Heatmaps of texture & roughness parameters and superficial & spatial features processed by mathematical statistics and machine learning models for different MCTS categories. A, unpaired t-student statistics with p-value <0.05 of all features for the selection of effective features in the cross-statistical algorithm. The X-axis shows 2484 superficial and spatial features from 27 parameters. The specific feature orders and names are listed in Fig. S5 (order is from left to right) and Data File 1 (Ref. [68]). The Y-axis shows 34 groups of MCTS comparisons in cell number, cell type, and different drug treatment categories. The specific comparison among different groups in different categories is listed in Fig. S5. B, the accuracy of supervised models for MCTS classifications by superficial and spatial parameters. C, the accuracy of unsupervised models for MCTS classifications by superficial and spatial parameters. Six supervised and six unsupervised models are used to classify nine MCTS categories by 27 texture and roughness parameters. The filtered heatmap (accuracy >0.5) in Fig. S6 illustrates the texture and roughness parameters contributing to the effective classification of MCTS categories across the 12 machine learning models. D, the weight/importance of features in superficial and spatial parameters for the cross-screening and composite-hyperparameter algorithms. The X-axis indicates the features within the parameters and the details are listed in Fig. S5 (order is from right to left). DT, decision tree model. GB, gradient boosting model. kNN, k-nearest neighbor model. LG, logistics model. NB, naïve bayes model. SVM, support vector machine model. KM, k-means model. BC, birch model. AH, agglomerative hierarchical model. MBK, mini batch k-means model. ST, spectral model. GM, Gaussian mixture model. O4, OVCAR-4 MCTS. O4_2-ME, OVCAR-4 MCTS treated by 2-ME with 1 µM, 10 µM, and 25 µM concentrations. O4_AZD, OVCAR-4 MCTS treated by AZD with 1 µM, 10 µM, and 25 µM concentrations. O4_R-ke, OVCAR-4 MCTS treated by R-ke with 1 µM, 10 µM, and 25 µM concentrations. O4_1 µM, OVCAR-4 MCTS treated with 1 µM 2-ME, AZD, and R-ke drugs. O4_10 µM, OVCAR-4 MCTS treated with 10 µM 2-ME, AZD, and R-ke drugs. O4_25 µM, OVCAR-4 MCTS treated with 25 µM 2-ME, AZD, and R-ke drugs. O8, OVCAR-8 MCTS with 5,000 and 50,000 cell numbers. Pan02H7, Panc02-H7 MCTS with different ratio mixtures of fibroblasts.