To address the problems of quality deterioration and taste decline associated with traditional high-temperature processing of ready-to-eat chicken liver, this study employed sous-vide technology to improve product quality. The effects of processing parameters on the quality characteristics and microstructure of chicken liver were systematically investigated. Particularly, the influence of different sous-vide durations (40, 45, 50, 55, and 60 min) at 80 °C on various quality indicators, including cooking loss, moisture content, water activity, color, pH, shear force, texture characteristics, electronic nose analysis, secondary structure, microstructure, and sensory attributes was examined. The results demonstrated that with increasing cooking time, the cooking loss and pH of chicken liver samples in the sous-vide group increased significantly, whereas the moisture content, water activity, and L^* and a^* values decreased significantly (P<0.05). The tenderness of the chicken liver initially decreased and then increased, whereas texture characteristics and sensory scores first increased and then decreased with extended processing time (P<0.05). Compared with the control group, the optimal processing time was determined to be 55 min, yielding the best overall quality. Fourier-transform infrared (FTIR) spectroscopy revealed that the secondary protein structure shifted from α-helix to β-sheet with longer processing times (P<0.05). Furthermore, an appropriate sous-vide duration (55 min) promoted a more orderly and compact reticular microstructure in the chicken liver. In conclusion, the optimal sous-vide process for ready-to-eat chicken liver, at 80 °C for 55 min. This paper provides certain technical support for the industrial production and quality improvement of low-temperature ready-to-eat chicken liver, and provides a new direction for the comprehensive utilization of livestock and poultry by-products.