The formation of self-assembled multilayer system on various substrates was studied using an N,N'-bis(ethyldihydrogenphosphate)-3,4,9,10-perylene (dicarbox imide) (EPPI), a well-known electron donor. The substrates used in this study were gold, silicon, quartz, and glass. In order to attach the EPPI to the substrates, a layer of an anchoring agent was initially formed on the substrates. Also zirconium was introduced between the adhesive layer and the EPPI layer in order to stabilize this multiplayer system. The characterization and self-assembling process of EPPI on a Zr-MUDP layer were elucidated using in-situ/ex-situ surface plasmon resonance (SPR), UV-Vis, and ellipsometry.