Peak-force Atomic Force Microscopy Research Articles

Regulating Grafting Density to Realize High-Areal-Capacity Silicon Submicroparticle Anodes Under Ultralow Binder Content.

Grafted biopolymer binders are demonstrated to improve the processability and cycling stability of the silicon (Si) nanoparticle anodes. However, there is little systematical exploration regarding the relationship between grafting density and performance of grafted binder for Si anodes, especially when Si particles exceed the critical breaking size. Herein, a series of guar gum grafted polyacrylamide (GP) binders with different grafting densities are designed and prepared to determine the optimal grafting density for maximizing the electrochemical performance of Si submicroparticle (SiSMP) anodes. Among various GP binders, GP5 with recommended grafting density demonstrates the strongest adhesion strength, best mechanical properties, and highest intrinsic ionic conductivity. These characteristics enable the SiSMP electrodes to sustain the electrode integrity and accelerate lithium-ion transport kinetics during cycling, resulting in high capacity and stable cyclability. The superior role of GP5 binder in enabling robust structure and stable interface of SiSMP electrodes is revealed through the PeakForce atomic force microscopy and in situ differential electrochemical mass spectrometry. Furthermore, the stable cyclabilities of high-loading SiSMP@GP5 electrode with ultralow GP5 content (1wt%) at high areal capacity as well as the good cyclability of Ah-level LiNi0.8Co0.1Mn0.1O2/SiSMP@GP5 pouch cell strongly confirms the practical viability of the GP5 binder.

Application of AFM on Identifying Mechanical Properties of Individual Minerals and Surface Properties of Crack with High Resolution in Shale

Improving the resolution and accuracy of the mechanical properties of organic-rich shale is very important. The results can reveal the mechanical properties of shale from micro scale and serve as a guide for the design of hydraulic fracture optimization parameters. This study introduced an advanced technique to obtain the mechanical properties of shale with high resolution (58.6 nm/pixel) by combining SEM, EDS, and Atomic Force Microscopy (AFM). To locate the target area in SEM and AFM accurately, a positioning technique that uses special distributions of pyrite was established. AFM PeakForce QNM mode was selected due to its advantages at capturing topography and mechanical properties in material. Results illustrated the ability of AFM to obtain the mechanical properties (modulus) of individual mineral components in shale, the detailed topography of crack, and mechanical properties of minerals in a specific area. In particular, the mechanical properties of minerals around crack explained the layered distribution of minerals around the fractures, and the cracks developed in the clay mineral layer was detected. This article demonstrates the great potential application of AFM in shale.

Nanoscale identification and characterization of the shear strength of minerals and organic matter in shale

Shales are composed of minerals and organic matter, whose individual properties are essential to determining the rock’s macroscopical deformation and strength. Scanning electron microscopy combined with electron energy dispersive spectroscopy (EDS) has been extensively used to evaluate composition, while peak-force atomic force microscopy (AFM) has been used on the determination of elastic modulus with nanometric resolution. Still, there is a need for tools to conduct an in-depth study of the minerals’ tribomechanical properties. Atomic force microscopy is a tool that can contribute to these studies, as it can simultaneously measure the tribomechanical properties and identify the phases. In this work, we propose using atomic force microscopy and energy dispersive spectroscopy to identify the shale components and to measure the in situ tribomechanical properties from the different phases. Friction images between the atomic force microscopy tip and the surface were acquired as a function of load. Minerals and organic matter were later identified by colocalized energy dispersive spectroscopy mapping. Then, the frictional characteristics of the major shale constituents were obtained by adjusting the Derjaguin-Muller-Toporov model to the selected components. Moreover, the identification of the different phases was performed. The results show that friction at the nanometer scale was observed to be higher for organic matter than for any other shale constituent, while shear strength was observed to be higher for quartz and lower for organic matter. These characteristics were used to differentiate shale constituents. It is shown that a careful comparison of friction can be used to differentiate the sulfite pyrite, tectosilicates (quartz, andesine, and albite), phyllosilicate biotite, and organic matter. The presented methodology gives novel information on friction properties in the nanoscale that are comparable to available centimetric characterization techniques contributing to the understanding of rock strength.

An Ion‐Conductive Grafted Polymeric Binder with Practical Loading for Silicon Anode with High Interfacial Stability in Lithium‐Ion Batteries

AbstractBinders are required to dissipate huge mechanical stress and enhance the lithium‐ion diffusion kinetics of silicon anodes during cycling. Herein, a stress‐distribution binder with high ionic conductivity (GG‐g‐PAM) is constructed by grating polyacrylamide (PAM) onto ion‐conductive guar gum (GG) backbone. The mechanical stress distribution toward the grafted PAM chain enables the effective stress dissipation of the GG‐g‐PAM binder, and thus maintains a stable electrode‐electrolyte interface during cycling. The stress dissipation ability of the GG‐g‐PAM binder is confirmed by PeakForce atomic force microscopy experiments and finite element simulations. In addition, lithium complexation sites provided by oxygen heteroatoms in GG of the GG‐g‐PAM binder construct the Li‐ion pathways for facilitating Li ionic diffusion in the Si anodes. The good cyclabilities of Ah‐level pouch cells based on Si nanoparticle anodes strongly confirm GG‐g‐PAM as a desirable binder for practical Si anodes.

Protein Nanopore Membranes Prepared by a Simple Langmuir-Schaefer Approach.

Filtration through membranes with nanopores is typically associated with high transmembrane pressures and high energy consumption. This problem can be addressed by reducing the respective membrane thickness. Here, a simple procedure is described to prepare ultrathin membranes based on protein nanopores, which exhibit excellent water permeance, two orders of magnitude superior to comparable, industrially applied membranes. Furthermore, incorporation of either closed or open protein nanopores allows tailoring the membrane's ion permeability. To form such membranes, the transmembrane protein ferric hydroxamate uptake protein component A (FhuA) or its open-pore variant are assembled at the air-water interface of a Langmuir trough, compressed to a dense film, crosslinked by glutaraldehyde, and transferred to various support materials. This approach allows to prepare monolayer or multilayer membranes with a very high density of protein nanopores. Freestanding membranes covering holes up to 5 μm in diameter are visualized by atomic force microscopy (AFM), helium ion microscopy, and transmission electron microscopy. AFM PeakForce quantitative nanomechanical property mapping (PeakForce QNM) demonstrates remarkable mechanical stability and elastic properties of freestanding monolayer membranes with a thickness of only 5nm. The new protein membrane can pave the way to energy-efficient nanofiltration.

Effects of Aging on Micromechanical Properties of Asphalt Binder Using AFM

The performance of an asphalt binder is closely related to its microstructure. Understanding the properties of asphalt binders at the nano/microscale may help to improve knowledge of their macroperformance. The emergence of atomic force microscopy (AFM) technology makes it possible to distinguish the microstructural and micromechanical properties of asphalt binder. In this paper, the effects of aging and modifiers on the micromechanical properties of asphalt binder were investigated by using AFM peak force quantitative nanomechanical mapping (PFQNM) mode. Four different virgin asphalt binders and one styrene-butadiene-styrene (SBS)-modified asphalt binder subjected to different aging conditions were considered. It is found that the variation of morphology of bee structures relates to the penetration grade. The Derjaguin-Muller-Toporov (DMT) modulus fluctuates violently inside the bee structure. With the increasing aging degree of asphalt binders, the DMT modulus for three phases (catana, peri, and para phases) all increase correspondingly. Moreover, the addition of modifier improves the microscopic properties of asphalt binder.

Spatial high resolution of actin filament organization by PeakForce atomic force microscopy

ObjectivesTo investigate the heterogeneous feature of actin filaments (ACFs) associated with the cellular membrane in HeLa and HCT‐116 cells at the nanoscale level.Materials and MethodsFluorescence microscopy coupled with atomic force microscopy (AFM) was used to identify and characterize ACFs of cells. The distribution of ACFs was detected by Fluor‐488‐phalloidin–labelled actin. The morphology of the ACFs was probed by AFM images. The spatial correlation of the microvilli and ACFs was explored with different forces of AFM loading on cells.ResultsIntricate but ordered structures of the actin cytoskeletons associated with cellular membrane were characterized and revealed. Two different layers of ACFs with distinct structural organizations were directly observed in HCT‐116 and HeLa cells. Bundle‐shaped ACFs protruding the cellular membrane forming the microvilli, and the network ACFs underneath the cellular membrane were resolved with high resolution under near‐physiological conditions. Approximately 14 nm lateral resolution was achieved when imaging single ACF beneath the cellular membrane. On the basis of the observed spatial distribution of the ultrastructure of the ACF organization, a model for this organization of ACFs was proposed.ConclusionsWe revealed the two layers of the ACF organization in Hela and HCT‐116 cells. The resolved heterogeneous structures at the nanoscale level provide a spatial view of the ACFs, which would contribute to the understanding of the essential biological functions of the actin cytoskeleton.

Nano-mechanical mapping of interdependent cell and ECM mechanics by AFM force spectroscopy

Extracellular matrix (ECM), as a dynamic component of the tissue, influences cell behavior and plays an important role in cell mechanics and tissue homeostasis. Reciprocally, this three-dimensional scaffold is dynamically, structurally and mechanically modified by cells. In the field of biophysics, the independent role of cell and ECM mechanics has been largely investigated; however, there is a lack of experimental data reporting the interdependent interplay between cell and ECM mechanics, measured simultaneously. Here, using Atomic Force Microscopy (AFM) we have characterized five different decellularized matrices diverse in their topography, ECM composition and stiffness and cultured them with normal and pathological fibroblasts (scar and Dupuytren’s). We investigated the change in topography and elasticity of these matrices due to cell seeding, by using AFM peak force imaging and mechanical mapping, respectively. We found normal fibroblasts soften these matrices more than pathological fibroblasts, suggesting that pathological fibroblasts are profoundly influencing tissue stiffening in fibrosis. We detected different ECM composition of decellularized matrices used here influences fibroblast stiffness, thus highlighting that cell mechanics not only depends on ECM stiffness but also on their composition. We used confocal microscopy to assess fibroblasts invasion and found pathological fibroblasts were invading the matrices deeper than normal fibroblasts.

Tunable Hybrid Biopolymeric Hydrogel Scaffolds Based on Atomic Force Microscopy Characterizations for Tissue Engineering.

Developing adequate biomaterials to engineer cell-scaffold interactions has become a promising way for physically regulating the biological behaviors of cells in the field of tissue engineering. Biopolymeric hydrogels have shown great merits as cellular scaffolds due to their biocompatible and biodegradable characteristics. In particular, the advent of atomic force microscopy (AFM) provides a powerful tool for characterizing native specimens at the micro/nanoscale, but utilizing AFM to investigate the detailed structures and properties of hydrogel scaffolds has been still scarce. In this paper, hybrid natural biopolymers are used to form hydrogel scaffolds which exhibit tunable structural and mechanical properties characterized by AFM peak force tapping imaging, and the applications of the formed hydrogel scaffolds in tissue engineering are studied. AFM morphological images showed that the cross-linking reactions of sodium alginate and gum arabic via calcium cations yielded the porous hydrogel scaffolds. By altering the component ratios, AFM mechanical images showed that the porous and mechanical properties (Young's modulus and adhesion force) of the hydrogel scaffolds were tunable. Next, the nanoscale structural and mechanical dynamics of the fabricated hydrogel scaffolds during the degradation process were revealed by AFM peak force tapping imaging. The experimental results on three different types of cells showed that the fabricated hydrogel scaffolds facilitate the formation of cellular spheroids. The research provides a novel idea to design tunable hydrogel scaffolds based on AFM characterizations for investigating cell-scaffold interactions, which will have potential impacts on tissue engineering.

Composite Nanostructures and Adhesion Analysis of Natural Plant Hydrogels Investigated by Atomic Force Microscopy.

Hydrogels designed by biomimetism and bioinspiration have been considered as a promising biomaterial for biomedical applications due to their biocompatible characteristics. Though biomimetic or bioinspired hydrogels have been widely studied by biochemical analysis, so far the detailed organizations and properties of native natural hydrogels are still not fully understood. The advent of atomic force microscopy (AFM) provides a potent tool for probing the biological samples in their living states with nanometer spatial resolution, which offers new possibilities for addressing the biological and material issues at the nanoscale. In this paper, AFM was utilized to characterize the structures and adhesive properties of the natural hydrogels produced by the carnivorous plant sundew. AFM morphological images of the mucilage secreted by three types of sundew show the composite nanostructures of the mucilage (e.g., nanoparticles, nanofibers, and porous polymeric networks), which are correlated with the mucilage's adhesive features revealed by AFM force measurements and peak force tapping mechanical imaging. The research demonstrates the prominent capabilities of AFM in characterizing the nanoscopic structures and properties of natural hydrogels with unprecedented spatial resolution, which is useful for understanding the underlying mechanisms guiding the behaviors of natural hydrogels and will potentially benefit the studies of biomimetic and bioinspired biomaterials.

Surface Properties of Alkoxysilane Layers Grafted in Supercritical Carbon Dioxide

Silicon oxide surface properties can be easily modified by grafting alkoxysilane molecules. Here, we studied the structure and the morphology of ultrathin layers prepared by the grafting of alkoxysilanes having different head groups (thiol, amine, and iodo) in supercritical carbon dioxide (CO2) on model plane silicon oxide surfaces. Several characterization techniques (X-ray reflectivity, water contact angle, X-ray photoelectron spectroscopy, and atomic force microscopy (AFM)) were used to determine the physicochemical properties of the layers prepared at different temperatures. Moreover, for the first time, AFM peak force measurements were used to delve deeper into the determination of the structure of these ultrathin alkoxysilane layers. The results show that the grafting temperature and the nature of the head group strongly affect the morphology and structure of the grafted layers. Dense monolayers are obtained with 3-(mercaptopropyl)trimethoxysilane at 60 °C, polycondensed layers are always prepared with [3-(aminoethylamino)propyl]trimethoxysilane, and a dense bilayer is synthesized with 3-(iodopropyl)triethoxysilane at 120 °C.

Nanoscale Multiparametric Imaging of Peptide-Assembled Nanofibrillar Hydrogels by Atomic Force Microscopy

Peptide-assembled hydrogels have emerged as a promising biomaterial for tissue engineering due to their controllable structures as well as biocompatible and biodegradable characteristics. However, the detailed situations of nanofibrillar mechanics during the formation of peptide-assembled hydrogels remain poorly understood. The advent of atomic force microscopy (AFM) provides a powerful tool for nanoscale characterizations and measurements of native biomaterials without pretreatments. In this paper, AFM peak force tapping multiparametric imaging was utilized to investigate the morphology and mechanics of individual nanofibrils during the gelation and degradation process of peptide-assembly hydrogels. Nanofibrillar hydrogels were fabricated by the self-assembly reactions of phenylalanine peptide molecules. AFM topography imaging distinctly visualized the diverse assembly behaviors of peptide-formed nanofibrils during the gelation and degradation of hydrogels, which were significantly correlated with the changes of nanofibrillar mechanics (Young's modulus, adhesion force, deformation, and persistence length) revealed by AFM mechanical imaging. The research provides a novel idea for nanoscale imaging and mechanical analysis of nanofibrillar behaviors in peptide-assembled hydrogels, which will have potential impacts on the characterization and design of supramolecular biomaterials.

Measurement of nanomechanical properties of DNA molecules by PeakForce atomic force microscopy based on DNA origami.

Characterization of the stiffness of thin DNA strands remains difficult. By constructing bilayer DNA molecules, we investigated their mechanical properties using AFM. Increased DNA thickness through DNA origami greatly reduced the substrate effect when measuring Young's modulus, thus providing a more accurate picture of the inherent nanomechanical properties of DNA.

AFM Peakforce QNM mode for measurement of nanosurface mechanical properties of Pt-cured silicones

Sylgard 184 is used for diverse applications including nanoimprinting, micro-electronic and mechanical systems (MEMS), soft robots, ice-phobic coatings and tactile and epidermal sensors. For such applications, understanding effects of processing conditions on nanoscale mechanical properties is essential for performance. It is surprising, then, that little attention has been paid to effects of cure temperature on surface and mechanical properties. The present work adds depth to understanding the complex relationships among hydrosilylation and little recognized autoxidation of SiH to SiOH and secondary network formation by condensation. Peakforce Quantitative Nanomechanical Mapping (PF-QNM), an atomic force microscopy (AFM) mode was used to provide a nanoscale map of deformation, adhesion, dissipation, modulus and morphology as a function of varying cure temperature and base-to-crosslinker ratio. The interaction of the SiOH rich oxide layer of the silicon AFM tip with the outermost Sylgard 184 silicone surface provided modulus and adhesion measurements that revealed important effects of cure conditions and composition. Results were interpreted in terms of a model for the concentration of near surface SiOH from autoxidation and changes due to SiOH condensation at high temperature cure. Moduli were found to be comparable with dynamic mechanical analysis (DMA), supporting PF-QNM as a tool for mechanical property measurements. The dependence of nanosurface mechanical properties on composition and network formation conditions provides guidance for future work using Pt-cured silicones.

Directly and quantitatively studying the interfacial interaction between SiO 2 and elastomer by using peak force AFM

Abstract The interfacial interaction between nanofillers and elastomer plays an important role in properties of elastomer nanocomposites. Up to now, most methods focus on studying the interphase of elastomer nanocomposites. The filler-elastomer interaction forces have not been directly and quantitatively characterized yet. In this study, the interaction forces at nano-Newton scale between elastomer matrix and silica (SiO 2 ) were directly and quantitatively characterized by using peak force quantitative nanomechanical mapping (PF-QNM) mode of atomic force microscopy (AFM). Three kinds of nitrile butadiene rubber (NBR) molecules with different molecular polarity were first grafted on the AFM tip by using click chemistry. The SiO 2 (or modified SiO 2 ) were pressed into flat films and worked as substrate. The interaction forces between these NBR layer grafted on AFM tip and SiO 2 (or modified SiO 2 ) substrate were directly and quantitatively obtained. The results show that as the acrylonitrile content of NBR increasing from 26 wt% to 41 wt%, the average interaction force of NBR/SiO 2 increases from 4.5 ± 1.0 nN to 7.7 ± 1.1 nN and that of NBR/m-SiO 2 increases from 6.2 ± 1.2 nN to 12.9 ± 1.0 nN. This study provides guidance in direct and quantitative characterization of filler-elastomer interactions at nano-Newton resolution.

Determination of Interfacial Amorphicity in Functional Powders.

The nature of the surfaces of particles of pharmaceutical ingredients, food powders, and polymers is a determining factor for their performance in for example tableting, powder handling, or mixing. Changes on the surface structure of the material will impact the flow properties, dissolution rate, and tabletability of the powder blend. For crystalline materials, surface amorphization is a phenomenon which is known to impact performance. Since it is important to measure and control the level of amorphicity, several characterization techniques are available to determine the bulk amorphous content of a processed material. The possibility of characterizing the degree of amorphicity at the surface, for example by studying the mechanical properties of the particles' surface at the nanoscale, is currently only offered by atomic force microscopy (AFM). The AFM PeakForce QNM technique has been used to measure the variation in energy dissipation (eV) at the surface of the particles which sheds light on the mechanical changes occurring as a result of amorphization or recrystallization events. Two novel approaches for the characterization of amorphicity are presented here. First, since particles are heterogeneous, we present a methodology to present the results of extensive QNM analysis of multiple particles in a coherent and easily interpreted manner, by studying cumulative distributions of dissipation data with respect to a threshold value which can be used to distinguish the crystalline and amorphous states. To exemplify the approach, which is generally applicable to any material, reference materials of purely crystalline α-lactose monohydrate and completely amorphous spray dried lactose particles were compared to a partially amorphized α-lactose monohydrate sample. Dissipation data are compared to evaluations of the lactose samples with conventional AFM and SEM showing significant topographical differences. Finally, the recrystallization of the surface amorphous regions in response to humidity was followed by studying the dissipation response of a well-defined surface region over time, which confirms both that dissipation measurement is a useful measure of surface amorphicity and that significant recrystallization occurs at the surface in response to humidity.

Rapid recognition and functional analysis of membrane proteins on human cancer cells using atomic force microscopy

Understanding the physicochemical properties of cell surface signalling molecules is important for us to uncover the underlying mechanisms that guide the cellular behaviors. Atomic force microscopy (AFM) has become a powerful tool for detecting the molecular interactions on individual cells with nanometer resolution. In this paper, AFM peak force tapping (PFT) imaging mode was applied to rapidly locate and visually map the CD20 molecules on human lymphoma cells using biochemically sensitive tips. First, avidin–biotin system was used to test the effectiveness of using PFT imaging mode to probe the specific molecular interactions. The adhesion images obtained on avidin-coated mica using biotin-tethered tips obviously showed the recognition spots which corresponded to the avidins in the simultaneously obtained topography images. The experiments confirmed the specificity and reproducibility of the recognition results. Then, the established procedure was applied to visualize the nanoscale organization of CD20s on the surface of human lymphoma Raji cells using rituximab (a monoclonal anti-CD20 antibody)-tethered tips. The experiments showed that the recognition spots in the adhesion images corresponded to the specific CD20–rituximab interactions. The cluster sizes of CD20s on lymphoma Raji cells were quantitatively analyzed from the recognition images. Finally, under the guidance of fluorescence recognition, the established procedure was applied to cancer cells from a clinical lymphoma patient. The results showed that there were significant differences between the adhesion images obtained on cancer cells and on normal cells (red blood cell). The CD20 distributions on ten cancer cells from the patient were quantified according to the adhesion images. The experimental results demonstrate the capability of applying PFT imaging to rapidly investigate the nanoscale biophysical properties of native membrane proteins on the cell surface, which is of potential significance in developing novel biomarkers for cancer diagnosis and drug development.

Effect of Actin Organization on the Stiffness of Living Breast Cancer Cells Revealed by Peak-Force Modulation Atomic Force Microscopy.

We study the correlation between cytoskeleton organization and stiffness of three epithelial breast cancer cells lines with different degrees of malignancy: MCF-10A (healthy), MCF-7 (tumorigenic/noninvasive), and MDA-MB-231 (tumorigenic/invasive). Peak-force modulation atomic force microscopy is used for high-resolution topography and stiffness imaging of actin filaments within living cells. In healthy cells, local stiffness is maximum where filamentous actin is organized as well-aligned stress fibers, resulting in apparent Young's modulus values up to 1 order of magnitude larger than those in regions where these structures are not observed, but these organized actin fibers are barely observed in tumorigenic cells. We further investigate cytoskeleton conformation in the three cell lines by immunofluorescence confocal microscopy. The combination of both techniques determines that actin stress fibers are present at apical regions of healthy cells, while in tumorigenic cells they appear only at basal regions, where they cannot contribute to stiffness as probed by atomic force microscopy. These results substantiate that actin stress fibers provide a dominant contribution to stiffness in healthy cells, while the elasticity of tumorigenic cells appears not predominantly determined by these structures. We also discuss the effects of the high-frequency indentations inherent to peak-force atomic force microscopy for the identification of mechanical cancer biomarkers. Whereas conventional low loading rate indentations (1 Hz) result in slightly differentiated average stiffness for each cell line, in high-frequency measurements (250 Hz) healthy cells are clearly discernible from both tumorigenic cells with an enhanced stiffness ratio; however, the two cancerous cell lines produced indistinguishable results.

Epithelial cell morphology and adhesion on diamond films deposited and chemically modified by plasma processes.

The authors show that nanocrystalline diamond (NCD) thin films prepared by microwave plasma enhanced chemical vapor deposition apparatus with a linear antenna delivery system are well compatible with epithelial cells (5637 human bladder carcinoma) and significantly improve the cell adhesion compared to reference glass substrates. This is attributed to better adhesion of adsorbed layers to diamond as observed by atomic force microscopy (AFM) beneath the cells. Moreover, the cell morphology can be adjusted by appropriate surface treatment of diamond by using hydrogen and oxygen plasma. Cell bodies, cytoplasmic rims, and filopodia were characterized by Peakforce AFM. Oxidized NCD films perform better than other substrates under all conditions (96% of cells adhered well). A thin adsorbed layer formed from culture medium and supplemented with fetal bovine serum (FBS) covered the diamond surface and played an important role in the cell adhesion. Nevertheless, 50-100 nm large aggregates formed from the RPMI medium without FBS facilitated cell adhesion also on hydrophobic hydrogenated NCD (increase from 23% to 61%). The authors discuss applicability for biomedical uses.

Quantitative imaging of diatoms by PeakForce atomic force microscopy

A novel approach to study nanomechanical properties such as deformation, adhesion or Young's modulus is presented for freshwater diatoms having sizes ranging from tens to hundreds of micrometers. Changes occurring in the environment, such as water acidity or temperature variations, have an impact on the diatoms population, as they can cause the death of certain species favoring simultaneously the growth of another. The nanomechanical properties were measured by means of atomic force microscopy (AFM) in a unique PeakForce tapping (PFT) mode. The observed changes of the Young's modulus measured in the frustule of Cyclotella meneghini diatom were in the range of 7–25 GPa, clearly confirming that, in this part of the diatom, degeneration occurs. The use of the PFT mode in the AFM allowed also for measuring the surface of diatoms exhibiting a very developed topography. This is a new methodology for quantitative measurements of nanomechanical properties allowing to monitor environmental changes. Copyright © 2014 John Wiley & Sons, Ltd.