Machine learning classification of normal and malignant cells on the basis of their viscoelastic properties

• Multifrequency AFM viscoelastic spectra classify prostate cells. • 19 mechanistic features incl. fT enable ML on small cohorts. • Fuzzy LAMDA achieved ∼75–79% accuracy with 40 cells/class. • Cancer cells show lower fT and higher loss tangent across frequencies. • Sensitivity prioritized; cell-level detection reached 79–81%. Cell mechanics, elasticity and viscoelasticity, are key markers of biological states like cancer. Atomic force microscopy (AFM) is ideal for such studies, but its low throughput limits large-scale use. Two solutions exist: automation for higher throughput, or high-density measurements for richer data. The latter enables machine learning (ML)-based classification, with viscoelastic parameters offering unique insights beyond static measures like Young’s modulus. This study used dynamic mechanical analysis (DMA) to classify cells, focusing on viscoelastic descriptors (storage/loss moduli) across frequencies. Normal (RWPE-1) and grade IV cancerous (PC3-GFP) prostate cells were probed at 1–200 Hz, generating 304 features per cell. The fuzzy logic-based LAMDA algorithm, trained on 19 selected features, classified cells using 40 samples per line. PC3-GFP cells showed higher deformability and heterogeneity, behaving more like viscous fluids at low frequencies. The model achieved 79% classification accuracy. Adding features improved performance, suggesting fewer training samples may suffice with rich datasets. A sensitivity-optimized threshold reduced false negatives in cancer detection. Combining viscoelastic analysis with ML effectively discriminates normal and malignant cells. Future work could refine training and integrate new features, though acquisition time remains a challenge. This approach offers a promising framework for mechanome-based diagnostics, with applications in cancer and stem cell research.