Free-standing activated carbon nanofibers (ACNF) were prepared through electrospinning combining with CO2 activation and then used for nonaqueous Li-O-2 battery cathodes. As-prepared ACNF based cathode was loosely packed with carbon nanofibers complicatedly overlapped. Owing to some micrometer-sized pores between individual nanofibers, relatively high permeability of O-2 across the cathode becomes feasible. Meanwhile, the mesopores introduced by CO2 activation act as additional nucleation sites for Li2O2 formation, leading to an increase in the density of Li2O2 particles along with a size decrease of the individual particles, and therefore, flake-like Li2O2 are preferentially formed. In addition, the free-standing structure of ACNF cathode eliminates the side reactions about PVDF. As a result, the Li-O-2 batteries with ACNF cathodes showed increased discharge capacities, reduced overpotentials, and longer cycle life in the case of full discharge and charge operation. This provides a novel pathway for the design of cathodes for Li-O-2 battery.