Space-resolved quantitative mechanical measurements of soft and supersoft materials by atomic force microscopy

Atomic force microscopy (AFM) has proven to be a valuable instrument to characterize quantitatively the mechanical and morphological properties of soft materials. For medium and hard samples (E>1 MPa), the nanomechanical accuracy of AFM is well established and ascertained. However, for soft samples, the experimental setup and data analysis for AFM are not yet firmly established. A calibration obtained for homogeneous samples with a Young’s modulus ranging from 100 Pa to a few kPa will prove its usefulness for nanomechanical AFM investigations of soft biological specimens, such as living cells and extracellular matrices. For this purpose, poly(N-isopropylacrylamide) (PNIPAM) hydrogels were synthesized in different methanol − water mixtures to produce a series of homogeneous samples with finely tunable mechanical properties. These samples allowed the comparison and validation of AFM force spectroscopy results using macroscopic and rheological techniques. In AFM measurements, the geometry of the indenter is fundamental to the model used for data interpretation; therefore, experiments were carried out using spherical micrometric and standard pyramidal sharp probes. Moreover, a PNIPAM gel embedded with hard microspheres was analyzed, which showed the capability of AFM for measuring the local mechanical properties of heterogeneous samples.