Nanomechanical characterization of soft nanomaterial using atomic force microscopy

Atomic force microscopy (AFM) is a promising method for generating high-spatial-resolution images, providing insightful perspectives on the nanomechanical attributes of soft matter, including cells, bacteria, viruses, proteins, and nanoparticles. AFM is widely used in biological and pharmaceutical sciences because it can scrutinize mechanical properties under physiological conditions. We comprehensively reviewed experimental techniques and fundamental mathematical models to investigate the mechanical properties, including elastic moduli and binding forces, of soft materials. To determine these mechanical properties, two-dimensional arrays of force-distance (f-d) curves are obtained through AFM indentation experiments using the force volume technique. For elasticity determination, models are divided into approach f-d curve-based models, represented by the Hertz model, and retract f-d curve-based models, exemplified by the Johnson-Kendall-Roberts and Derjaguin-M & uuml;ller-Toporov models. Especially, the Chen, Tu, and Cappella models, developed from the Hertz model, are used for thin samples on hard substrates. Additionally, the establishment of physical or chemical bonds during indentation experiments, observable in retract f-d curves, is crucial for the adhesive properties of samples and binding affinity between antibodies (receptors) and antigens (ligands). Chemical force microscopy, single-molecule force spectroscopy, and singlecell force spectroscopy are primary AFM methods that provide a comprehensive view of such properties through retract curve analysis. Furthermore, this paper, structured into key thematic sections, also reviews the exemplary application of AFM across multiple scientific disciplines. Notably, cancer cells are softer than healthy cells, although more sophisticated investigations are required for prognostic applications. AFM also investigates how bacteria adapt to antibiotics, addressing antimicrobial resistance, and reveals that stiffer virus capsids indicate reduced infectivity, aiding in the development of new strategies to combat viral infections. Moreover, AFM paves the way for innovative therapeutic approaches in designing effective drug delivery systems by providing insights into the physical properties of soft nanoparticles and the binding affinity of target moieties. Our review provides researchers with representative studies applying AFM to a wide range of cross-disciplinary research.