Local fatigue experiments on microscale testing geometries offer the opportunity to isolate microstructure contributions to mechanical deformation behavior. In contrast to macroscopic fatigue testing, it is therefore possible to characterize the effect of individual features of interest independently. In this study Accumulative Roll Bonding (ARB) processed copper sheets with a bimodal microstructure were analyzed. Micropillars were milled by FIB structuring, producing testing geometries in individual layers of the material. Due to the bimodal microstructure, the different microsamples in each layer show varying grain sizes, which results in a change of the respective fatigue properties. Additionally micropillars at the interface were prepared to study interface contributions on the fatigue behavior. The investigations were executed by a novel approach that combines dynamic nanoindentation and micropillar compression. With this technique the high cycle fatigue range is easily accessible for microscale samples. Observation of the underlying deformation processes was performed by recording SEM micrographs of the deformed samples. Continuative investigations were realized by FIB milling of the deformed samples to create a cross-sectional view of the deformed microstructure.