Partial and total splenectomies are associated with a high risk of substantial blood loss. Lasers operating at wavelengths strongly absorbed by water have the potential to improve hemostasis and cut while providing a narrow zone of thermal damage. The aim of this study is to compare a thulium-doped fiber laser (TDFL) emitting a wavelength of 1940 nm and a diode laser (DL) operating at 1470 nm for spleen surgery in a pig model. A partial splenectomy and spleen incisions were made in 12 animals using the two laser devices. The hemostasis was evaluated visually during surgeries. Post-mortem and histopathological evaluations were done on days 0, 7, and 14 following surgery. Neither TDFL nor DL caused bleeding on day 0 or delayed bleeding. On day 14, pale streaks at the site of incision were slightly wider after cutting with DL than with TDFL. Histological analysis revealed a carbonized zone with exudation and a deeper zone of thermal tissue damage on day 0. The width of the thermal changes was 655.26 ± 107.70 μm for TDFL and 1413.37 ± 111.85 μm for DL. On day 7, a proliferation of fibroblasts and splenocytes was visible, as well as a formation of multinucleated giant cells adjacent to the residues of carbonization. The zone of thermal damage was broader for DL (1157.5 ± 262.77 μm) than for TDFL (682.22 ± 116.58 μm). On day 14, cutting sites were filled with connective and granulation tissues with the residues of carbonization. The zone of thermal damage was narrower for TDFL (761.65 ± 34.3 μm) than for DL (1609.82 ± 202.22 μm). Thus, both lasers are efficient in spleen surgery, providing good hemostasis. However, TDFL produces a narrower zone of thermal damage, which suggests its better efficiency for spleen surgery, especially when performing more precise procedures.