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Here, we report a comprehensive study that combines in situ scanning electron microscopy experiments and atomistic simulations to quantify the effect of crystal size on the transformation in deformation modes in a-axis oriented Mg single crystals at room temperature. The experimental results indicate that the deformation is dominated by the nucleation and propagation of tensile twins. The stress required for twin propagation was found to increase with decreasing sample size, showing a typical “smaller is stronger” behavior. Furthermore, an anomalous increase in strain hardening is first reported for microcrystals having diameters larger than ∼18 μm, which is induced by twin-twin and dislocation-twin interactions. The hardening rate gradually decreases toward the bulk response as the microcrystal size increases. Below 18 μm, deformation is dominated by the nucleation and propagation of a single tensile twin …