Atomic force microscopy was used to characterize the structural evolution of the V2O5(001) surface during the electrochemical cycling of lithium. With Li insertion, nanometer-scale pits develop at the V2O5(001) surface. The pits first appear as the composition of the crystal approaches Li0.0006V2O5. Pit nucleation and growth continue through further discharge, resulting in a micro-porous (001) surface morphology. During subsequent Li extraction, cracks develop along the V2O5 <010> axis. Surface regions in the vicinity of these cracks “swell” during ensuing lithiation reactions, suggesting that the cracks locally facilitate Li uptake.