The distribution of lectin-binding sites in oxyntic and chief cells of isolated rabbit gastric glands was determined with seven fluoresceinated lectins, to ascertain which lectins might best be used in the biochemical characterization of cell membranes and glycoproteins of these two cell types. Oxyntic cell canaliculi were labeled by wheat germ, Helix pomatia, and peanut lectins, suggesting a predominance of N-acetylhexosamines. Tubulovesicles were heavily stained by wheat germ, Helix pomatia, and Ricinus communis I lectins, indicative of N-acetylhexosamine- and galactose-containing glycoconjugates. Diffuse oxyntic cell cytoplasmic staining was observed with the mannose-binding lectin concanavalin A. This lectin, along with wheat germ, soybean, Helix pomatia, and Ricinus communis I lectins, bound to oxyntic cell basolateral membranes, indicating mannose, N-acetylhexosamine, and galactose residues. Chief cell apical membranes were labeled with peanut, Ricinus communis I, Helix pomatia, and Ulex europaeus lectins, suggesting a predominance of N-acetylhexosamine, galactose, and fucose residues. None of the lectins demonstrated any significant affinity for chief cell cytoplasm or basolateral membrane. Ulex europaeus agglutinin-binding sites were additionally concentrated in lateral intercellular spaces. The results of this study indicate that the range of utility of isolated rabbit gastric glands can be expanded to include histochemical work. In addition, the data suggest the applicability of lectin affinity chromatography in the isolation and characterization of oxyntic and chief cell membranes and glycoproteins.