The dynamic evolution of microbubbles under ultrasonic excitation is fundamental to applications ranging from targeted drug delivery to acoustic cleaning. This study employs a synchronous high-speed microscopic imaging system to systematically investigate the size-dependent stability and fragmentation of air microbubbles (R0 = 25–82.5 μm) in a free field at a driving frequency of 16.6 kHz. Our results demonstrate a clear mechanistic transition from stable radial oscillations to complex surface instabilities and, eventually, deterministic fragmentation. Smaller bubbles (R0 < 55 μm) exhibit long-term stability, transitioning through higher-order surface modes (n = 3 to n = 4) as surface energy accumulates. In contrast, larger bubbles (R0 > 60 μm) undergo violent non-spherical deformations characterized by centripetal necking and high-speed micro-jetting. Notably, we identify an inverse relationship between initial radius and fragmentation onset time, with larger bubbles reaching instability thresholds significantly earlier. Furthermore, a transition from stochastic breakup to bimodal, volume-symmetric splitting was observed as R0 increased, where daughter bubbles reached comparable volumes. These findings provide a theoretical and empirical basis for the controlled generation of monodisperse microbubble clouds, offering significant potential for enhancing the efficacy of ultrasonic contrast agents and therapeutic cavitation.