Following incubation in an isotonic saline solution containing 20 mM EDTA, human oral mucosa may be separated into its epithelial and connective tissue components. Ultrastructural study of the separated tissues reveals that the plane of separation is through the lamina lucida. Hemidesmosomal structure is altered by the separation process: the peripheral density is absent but a fine, generally filamentous material remains associated with the outer membrane leaflet of the hemidesmosome. Desmosome structure is not altered. An intact lamina densa remains attached to the connective tissue fragment. Oral mucosa incubated in EDTA-saline containing calcium, or its return to a divalent cation-supplemented medium after treatment with EDTA, prevents separation. By maintaining the structural integrity of the hemidesmosome, divalent cations appear to play a principal role in uniting oral mucosal epithelium to the lamina propria.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/49659/1/1001330406_ftp.pd