Thus, the TOLC will offer a unique opportunity to investigate longitudinal studies of a wide range of behavior in an unrestricted walking Drosophila .

Navigation of a Freely Walking Fruit Fly in Infinite Space Using a Transparent Omnidirectional Locomotion Compensator (TOLC)