Kuroshio is an important western boundary current system in the North Pacific subtropical gyre. Mesoscale eddies play an important role in Kuroshio path variations, which significantly affect fisheries, marine navigation, and climate in regions along the Kuroshio path. However, the direct physical impacts of the Kuroshio variabilities on mixing dynamics within the water columns off the southern coast of Japan remain unclear due to the lack of observational records. The radiocarbon (Δ14C) in dissolved inorganic carbon (DIC) of seawater has been used as a reliable tracer of water circulation and mixing processes. Here, we present the high‐resolution dissolved inorganic radiocarbon (DIC ∆14C) measurements for water samples collected during the Kuroshio large‐meander (LM) period in March 2022 to observe the mixing dynamics within the Kuroshio area and elucidate the controls of mixing processes. In the present study, horizontal variations (12–175‰) in DIC Δ14C values were observed between 400 and 1,000 m and were attributed mainly to changes in the depth of isopycnal surfaces associated with mesoscale eddies and the position of the Kuroshio axis. Furthermore, fluctuations in DIC Δ14C values (10–25‰) were observed on the same isopycnal surfaces ( 25.5–27.0), suggesting the occurrence of diapycnal mixing. By comparing this newly obtained data with values previously reported at similar locations, variations in penetration depths of high bomb 14C signals from past observations are found to be mainly caused by changes in the depth of isopycnal surfaces associated with mesoscale eddies and Kuroshio path variations.