Self-assembled monolayers (SAMs) have attracted attention due to their chemical and structural properties providing numerous new applications such as molecular electronics and electrochemistry. SAMs were optimized by experimental techniques including Scanning Tunneling Microscope (STM), Scanning Tunnelling Spectroscopy (STS), X-Ray Photoelectron Spectroscopy (XPS), Polarization-Modulation Infrared Reflection-Absorption Spectroscopy (PM-IRRAS), Cyclic Voltammetry (CV), Electrochemical Impedance Spectroscopy (EIS). The first part of this dissertation deals with the dynamic behaviour of decanethiol SAMs. The dynamic behaviour alkanethiols SAM is unique for its configuration giving indirect evidence for the structural ordering within the formation. Structural stability of decanethiol (decanesulfonates) SAMs were investigated by space- and time- resolved STM. Decanesulfonate phase shows less dynamic behaviour and is structurally more stable compared to the decanethiol phases. The second part of this dissertation describes the binding properties of alkyne molecules adsorbed on gold. Alkyne oxidation occurs at ambient conditions but it is found that unlike thiols, ordered alkyne SAM structure has still chemical bonding between carbon and gold. Alkyne SAMs are good candidates for the ambient molecular electronics application. The last part of this dissertation presents the study of a monolayer of a Ru(II) complex which is prepared on gold substrate. With the help of STM and XPS methods, the ordered structure and binding properties of the CS28 molecules were characterized by providing a deeper insight into the carboxyl and sulfur groups binding affinity to gold substrate. CV and EIS methods were used to compare the adsorption properties and charge transfer process with the bare gold substrate and SAMs.