Siberian Federal University

The cell mechanical assay has emerged as a powerful approach to studying cellular behav- ior and protein dynamics. This work presents the novel protocol that combines cell nanomechanical assay with rapid protein expression profiling, enabling comprehensive insight into cellular responses. The new protocol leverages advanced techniques in atomic force microscopy (AFM) to measure the me- chanical properties of individual cells, while simultaneously utilizing a laser scanning microscopy for the high-throughput quantification of protein expression levels. This dual-assay method allows researchers to elucidate the relationship between cellular mechanical properties and protein dynamics, uncovering critical insights into cellular physiology and pathophysiology. The effectiveness of the protocol was vali- dated through experiments with cancer cells, showcasing its potential in colocalization of wnt3a ligand molecule and actin cytoskeleton with Young’s modulus patterns of the cell. Our findings indicate that this integrated approach not only enhances the accuracy of cellular assessments but also accelerates the understanding of cellular mechanisms at the nanoscale. This protocol holds promise for applications in drug development, diagnostics, and personalized medicine, offering a new lens through which we can explore the intricate interplay between cellular mechanics and protein expression