In this work, we investigated the evolution of the graphite-water interface in a nitrogen atmosphere by using frequency-modulation atomic force microscopy (FM-AFM). A highly ordered pyrolytic graphite (HOPG) sample was immersed in pre-degassed water and subsequently placed in the nitrogen environment. The dissolved nitrogen molecules diffused in water and self-assembled into ordered row-like structural domains at the interface. Nucleation and growth processes of the domains were observed. When the coverage of the ordered structure surpassed 50%, small clusters began to appear on the ordered structure. The number density of the clusters increases as the coverage of the ordered structures increases and these clusters may hop on the ordered structures. A model is proposed to explain the evolution of the nitrogen molecule self-assembly process at the interface. The observation of nitrogen clusters may shed light on the nature and nucleation of the so-called nanobubbles at hydrophobic-water interfaces. (C) 2014 Elsevier B.V. All rights reserved.