Self-assembled monolayer (SAM) represents one of the methods to precisely modify surface structures in the nanoscale dimension. It has opened up a new era of exploration and has a profound impact on sensors and biosensors due to its unique properties. Different self-assemblies will be considered in this chapter: SAM of metallic nanoparticles on polymeric film, SAM of surfactant on polymeric film, and SAM of S-containing compounds on nanometallic films. The main goal of this chapter is to present comprehensive collection of the recent achievements in this area. Several issues will be discussed including the morphology, sensitivity, selectivity, stability, and electrochemical properties of the sensor.