Montserrat Calleja

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.

PATTERNING OF SILICON SURFACES WITH NONCONTACT ATOMIC FORCE MICROSCOPY : FIELD-INDUCED FORMATION OF NANOMETER-SIZE WATER BRIDGES

Nanometer-size water bridges have been used to confine the oxidation of silicon surfaces with a noncontact atomic force microscope. The formation of a water bridge between two surfaces separated by a gap of a few nanometers is driven by the application of an electrical field. Once a liquid bridge is formed, its length and neck diameter can be modified by changing the tip-sample separation. The liquid bridge provides the ionic species and the spatial confinement to pattern Si(100) surfaces in noncontact force microscopy. The method is applied to write arrays of several thousands dots with a periodicity of 40 nm and an average width of 10 nm.

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.

Local oxidation of silicon surfaces by dynamic force microscopy: Nanofabrication and water bridge formation

Local oxidation of silicon surfaces by atomic force microscopy is a very promising lithographic approach at nanometer scale. Here, we study the reproducibility, voltage dependence, and kinetics when the oxidation is performed by dynamic force microscopy modes. It is demonstrated that during the oxidation, tip and sample are separated by a gap of a few nanometers. The existence of a gap increases considerably the effective tip lifetime for performing lithography. A threshold voltage between the tip and the sample must be applied in order to begin the oxidation. The existence of a threshold voltage is attributed to the formation of a water bridge between tip and sample. It is also found that the oxidation kinetics is independent of the force microscopy mode used (contact or noncontact).

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.

Origin of the response of nanomechanical resonators to bacteria adsorption

Resonant microcantilevers are being actively investigated as sensitive mass sensors for biological detection. By performing experiments of adsorption of the bacteria Escherichia coli on singly clamped microcantilevers, we demonstrate that the effect of the added mass is not the only and may not be the main origin of the response of these sensors. The experiments show that the magnitude and sign of resonance frequency shift both depend critically on the distribution of the adsorbed bacterial cells on the cantilever. We relate this behavior to the added mass that shifts the resonance to lower frequencies and the higher effective flexural rigidity of the cantilever due to the bacteria stiffness that shifts the resonance to higher frequencies. Both effects can be uncoupled by positioning the cells where each effect dominates, near the free cantilever end for measuring the added mass or near the clamping for measuring the increase of flexural rigidity.

Nano-oxidation of silicon surfaces by noncontact atomic-force microscopy: Size dependence on voltage and pulse duration

Local oxidation of silicon surfaces by noncontact atomic-force microscopy is an emerging and promising method for patterning surfaces at the nanometer scale due to its very precise control of the feature size. Here, we study the voltage and pulse duration conditions to generate a motive of a given height with the minimum lateral size. We find that for a fixed tip–sample separation, the combination of short pulses and relatively high voltages (∼20 V) produces the highest height:width ratio. The application of relatively high voltages produces a fast growth rate in the vertical direction while the lateral diffusion of oxyanions is inhibited for short pulses. The above results are applied to generate lines of tens of microns in length with an average width at half maximum of about 10 nm.

Photothermal excitation of microcantilevers in liquids

We report the selective excitation of the flexural modes of microcantilevers in aqueous solutions, by applying the photothermal excitation technique. The experiments show that a particular vibration mode can be efficiently excited by focusing the intensity-modulated laser beam on regions of high curvature of the vibration shape. In addition, the resulting resonant peaks in liquid appear distorted by an amplitude component that decreases with the frequency. This distortion produces a shift of the resonance to lower frequencies. A theoretical model based on the transformation of optical energy into mechanical energy via an intermediate thermal stage is proposed to interpret the experimental results. The theory shows that the driven oscillation of the cantilever depends on the curvature of the eigenmode at the excitation position and the heating induced by the excitation laser, which decreases with the frequency. The results reported here set the basis for efficient excitation of high vibration modes in liqu...

Microfabricated photoplastic cantilever with integrated photoplastic/carbon based piezoresistive strain sensor

We present an SU-8 micrometer sized cantilever strain sensor with an integrated piezoresistor made of a conductive composite of SU-8 polymer and carbon black particles. The composite has been developed using ultrasonic mixing. Cleanroom processing of the polymer composite has been investigated and it has been shown that it is possible to pattern the composite by standard UV photolithography. The composite material has been integrated into an SU-8 microcantilever and the polymer composite has been demonstrated to be piezoresistive with gauge factors around 15–20. Since SU-8 is much softer than silicon and the gauge factor of the composite material is relatively high, this polymer based strain sensor is more sensitive than a similar silicon based cantilever sensor.

Low-noise polymeric nanomechanical biosensors

A sensor device based on a single polymer cantilever and optical readout has been developed for detection of molecular recognition reactions without the need of a reference cantilever for subtraction of unspecific signals. Microcantilevers have been fabricated in the photoresist SU-8 with one surface passivated with a thin fluorocarbon layer. The SU-8 surface is sensitized with biological receptors by applying silanization methods, whereas the fluorocarbon surface remains inert to these processes. The thermal and mechanical properties of the chosen materials allow overcoming the main limitations of gold-coated silicon cantilevers: the temperature, pH, and ionic strength cross sensitivities. This is demonstrated by comparing the response of SU-8 cantilevers and that of gold-coated silicon nitride cantilevers to variations in temperature and pH. The sensitivity of the developed polymeric nanomechanical sensor is demonstrated by real-time detection of the human growth hormone with sensitivity in differential...