The objective of this study was to determine the potential of multispectral imaging (MSI) data recorded in the visible and near infrared electromagnetic regions to predict the structural features of intramuscular connective tissue, the proportion of intramuscular fat (IMF), and some characteristic parameters of muscle fibers involved in beef sensory quality. In order to do this, samples from three muscles (Longissimus thoracis, Semimembranosus and Biceps femoris) of animals belonging to three breeds (Aberdeen Angus, Limousine, and Blonde d’Aquitaine) were used (120 samples). After the acquisition of images by MSI and segmentation of their morphological parameters, a back propagation artificial neural network (ANN) model coupled with partial least squares was applied to predict the muscular parameters cited above. The results presented a high accuracy and are promising (R2 test > 0.90) for practical applications. For example, considering the prediction of IMF, the regression model giving the best ANN model exhibited R2P = 0.99 and RMSEP = 0.103 g × 100 g−1 DM.