Grain refinement is one of the methods applied to strengthen metallic materials, and various peculiar mechanical properties have been reported to be expressed when the grain size is reduced to less than submicron dimensions. This is considered to be due to a change in the behavior of dislocations that are associated with plastic deformation. In situ synchrotron radiation measurements of microstructural changes during deformation in face-centered cubic (fcc) metals with grain sizes of 20 μm to 5 nm were performed to systematically investigate the effects of grain size on dislocation behavior during plastic deformation. In pure aluminum with grain sizes of 20 to 3 μm, the dislocation density during plastic deformation was approximately 1014 m−2, regardless of the grain size. However, when the grain size was less than 3 μm, the dislocation density increased monotonically in proportion to the grain size to the power of -1. Furthermore, in a nickel alloy with a grain size of less than 10 nm, this relationship was no longer satisfied, and the results suggested that deformation progresses due to partial dislocations. In materials with a grain size of less than 1 μm, the dislocation density after unloading became much smaller than that during loading.