Characterization of high-intensity focused ultrasound (HIFU) systems using ex vivo tissues is an important part of the preclinical testing for new HIFU devices. In ex vivo characterization, the lesion volume produced by the absorption of HIFU energy is quantified as operational parameters are varied. This paper examines the three methods used for lesion-volume quantification: histology, magnetic resonance (MR) imaging, and numerical calculations. The methods were studied in the context of a clinically relevant problem for HIFU procedures—that of quantifying the change in the lesion volume with changing sonication time. The lesion volumes of sonicated samples of porcine liver were determined using the three methods, at focal intensities ranging from to and sonication times between 20 s and 40 s. It was found that histology consistently yielded lower lesion volumes than the other two methods, and the calculated values were below magnetic resonance imaging (MRI) at high applied energies. Still, the three methods agreed with each other to within a difference for all of the experiments. Increasing the sonication time produced much larger changes in the lesion volume than increasing the acoustic intensity, for the same total energy expenditure, at lower energy (less than 1000 J) levels. At higher energy levels, (around 1500 J), increasing the sonication time and increasing the intensity produced roughly the same change in the lesion volume for the same total energy expenditure.