Interfibrillar bonding of collagen fibrils in tissue grown from rabbit chondrocytes in culture was examined by a variety of electron microscopy techniques. Interfibrillar bonding is expected to increase tissue strength and may be a desirable feature in engineered cartilage and other soft tissues. The apparent bonding evident by scanning electron microscopy, using standard chemical fixation processing, is suspected to be artifact due to drying. The goal of this article was to establish the existence of interribrillar bonding, apart from any processing artifacts. Specimens prepared by transmission electron microscopy, scanning electron microscopy (SEM) after notching and fixing under load, and cryo-SEM all showed evidence of bonding, supporting the existence of bonding in the unprocessed tissue. Exclusion from the bond space of gold particles labeled to decorin further supported the existence of natural bonds. Artifactual bonding may still be occurring with some of the methods used, but interfibrillar bonds exist in natural tissue. The bond distance was estimated to be 7-14 nm. Demonstration of the existence of these bonds supports further study of their mechanism and effect on tissue properties.