The acidophilic and chemolithotrophic Thiobacillus ferrooxidans bacteria grown in the presence of soluble ferrous ions and solid substrates (sulfur, pyrite and chalcopyrite) as their energy source have been used in the investigations. The cell surfaces are characterized by zeta-potential, contact angle, FT-IR and FT-Raman studies. The adsorption isotherms of the cells on pyrite and chalcopyrite indicate that the adsorption begins on pyrite at a much lower equilibrium cell population than on chalcopyrite. The isotherms tend to level off at a cell adsorption density corresponding to a horizontal monolayer coverage when the geometrical dimensions of the cells are considered. In the presence of cells, the xanthate flotation of pyrite completely depressed, whereas chalcopyrite flotation is unaffected. The results suggest that chalcopyrite can be selectively floated from pyrite, also at a neutral pH region, in the presence of T. ferrooxidans cells