An in vitro model consisting of enamel from extracted human molars, suspended from wires in inoculated culture tubes, was used to study the adhesion of bacteria to enamel. Under conditions in which there was no macroscopically visible plaque formation, electron micrographs showed no bacterial deposits on the enamel surface. In samples where Streptococcus mutans attached to enamel, an extracellular, pellicle-like material was associated with the bacteria adjacent to the enamel. This material appeared to bind to the enamel surface and to mediate bacterial attachment. Membrane-filtered (Millipore Corp.) saliva deposited a thin surface layer on the enamel, but there were no observable alterations of S. mutans attachment to enamel pretreated with saliva. It was noted that Bratthall serotype c and e strains of S. mutans, when grown in glucose-containing medium, attached, although less tenaciously, to enamel and nichrome wires. Chemical and gas chromatographic analyses of cell-associated materials formed by serotype c and e strains cultured in glucose-containing medium revealed low amounts of glucose-positive material and no polymer linkages characteristic of glucan; yet the same strains cultured in sucrose-containing medium had relatively high amounts of glucose-positive material, with polymer linkages-characteristic of glucan. Serotype a, b, and d strains could attach only in sucrose-containing media.