The thickness of the metal-transducer nano-film is an essential parameter for high-pressure time-domain-thermoreflectance (TDTR) measurements. In this article, an accurate method was proposed to determine the transducer thickness in high-pressure conditions using the pressure-volume equation of state combined with an image processing method. Both the elastic and plastic deformation of the sample squeezed in diamond anvil cells were considered in this method. High-pressure TDTR measurements on thermal conductivity of MgO and mica were further taken for comparing the influence from different thickness-characterization methods up to ∼18 GPa, and the proposed method accurately captured the effect of plastic deformation on thermal conductivity for the first time. This work can not only help achieve more accurate TDTR measurements under high-pressure but also provide valuable guidance for the diamond anvil cell application in nanoscale research.