AbstractSingle crystal perovskites have garnered significant attention as potential replacements for existing absorber layer materials. Despite the extensive investigations on their photoinduced charge‐carriers dynamics, most of the time‐resolved techniques focus on bulk properties, neglecting surface characteristic which plays a crucial role for their optoelectronic performance. Herein, 4D ultrafast scanning electron microscopy (4D‐USEM) is utilized to probing the photogenerated carrier transport at the first few nanometers, alongside density functional theory (DFT) to track both defect centers and ions migration. Two compositions of mixed cation are investigated: FA_0.6MA_0.4PbI₃ and FA_0.4MA_0.6PbI₃, interestingly, the former displays a longer lifetime compared to the latter due the presence of a higher surface‐defect centers. DFT calculations fully support that revealing samples with higher FA content have a lower energy barrier for iodide ions to migrate from the bulk to top layer, assisting in passivating surface vacancies, and a higher energy diffusion barrier to escape from surface to vacuum, resulting in fewer vacancies and longer‐lived hole–electron pairs. These findings manifest the influence of cation selection on charge carrier transport and formation of defects, and emphasize the importance of understanding ion migrations role in controlling surface vacancies to assist engineering high‐performance optoelectronic devices based on single crystal perovskites.