Since the "invention" of free flight, the key question is whether airborne self-separation can safely accommodate very high traffic demand. The aim of this paper is to answer this question for en route airspace. Therefore, an advanced airborne self-separation concept of operation is evaluated on safety risk at very high traffic demands. The advanced airborne self-separation concept of operations considered is of the trajectory-based operation type, in the sense that each aircraft manages a conflict-free four-dimensional trajectory intent and broadcasts this to the other aircraft. Complementary to this trajectory-based operation layer, each aircraft makes use of a short-term conflict detection and resolution layer that aims to resolve any remaining problems, such as significant deviations from fourdimensional intents due to wind prediction errors. Safety risk analysis is conducted using advanced techniques in agent-based modeling and rare-event Monte Carlo simulation. The results obtained show that the advanced selfseparation concept of operations considered has a remarkably good collaboration between the trajectory-based operation and the tactical resolution layers: as a result of which, it can safely accommodate very high en route traffic demands.