In this study, the estimation of ultrasound parameters is evaluated for in vivo differentiation of intraocular melanomas. For this purpose, both tissue and image parameters of the ultrasound signal are considered. These parameters comprised, respectively, the frequency dependent attenuation and backscattering coefficient of the melanoma tissue, and the first and second-order statistics of the amplitude-modulated and phase-derivative images of the melanomas. A diffraction correction procedure has been applied prior to the estimation of the parameters to correct the ultrasound signals for the echographic equipment used and for the various distances of the region-of-interest to the transducer. In addition, a pre-processing to select a homogeneous region from the tumours was implemented to obtain consistent estimates of the ultrasound parameters, because the accuracy and the precision of the parameters would be greatly reduced by the inhomogeneity of the melanoma tissue. The estimation methods are evaluated by means of the accuracy and precision of the parameters estimated from simulated ultrasound data and data obtained from a tissue-mimicking phantom. The mutual correlations of the parameters are discussed for the ultrasound data obtained from the melanomas. This study enabled a preselection of the independent ultrasound parameters that could be used in a discriminant analysis to perform a differentiation of intraocular melanomas. The sensitivity and specificity of differentiating spindle cell type from mixed-epitheloid meleanomas were 92 and 89 percent, respectively.