A human-in-the-loop simulation experiment was designed and conducted to evaluate an airborne self-separation concept. The activity supports the National Aeronautics and Space Administration s (NASA) research focus on function allocation for separation assurance. The objectives of the experiment were twofold: (1) use experiment design features in common with a companion study of ground-based automated separation assurance to promote comparability, and (2) assess agility of self-separation operations in managing trajectory-changing events in high traffic density, en-route operations with arrival time constraints. This paper describes the experiment and presents initial results associated with subjective workload ratings and group discussion feedback obtained from the experiment s commercial transport pilot participants