The aim of this study was to develop a system to classify the marbling of beef using the hyperspectral imaging technology. The Japanese standard classification of the degree of marbling of beef was used as reference and twelve standards were digitized to obtain the parameters of shape and spatial distribution of marbling of each class. A total of 35 samples M. longissmus dorsi muscle were scanned by the hyperspectral imaging system of 400–1000 nm in reflectance mode. The wavelength of 528 nm was selected to segment the sample and the background, and 440 nm was used for classified the samples. Processing algorithms on image, based on decision tree method, were used in the region of interest obtaining a classification error of 0.08% in the building stage. The results showed that the proposed technique has a great potential, as a non-destructive and fast technique, that can be used to classify beef with respect to the degree of marbling.