The dynamics of the mid-channel bars (MCBs) in the Three Gorges Reservoir (TGR) were substantially impacted by the large water-level changes due to the impoundments of the TGR. However, it is still not clear how the morphology of the MCBs changed under the influence of water level and hydrological regime changes induced by the impoundments and operation of the TGR. In this work, the MCBs in the TGR were retrieved using Landsat remote sensing images from 1989 to 2019, and the spatio-temporal variations in the number, area, morphology and location of the MCBs during different impoundment periods were investigated. The results showed that the number and area of MCBs changed dramatically with water-level changes, and the changes were dominated by MCBs with an area less than 0.03 km² and larger than 1 km². The area of MCBs decreased progressively with the rising water level, and the number generally showed a decreasing trend, with the minimum number occurring at the third stage when the water level reached 139 m, resulting in the maximum average area at this period. The ratio of length to width of the MCBs generally decreased with the changes in hydrological and sediment regimes, leading to a shape adjustment from narrow–long to relatively short–round with the rising of the water level. The water impoundments of the TGR led to the migration of the dominant area from the upper section to the middle section of the TGR and resulted in a more even distribution of MCBs in the TGR. The results improve our understanding of the mechanisms of the development of MCBs in the TGR under the influence of water impoundment coupled with the annually cyclic hydrological regime and longer periods of inundation and exposure.