Light and electron microscopic immunocytochemistry was used to localize S-100 protein. It was demonstrated in chondrocytes of the proliferative and hypertrophic zones but none could be found in the fibrous articular layer. The staining intensity for S-100 protein was stronger in the hypertrophic cells than in chondrocytes from the proliferative zone. Ultrastructurally, immunoreactive gold particles were detected in the cytosolic region, closely apposed to the profiles of endoplasmic reticulum with occasional dense aggregations. A few gold particles were seen on the nuclear chromatin and on condensing vacuoles of Golgi complexes. Because of the calcium-binding and possibly fatty acid-binding properties of the protein molecule, the immunocytochemical expression of S-100 protein in the chondrocytes may be involved in cartilage cell differentiation, metabolism and mineralization.