Caves with dim natural light, and lighted hypogean environments, have been found to host phototrophic microorganisms from various taxonomic groups. These microorganisms group themselves into assemblies known as communities or biofilms, which are associated with rock surfaces. In this work, the phototrophic biofilms that colonise speleothems, walls and floors in three tourist caves (Spain) were studied. Confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM) were used to study these organisms and acquire three-dimensional data on their biofilm structure. CLSM was used in a multi-channel mode whereby the different channels map individual biofilm components. Cyanobacteria, green microalgae, diatoms, mosses and lichens were found to be grouped as biofilms that differed according to the sampling sites. The biofilms were classified into six types regarding their environmental conditions. These types were defined by their constituent organisms, the thickness of their photosynthetic layers and their structure. Light-related stress is associated with lower biofilm thickness and species diversity, as is low humidity, and, in the case of artificially illuminated areas, the duration of light exposure.