Ricardo Garcia

Dynamic atomic force microscopy methods

Abstract In this report we review the fundamentals, applications and future tendencies of dynamic atomic force microscopy (AFM) methods. Our focus is on understanding why the changes observed in the dynamic properties of a vibrating tip that interacts with a surface make possible to obtain molecular resolution images of membrane proteins in aqueous solutions or to resolve atomic-scale surface defects in ultra high vacuum (UHV). Our description of the two major dynamic AFM modes, amplitude modulation atomic force microscopy (AM-AFM) and frequency modulation atomic force microscopy (FM-AFM) emphasises their common points without ignoring the differences in experimental set-ups and operating conditions. Those differences are introduced by the different feedback parameters, oscillation amplitude in AM-AFM and frequency shift and excitation amplitude in FM-AFM, used to track the topography and composition of a surface. The theoretical analysis of AM-AFM (also known as tapping-mode) emphasises the coexistence, in many situations of interests, of two stable oscillation states, a low and high amplitude solution. The coexistence of those oscillation states is a consequence of the presence of attractive and repulsive components in the interaction force and their non-linear dependence on the tip–surface separation. We show that key relevant experimental properties such as the lateral resolution, image contrast and sample deformation are highly dependent on the oscillation state chosen to operate the instrument. AM-AFM allows to obtain simultaneous topographic and compositional contrast in heterogeneous samples by recording the phase angle difference between the external excitation and the tip motion (phase imaging). Significant applications of AM-AFM such as high-resolution imaging of biomolecules and polymers, large-scale patterning of silicon surfaces, manipulation of single nanoparticles or the fabrication of single electron devices are also reviewed. FM-AFM (also called non-contact AFM—NC-AFM) has achieved the long-standing goal of true atomic resolution with AFM in UHV. Our analysis starts with a discussion of the relation between frequency shifts and tip–surface interactions, emphasising the ability of perturbation theory to describe the measured frequency shift. We discuss the role of short-range chemical interactions in the atomic contrast, with particular attention to semiconductor and ionic (alkali halides and oxides) surfaces. Also included is a detailed quantitative comparison between theoretical simulations and experiment. Inversion procedures, the determination of the tip–sample interaction from the frequency shift versus distance curves above specific sites, are also reviewed. We finish with a discussion of the optimal range of experimental operation parameters, and the use of damping (excitation amplitude) as a source of atomic contrast, including the possible interpretation in terms of microscopic dissipation mechanisms.

Biomechanical Remodeling of the Microenvironment by Stromal Caveolin-1 Favors Tumor Invasion and Metastasis

Mechanotransduction is a key determinant of tissue homeostasis and tumor progression. It is driven by intercellular adhesions, cell contractility, and forces generated within the microenvironment and is dependent on extracellular matrix composition, organization, and compliance. We show that caveolin-1 (Cav1) favors cell elongation in three-dimensional cultures and promotes Rho- and force-dependent contraction, matrix alignment, and microenvironment stiffening through regulation of p190RhoGAP. In turn, microenvironment remodeling by Cav1 fibroblasts forces cell elongation. Cav1-deficient mice have disorganized stromal tissue architecture. Stroma associated with human carcinomas and melanoma metastases is enriched in Cav1-expressing carcinoma-associated fibroblasts (CAFs). Cav1 expression in breast CAFs correlates with low survival, and Cav1 depletion in CAFs decreases CAF contractility. Consistently, fibroblast expression of Cav1, through p190RhoGAP regulation, favors directional migration and invasiveness of carcinoma cells in vitro. In vivo, stromal Cav1 remodels peri- and intratumoral microenvironments to facilitate tumor invasion, correlating with increased metastatic potency. Thus, Cav1 modulates tissue responses through force-dependent architectural regulation of the microenvironment.

The emergence of multifrequency force microscopy

In atomic force microscopy a cantilever with a sharp tip attached to it is scanned over the surface of a sample, and information about the surface is extracted by measuring how the deflection of the cantilever - which is caused by interactions between the tip and the surface - varies with position. In the most common form of atomic force microscopy, dynamic force microscopy, the cantilever is made to vibrate at a specific frequency, and the deflection of the tip is measured at this frequency. But the motion of the cantilever is highly nonlinear, and in conventional dynamic force microscopy, information about the sample that is encoded in the deflection at frequencies other than the excitation frequency is irreversibly lost. Multifrequency force microscopy involves the excitation and/or detection of the deflection at two or more frequencies, and it has the potential to overcome limitations in the spatial resolution and acquisition times of conventional force microscopes. Here we review the development of five different modes of multifrequency force microscopy and examine its application in studies of proteins, the imaging of vibrating nanostructures, measurements of ion diffusion and subsurface imaging in cells.

Nano-chemistry and scanning probe nanolithographies.

The development of nanometer-scale lithographies is the focus of an intense research activity because progress on nanotechnology depends on the capability to fabricate, position and interconnect nanometer-scale structures. The unique imaging and manipulation properties of atomic force microscopes have prompted the emergence of several scanning probe-based nanolithographies. In this tutorial review we present the most promising probe-based nanolithographies that are based on the spatial confinement of a chemical reaction within a nanometer-size region of the sample surface. The potential of local chemical nanolithography in nanometer-scale science and technology is illustrated by describing a range of applications such as the fabrication of conjugated molecular wires, optical microlenses, complex quantum devices or tailored chemical surfaces for controlling biorecognition processes.

Advanced scanning probe lithography

The nanoscale control afforded by scanning probe microscopes has prompted the development of a wide variety of scanning-probe-based patterning methods. Some of these methods have demonstrated a high degree of robustness and patterning capabilities that are unmatched by other lithographic techniques. However, the limited throughput of scanning probe lithography has prevented its exploitation in technological applications. Here, we review the fundamentals of scanning probe lithography and its use in materials science and nanotechnology. We focus on robust methods, such as those based on thermal effects, chemical reactions and voltage-induced processes, that demonstrate a potential for applications.

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.

Lane-Change Fuzzy Control in Autonomous Vehicles for the Overtaking Maneuver

The automation of the overtaking maneuver is considered to be one of the toughest challenges in the development of autonomous vehicles. This operation involves two vehicles (the overtaking and the overtaken) cooperatively driving, as well as the surveillance of any other vehicles that are involved in the maneuver. This operation consists of two lane changes-one from the right to the left lane of the road, and the other is to return to the right lane after passing. Lane-change maneuvers have been used to move into or out of a circulation lane or platoon; however, overtaking operations have not received much coverage in the literature. In this paper, we present an overtaking system for autonomous vehicles equipped with path-tracking and lane-change capabilities. The system uses fuzzy controllers that mimic human behavior and reactions during overtaking maneuvers. The system is based on the information that is supplied by a high-precision Global Positioning System and a wireless network environment. It is able to drive an automated vehicle and overtake a second vehicle that is driving in the same lane of the road.

Compositional mapping of surfaces in atomic force microscopy by excitation of the second normal mode of the microcantilever

In this letter we study the tip motion of a rectangular microcantilever in the proximity of a surface. The microcantilever has a hemispherical tip attached at its free end. The theoretical simulations led us to propose a method for mapping simultaneously the topography and the chemical composition of a sample surface in noncontact AM‐AFM. The method consists of exciting the first two modes of the microcantilever. The output signal of the first mode is used to image the topography of the sample while the second mode is used to map changes in the composition of the atoms or molecules under the tip. The simulations were performed by modeling the three dimensional microcantilever as a rectangular beam and applying the Newton equation. 25 Then the dynamic deflection function w(x,t) is described by

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.