Microcapsules containing rejuvenator are particles used to increase the self-healing capability of asphalt. To date, no reports focus on studying their behaviors in asphalt binders between the aggregates. The purpose of this work was to directly observe the states of self-healing microcapsules in asphalt binders. Asphalt samples were prepared by mixing bitumen and various weight contents of microcapsules. Experimental tests were carried out to observe the morphology, integrity, distribution, thermal stability, interface bonding and triggered rupture of microcapsules in asphalt binders. Fluorescence microscope morphologies and X-ray computed tomography images showed that microcapsules survived in asphalt resisting high temperature and strong agitation without premature damage. At the same time, microcapsules were homogenously dispersed in asphalt binders avoiding particles aggregation and adhesion. A circular heating-cooling process was used to simulate temperature changes in the natural environment. It was found that microcapsules still kept a stable state after an extreme temperature change. In addition, interface debonding phenomenon did not appear. Microscopic observation results reflected that microcapsules could be pierced by microcracks in the asphalt binder and the encapsulated rejuvenator flowed out under the force of capillary action. All the above conclusions indicate that microcapsules containing rejuvenator meet the application conditions and play the role of self-healing material in asphalt binders.