Biodeterioration damage is an important issue in conservation and restoration of built heritage, especially when ceramic materials are used. Biological colonization of ceramic roofing tiles by lichens is a common phenomenon. However, there are no reports to date of lichens removal from unglazed roofing tiles for conservation purposes. This paper for the first time reveals the results of a combined procedure undertaken to assess the removal of lichens on different kinds of unglazed ceramic roofing tiles by treatments based on both dual sequential laser irradiation and treatment using Acticide® CF biocide. Three species of lichens were identified: Verrucaria nigrescens, Calogaya decipiens and Pyrenodesmia teicholyta. The chemical and mineralogical composition of roofing tiles were characterized by X-ray fluorescence (XRF) spectrometry, optical polarized petrographic microscopy, and X-ray diffraction (XRD). Laser irradiation was accomplished by applying sequences of nanosecond laser pulses at two wavelengths (1064 and 266 nm). After dual sequential laser irradiation a biocide was applied. To assess the combined effect of both treatments several techniques were used, including stereo and fluorescence (FM) microscopies, scanning (SEM) and transmission (TEM) electron microscopies, and FT-Raman spectroscopy. Chemical composition of the analyzed roofing tiles was shown as a relevant factor regarding the degree of interaction between the biological colonization and the substrate, and hence, the bioweathering effect. The combined procedure has proved to be very effective in removing some lichen thalli and damaging completely the bionts in all species.