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In this paper, we report the successful combination of macroscopic uniaxial tensile testing of bulk specimen combined with In situ dislocation-scale observations of the evolution of deformation microstructures during loading at several stress states. The dislocation-scale observations were performed by Accurate Electron Channeling Contrast Imaging in order to follow the defects evolution and their interactions with grain boundaries for several regions of interest during macroscopic loading. With this novel in situ procedure, the slip systems governing the deformation in polycrystalline bulk β-Ti21S are tracked during the macroscopic uniaxial tensile test. For instance, curved slip lines that are associated with “pencil glide” phenomenon and tangled dislocation networks are evidenced.