Measuring degeneracy and broken-symmetry states of a system at nanoscale requires extremely high energy and spatial resolution, which has so far eluded direct observation. Here, we realize measurement of the degeneracy and subtle broken-symmetry states of graphene at nanoscale for the first time. By using edge-free graphene quantum dots, we are able to measure valley splitting and valley-contrasting spin splitting of graphene at the single-electron level. Our experiments detect large valley splitting around atomic defects of graphene due to the coexistence of sublattice symmetry breaking and time reversal symmetry breaking. Large valley-contrasting spin splitting induced by enhanced spin-orbit coupling around the defects is also observed. These results reveal unexplored exotic electronic states in graphene at nanoscale induced by the atomic defects.
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