We introduce new realistic three-dimensional tissue phantoms which can help to understand the optical properties of human epithelium as well as the optical signatures associated with the dysplasia to carcinoma sequence. The phantoms are based on a step by step multilayer reconstitution of the epithelial tissue using main components characteristic for the human epithelium. Each consecutive step is aimed to increase the similarity between real tissue and a phantom. We began by modeling the stromal layer which predominantly consists of a network of collagen bundles. Phantoms consisting of a collagen matrix alone and in the presence of embedded cervical cells were created. Their morphology and fluorescence properties were studied and were compared with those of cervical epithelium. We show that the phantoms resemble the microstructure and the optical properties of the human epithelial tissue. We also demonstrate that the proposed phantoms provide an opportunity to study changes in optical properties of different tissue components as a result of their interactions with each other or exogenous factors. © 2002 Society of Photo-Optical Instrumentation Engineers.