Using a specific polyclonal antibody against type VI collagen, the distribution of type VI collagen in the bovine cornea was investigated by indirect immunofluorescence microscopy and immunoelectron microscopy. By immunofluorescence, type VI collagen was detectable throughout the stroma. In addition to the uniform but relatively weak signals, a laminar pattern similar to the arrangement of stromal lamellae was clearly seen. Consistent with the immunofluorescent pattern, immunoelectron micrographs showed strong linear reactions located in the interlamellar spaces. The reactions were also observed in the pericellular region of the keratocytes. Epithelium, Descemet’s membrane and endothelium were negative.