Preventing chaotic motion in tapping-mode atomic force microscope

During the last 30 years the Atomic Force Microscopy became the most powerful tool for surface probing in atomic scale. The Tapping-Mode Atomic Force Microscope is used to generate high quality accurate images of the samples surface. However, in this mode of operation the microcantilever frequently presents chaotic motion due to the nonlinear characteristics of the tip-sample forces interactions, degrading the image quality. This kind of irregular motion must be avoided by the control system. In this work, the tip-sample interaction is modelled considering the Lennard-Jones potentials and the two-term Galerkin aproximation. Additionally, the State Dependent Ricatti Equation and Time-Delayed Feedback Control techniques are used in order to force the Tapping-Mode Atomic Force Microscope system motion to a periodic orbit, preventing the microcantilever chaotic motion

Influence of salinity on the interannual heat storage trends in the Atlantic estimated from altimeters and Pilot Research Moored Array in the Tropical Atlantic data

Changes in the oceanic heat storage (HS) can reveal important evidences of climate variability related to ocean heat fluxes. Specifically, long-term variations in HS are a powerful indicator of climate change as HS represents the balance between the net surface energy flux and the poleward heat transported by the ocean currents. HS is estimated from sea surface height anomaly measured from the altimeters TOPEX/Poseidon and Jason 1 from 1993 to 2006. To characterize and validate the altimeter-based HS in the Atlantic, we used the data from the Pilot Research Moored Array in the Tropical Atlantic (PIRATA) array. Correlations and rms differences are used as statistical figures of merit to compare the HS estimates. The correlations range from 0.50 to 0.87 in the buoys located at the equator and at the southern part of the array. In that region the rms differences range between 0.40 and 0.51 x 10(9) Jm(-2). These results are encouraging and indicate that the altimeter has the precision necessary to capture the interannual trends in HS in the Atlantic. Albeit relatively small, salinity changes can also have an effect on the sea surface height anomaly. To account for this effect, NCEP/GODAS reanalysis data are used to estimate the haline contraction. To understand which dynamical processes are involved in the HS variability, the total signal is decomposed into nonpropagating basin-scale and seasonal (HS(l)) planetary waves, mesoscale eddies, and small-scale residual components. In general, HS(l) is the dominant signal in the tropical region. Results show a warming trend of HS(l) in the past 13 years almost all over the Atlantic basin with the most prominent slopes found at high latitudes. Positive interannual trends are found in the halosteric component at high latitudes of the South Atlantic and near the Labrador Sea. This could be an indication that the salinity anomaly increased in the upper layers during this period. The dynamics of the South Atlantic subtropical gyre could also be subject to low-frequency changes caused by a trend in the halosteric component on each side of the South Atlantic Current.

Formation of an extended CoSi2 thin nanohexagons array coherently buried in silicon single crystal

A Co-doped silica film was deposited on the surface of a Si(100) wafer and isothermally annealed at 750 degrees C to form spherical Co nanoparticles embedded in the silica film and a few atomic layer thick CoSi2 nanoplatelets within the wafer. The structure, morphology, and spatial orientation of the nanoplatelets were characterized. The experimental results indicate that the nanoplatelets exhibit hexagonal shape and a uniform thickness. The CoSi2 nanostructures lattice is coherent with the Si lattice, and each of them is parallel to one of the four planes belonging to the {111} crystallographic form of the host lattice. (C) 2012 American Institute of Physics. [doi:10.1063/1.3683493]

Phase-Locked loops lock-in range in Frequency Modulated-Atomic Force Microscope nonlinear control system

Since the mid 1980s the Atomic Force Microscope is one the most powerful tools to perform surface investigation, and since 1995 Non-Contact AFM achieved true atomic resolution. The Frequency-Modulated Atomic Force Microscope (FM-AFM) operates in the dynamic mode, which means that the control system of the FM-AFM must force the micro-cantilever to oscillate with constant amplitude and frequency. However, tip-sample interaction forces cause modulations in the microcantilever motion. A Phase-Locked loop (PLL) is used to demodulate the tip-sample interaction forces from the microcantilever motion. The demodulated signal is used as the feedback signal to the control system, and to generate both topographic and dissipation images. As a consequence, a proper design of the PLL is vital to the FM-AFM performance. In this work, using bifurcation analysis, the lock-in range of the PLL is determined as a function of the frequency shift (Q) of the microcantilever and of the other design parameters, providing a technique to properly design the PLL in the FM-AFM system. (C) 2011 Elsevier B.V. All rights reserved.

MSX2 copy number increase and craniosynostosis: copy number variation detected by array comparative genomic hybridization

Fleury Group

Microelectrode array in mixed alkanethiol self-assembled monolayers: Electrochemical studies

We report an efficient alternative to obtain recessed microelectrodes device on gold electrode surface, in which mixed self-assembled monolayer of long and short carbon alkanethiol chains was used for this purpose. Development of the modified electrodes included the chemical adsorption of 11-mercaptoundecanoic acid and 2-mercaptoethanol solution, as well as their mixtures, on gold surface, resulting in the final mixed self-assembled monolayer configuration. For comparison, the electrochemical performance of self-assembled monolayer of 11-mercaptoundecanoic acid. 3-mercaptopropionic acid, 4-mercapto-1-butanol and 6-mercapto-1-hexanol modified electrodes was also investigated. It was verified that, in the mixed self-assembled monolayer, the 11-mercaptoundecanoic acid acts as a barrier for electron transfer while the short alkanethiol chair is deposited in an island-like shape through which electrons can be freely transferred to ions in solution, allowing electrochemical reactions to occur. The performance of the modified electrodes toward microelectrode behavior was investigated via cyclic voltammetry and electrochemical impedance spectroscopy measurements using [Fe(CN)(6)](3-/4-) redox couple as a probe. In this case, sigmoidal voltammetric responses were obtained, very similar to those observed for microelectrodes. Such behavior reinforces the proposition of electron transfer through the short alkanethiol chain layer and surface blockage by the long chain one. Electrochemical impedance results allowed calculated the mean radius value of each microelectrode disks of 3.8 mu m with about 22 mu m interval between them. The microelectrode environment provided by the mixed self-assembled monolayer can be conveniently used to provide an efficient catalytic conversion in biosensing applications. (C) 2012 Elsevier Ltd. All rights reserved.

Development of an AMR-ACB Array for Gastrointestinal Motility Studies

The association between anisotropic magnetoresistive (AMR) sensor and AC biosusceptometry (ACB) to evaluate gastrointestinal motility is presented. The AMR-ACB system was successfully characterized in a bench-top study, and in vivo results were compared with those obtained by means of simultaneous manometry. Both AMR-ACB and manometry techniques presented high temporal cross correlation between the two periodicals signals (R = 0.9 +/- 0.1; P < 0.05). The contraction frequencies using AMR-ACB were 73.9 +/- 7.6 mHz and using manometry were 73.8 +/- 7.9 mHz during the baseline (r = 98, p < 0.05). The amplitude of contraction using AMR-ACB was 396 +/- 108 mu T.s and using manometry were 540 +/- 198 mmHg.s during the baseline. The amplitudes of signals for AMR-ACB and manometric recordings were similarly increased to 86.4% and 89.3% by neostigmine, and also decreased to 27.2% and 21.4% by hyoscine butylbromide in all animals, respectively. The AMR-ACB array is nonexpensive, portable, and has high-spatiotemporal resolution to provide helpful information about gastrointestinal tract.

A complete CMOS UWB timed-array transmitter with a 3D Vivaldi antenna array for electronic high-resolution beam spatial scanning

We present a new Ultra Wide Band (UWB) Timed- Array Transmitter System with Beamforming capability for high-resolution remote acquisition of vital signals. The system consists of four identical channels, where each is formed of a serial topology with three modules: programmable delay circuit (PDC or τ), a novel UWB 5th Gaussian Derivative order pulse generator circuit (PG), and a planar Vivaldi antenna. The circuit was designed using 0.18μm CMOS standard process and the planar antenna array was designed with filmconductor on Rogers RO3206 substrate. Spice simulations results showed the pulse generation with 104 mVpp amplitude and 500 ps width. The power consumption is 543 μW, and energy consumption 0.27 pJ per pulse using a 2V power supply at a pulse repetition rate (PRR) of 100 MHz. Electromagnetic simulations results, using CST Microwave (MW) Studio 2011, showed the main lobe radiation with a gain maximum of 13.2 dB, 35.5º x 36.7º angular width, and a beam steering between 17º and -11º for azimuthal (θ) angles and 17º and -18º for elevation (φ) angles at the center frequency of 6 GHz

Application of FTIR microscopy to cultural heritage materials

Research in art conservation has been developed from the early 1950s, giving a significant contribution to the conservation-restoration of cultural heritage artefacts. In fact, only through a profound knowledge about the nature and conditions of constituent materials, suitable decisions on the conservation and restoration measures can thus be adopted and preservation practices enhanced. The study of ancient artworks is particularly challenging as they can be considered as heterogeneous and multilayered systems where numerous interactions between the different components as well as degradation and ageing phenomena take place. However, difficulties to physically separate the different layers due to their thickness (1-200 µm) can result in the inaccurate attribution of the identified compounds to a specific layer. Therefore, details can only be analysed when the sample preparation method leaves the layer structure intact, as for example the preparation of embedding cross sections in synthetic resins. Hence, spatially resolved analytical techniques are required not only to exactly characterize the nature of the compounds but also to obtain precise chemical and physical information about ongoing changes. This thesis focuses on the application of FTIR microspectroscopic techniques for cultural heritage materials. The first section is aimed at introducing the use of FTIR microscopy in conservation science with a particular attention to the sampling criteria and sample preparation methods. The second section is aimed at evaluating and validating the use of different FTIR microscopic analytical methods applied to the study of different art conservation issues which may be encountered dealing with cultural heritage artefacts: the characterisation of the artistic execution technique (chapter II-1), the studies on degradation phenomena (chapter II-2) and finally the evaluation of protective treatments (chapter II-3). The third and last section is divided into three chapters which underline recent developments in FTIR spectroscopy for the characterisation of paint cross sections and in particular thin organic layers: a newly developed preparation method with embedding systems in infrared transparent salts (chapter III-1), the new opportunities offered by macro-ATR imaging spectroscopy (chapter III-2) and the possibilities achieved with the different FTIR microspectroscopic techniques nowadays available (chapter III-3). In chapter II-1, FTIR microspectroscopy as molecular analysis, is presented in an integrated approach with other analytical techniques. The proposed sequence is optimized in function of the limited quantity of sample available and this methodology permits to identify the painting materials and characterise the adopted execution technique and state of conservation. Chapter II-2 describes the characterisation of the degradation products with FTIR microscopy since the investigation on the ageing processes encountered in old artefacts represents one of the most important issues in conservation research. Metal carboxylates resulting from the interaction between pigments and binding media are characterized using synthesised metal palmitates and their production is detected on copper-, zinc-, manganese- and lead- (associated with lead carbonate) based pigments dispersed either in oil or egg tempera. Moreover, significant effects seem to be obtained with iron and cobalt (acceleration of the triglycerides hydrolysis). For the first time on sienna and umber paints, manganese carboxylates are also observed. Finally in chapter II-3, FTIR microscopy is combined with further elemental analyses to characterise and estimate the performances and stability of newly developed treatments, which should better fit conservation-restoration problems. In the second part, in chapter III-1, an innovative embedding system in potassium bromide is reported focusing on the characterisation and localisation of organic substances in cross sections. Not only the identification but also the distribution of proteinaceous, lipidic or resinaceous materials, are evidenced directly on different paint cross sections, especially in thin layers of the order of 10 µm. Chapter III-2 describes the use of a conventional diamond ATR accessory coupled with a focal plane array to obtain chemical images of multi-layered paint cross sections. A rapid and simple identification of the different compounds is achieved without the use of any infrared microscope objectives. Finally, the latest FTIR techniques available are highlighted in chapter III-3 in a comparative study for the characterisation of paint cross sections. Results in terms of spatial resolution, data quality and chemical information obtained are presented and in particular, a new FTIR microscope equipped with a linear array detector, which permits reducing the spatial resolution limit to approximately 5 µm, provides very promising results and may represent a good alternative to either mapping or imaging systems.

DNA-Array-Technologie

Es wurde ein genomischer DNA-Array der Modellpflanze Arabidopsis thaliana mit einer 13.800 EST-Klone umfassenden cDNA-Bibliothek entwickelt und in der Genexpressionsanalyse der pflanzlichen Pathogenabwehr eingesetzt. Mittels PCR-Amplifikation sind 13.000 PCR-Produkte der cDNA-Fragmente hergestellt worden, mit denen 66 genomische Arabidopsis-Arrays auf Nylon und Polypropylen als Trägermaterial hergestellt werden konnten. Die Validierung mit Fluoreszenz- und Radiaktivhybridisierung sowie der Vergleich von drei Normalisierungsmethoden führte zu reproduzierbaren Ergebnissen bei hohem Korrelationskoeffizienten. Die etablierte DNA-Array-Technologie wurde zur Genexpressionsanalyse der pathogeninduzierten Abwehrmechanismen der Pflanze Arabidopsis thaliana in den ersten 24 Stunden nach Infektion mit dem avirulenten Bakterium Pseudomonas syringae pv. tomato eingesetzt. In einer Auswahl von 75 Genen der Stoffwechselwege Glycolyse, Citrat-Cyclus, Pentosephosphat-Cyclus und Glyoxylatmetabolismus konnte für 25 % der Gene, im Shikimat-, Tryptophan- und Phenylpropanoidsyntheseweg für 60 % der Gene eine erhöhte Transkriptionsrate nachgewiesen werden. Die Ergebnisse dieser Arbeit stimmen mit experimentellen Daten verschiedener unabhängiger Studien zur pflanzlichen Pathogenantwort überein. Darüberhinaus sind erstmals Transkriptionsprofile von bisher auf Transkriptionsebene nicht untersuchten Genen erstellt worden. Diese Ergebnisse bestätigen die transkriptionelle Aktivierung ganzer Stoffwechselwege und gewähren erstmals einen Einblick in die koordinierte differentielle Transkription ganzer Stoffwechselwege während der Pathogenabwehr.

Ruolo dei glicosaminoglicani su struttura e funzione dei crimps nei tendini e nei legamenti

The biomechanical roles of both tendons and ligaments are fulfilled by extracellular matrix of these tissues. In particular, tension is mainly transmitted and resisted by fibrous proteins (collagen, elastin), whereas compressive load is absorbed by water-soluble glycosaminoglycans (GAGs). GAGs spanning the interfibrillar spaces and interacting with fibrils also seem to play a part in transmitting and resisting tensile stresses. Apart from different functional roles and collagen array, tendons and ligaments share the same basic structure showing periodic undulations of collagen fibers or crimps. Each crimp is composed of many knots of each single fibril or fibrillar crimps. Fibrillar and fiber crimps act as shock absorbers during the initial elongation of both tendons and ligaments and assist the elastic recoil of fibrils and fibers when the tensile stress is removed. The aim of this thesis was to evaluate whether GAGs directly affect the 3D microstructural integrity of fibrillar crimp and fiber crimps in both tendons and ligaments. Achilles tendons and medial collateral ligaments of the knee from eight female Sprague-Dawley rats (90 days old) were digested with chondroitinase ABC to remove GAGs and observed under a scanning electron microscope (SEM). In addition, isolated fibrils from these tissues obtained by mechanical homogenization were analyzed by a transmission electron microscope (TEM). Both samples digested with chondroitinase ABC or mechanically disrupted still showed crimps and fibrillar crimps comparable to tissues with a normal GAGs content. All fibrils in the fibrillar crimp region always twisted leftwards, thus changing their running plane, and then sharply bent, changing their course on a new plane. These data suggest that GAGs do not affect structural integrity or fibrillar crimps functions that seem mainly related to the local fibril leftward twisting and the alternating handedness of collagen from a molecular to a supramolecular level.

Continuos Flow Single Cell Separation into Open Microwell Arrays

A novel design based on electric field-free open microwell arrays for the automated continuous-flow sorting of single or small clusters of cells is presented. The main feature of the proposed device is the parallel analysis of cell-cell and cell-particle interactions in each microwell of the array. High throughput sample recovery with a fast and separate transfer from the microsites to standard microtiter plates is also possible thanks to the flexible printed circuit board technology which permits to produce cost effective large area arrays featuring geometries compatible with laboratory equipment. The particle isolation is performed via negative dielectrophoretic forces which convey the particles’ into the microwells. Particles such as cells and beads flow in electrically active microchannels on whose substrate the electrodes are patterned. The introduction of particles within the microwells is automatically performed by generating the required feedback signal by a microscope-based optical counting and detection routine. In order to isolate a controlled number of particles we created two particular configurations of the electric field within the structure. The first one permits their isolation whereas the second one creates a net force which repels the particles from the microwell entrance. To increase the parallelism at which the cell-isolation function is implemented, a new technique based on coplanar electrodes to detect particle presence was implemented. A lock-in amplifying scheme was used to monitor the impedance of the channel perturbed by flowing particles in high-conductivity suspension mediums. The impedance measurement module was also combined with the dielectrophoretic focusing stage situated upstream of the measurement stage, to limit the measured signal amplitude dispersion due to the particles position variation within the microchannel. In conclusion, the designed system complies with the initial specifications making it suitable for cellomics and biotechnology applications.

High resolution study of local stress inside alumina - micro mechanical analysis using laser scanning confocal microscope

The aim of this work is to measure the stress inside a hard micro object under extreme compression. To measure the internal stress, we compressed ruby spheres (a-Al2O3: Cr3+, 150 µm diameter) between two sapphire plates. Ruby fluorescence spectrum shifts to longer wavelengths under compression and can be related to the internal stress by a conversion coefficient. A confocal laser scanning microscope was used to excite and collect fluorescence at desired local spots inside the ruby sphere with spatial resolution of about 1 µm3. Under static external loads, the stress distribution within the center plane of the ruby sphere was measured directly for the first time. The result agreed to Hertz’s law. The stress across the contact area showed a hemispherical profile. The measured contact radius was in accord with the calculation by Hertz’s equation. Stress-load curves showed spike-like decrease after entering non-elastic phase, indicating the formation and coalescence of microcracks, which led to relaxing of stress. In the vicinity of the contact area luminescence spectra with multiple peaks were observed. This indicated the presence of domains of different stress, which were mechanically decoupled. Repeated loading cycles were applied to study the fatigue of ruby at the contact region. Progressive fatigue was observed when the load exceeded 1 N. As long as the load did not exceed 2 N stress-load curves were still continuous and could be described by Hertz’s law with a reduced Young’s modulus. Once the load exceeded 2 N, periodical spike-like decreases of the stress could be observed, implying a “memory effect” under repeated loading cycles. Vibration loading with higher frequencies was applied by a piezo. Redistributions of intensity on the fluorescence spectra were observed and it was attributed to the repopulation of the micro domains of different elasticity. Two stages of under vibration loading were suggested. In the first stage continuous damage carried on until certain limit, by which the second stage, e.g. breakage, followed in a discontinuous manner.

Surface-plasmon optical and electrochemical characterization of biofunctional surface architectures

The development and characterization of biomolecule sensor formats based on the optical technique Surface Plasmon Resonance (SPR) Spectroscopy and electrochemical methods were investigated. The study can be divided into two parts of different scope. In the first part new novel detection schemes for labeled targets were developed on the basis of the investigations in Surface-plamon Field Enhanced Spectroscopy (SPFS). The first one is SPR fluorescence imaging formats, Surface-plamon Field Enhanced Fluorescence Microscopy (SPFM). Patterned self assembled monolayers (SAMs) were prepared and used to direct the spatial distribution of biomolecules immobilized on surfaces. Here the patterned monolayers would serve as molecular templates to secure different biomolecules to known locations on a surface. The binding processed of labeled target biomolecules from solution to sensor surface were visually and kinetically recorded by the fluorescence microscope, in which fluorescence was excited by the evanescent field of propagating plasmon surface polaritons. The second format which also originates from SPFS technique, Surface-plamon Field Enhanced Fluorescence Spectrometry (SPFSm), concerns the coupling of a fluorometry to normal SPR setup. A spectrograph mounted in place of photomultiplier or microscope can provide the information of fluorescence spectrum as well as fluorescence intensity. This study also firstly demonstrated the analytical combination of surface plasmon enhanced fluorescence detection with analyte tagged by semiconducting nano- crystals (QDs). Electrochemically addressable fabrication of DNA biosensor arrays in aqueous environment was also developed. An electrochemical method was introduced for the directed in-situ assembly of various specific oligonucleotide catcher probes onto different sensing elements of a multi-electrode array in the aqueous environment of a flow cell. Surface plasmon microscopy (SPM) is utilized for the on-line recording of the various functionalization steps. Hybridization reactions between targets from solution to the different surface-bound complementary probes are monitored by surface-plasmon field-enhanced fluorescence microscopy (SPFM) using targets that are either labeled with organic dyes or with semiconducting quantum dots for color-multiplexing. This study provides a new approach for the fabrication of (small) DNA arrays and the recording and quantitative evaluation of parallel hybridization reactions. In the second part of this work, the ideas of combining the SP optical and electrochemical characterization were extended to tethered bilayer lipid membrane (tBLM) format. Tethered bilayer lipid membranes provide a versatile model platform for the study of many membrane related processes. The thiolipids were firstly self-assembled on ultraflat gold substrates. Fusion of the monolayers with small unilamellar vesicles (SUVs) formed the distal layer and the membranes thus obtained have the sealing properties comparable to those of natural membranes. The fusion could be monitored optically by SPR as an increase in reflectivity (thickness) upon formation of the outer leaflet of the bilayer. With EIS, a drop in capacitance and a steady increase in resistance could be observed leading to a tightly sealing membrane with low leakage currents. The assembly of tBLMs and the subsequent incorporation of membrane proteins were investigated with respect to their potential use as a biosensing system. In the case of valinomycin the potassium transport mediated by the ion carrier could be shown by a decrease in resistance upon increasing potassium concentration. Potential mediation of membrane pores could be shown for the ion channel forming peptide alamethicin (Alm). It was shown that at high positive dc bias (cis negative) Alm channels stay at relatively low conductance levels and show higher permeability to potassium than to tetramethylammonium. The addition of inhibitor amiloride can partially block the Alm channels and results in increase of membrane resistance. tBLMs are robust and versatile model membrane architectures that can mimic certain properties of biological membranes. tBLMs with incorporated lipopolysaccharide (LPS) and lipid A mimicking bacteria membranes were used to probe the interactions of antibodies against LPS and to investigate the binding and incorporation of the small antimicrobial peptide V4. The influence of membrane composition and charge on the behavior of V4 was also probed. This study displays the possibility of using tBLM platform to record and valuate the efficiency or potency of numerous synthesized antimicrobial peptides as potential drug candidates.