Álvaro San Paulo

High-resolution imaging of antibodies by tapping-mode atomic force microscopy: attractive and repulsive tip-sample interaction regimes.

A force microscope operated with an amplitude modulation feedback (usually known as tapping-mode atomic force microscope) has two tip-sample interaction regimes, attractive and repulsive. We have studied the performance of those regimes to imaging single antibody molecules. The attractive interaction regime allows determination of the basic morphologies of the antibodies on the support. More importantly, this regime is able to resolve the characteristic Y-shaped domain structure of antibodies and the hinge region between domains. Imaging in the repulsive interaction regime is associated with the irreversible deformation of the molecules. This causes a significant loss in resolution and contrast. Two major physical differences distinguish the repulsive interaction regime from the attractive interaction regime: the existence of tip-sample contact and the strength of the forces involved.

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.

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.

Amplitude curves and operating regimes in dynamic atomic force microscopy

The experimental dependence of the amplitude on the average tip-sample distance has been studied to understand the operation of an atomic force microscope with an amplitude modulation feedback. The amplitude curves can be classified in three major groups according to the existence or not of a local maximum and how the maximum is reached (steplike discontinuities vs. smooth transitions). A model describing the cantilever motion as a forced nonlinear oscillator allows to associate the features observed in the amplitude curves with the tip-sample interaction force. The model also allows to define two elemental tip-sample interaction regimes, attractive and repulsive. The presence of a local maximum in the amplitude curves is related to a transition between the attractive and the repulsive regime.

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 Youngs 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.

Optomechanics with silicon nanowires by harnessing confined electromagnetic modes.

The optomechanical coupling that emerges in an optical cavity in which one of the mirrors is a mechanical resonator has allowed sub-Kelvin cooling with the prospect of observing quantum phenomena and self-sustained oscillators with very high spectral purity. Both applications clearly benefit from the use of the smallest possible mechanical resonator. Unfortunately, the optomechanical coupling largely decays when the size of the mechanical system is below the light wavelength. Here, we propose to exploit the optical resonances associated to the light confinement in subwavelength structures to circumvent this limitation, efficiently extending optomechanics to nanoscale objects. We demonstrate this mechanism with suspended silicon nanowires. We are able to optically cool the mechanical vibration of the nanowires from room temperature to 30-40 K or to obtain regenerative mechanical oscillation with a frequency stability of about one part per million. The reported optomechanical phenomena can be exploited for developing cost-optimized mass sensors with sensitivities in the zeptogram range.

Amplitude, deformation and phase shift in amplitude modulation atomic force microscopy: a numerical study for compliant materials

Numerical simulations were applied to investigate the motion of a tip interacting with a compliant sample. The dependence of the amplitude, deformation, contact time and phase shift on the mechanical properties of the sample, free oscillation amplitude and cantilever force constant were investigated. The compliance and surface adhesion energy favour the formation of an adhesion neck between tip and surface. The neck modifies the tip motion when its length is comparable to the free oscillation amplitude. The simulations also show that the tip may oscillate fully indented on the sample if large force constant cantilevers and amplitudes are used. A good compromise between stability and resolution is achieved by using low force constant cantilevers and large oscillation amplitudes. The agreement obtained between theory and experimental data supports the conclusions of the model.

High-sensitivity linear piezoresistive transduction for nanomechanical beam resonators

Highly sensitive conversion of motion into readable electrical signals is a crucial and challenging issue for nanomechanical resonators. Efficient transduction is particularly difficult to realize in devices of low dimensionality, such as beam resonators based on carbon nanotubes or silicon nanowires, where mechanical vibrations combine very high frequencies with miniscule amplitudes. Here we describe an enhanced piezoresistive transduction mechanism based on the asymmetry of the beam shape at rest. We show that this mechanism enables highly sensitive linear detection of the vibration of low-resistivity silicon beams without the need of exceptionally large piezoresistive coefficients. The general application of this effect is demonstrated by detecting multiple-order modes of silicon nanowire resonators made by either top-down or bottom-up fabrication methods. These results reveal a promising approach for practical applications of the simplest mechanical resonators, facilitating its manufacturability by very large-scale integration technologies.

Silicon nanowires: where mechanics and optics meet at the nanoscale

Mechanical transducers based on nanowires promise revolutionary advances in biological sensing and force microscopy/spectroscopy. A crucial step is the development of simple and non-invasive techniques able to detect displacements with subpicometer sensitivity per unit bandwidth. Here, we design suspended tapered silicon nanowires supporting a range of optical resonances that confine and efficiently scatter light in the visible range. Then, we develop an optical method for efficiently coupling the evanescent field to the regular interference pattern generated by an incoming laser beam and the reflected beam from the substrate underneath the nanowire. This optomechanical coupling is here applied to measure the displacement of 50 nm wide nanowires with sensitivity on the verge of 1 fm/Hz1/2 at room temperature with a simple laser interferometry set-up. This method opens the door to the measurement of the Brownian motion of ultrashort nanowires for the detection of single biomolecular recognition events in liquids, and single molecule spectroscopy in vacuum.

Detection of nanomechanical vibrations by dynamic force microscopy in higher cantilever eigenmodes

The authors present a method based on dynamic force microscopy to characterize subnanometer-scale mechanical vibrations in resonant micro- and nanoelectromechanical systems. The method simultaneously employs the first eigenmode of the microscope cantilever for topography imaging and the second eigenmode for the detection of the resonator vibration. Here, they apply this scheme for the characterization of a 1.6GHz film bulk acoustic resonator, showing that it overcomes the main limitations of acoustic imaging in contact-mode atomic force microscopy. The method provides nanometer-scale lateral resolution on arbitrarily high resonant frequency systems, which makes it applicable to a wide diversity of electromechanical systems.