Atomic Force Microscope Cantilever Research Articles

In situ measurement of cell stiffness of Arabidopsis roots growing on a glass micropillar support by atomic force microscopy.

Atomic force microscopy (AFM) can measure the mechanical properties of plant tissue at the cellular level, but for in situ observations, the sample must be held in place on a rigid support and it is difficult to obtain accurate data for living plants without inhibiting their growth. To investigate the dynamics of root cell stiffness during seedling growth, we circumvented these problems by using an array of glass micropillars as a support to hold an Arabidopsis thaliana root for AFM measurements without inhibiting root growth. The root elongated in the gaps between the pillars and was supported by the pillars. The AFM cantilever could contact the root for repeated measurements over the course of root growth. The elasticity of the root epidermal cells was used as an index of the stiffness. By contrast, we were not able to reliably observe roots on a smooth glass substrate because it was difficult to retain contact between the root and the cantilever without the support of the pillars. Using adhesive to fix the root on the smooth glass plane overcame this issue, but prevented root growth. The glass micropillar support allowed reproducible measurement of the spatial and temporal changes in root cell elasticity, making it possible to perform detailed AFM observations of the dynamics of root cell stiffness.

High-resolution measurement of atomic force microscope cantilever resonance frequency

The atomic force microscope (AFM) is widely used in a wide range of applications due to its high scanning resolution and diverse scanning modes. In many applications, there is a need for accurate and precise measurement of the vibrational resonance frequency of a cantilever. These frequency shifts can be related to changes in mass of the cantilever arising from, e.g., loss of fluid due to a nanolithography operation. A common method of measuring resonance frequency examines the power spectral density of the free random motion of the cantilever, commonly known as a thermal. While the thermal is capable of reasonable measurement resolution and speed, some applications are sensitive to changes in the resonance frequency of the cantilever, which are small, rapid, or both, and the performance of the thermal does not offer sufficient resolution in frequency or in time. In this work, we describe a method based on a narrow-range frequency sweep to measure the resonance frequency of a vibrational mode of an AFM cantilever and demonstrate it by monitoring the evaporation of glycerol from a cantilever. It can be seamlessly integrated into many commercial AFMs without additional hardware modifications and adapts to cantilevers with a wide range of resonance frequencies. Furthermore, this method can rapidly detect small changes in resonance frequency (with our experiments showing a resolution of ∼0.1 Hz for cantilever resonances ranging from 70 kHz to 300 kHz) at a rate far faster than with a thermal. These attributes are particularly beneficial for techniques such as dip-pen nanolithography.

Abstract 13652: An Innate YAP/TAZ Mechanosensing Deficit in Hutchinson-Gilford Progeria Syndrome

Hutchinson-Gilford Progeria Syndrome (HGPS) is a disease of accelerated aging causing death in the mid-teens from myocardial infarction or stroke. The disease is caused by a point mutation in the gene encoding lamin-A. The mutated scaffolding protein is aberrantly farnesylated inducing a constellation of defects included nuclear abnormalities, genomic damage, and rapid senescence. Therapy targeting the abnormal farnesylation provides a modest extension of life, thus new insights and therapeutic approaches are urgently needed for these children. Consistent with previous morphological observations and new studies implicating YAP/TAZ mechanobiology as an important mechanical pathway for endothelial cell (EC) health under shear stress, we hypothesized that HGPS ECs have an innate mechanical disturbance rendering them unable to respond to external, atheroprotective cues. We used a microfluidic vessel-on-a-chip with channel geometries and fluid flow to precisely model the hemodynamic stimuli present in vasculature as we have previously described. We cultured iPSC-derived HGPS ECs in this system to study mechanoresponse to shear stress and YAP/TAZ signaling. HGPS ECs manifest a rounded, flattened appearance characteristic of senescent ECs, are unable to align in response to flow, and have aberrant YAP/TAZ activity despite unidirectional laminar flow. To explore the physical underpinnings of such biochemical disturbances, we used atomic force microscopy (AFM) to precisely characterize the shape of individual HGPS cells, and their deformation to a controlled force applied by the AFM cantilever. Preliminary measurements confirmed that HGPS cells have a reduced profile and are compositely stiffer (nuclear modulus + cytoskeletal modulus) than cells derived from the unaffected parent of the child. These data provide evidence of altered biophysical properties of senescent cells which we term “mechanical aging,” which is associated with aberrant signaling in response to hemodynamic stimuli. Further characterization of mechanical aging may lead to new therapeutic approaches for HGPS and other age-related diseases.

Dynamic analysis of flexural vibration mode of an atomic force microscope cantilever with a sidewall probe in liquid.

In this paper, dynamic behavior and the resonance frequencies of flexural vibration modes of an atomic force microscope cantilever with sidewall probe immersed in liquid to surface stiffness variations have been investigated and a closed-form expression is derived. Using numerical analysis, the flexural resonance frequencies of microcantilever immersed in liquid are calculated and the results are compared with the air environment. Then, the effect of sidewall length and normal stiffness on the frequency is investigated. Moreover, the surface-coupled effect between the cantilever and simple surface when the cantilever is close to the surface is considered and its effect on the quality factors of the first to fourth modes is studied.

Determination of Hamaker constant for CNT and AFM probe configuration inside SEM

Mounting the atomic force microscope (AFM) probe on the gripper of the robotic nanoscale manipulators system enables the combination of the AFM and scanning electron microscope (SEM). The AFM–SEM-based method of determining Hamaker constant for carbon nanotube (CNT) and AFM cantilever in the vacuum condition of SEM, which is completed through a robotic nanoscale manipulators system, is presented for the first time. During Hamaker constant determining experiment, the elastic force is measured utilising integrated AFM–SEM imaging technique through detecting the deflection of AFM cantilever that actually equals van der Waals adhesion force. The van der Waals attraction force between CNT and AFM cantilever is calculated through the modelling of a cylindrical nanotube and a flat gold-coated surface. The experiment results indicate that Hamaker's constant measured are comparable to typical values published by previous researchers, being of the order of 100 zJ, which indicates that the present approach is valid and reliable.

Quantitative characterization of dielectric properties of nanoparticles using electrostatic force microscopy

We use a new method based on Electrostatic Force Microscopy (EFM) to perform quantitative measurements of the dielectric constants of individual carboxylated as well as non-functionalized polystyrene nanospheres. The EFM data record the oscillation phase of an atomic force microscope (AFM) cantilever as a function of the AFM tip position. In our experiments, the relative dielectric constant of the sample is measured from the EFM phase shifts vs the tip–surface separation, according to a simple analytical model describing the tip–surface interactions. We perform a comprehensive study of how the dielectric constant depends on the sphere diameter for both types of nanospheres. Our results demonstrate that the experimental method has a high-resolution for measuring the dielectric constant of nano/microbeads and other nanoscale materials and is simple to implement on standard atomic force microscopes. This non-invasive technique can be applied to measure the electrical properties of colloidal particles, polymers, interphases, and polymer nanocomposites.

Modeling of non-contact atomic force microscope with two-term excitations

The goal of this article is to study the dynamical behavior of atomic force microscope cantilever in its non-contact mode of operation. The lumped parameter model is used to construct the mathematical model of the cantilever. The tip of the cantilever is excited by two harmonic terms and is in interaction with the sample surface. The Van der Waals force, tip-sample interaction force, makes the system nonlinear. Using multiple scales method, the frequency response equation is found. The effects on the amplitude of excitations, the damping coefficient, and initial sample – tip distance is studied and presented as the results.

Quantitative characterization of dielectric properties of polymer fibers and polymer composites using electrostatic force microscopy

We use a new method based on electrostatic force microscopy (EFM) to perform quantitative measurements of the dielectric constants of individual electrospun nanofibers of poly(L-lactic acid) (PLLA), as well as composite fibers of PLLA with embedded multiwall carbon nanotubes (MWCNT-PLLA). The EFM data record the oscillation phase of an atomic force microscope (AFM) cantilever as a function of the AFM tip position. In our experiments the relative dielectric constants ϵ of the sample are measured from the EFM phase shifts vs. the tip-surface separation, according to a simple analytical model describing the tip-surface interactions. We perform a comprehensive study of how the dielectric constant depends on the fiber diameter for both electrospun PLLA and MWCNT/PLLA fiber composites. Our measurements show that EFM can distinguish between dielectric properties of PLLA fibers and fiber composites with different diameters. Dielectric constants of both PLLA and MWCNT-PLLA composite fibers decrease with increasing fiber diameter. In the limit of large fiber diameters (D > 100 nm), we measure dielectric constants in the range: ϵ = 3.4–3.8, similar to the values obtained for unoriented PLLA films: ϵfilm = 2.4–3.8. Moreover, the dielectric constants of the small diameter MWCNT-PLLA composites are significantly larger than the corresponding values obtained for PLLA fibers. For MWCNT-PLLA nanofiber composites of small diameters (D < 50 nm), ϵ approaches the values measured for neat MWCNT: ϵCN = 12 ± 2. These results are consistent with a simple fiber structural model that shows higher polarizability of thinner fibers, and composites that contain MWCNTs. The experimental method has a high-resolution for measuring the dielectric constant of soft materials, and is simple to implement on standard atomic force microscopes. This non-invasive technique can be applied to measure the electrical properties of polymers, interphases, and polymer nanocomposites.

Nanomechanical sampling of material for nanoscale mass spectrometry chemical analysis.

The ability to spatially resolve the chemical distribution of compounds on a surface is important in many applications ranging from biological to material science. To this extent, we have recently introduced a hybrid atomic force microscopy (AFM)-mass spectrometry (MS) system for direct thermal desorption and pyrolysis of material with nanoscale chemical resolution. However, spatially resolved direct surface heating using local thermal desorption becomes challenging on material surfaces with low melting points, because the material will undergo a melting phase transition due to heat dissipation prior to onset of thermal desorption. Therefore, we developed an approach using mechanical sampling and collection of surface materials on an AFM cantilever probe tip for real-time analysis directly from the AFM tip. This approach allows for material to be concentrated directly onto the probe for subsequent MS analysis. We evaluate the performance metrics of the technique and demonstrate localized MS sampling from a candelilla wax matrix containing UV stabilizers avobenzone and oxinoxate from areas down to 250nm × 250nm. Overall, this approach removes heat dissipation into the bulk material allowing for a faster desorption and concentration of the gas phase analyte from a single heating pulse enabling higher signal levels from a given amount of material in a single sampling spot.Graphical abstract.

Nano polarimetry: enhanced AFM-NSOM triple-mode polarimeter tip

A novel application of a combined and enhanced NSOM-AFM tip-photodetector system resulted in a nanoscale Polarimeter, generated by four different holes, each sharing a different shape, and enabling that the four photonic readouts forming the tip will be the four Stokes coefficients, this in order to place the polarization state in the Poincare sphere. The new system has been built on standard Atomic Force Microscope (AFM) cantilever, in order to serve as a triple-mode scanning system, sharing complementary scanning topography, optical data analysis and polarization states. This new device, which has been designed and simulated using Comsol Multi-Physics software package, consists in a Platinum-Silicon drilled conical photodetector, sharing subwavelength apertures, and has been processed using advanced nanotechnology tools on a commercial silicon cantilever. After a comparison study of drilled versus filled tips advantages, and of several optics phenomena such as interferences, the article presents the added value of multiple-apertures scanning tip for nano-polarimetry.

Tribological characteristics of atomic-scale niobium diselenide grown via chemical vapor deposition

The nanoscale tribological characteristics of atomic-layered niobium diselenide (NbSe2) were investigated using atomic force microscopy (AFM). Two-dimensional NbSe2 atomic layers were produced on a weakly-adherent silicon wafer using two different methods: (1) mechanical exfoliation and (2) controlled synthesis via chemical vapor deposition. Using an AFM cantilever tip, a normal force was applied on the NbSe2 atomic-scale thin films in order to measure friction force. The as-synthesized NbSe2 thin films exhibited atomic lattice stick-slip friction, with the thinnest sheets showing a sliding length-dependent increase in static friction. This tendency is attributed to the increased susceptibility of the thinner chemically-synthesized NbSe2 sheets toward out-of-plane elastic deformation.

Lateral force calibration for atomic force microscope cantilevers using a suspended nanowire

Friction measurement via atomic force microscope (AFM) relies on accurate calibration for the torsional spring constant of the AFM cantilever and its lateral deflection sensitivity. Here we describe a method that employs a suspended nanowire (SNW) as a reference beam to quantify the torsional spring constant of AFM cantilevers. Based on the fact that a uniform SNW with cylindrical symmetry has an identical spring constant when bent in any direction perpendicular to its axis, the spring constant of the SNW in a normal direction is determined by an AFM cantilever with a known normal spring constant, and is subsequently used as a force transfer standard to calibrate the torsional spring constant of the AFM cantilever. The lateral deflection sensitivity can be accurately measured by pushing the AFM tip laterally on the groove edge. The calibration result is compared to the well-known diamagnetic lateral force calibrator method and shows an uncertainty of 15% or better. The presented method is applicable for the lateral force calibration of AFM cantilevers in a wide range of instruments including inverted configurations and in an ultrahigh vacuum.

Long-Range Ordered Water Correlations between A–T/C–G Nucleotides

Summary The interactions between complementary base pairs are crucial to the helical duplex structure of DNA. Although base-base interactions have been widely reported, the study of long-range interactions in the base-pairing processes is still a major challenge in this field. Here, we first study the long-range interactions between the base pairs by atomic force microscopy-based single-molecule force spectroscopy (SMFS). The SMFS results of A–T/C–G imply that there are weak long-range interactions between them, which have a range of 15–25 nm. Raman spectroscopy results imply that these weak interactions can be attributed to multiplex hydrogen bond interaction of ordered water structure between A–T/C–G nucleotides. In addition, the theoretical calculations deduce that the ideal structure of these water molecules exhibits a specific helical structure. Our findings might open up a new understanding of biological assembly processes and provide helpful strategies for bio-nanotechnology.

Measurement of the Poisson’s ratio and Young’s modulus of an isotropic material with T-shape contact resonance atomic force microscopy

The Poisson’s ratio and the Young’s modulus play an important role in the characterization of nanomaterial mechanical properties. They are the vital parameters of understanding nanoscale material behavior. Here we report a method of quantitatively determining the values of the Poisson’s ratio and the Young’s modulus with a T-shape contact resonance atomic force microscopy. Unlike the cantilever of a traditional atomic force microscopy, the flexural and torsional modes of the T-shape cantilever are simultaneously excited and coupled in the contact mode. Through the analysis, the bifurcation of the coupled contact resonance frequencies is found in higher modes with the increasing contact stiffness. More importantly, the frequency bifurcation point can be used to decouple the Poisson’s ratio and the Young’s modulus, which leads to the determination of their separate values. In contrast to the previous methods, in which the Poisson’s ratio and the Young’s modulus are intrinsically coupled and there is no effective way of decoupling, the method presented in this study offers a new way of decoupling and determining these two parameters. This efficient and accurate method can be of significant help to the characterization of various nanomaterials.

Indenting soft samples (hydrogels and cells) with cantilevers possessing various shapes of probing tip

The identification of cancer-related changes in cells and tissues based on the measurements of elastic properties using atomic force microscopy (AFM) seems to be approaching clinical application. Several limiting aspects have already been discussed; however, still, no data have shown how specific AFM probe geometries are related to the biomechanical evaluation of cancer cells. Here, we analyze and compare the nanomechanical results of mechanically homogenous polyacrylamide gels and heterogeneous bladder cancer cells measured using AFM probes of various tip geometry, including symmetric and non-symmetric pyramids and a sphere. Our observations show large modulus variability aligned with both types of AFM probes used and with the internal structure of the cells. Altogether, these results demonstrate that it is possible to differentiate between compliant and rigid samples of kPa elasticity; however, simultaneously, they highlight the strong need for standardized protocols for AFM-based elasticity measurements if applied in clinical practice including the use of a single type of AFM cantilever.

Calibration of T-shaped atomic force microscope cantilevers using the thermal noise method.

The tip-sample interaction force measurements in atomic force microscopy (AFM) provide information about materials' properties with nanoscale resolution. The T-shaped cantilevers used in Torsional-Harmonic AFM allow measuring the rapidly changing tip-sample interaction forces using the torsional (twisting) deflections of the cantilever due to the off-axis placement of the sharp tip. However, it has been difficult to calibrate these cantilevers using the commonly used thermal noise-based calibration method as the mechanical coupling between flexural and torsional deflections makes it challenging to determine the deflection sensitivities from force-distance curves. Here, we present thermal noise-based calibration of these T-shaped AFM cantilevers by simultaneously analyzing flexural and torsional thermal noise spectra, along with deflection signals during a force-distance curve measurement. The calibration steps remain identical to the conventional thermal noise method, but a computer performs additional calculations to account for mode coupling. We demonstrate the robustness of the calibration method by determining the sensitivity of calibration results to the laser spot position on the cantilever, to the orientation of the cantilever in the cantilever holder, and by repeated measurements. We validated the quantitative force measurements against the known unfolding force of a protein, the I91 domain of titin, which resulted in consistent unfolding force values among six independently calibrated cantilevers.

Direct Observation of Cell Surface Sialylation by Atomic Force Microscopy Employing Boronic Acid–Sialic Acid Reversible Interaction

Tracing cell surface sialylation dynamics at a scale of the glycolipoprotein microdomain (lipid rafts) formations remains an intriguing challenge of cellular biology. Here, we demonstrate that this goal is accessible, taking advantage of a boronic acid (BA)-based reversible molecular recognition chemistry. A BA-end-functionalized poly(ethylene glycol) was decorated onto an atomic force microscopy (AFM) cantilever, which provided a dynamic and sialic acid (SA)-specific imaging mode. Using this technique, we were able to heat map the SA expression levels not only on protein-decorated substrates but also directly on the cell surfaces, with a submicrometer scale resolution that may be relevant to that of the lipid rafts formation. The SA specificity and the binding reversibility of the probe were confirmed from its pH-dependent characteristics and an inhibition assay using free state SA. This finding may provide a noninvasive means for assessing a variety of SA-involved glycosylation dynamics spanning from physiology to pathology.

High-Precision Cutting Edge Radius Measurement of Single Point Diamond Tools Using an Atomic Force Microscope and a Reverse Cutting Edge Artifact

This paper presents a measurement method for high-precision cutting edge radius of single point diamond tools using an atomic force microscope (AFM) and a reverse cutting edge artifact based on the edge reversal method. Reverse cutting edge artifact is fabricated by indenting a diamond tool into a soft metal workpiece with the bisector of the included angle between the tool’s rake face and clearance face perpendicular to the workpiece surface on a newly designed nanoindentation system. An AFM is applied to measure the topographies of the actual and the reverse diamond tool cutting edges. With the proposed edge reversal method, a cutting edge radius can be accurately evaluated based on two AFM topographies, from which the convolution effect of the AFM tip can be reduced. The accuracy of the measurement of cutting edge radius is significantly influenced by the geometric accuracy of reverse cutting edge artifact in the proposed measurement method. In the nanoindentation system, the system operation is optimized for achieving high-precision control of the indentation depth of reverse cutting edFigurege artifact. The influence of elastic recovery and the AFM cantilever tip radius on the accuracy of cutting edge radius measurement are investigated. Diamond tools with different nose radii are also measured. The reliability and capability of the proposed measurement method for cutting edge radius and the designed nanoindentation system are demonstrated through a series of experiments.

NONLINEAR VIBRATIONS OF THE IMMERSED DAGGER-SHAPED ATOMIC FORCE MICROSCOPE CANTILEVER IN DIFFERENT LIQUIDS STUDIED BY EXPERIMENTAL AND THEORETICAL METHODS

In this paper, the nonlinear dynamic behavior of an immersed dagger-shaped atomic force microscope cantilever in different liquids has been investigated for the first time. The Timoshenko beam theory, which considers rotatory inertia and shear deformation effects, has been used for modeling the cantilever. The nonlinear tip-sample interaction force has been modeled by using the Hertzian contact theory. Water, methanol, acetone, and carbon tetrachloride are considered as the immersion environments. In most cases, the softening behavior has been observed for the cantilever. The resonant frequency is found to decrease with an increase in the liquid viscosity. The experimental results have been compared with the theoretical predictions and are found to be in good agreement.

Dynamic analysis of atomic force microscope in tapping mode

The Atomic force microscope (AFM) is an enormously valuable tool in a wide variety of applications because of its function to depict in different mediums with the sub-nano-meter resolution, and also manipulating objects with nano-meter-scale features and measuring forces with better than pico-newton resolution. In this paper, the lumped parameter model is used to construct the mathematical model of the AFM cantilever. The cantilever tip, excited by the harmonic external force, is under the influence of the tip-sample interaction force. Since, we consider the AFM in the operation mode named dynamic contact mode, the Deryagin-Muller-Toporov (DMT) force is considered as the interaction force between the cantilever tip and the surface of the sample. DMT force causes non-linearity. To solve the equation of motion, the Van der Pol method is used to obtain the frequency response equation to investigate the non-linearity effect as well as the amplitude of the excitation on the response. The stability of steady state motion is investigated.