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Biofilms are formed by microbial cells growing onto solid surfaces embedded in a polymer matrix, mainly consisting of eso-polysaccharides (EPS), produced by the cells themselves. Biofilms are not tissue, but aggregates, defined as a city of microbes, with a complex morphology including canyons and streamers, whose function is not totally identified yet. The organization, growth and morphology of these aggregates depends strongly on the type of substrate and on the various compounds allowing the communication among cells and quorum sensing along complex gradients. The importance of biofilms of fungal, bacterial and mixed origin depends on their increased resistance to antibiotics, anti-fungal drugs and extreme conditions. Furthermore, biofilms are almost ubiquitous and their formation has been demonstrated to be of primary importance to determine the success of the microorganisms able to form it.