PeakForce Tapping Atomic Force Microscopy Research Articles

Direct Observation of Contact Electrification Effects at Nanoscale Using Scanning Probe Microscopy

AbstractIn the last decade, contact electrification has gained attention for its potential in energy harvesting, addressing fundamental physics. Scanning probe microscopy (SPM) has evolved as a powerful platform, enabling the in situ characterization/manipulation of a sample's tribological/electrical properties at nanoscale. However, although both the sliding and tapping modes are available in the energy harvesting, the lateral sliding using contact mode SPM has predominantly been employed in triboelectric study. In this work, contact electrification on polydimethylsiloxane is investigated, using peak force tapping atomic force microscopy (PF‐AFM). As PF‐AFM quasi‐discretely offers vertical tapping motions of the probe with a regulated force amplitude, resembling a vertical‐type triboelectric nanogenerator, while minimizing lateral forces. The subsequent surface potential measurements reveal that the generated tribocharge is influenced by both the effective work function difference and energy dissipation at the interface. Furthermore, the accumulation of transferred charges is explored by measuring tip‐sample current during the PF‐AFM operation, showing the comparable results with the surface potential measurement. The results can be attributed to the contact potential difference assisted by the energy dissipation at the interface. This study offers an advanced opportunity to understand and study charge generation behavior based on surface properties without damaging the sample.

Boxcar Averaging Scanning Nonlinear Dielectric Microscopy.

Scanning nonlinear dielectric microscopy (SNDM) is a near-field microwave-based scanning probe microscopy method with a wide variety of applications, especially in the fields of dielectrics and semiconductors. This microscopy method has often been combined with contact-mode atomic force microscopy (AFM) for simultaneous topography imaging and contact force regulation. The combination SNDM with intermittent contact AFM is also beneficial for imaging a sample prone to damage and using a sharp microscopy tip for improving spatial resolution. However, SNDM with intermittent contact AFM can suffer from a lower signal-to-noise (S/N) ratio than that with contact-mode AFM because of the shorter contact time for a given measurement time. In order to improve the S/N ratio, we apply boxcar averaging based signal acquisition suitable for SNDM with intermittent contact AFM. We develop a theory for the S/N ratio of SNDM and experimentally demonstrate the enhancement of the S/N ratio in SNDM combined with peak-force tapping (a trademark of Bruker) AFM. In addition, we apply the proposed method to the carrier concentration distribution imaging of atomically thin van der Waals semiconductors. The proposed method clearly visualizes an anomalous electron doping effect on few-layer Nb-doped MoS2. The proposed method is also applicable to other scanning near-field microwave microscopes combined with peak-force tapping AFM such as scanning microwave impedance microscopy. Our results indicate the possibility of simultaneous nanoscale topographic, electrical, and mechanical imaging even on delicate samples.

Elastic modulus mapping for bovine cortical bone from submillimeter- to submicron-scales using PeakForce Tapping atomic force microscopy

Bone is a composite material consisting of organic and inorganic components that are organized into hierarchical structures to provide load-bearing functions. This paper presents the results of PeakForce Tapping atomic force microscopy (AFM) scans on cut and polished bovine cortical bone specimens that were submerged in water. The elastic modulus map and surface morphology were obtained for various bone hierarchical structures from submillimeter- to submicron-scales. The elastic modulus of osteons (20.51 ± 6.85 GPa) was slightly lower than the interstitial bone (21.87 ± 5.48 GPa); they were both much greater than that of the cement lines (7.49 ± 4.23 GPa). The elastic modulus in the lamella structures varied periodically from higher values in thick sub-lamellae (21.49 ± 6.58 GPa) to the lower values in thin sub-lamellae (9.67 ± 2.69 GPa). The results also show relatively softer mineralized collagen fibril bundle arrays (12.94 ± 2.71 GPa) embedded in harder matrix materials (28.39 ± 5.75 GPa). The variations in the elastic modulus suggest different degrees of mineralization or different fibril orientations. The histograms of elastic modulus indicate the dominating compositions or dominating fibril orientations.

Surface properties of thermally treated European beech wood studied by PeakForce Tapping atomic force microscopy and Fourier-transform infrared spectroscopy

AbstractNatural constituents of wood cell-wall layers are affected in various ways by thermal treatment. This study investigated the effect of high-temperature treatment on the properties of cell-wall layers. The properties were studied using PeakForce quantitative nanomechanical mapping and Fourier-transform infrared spectroscopy (FTIR). European beech wood was thermally treated at 200 °C for 1, 3, and 5 h in an oxidizing atmosphere. Modulus of elasticity, adhesion force, and roughness of the secondary S2layer and the compound middle lamella (CML) were determined using atomic force microscopy (AFM). Results showed that both the S2layer and CML were affected by thermal treatment. Stiffening of the S2layer was caused by increased crystallinity of the cellulose-dominated component, having peaked after 1 h of treatment. The degradation thereafter resulted in a decrease of the S2as well as the CML stiffness. An increase of CML roughness after 3 h of treatment was associated with the effect of thermal degradation on CML integrity. The analysis suggested that the reduction in syringyl lignin is potentially associated with an increase in adhesion of cell-wall layers.

Single molecule analysis reveals monomeric XPA bends DNA and undergoes episodic linear diffusion during damage search

Nucleotide excision repair (NER) removes a wide range of DNA lesions, including UV-induced photoproducts and bulky base adducts. XPA is an essential protein in eukaryotic NER, although reports about its stoichiometry and role in damage recognition are controversial. Here, by PeakForce Tapping atomic force microscopy, we show that human XPA binds and bends DNA by ∼60° as a monomer. Furthermore, we observe XPA specificity for the helix-distorting base adduct N-(2’-deoxyguanosin-8-yl)-2-acetylaminofluorene over non-damaged dsDNA. Moreover, single molecule fluorescence microscopy reveals that DNA-bound XPA exhibits multiple modes of linear diffusion between paused phases. The presence of DNA damage increases the frequency of pausing. Truncated XPA, lacking the intrinsically disordered N- and C-termini, loses specificity for DNA lesions and shows less pausing on damaged DNA. Our data are consistent with a working model in which monomeric XPA bends DNA, displays episodic phases of linear diffusion along DNA, and pauses in response to DNA damage.

Adhesion Effect on the Hyperfine Frequency Shift of an Alkali Metal Vapor Cell with Paraffin Coating Using Peak-Force Tapping AFM

We have investigated the adhesion effect on the hyperfine frequency shift of an alkali metal vapor cell with paraffin coating using the peak-force tapping AFM (atomic force microscopy) technique by developing a uniform and high-quality paraffin coating method. We observed a relatively uniform temperature field on the substrate can be obtained theoretically and experimentally with the closed-type previse temperature-controlled evaporation method. The roughness and adhesion of the coating surface as low as 0.8 nm and 20 pN were successfully obtained, respectively. Furthermore, the adhesion information dependence of the topography was investigated from the force spectroscopy, which indicates that the adhesion force jumped on the edge of the particles and stepped but remained constant above the particles and steps regardless of their height for paraffin coating. Finally, we can evaluate the relaxation and the hyperfine frequency shift of an alkali metal vapor cell through accurately calculating the surface adsorption energy of the paraffin coating from peak-force tapping information. This finding is crucial for improving the sensitivity of the atomic sensors through directly analyzing the adhesion effect of the paraffin coating films instead of measuring the relaxation times.

Atomic force microscopy of adsorbed proteoglycan mimetic nanoparticles: Toward new glycocalyx-mimetic model surfaces

Blood vessels present a dense, non-uniform, polysaccharide-rich layer, called the endothelial glycocalyx. The polysaccharides in the glycocalyx include polyanionic glycosaminoglycans (GAGs). This polysaccharide-rich surface has excellent and unique blood compatibility. We report new methods for preparing and characterizing dense GAG surfaces that can serve as models of the vascular endothelial glycocalyx. The GAG-rich surfaces are prepared by adsorbing heparin or chondroitin sulfate-containing polyelectrolyte complex nanoparticles (PCNs) to chitosan-hyaluronan polyelectrolyte multilayers (PEMs). The surfaces are characterized by PeakForce tapping atomic force microscopy, both in air and in aqueous pH 7.4 buffer, and by PeakForce quantitative nanomechanics (PF-QNM) mode with high spatial resolution. These new surfaces provide access to heparin-rich or chondroitin sulfate-rich coatings that mimic both composition and nanoscale structural features of the vascular endothelial glycocalyx.

Nematode Epicuticle Visualisation by PeakForce Tapping Atomic Force Microscopy.

The free-living soil nematode Caenorhabditis elegans has become an iconic experimental model animal in biology. This transparent animal can be easily imaged using optical microscopy to visualise its organs, tissues, single cells and subcellular events. The epicuticle of C. elegans nematodes has been studied at nanoscale using transmission and scanning (SEM) electron microscopies. As a result, imaging artefacts can appear due to embedding the worms into resins or coating the worms with a conductive gold layer. In addition, fixation and contrasting may also damage the cuticle. Conventional tapping mode atomic force microscopy (AFM) can be applied to image the cuticle of the dried nematodes in air, however this approach also suffers from imaging defects. Ideally, the nematodes should be imaged under conditions resembling their natural environment. Recently, we reported the use of PeakForce Tapping AFM mode for the successful visualisation and numerical characterisation of C. elegans nematode cuticle both in air and in liquid ( Fakhrullina et al., 2017 ). We imaged the principal nematode surface structures and characterised mechanical properties of the cuticle. This protocol provides the detailed description of AFM imaging of liquid-immersed C. elegans nematodes using PeakForce Tapping atomic force microscopy.

Nanomechanical properties of composite protein networks of erythroid membranes at lipid surfaces

Erythrocyte membranes have been particularly useful as a model for studies of membrane structure and mechanics. Native erythroid membranes can be electroformed as giant unilamellar vesicles (eGUVs). In the presence of ATP, the erythroid membrane proteins of eGUVs rearrange into protein networks at the microscale. Here, we present a detailed nanomechanical study of individual protein microfilaments forming the protein networks of eGUVs when spread on supporting surfaces. Using Peak Force tapping Atomic Force Microscopy (PF-AFM) in liquid environment we have obtained the mechanical maps of the composite lipid-protein networks supported on solid surface. In the absence of ATP, the protein pool was characterized by a Young’s Modulus Epool≈5–15MPa whereas the complex filaments were found softer after protein supramolecular rearrangement; Efil≈0.4MPa. The observed protein softening and reassembling could be relevant for understanding the mechanisms of cytoskeleton reorganization found in pathological erythrocytes or erythrocytes that are affected by biological agents.

Epoxy-acrylic core-shell particles by seeded emulsion polymerization

We developed a novel method for synthesizing epoxy-acrylic hybrid latexes. We first prepared an aqueous dispersion of high molecular weight solid epoxy prepolymers using a mechanical dispersion process at elevated temperatures, and we subsequently used the epoxy dispersion as a seed in the emulsion polymerization of acrylic monomers comprising methyl methacrylate (MMA) and methacrylic acid (MAA). Advanced analytical techniques, such as scanning transmission X-ray microscopy (STXM) and peak force tapping atomic force microscopy (PFT-AFM), have elucidated a unique core-shell morphology of the epoxy-acrylic hybrid particles. Moreover, the formation of the core-shell morphology in the seeded emulsion polymerization process is primarily attributed to kinetic trapping of the acrylic phase at the exterior of the epoxy particles. By this new method, we are able to design the epoxy and acrylic polymers in two separate steps, and we can potentially synthesize epoxy-acrylic hybrid latexes with a broad range of compositions.

Controlled immobilization of His-tagged proteins for protein-ligand interaction experiments using Ni²⁺-NTA layer on glass surfaces.

Gold surfaces functionalized with nickel-nitrilotriacetic acid (Ni²⁺-NTA) as self-assembled monolayers (SAM) to immobilize histidine (His)-tagged biomolecules are broadly reported in the literature. However, the increasing demand of using microfluidic systems and biosensors takes more and more advantage on silicon technology which provides dedicated glass surfaces and substantially allows cost and resource savings. Here we present a novel method for the controlled oriented immobilization of His-tagged proteins on glass surfaces functionalized with a Ni²⁺-NTA layer. Exemplarily, the protein pattern morphology after immobilization on the Ni²⁺-NTA layer of His6-tagged soluble receptor for advanced glycation endproducts (sRAGE) was investigated and compared to non-oriented immobilization of sRAGE on amino SAM by using scanning electron microscopy (SEM). Moreover, we demonstrated interaction of immobilized sRAGE with three structurally different ligands, S100A12, S100A4, and glycated low density lipoproteins (glycLDL), by means of peak-force tapping atomic force microscopy (PF-AFM). We showed a clear discrimination of different protein-ligand orientations by differential height measurements.

Investigation into local cell mechanics by atomic force microscopy mapping and optical tweezer vertical indentation

Investigating the mechanical properties of cells could reveal a potential source of label-free markers of cancer progression, based on measurable viscoelastic parameters. The Young’s modulus has proved to be the most thoroughly studied so far, however, even for the same cell type, the elastic modulus reported in different studies spans a wide range of values, mainly due to the application of different experimental conditions. This complicates the reliable use of elasticity for the mechanical phenotyping of cells. Here we combine two complementary techniques, atomic force microscopy (AFM) and optical tweezer microscopy (OTM), providing a comprehensive mechanical comparison of three human breast cell lines: normal myoepithelial (HBL-100), luminal breast cancer (MCF-7) and basal breast cancer (MDA-MB-231) cells. The elastic modulus was measured locally by AFM and OTM on single cells, using similar indentation approaches but different measurement parameters. Peak force tapping AFM was employed at nanonewton forces and high loading rates to draw a viscoelastic map of each cell and the results indicated that the region on top of the nucleus provided the most meaningful results. OTM was employed at those locations at piconewton forces and low loading rates, to measure the elastic modulus in a real elastic regime and rule out the contribution of viscous forces typical of AFM. When measured by either AFM or OTM, the cell lines’ elasticity trend was similar for the aggressive MDA-MB-231 cells, which were found to be significantly softer than the other two cell types in both measurements. However, when comparing HBL-100 and MCF-7 cells, we found significant differences only when using OTM.

Observation of elastic modulus inhomogeneities in thermosetting epoxies using AFM – Discerning facts and artifacts

Observations of a nanometer-scale nodular morphology on differently prepared surfaces of thermosets have frequently been interpreted as a sign for an inhomogeneous molecular network, which would result in an inhomogeneous modulus distribution within those thermosets. In order to test this hypothesis, the Peak-Force Tapping atomic force microscopy (AFM) mode was used on fracture surfaces and ultramicrotome cuts of epoxy and other polymers using differently sharp AFM probes.The nodular morphology is quite likely caused by an AFM artifact, which also seems to cause an apparently inhomogeneous modulus distribution; a variation in the tip–sample contact area could explain this effect. Smooth surfaces are necessary in order to reduce the contribution from this artifact. Ultramicrotome cutting currently seems to be the most appropriate surface preparation technique for the measurement of modulus distribution at the nanometer scale.All investigated materials seem to be homogeneous on a scale on the order of 10 nm–1000 nm. If modulus inhomogeneities are present, their amount or their lateral size is too small to be unambiguously measurable with this technique. From this data, it seems unlikely that epoxy exhibits an inhomogeneous molecular network.

New understanding of microstructure formation of the rubber phase in thermoplastic vulcanizates (TPV)

The breakup of the rubber phase in an ethylene-propylene-diene monomer (EPDM)/polypropylene (PP) blend at the early stage of dynamic vulcanization is similar to that in an unvulcanized EPDM/PP blend because of the low crosslink density of the EPDM phase. In this work, the minimum size of the rubber phase in the unvulcanized EPDM/PP blend was first calculated by using the critical breakup law of viscoelastic droplets in a matrix. The calculated results showed that the minimum size of the rubber phase in the unvulcanized blend was in the nanometer scale (25-46 nm), not the micrometer scale as reported in many works. Meanwhile, the actual size of the rubber phase in the thermoplastic vulcanizate (TPV) at both the early stage and the final stage of dynamic vulcanization was observed by using peak force tapping atomic force microscopy (PF-AFM). The results indicated that the EPDM phase indeed broke up into nanoparticles at the early stage of dynamic vulcanization, in good agreement with the calculated results. More interestingly, we first revealed that the micrometer-sized rubber particles commonly observed in TPV were actually the agglomerates of rubber nanoparticles with diameters between 40 and 60 nm. The mechanism for the formation of rubber nanoparticles and their agglomerates during dynamic vulcanization was then discussed. Our work provides guidance to control the microstructure of the rubber phase in TPV to prepare high performance TPV products for a wide range of applications in the automobile and electronic industries.

Comparison of the physical characteristics of chlorosomes from three different phyla of green phototrophic bacteria

Chlorosomes, the major antenna complexes in green sulphur bacteria, filamentous anoxygenic phototrophs, and phototrophic acidobacteria, are attached to the cytoplasmic side of the inner cell membrane and contain thousands of bacteriochlorophyll (BChl) molecules that harvest light and channel the energy to membrane-bound reaction centres. Chlorosomes from phototrophs representing three different phyla, Chloroflexus (Cfx.) aurantiacus, Chlorobaculum (Cba.) tepidum and the newly discovered “Candidatus (Ca.) Chloracidobacterium (Cab.) thermophilum” were analysed using PeakForce Tapping atomic force microscopy (PFT-AFM). Gentle PFT-AFM imaging in buffered solutions that maintained the chlorosomes in a near-native state revealed ellipsoids of variable size, with surface bumps and undulations that differ between individual chlorosomes. Cba. tepidum chlorosomes were the largest (133×57×36nm; 141,000nm3 volume), compared with chlorosomes from Cfx. aurantiacus (120×44×30nm; 84,000nm3) and Ca. Cab. thermophilum (99×40×31nm; 65,000nm3). Reflecting the contributions of thousands of pigment–pigment stacking interactions to the stability of these supramolecular assemblies, analysis by nanomechanical mapping shows that chlorosomes are highly stable and that their integrity is disrupted only by very strong forces of 1000–2000pN. AFM topographs of Ca. Cab. thermophilum chlorosomes that had retained their attachment to the cytoplasmic membrane showed that this membrane dynamically changes shape and is composed of protrusions of up to 30nm wide and 6nm above the mica support, possibly representing different protein domains. Spectral imaging revealed significant heterogeneity in the fluorescence emission of individual chlorosomes, likely reflecting the variations in BChl c homolog composition and internal arrangements of the stacked BChls within each chlorosome.

On the use of peak-force tapping atomic force microscopy for quantification of the local elastic modulus in hardened cement paste

A surface of epoxy-impregnated hardened cement paste was investigated using a novel atomic force microscopy (AFM) imaging mode that allows for the quantitative mapping of the local elastic modulus. The analyzed surface was previously prepared using focussed ion beam milling. The same surface was also characterized by electron microscopy and energy-dispersive X-ray spectroscopy. We demonstrate the capability of this quantitative nanomechanical mapping to provide information on the local distribution of the elastic modulus (from about 1 to about 100 GPa) with a spatial resolution in the range of decananometers, that corresponds to that of low-keV back-scattered electron imaging. Despite some surface roughness which affects the measured nanomechanical properties it is shown that topography, adhesion and Young's modulus can be clearly distinguished. The quantitative mapping of the local elastic modulus is able to discriminate between phases in the cement paste microstructure that cannot be distinguished from the corresponding back-scattered electron images.