Laser removal of biodeteriogen layers warrants detailed studies due to the advantages it brings with respect to mechanical elimination or the use of biocides. We have investigated elimination of biological crusts on dolomite stones from heritage sites in central Spain. The samples were colonized by epilithic crustose lichens of different species, such as Caloplaca sp. and Verrucaria nigrescens. A comparative study was carried out by applying infrared (1064 nm) and ultraviolet (355 nm) nanosecond laser pulses and sequences pulses of the two wavelengths using a Q-switched Nd: YAG system. To detect anatomical and ultrastructural damage to the lichens, and to assess possible morphological and chemical changes on the underlying stone induced by laser irradiation, we used stereomicroscopy, scanning electron microscopy with backscattered electron imaging and Fourier transform Raman spectroscopy. The optimal conditions for removal of the colonization crust, while ensuring preservation of the lithic substrate, were obtained for dual infrared-ultraviolet sequential irradiation. (C) 2015 Elsevier B.V. All rights reserved.