Atomic force microscopy allows for the acquisition of depth-resolved nanomechanical properties deduced from maps of force–distance curves recorded on cancerous epithelial breast cells.
In the atomic force microscopy (AFM) investigation of soft polymers and liquids, the tip-sample interaction is dominated by long-range van der Waals forces, capillary forces and adhesion. Furthermore, the tip can indent several tens of nanometres into the surface, and it can pull off a polymer filament from the surface. Therefore, measuring the unperturbed shape of a polymeric fluid can be challenging. Here, we study the tip-sample interaction with polystyrene droplets swollen in chloroform vapour, where we can utilize the solvent vapour concentration to adjust the specimen's mechanical properties from a stiff solid to a fluid film. With the same AFM tip, we use two different AFM force spectroscopy methods to measure three-dimensional (3D) depth profiles of the tip-sample interaction: force-distance (FD) curves and amplitude-phase-distance (APD) curves. The 3D depth profiles reconstructed from FD and APD measurements provide detailed insight into the tip-sample interaction mechanism for a fluid polymer solution. The fluid's intrinsic relaxation time, which we measure with an AFM-based step-strain experiment, is essential for understanding the tip-sample interaction mechanism. Furthermore, measuring 3D depth profiles and using APD data to reconstruct the unperturbed surface comprise a versatile methodology for obtaining accurate dimensional measurements of fluid and gel-like objects on the nanometre scale.
Connective tissue displays a large compositional and structural complexity that involves multiple length scales. In particular, on the molecular and the nanometer level, the elementary processes that determine the biomechanics of collagen fibrils in connective tissues are still poorly understood. Here, we use atomic force microscopy (AFM) to determine the three-dimensional (3D) depth profiles of the local nanomechanical properties of collagen fibrils and their embedding interfibrillar matrix in native (unfixed), hydrated Achilles tendon of sheep and chickens. AFM imaging in air with controlled humidity preserves the tissue’s water content, allowing the assembly of collagen fibrils to be imaged in high resolution beneath an approximately 5–10 nm thick layer of the fluid components of the interfibrillar matrix. We collect pointwise force–distance (FD) data and amplitude–phase–distance (APD) data, from which we construct 3D depth profiles of the local tip–sample interaction forces. The 3D images reveal the nanomechanical morphology of unfixed, hydrated collagen fibrils in native tendon with a 0.1 nm depth resolution and a 10 nm lateral resolution. We observe a diversity in the nanomechanical properties among individual collagen fibrils in their adhesive and in their repulsive, viscoelastic mechanical response as well as among the contact points between adjacent collagen fibrils. This sheds new light on the role of interfibrillar bonds and the mechanical properties of the interfibrillar matrix in the biomechanics of tendon.
We investigate the surface and subsurface morphology of stearylamine-modified graphene oxide sheets and polystyrene-grafted functionalized graphene oxide sheets through atomic force microscopy (AFM) operated in multi-set point intermittent contact (MUSIC) mode. This allows a depth-resolved mapping of the nanomechanical properties of the top surface layer of the functionalized graphene oxide sheets. On the surface of stearylamine-functionalized graphene oxide sheets, we can distinguish areas of hydrophilic graphene oxide from hydrophobic areas functionalized with stearylamine. We find that every sheet of graphene oxide is functionalized with stearylamine on both sides of the sheet. The exposure of polystyrene-grafted functionalized graphene oxide to chloroform vapor during the AFM measurement causes a selective swelling and a softening of the polystyrene envelope. The depth-resolved mapping of the tip−sample interaction allows the shape of the folded and wrinkled graphene oxide sheets within the polystyrene envelope to be imaged; furthermore, it allows the thickness of the swollen polystyrene envelope to be measured. This yields the swelling degree, the grafting density, and the average chain conformation of the grafted polystyrene chains, which we find to be in the brush regime. Our work demonstrates a versatile methodology for imaging and characterizing functionalized and polymer-grafted two-dimensional materials on the nanometer scale.
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