The　two-dimensional　(2D)　assembly　of　the　palladium　nanoparticles　(Pd　NPs)　was　studied　in　this　work.　The　cubic　Pd　NPs　were　successfully　synthesized　and　assembled　on　mica　and　silicon　wafer　in　the　dip-coating　way.　The　morphology　of　the　Pd　NPs　and　the　topography　of　the　Pd　NPs　assembly　on　the　substrates　were　characterized　with　transmission　electron　microscopy　(TEM)　and　atomic　force　microscopy　(AFM).　In　the　process　of　the　fabrication,　the　excess　cetyltrimethylammonium　bromide　(CTAB)　was　removed　with　the　＂deposition-redispersion＂　strategy,　the　UV-vis　spectra　and　zeta-potential　of　the　Pd　NPs　colloid　were　measured.　It　was　found　that　the　assembly　and　AFM　characterization　of　the　Pd　NPs　were　affected　negatively　by　the　presence　of　excess　CTAB.　The　hydrophilic　property　of　the　substrate　is　the　crucial　factor　to　control　the　2D　assembly　of　the　Pd　NPs.　Compared　with　the　washed　silicon　wafer,　mica　is　ultra-hydrophilic　and　can　attract　more　Pd　NPs.