The accurate detection of malignant tissue during colorectal surgery impacts operation outcome. The non-invasive spectral imaging combined with machine learning (ML) methods showed to be promising for tumor identification. However, large spectral range implies large computing time. To reduce the number of features, ML methods (e.g. logistic regression and convolutional neuronal network CNN) were evaluated based on four physiological tissue parameters to automatically classify cancer and healthy mucosa in resected colon tissue. A ROC AUC of 0.81 was achieved with the CNN. This study shows that the use of only specific wavelengths bands can detect cancer.Clinical Relevance— These outcomes support the possibility to automatically classify colon tumor based on physiological parameters calculated using only specific wavelength bands. Hence, future image-guided colorectal surgeries can be performed with real-time multispectral imaging.