The “Drop casting” and “spin-coating” techniques are frequently utilized to manufacture nanostructure materials like TiO2, in which the modifying layer comprises nanoparticles (NPs). The study aims to use spin-coating and Drop-casting techniques to deposit highly uniform and good adhesion nanoparticles. TiO2 NPs have been successfully deposited using the spin-coating technique which has smoother films with discernible crystallinity used for antibacterial activity. The structural, morphological, and optical properties of TiO2 NPs prepared through Drop-casting and spin-coating techniques were studied using XRD, SEM, AFM, and UV-Vis, respectively. XRD patterns of TiO2 NPs which were prepared by Drop-casting and spin-coating techniques and the solution prepared with concentrations of (60,120) g/L was the monoclinic phase of TiO2 nanoparticle. The average crystalline size was increased with increasing concentrations. The films have a smooth surface, a high density, and excellent adhesion to the glass substrate, according to SEM and AFM’s observation of the images. Additionally, because of the growth in particle size and increasing solution concentrations, the surface roughness for synthesized samples increased. UV-Vis illustrated the allowed direct electronic transition and the value of band gaps increased with increasing concentrations of TiO2 NPs. Finally, all the modification characterizations via the spin-coating technique contribute to knowing the effect of TiO2 NPs concentrations on the antimicrobial activity which was increased with increasing NPs concentration.