In fiber-reinforced polymer composites, the fiber-matrix interface controls stress transfer mechanisms, thereby affecting mechanical performance. Interfacial properties are often extracted via single-fiber composite tests. In these tests, the load is transferred from the polymer to the fiber through interfacial shear stresses, necessitating the evaluation of interfacial shear properties. To adopt these properties in the design of industrially relevant composites, one must assume that the damage mechanisms in single-fiber composites are representative of those in multi-fiber composites, consisting of highly aligned, unidirectional plies with high fiber volume fractions. That assumption, however, has never been validated. In this paper, the real-time damage development is monitored in single-fiber and multi-fiber composites using in situ X-ray holo-tomography at 150-nm pixel size. The technique enables the first-ever 3D detection of longitudinal interfacial debonding in carbon and glass single-fiber composites. This mechanism is not detected in multi-fiber composite specimens, suggesting that single-fiber composites are intrinsically unrepresentative of realistic composite behavior.