This paper reports a study on the assembly of gold nanoparticles via a tetradentate organosulfur ligand, tetra[(methylthio)methyl] silane. We have characterized the evolution of the assembly from individual nanoparticles to spheres (30 ∼ 160 nm) of linked nanoparticles using UV-Visible, TEM, and AFM techniques. We have also demonstrated that the assemblies could be effectively disassembled via manipulating the ligand chemistry. Intriguing assembly-substrate interactions were observed, which could be related to interfacial hydrophobicity. Implications of these findings to the development of abilities in interfacial manipulation of the nanostructures are also discussed.