Javier Tamayo

EFFECTS OF ELASTIC AND INELASTIC INTERACTIONS ON PHASE CONTRAST IMAGES IN TAPPING-MODE SCANNING FORCE MICROSCOPY

The dependence of phase contrast in tapping-mode scanning force microscopy on elastic and inelastic interactions is studied. The cantilever–tip ensemble is simulated as a driven, damped harmonic oscillator. It is found that for tip–sample elastic interactions, phase contrast is independent of the sample’s elastic properties. However, phase contrast associated with elastic modulus variations are observed if viscous damping or adhesion energy hysteresis is considered during tip–sample contact. The phase shift versus tip–sample equilibrium separation was measured for a compliant material (polypropylene) and for a stiff sample (mica). The agreement obtained between theory and experiment supports the conclusions derived from the model. These results emphasize the relevance of energy dissipating processes at the nanometer scale to explain phase contrast imaging in tapping-mode force microscopy.

Relationship between phase shift and energy dissipation in tapping-mode scanning force microscopy

Force curves taken during a load–unload cycle show the presence of a hysteresis loop. The area enclosed by the loop is used to measure the energy dissipated by the tip-sample interaction in tapping-mode scanning force microscopy. The values of the energy loss obtained from force curves are compared with the results derived from a model based on phase shift measurements. The agreement obtained between both methods demonstrates that for the same operating conditions, the higher the phase shift the larger the amount of energy dissipated by the tip-sample interaction. It also confirms the prediction that phase-contrast images can only arise if there are tip-sample inelastic interactions.

Label-free detection of DNA hybridization based on hydration-induced tension in nucleic acid films

The properties of water at the nanoscale are crucial in many areas of biology, but the confinement of water molecules in sub-nanometre channels in biological systems has received relatively little attention. Advances in nanotechnology make it possible to explore the role played by water molecules in living systems, potentially leading to the development of ultrasensitive biosensors. Here we show that the adsorption of water by a self-assembled monolayer of single-stranded DNA on a silicon microcantilever can be detected by measuring how the tension in the monolayer changes as a result of hydration. Our approach relies on the microcantilever bending by an amount that depends on the tension in the monolayer. In particular, we find that the tension changes dramatically when the monolayer interacts with either complementary or single mismatched single-stranded DNA targets. Our results suggest that the tension is mainly governed by hydration forces in the channels between the DNA molecules and could lead to the development of a label-free DNA biosensor that can detect single mutations. The technique provides sensitivity in the femtomolar range that is at least two orders of magnitude better than that obtained previously with label-free nanomechanical biosensors and with label-dependent microarrays.

Chemical sensors and biosensors in liquid environment based on microcantilevers with amplified quality factor

A new technique is presented for bio/chemical sensors, based on microcantilevers, for detection in liquid environment. The low quality factor of the cantilever in liquid is increased up to three orders of magnitude by using Q-control. This enables AC detection that is immune to the long-term drift of the DC cantilever response in liquids, and to temperature variations. This technique has been applied for the detection of ethanol in aqueous solution by using the microbalance method, and for antibody/antigen recognition by the surface stress method. The results show the feasibility and very high sensitivity of these novel devices.

Nanomechanical mass sensing and stiffness spectrometry based on two-dimensional vibrations of resonant nanowires.

One-dimensional nanomechanical resonators based on nanowires and nanotubes have emerged as promising candidates for mass sensors. When the resonator is clamped at one end and the atoms or molecules being measured land on the other end (which is free to vibrate), the resonance frequency of the device decreases by an amount that is proportional to the mass of the atoms or molecules. However, atoms and molecules can land at any position along the resonator, and many biomolecules have sizes that are comparable to the size of the resonator, so the relationship between the added mass and the frequency shift breaks down. Moreover, whereas resonators fabricated by top-down methods tend to vibrate in just one dimension because they are usually shaped like diving boards, perfectly axisymmetric one-dimensional nanoresonators can support flexural vibrations with the same amplitude and frequency in two dimensions. Here, we propose a new approach to mass sensing and stiffness spectroscopy based on the fact that the nanoresonator will enter a superposition state of two orthogonal vibrations with different frequencies when this symmetry is broken. Measuring these frequencies allows the mass, stiffness and azimuthal arrival direction of the adsorbate to be determined.

Development of nanomechanical biosensors for detection of the pesticide DDT.

We report the use of a novel technique for detection of the organochlorine insecticide compound dichlorodiphenyltrichloroethane (DDT) by measuring the nanometer-scale bending of a microcantilever produced by differential surface stress. A synthetic hapten of the pesticide conjugated with bovine serum albumin (BSA) was covalently immobilised on the gold-coated side of the cantilever by using thiol self assembled monolayers. The immobilisation process is characterised by monitoring the cantilever deflection in real-time. Then specific detection is achieved by exposing the cantilever to a solution of a specific monoclonal antibody to the DDT hapten derivative. The specific binding of the antibodies on the cantilever sensitised side is measured with nanomolar sensitivity. Direct detection is proved by performing competitive assays, in which the cantilever is exposed to a mixed solution of the monoclonal antibody and DDT. The future prospects and limitations to be overcome for the application of nanomechanical sensors for pesticide detection are discussed.

Piconewton regime dynamic force microscopy in liquid

In this work, a cantilever in a fluid is driven by a mixed signal composed of a standard driving signal and a feedback signal consisting of the amplified and phase shifted oscillation signal. This mimics the oscillation of a cantilever with a quality factor up to three orders of magnitude higher than its natural Q (∼1). This technique allows the identification of the resonance frequency of the cantilever by mechanical excitation of the fluid. The improved sensitivity has been checked by imaging a very soft sample of 1% agarose gel in the dynamic mode. A force smaller than 50 pN could be applied to the sample, improving the spatial resolution and the phase contrast significantly. This technique provides a major improvement in atomic force microscopy/spectroscopy in liquids.

Effect of the adsorbate stiffness on the resonance response of microcantilever sensors

The authors present a theoretical model to predict the resonance frequency shift due to molecule adsorption on micro- and nanocantilevers. They calculate the frequency shift experienced by cantilevers made of either silicon or the polymer SU-8, when two adsorbates, myosin protein and an alkanethiol, are attached to the cantilever surface. They demonstrate that the effect of the adsorbate stiffness can be comparable or even larger than the mass effect, producing positive frequency shifts. The results provide methods for decoupling both opposite effects and routes for the design of resonators with high sensitivity to molecule adsorption based on either stiffness or mass effects.

High-Q Dynamic Force Microscopy in Liquid and Its Application to Living Cells

We present a new dynamic force microscopy technique for imaging in liquids in the piconewton regime. The low quality factor (Q) of the cantilever is increased up to three orders of magnitude by the implementation of a positive feedback control. The technique also includes a phase-locked loop unit to track the resonance of the cantilever. Experiments and computer simulations indicate that the tip-sample forces are below 100 pN, about two orders of magnitude lower than in conventional tapping mode atomic force microscopy. Furthermore, the spectroscopic ability is greatly enhanced. Either the phase shift or the resonant frequency shows a high sensitivity to variations in either the energy dissipation or conservative interactions between the tip and the sample, respectively. The potential of this technique is demonstrated by imaging living cells.

Phase contrast and surface energy hysteresis in tapping mode scanning force microsopy

Phase imaging is one of the most attractive features of tapping mode scanning force microscopy operation. In this paper we analyse the relationship between phase contrast imaging and the energy loss due to tip-sample interaction forces. An analytical relationship is obtained between the phase shift and the energy loss. Experiments performed on graphite are in agreement with the analytical expression.