Atomic force microscopy of biomimetic systems

This thesis was driven by the ambition to create suitable model systems that mimic complex processes in nature, like intramolecular transitions, such as unfolding and refolding of proteins, or intermolecular interactions between different cell compo-nents. Novel biophysical approaches were adopted by employing atomic force mi-croscopy (AFM) as the main measurement technique due to its broad diversity. Thus, high-resolution imaging, adhesion measurements, and single-molecule force distance experiments were performed on the verge of the instrumental capabilities. As first objective, the interaction between plasma membrane and cytoskeleton, me-diated by the linker protein ezrin, was pursued. Therefore, the adsorption process and the lateral organization of ezrin on PIP2 containing solid-supported membranes were characterized and quantified as a fundament for the establishment of a biomimetic model system. As second component of the model system, actin filaments were coated on functionalized colloidal probes attached on cantilevers, serving as sensor elements. The zealous endeavor of creating this complex biomimetic system was rewarded by successful investigation of the activation process of ezrin. As a result, it can be stated that ezrin is activated by solely binding to PIP2 without any further stimulating agents. Additional cofactors may stabilize and prolong the active conformation but are not essentially required for triggering ezrin’s transformation into an active conformation. In the second project, single-molecule force distance experiments were performed on bis-loop tetra-urea calix[4]arene-catenanes with different loading rates (increase in force per second). These macromolecules were specifically designed to investigate the rupture and rejoining mechanism of hydrogen bonds under external load. The entangled loops of capsule-like molecules locked the unbound state of intramolecular hydrogen bonds mechanically, rendering a rebinding observable on the experimental time scale. In conjunction with Molecular Dynamics simulations, a three-well potential of the bond rupture process was established and all kinetically relevant parameters of the experiments were determined by means of Monte Carlo simulations and stochastic modeling. In summary, it can be stated that atomic force microscopy is an invaluable tool to scrutinize relevant processes in nature, such as investigating activation mechanisms in proteins, as shown by analysis of the interaction between F-actin and ezrin, as well as exploring fundamental properties of single hydrogen bonds that are of paramount interest for the complete understanding of complex supramolecular structures.

Measurement of the forward-backward asymmetry of Z boson decays in electron-positron pairs with the ATLAS detector in proton-proton collisions at root out s = 7TeV at the LHC

In this thesis the measurement of the effective weak mixing angle wma in proton-proton collisions is described. The results are extracted from the forward-backward asymmetry (AFB) in electron-positron final states at the ATLAS experiment at the LHC. The AFB is defined upon the distribution of the polar angle between the incoming quark and outgoing lepton. The signal process used in this study is the reaction pp to zgamma + X to ee + X taking a total integrated luminosity of 4.8\,fb^(-1) of data into account. The data was recorded at a proton-proton center-of-mass energy of sqrt(s)=7TeV. The weak mixing angle is a central parameter of the electroweak theory of the Standard Model (SM) and relates the neutral current interactions of electromagnetism and weak force. The higher order corrections on wma are related to other SM parameters like the mass of the Higgs boson.rnrnBecause of the symmetric initial state constellation of colliding protons, there is no favoured forward or backward direction in the experimental setup. The reference axis used in the definition of the polar angle is therefore chosen with respect to the longitudinal boost of the electron-positron final state. This leads to events with low absolute rapidity have a higher chance of being assigned to the opposite direction of the reference axis. This effect called dilution is reduced when events at higher rapidities are used. It can be studied including electrons and positrons in the forward regions of the ATLAS calorimeters. Electrons and positrons are further referred to as electrons. To include the electrons from the forward region, the energy calibration for the forward calorimeters had to be redone. This calibration is performed by inter-calibrating the forward electron energy scale using pairs of a central and a forward electron and the previously derived central electron energy calibration. The uncertainty is shown to be dominated by the systematic variations.rnrnThe extraction of wma is performed using chi^2 tests, comparing the measured distribution of AFB in data to a set of template distributions with varied values of wma. The templates are built in a forward folding technique using modified generator level samples and the official fully simulated signal sample with full detector simulation and particle reconstruction and identification. The analysis is performed in two different channels: pairs of central electrons or one central and one forward electron. The results of the two channels are in good agreement and are the first measurements of wma at the Z resonance using electron final states at proton-proton collisions at sqrt(s)=7TeV. The precision of the measurement is already systematically limited mostly by the uncertainties resulting from the knowledge of the parton distribution functions (PDF) and the systematic uncertainties of the energy calibration.rnrnThe extracted results of wma are combined and yield a value of wma_comb = 0.2288 +- 0.0004 (stat.) +- 0.0009 (syst.) = 0.2288 +- 0.0010 (tot.). The measurements are compared to the results of previous measurements at the Z boson resonance. The deviation with respect to the combined result provided by the LEP and SLC experiments is up to 2.7 standard deviations.

Revealing atomic-scale details in the force fields above carbonate surfaces

Diese Arbeit stellt eine ausführliche Studie fundamentaler Eigenschaften der Kalzit CaCO3(10.4) und verwandter Mineraloberflächen dar, welche nicht nur durch die Verwendung von Nichtkontakt Rasterkraftmikroskopie, sondern hauptsächlich durch die Messung von Kraftfeldern ermöglicht wurde. Die absolute Oberflächenorientierung sowie der hierfür zugrundeliegende Prozess auf atomarer Skala konnten erfolgreich für die Kalzit (10.4) Oberfläche identifiziert werden.rnDie Adsorption chiraler Moleküle auf Kalzit ist relevant im Bereich der Biomineralisation, was ein Verständnis der Oberflächensymmetrie unumgänglich macht. Die Messung des Oberflächenkraftfeldes auf atomarer Ebene ist hierfür ein zentraler Aspekt. Eine solche Kraftkarte beleuchtet nicht nur die für die Biomineralisation wichtige Wechselwirkung der Oberfläche mit Molekülen, sondern enthält auch die Möglichkeit, Prozesse auf atomarer Skala und damit Oberflächeneigenschaften zu identifizieren.rnDie Einführung eines höchst flexiblen Messprotokolls gewährleistet die zuverlässige und kommerziell nicht erhältliche Messung des Oberflächenkraftfeldes. Die Konversion der rohen ∆f Daten in die vertikale Kraft Fz ist jedoch kein trivialer Vorgang, insbesondere wenn Glätten der Daten in Frage kommt. Diese Arbeit beschreibt detailreich, wie Fz korrekt für die experimentellen Bedingungen dieser Arbeit berechnet werden können. Weiterhin ist beschrieben, wie Lateralkräfte Fy und Dissipation Γ erhalten wurden, um das volle Potential dieser Messmethode auszureizen.rnUm Prozesse auf atomarer Skala auf Oberflächen zu verstehen sind die kurzreichweitigen, chemischen Kräfte Fz,SR von größter Wichtigkeit. Langreichweitige Beiträge müssen hierzu an Fz angefittet und davon abgezogen werden. Dies ist jedoch eine fehleranfällige Aufgabe, die in dieser Arbeit dadurch gemeistert werden konnte, dass drei unabhängige Kriterien gefunden wurden, die den Beginn zcut von Fz,SR bestimmen, was für diese Aufgabe von zentraler Bedeutung ist. Eine ausführliche Fehleranalyse zeigt, dass als Kriterium die Abweichung der lateralen Kräfte voneinander vertrauenswürdige Fz,SR liefert. Dies ist das erste Mal, dass in einer Studie ein Kriterium für die Bestimmung von zcut gegeben werden konnte, vervollständigt mit einer detailreichen Fehleranalyse.rnMit der Kenntniss von Fz,SR und Fy war es möglich, eine der fundamentalen Eigenschaften der CaCO3(10.4) Oberfläche zu identifizieren: die absolute Oberflächenorientierung. Eine starke Verkippung der abgebildeten Objekte

DensiProbe Spine: a novel instrument for intraoperative measurement of bone density in transpedicular screw fixation

STUDY DESIGN.: Cadaver study. OBJECTIVE.: To determine bone strength in vertebrae by measuring peak breakaway torque or indentation force using custom-made pedicle probes. SUMMARY OF BACKGROUND DATA.: Screw performance in dorsal spinal instrumentation is dependent on bone quality of the vertebral body. To date no intraoperative measuring device to validate bone strength is available. Destructive testing may predict bone strength in transpedicular instrumentations in osteoporotic vertebrae. Insertional torque measurements showed varying results. METHODS.: Ten human cadaveric vertebrae were evaluated for bone mineral density (BMD) measurements by quantitative computed tomography. Peak torque and indentation force of custom-made probes as a measure for mechanical bone strength were assessed via a transpedicular approach. The results were correlated to regional BMD and to biomechanical load testing after pedicle screw implementation. RESULTS.: Both methods generated a positive correlation to failure load of the respective vertebrae. The correlation of peak breakaway torque to failure load was r = 0.959 (P = 0.003), therewith distinctly higher than the correlation of indentation force to failure load, which was r = 0.690 (P = 0.040). In predicting regional BMD, measurement of peak torque also performed better than that of indentation force (r = 0.897 [P = 0.002] vs. r = 0.777 [P = 0.017]). CONCLUSION.: Transpedicular measurement of peak breakaway torque is technically feasible and predicts reliable local bone strength and implant failure for dorsal spinal instrumentations in this experimental setting.

Sonographic Assessment of Fetal Cardiac Function: Introduction and Direct Measurement of Cardiac Function

Noninvasive blood flow measurements based on Doppler ultrasound studies are the main clinical tool for studying the cardiovascular status in fetuses at risk for circulatory compromise. Usually, qualitative analysis of peripheral arteries and, in particular clinical situations such as severe growth restriction or volume overload, also of venous vessels close to the heart or of flow patterns in the heart are being used to gauge the level of compensation in a fetus. Quantitative assessment of the driving force of the fetal circulation, the cardiac output, however, remains an elusive goal in fetal medicine. This article reviews the methods for direct and indirect assessment of cardiac function and explains new clinical applications. Part 1 of this review describes the concept of cardiac function and cardiac output and the techniques that have been used to quantify output. Part 2 summarizes the use of arterial and venous Doppler studies in the fetus and gives a detailed description of indirect measures of cardiac function (like indices derived from the duration of segments of the cardiac cycle) with current examples of their application.

Second-Generation Orthopaedic Measurement Device for in-vivo Assessment of Bone Quality

There are two main types of bone in the human body, trabecular and cortical bone. Cortical bone is primarily found on the outer surface of most bones in the body while trabecular bone is found in vertebrae and at the end of long bones (Ross 2007). Osteoporosis is a condition that compromises the structural integrity of trabecular bone, greatly reducing the ability of the bone to absorb energy from falls. The current method for diagnosing osteoporosis and predicting fracture risk is measurement of bone mineral density. Limitations of this method include dependence on the bone density measurement device and dependence on type of test and measurement location (Rubin 2005). Each year there are approximately 250,000 hip fractures in the United States due to osteoporosis (Kleerekoper 2006). Currently, the most common method for repairing a hip fracture is a hip fixation surgery. During surgery, a temporary guide wire is inserted to guide the permanent screw into place and then removed. It is believed that directly measuring this screw pullout force may result in a better assessment of bone quality than current indirect measurement techniques (T. Bowen 2008-2010, pers. comm.). The objective of this project is to design a device that can measure the force required to extract this guide wire. It is believed that this would give the surgeon a direct, quantitative measurement of bone quality at the site of the fixation. A first generation device was designed by a Bucknell Biomedical Engineering Senior Design team during the 2008- 2009 Academic Year. The first step of this project was to examine the device, conduct a thorough design analysis, and brainstorm new concepts. The concept selected uses a translational screw to extract the guide wire. The device was fabricated and underwent validation testing to ensure that the device was functional and met the required engineering specifications. Two tests were conducted, one to test the functionality of the device by testing if the device gave repeatable results, and the other to test the sensitivity of the device to misalignment. Guide wires were extracted from 3 materials, low density polyethylene, ultra high molecular weight polyethylene, and polypropylene and the force of extraction was measured. During testing, it was discovered that the spring in the device did not have a high enough spring constant to reach the high forces necessary for extracting the wires without excessive deflection of the spring. The test procedure was modified slightly so the wires were not fully threaded into the material. The testing results indicate that there is significant variation in the screw pullout force, up to 30% of the average value. This significant variation was attributed to problems in the testing and data collection, and a revised set of tests was proposed to better evaluate the performance of the device. The fabricated device is a fully-functioning prototype and further refinements and testing of the device may lead to a 3rd generation version capable of measuring the screw pullout force during hip fixation surgery.

Clinical retention force development of double crowns

This study deals with the development of the retentive forces of double crowns intraorally measured. Twenty-five combined fixed-removable prostheses with a total of 84 double crowns were included in the study. The intraoral measurement was performed at 72 defined measuring points directly adjacent to the double crowns of the dentures. The measurement was performed 4-6 weeks (baseline), 6 months (recall 1), and 18 months (recall 2) after the insertion of the restoration. A specifically designed measuring device was used. The median values for the single measuring points reached 4.705 N at the baseline, 5.190 N after 6 months, and 3.740 N after 18 months. The measured values were analyzed according to differences between the median retention forces at the three defined points in time. The statistical analysis of the median values showed no statistical difference for the retention force change after 6 months but for the decrease until the second recall (Mann-Whitney test). The retention force per denture was calculated by a summation of the single measuring points. At the baseline, 12.9 N was reached. The forces did only decrease slightly and were not statistically significant. The results indicate that retention force values of double crowns, measured intraorally at the patient, do not relevantly change clinically within the first 1.5 years. Within the limitations of this study, it can be stated that wear does not influence the retentive forces of double crowns within the first 18 months. After this period the retention force should be still sufficient for denture retention.

Impact of measurement errors on the determination of the linear modulus of human meniscal attachments

For the development of meniscal substitutes and related finite element models it is necessary to know the mechanical properties of the meniscus and its attachments. Measurement errors can falsify the determination of material properties. Therefore the impact of metrological and geometrical measurement errors on the determination of the linear modulus of human meniscal attachments was investigated. After total differentiation the error of the force (+0.10%), attachment deformation (−0.16%), and fibre length (+0.11%) measurements almost annulled each other. The error of the cross-sectional area determination ranged from 0.00%, gathered from histological slides, up to 14.22%, obtained from digital calliper measurements. Hence, total measurement error ranged from +0.05% to −14.17%, predominantly affected by the cross-sectional area determination error. Further investigations revealed that the entire cross-section was significantly larger compared to the load-carrying collagen fibre area. This overestimation of the cross-section area led to an underestimation of the linear modulus of up to −36.7%. Additionally, the cross-sections of the collagen-fibre area of the attachments significantly varied up to +90% along their longitudinal axis. The resultant ratio between the collagen fibre area and the histologically determined cross-sectional area ranged between 0.61 for the posterolateral and 0.69 for the posteromedial ligament. The linear modulus of human meniscal attachments can be significantly underestimated due to the use of different methods and locations of cross-sectional area determination. Hence, it is suggested to assess the load carrying collagen fibre area histologically, or, alternatively, to use the correction factors proposed in this study.

DensiProbe Spine: an intraoperative measurement of bone quality in spinal instrumentation. A clinical feasibility study

BACKGROUND CONTEXT A new device, DensiProbe, has been developed to provide surgeons with intraoperative information about bone strength by measuring the peak breakaway torque. In cases of low bone quality, the treatment can be adapted to the patient's condition, for example, by improving screw-anchorage with augmentation techniques. PURPOSE The objective of this study was to investigate the feasibility of DensiProbe Spine in patients undergoing transpedicular fixation. STUDY DESIGN Prospective feasibility study on consecutive patients. PATIENT SAMPLE Fourteen women and 16 men were included in this study. OUTCOME MEASURES Local and general bone quality. METHODS These consecutive patients scheduled for transpedicular fixation were evaluated for bone mineral density (BMD), which was measured globally by dual-energy X-ray absorptiometry and locally via biopsies using quantitative microcomputed tomography. The breakaway torque force within the vertebral body was assessed intraoperatively via the transpedicular approach with the DensiProbe Spine. The results were correlated with the areal BMD at the lumbar spine and the local volumetric BMD (vBMD) and a subjective impression of bone strength. The feasibility of the method was evaluated, and the clinical and radiological performance was evaluated over a 1-year follow-up. This study was funded by an AO Spine research grant; DensiProbe was developed at the AO Research Institute Davos, Switzerland; the AO Foundation is owner of the intellectual property rights. RESULTS In 30 patients, 69 vertebral levels were examined. The breakaway torque consistently correlated with an experienced surgeon's quantified impression of resistance as well as with vBMD of the same vertebra. Beyond a marginal prolongation of surgery time, no adverse events related to the usage of the device were observed. CONCLUSIONS The intraoperative transpedicular measurement of the peak breakaway torque was technically feasible, safe, and reliably predictive of local vBMD during dorsal spinal instrumentations in a clinical setting. Larger studies are needed to define specific thresholds that indicate a need for the augmentation or instrumentation of additional levels.

A new application of emulsions to measure the gravitational force on antihydrogen

We propose to build and operate a detector based on the emulsion film technology for the measurement of the gravitational acceleration on antimatter, to be performed by the AEgIS experiment (AD6) at CERN. The goal of AEgIS is to test the weak equivalence principle with a precision of 1% on the gravitational acceleration g by measuring the vertical position of the annihilation vertex of antihydrogen atoms after their free fall while moving horizontally in a vacuum pipe. With the emulsion technology developed at the University of Bern we propose to improve the performance of AEgIS by exploiting the superior position resolution of emulsion films over other particle detectors. The idea is to use a new type of emulsion films, especially developed for applications in vacuum, to yield a spatial resolution of the order of one micron in the measurement of the sag of the antihydrogen atoms in the gravitational field. This is an order of magnitude better than what was planned in the original AEgIS proposal.

Load Measurement on Foundations of Rockfall Protection Systems

Rockfall protection barriers are connected to the ground using steel cables fixed with anchors and foundations for the steel posts. It is common practice to measure the forces in the cables, while to date measurements of forces in the foundations have been inadequately resolved. An overview is presented of existing methods to measure the loads on the post foundations of rockfall protection barriers. Addressing some of the inadequacies of existing approaches, a novel sensor unit is presented that is able to capture the forces acting on post foundations in all six degrees of freedom. The sensor unit consists of four triaxial force sensors placed between two steel plates. To correctly convert the measurements into the directional forces acting on the foundation a special in-situ calibration procedure is proposed that delivers a corresponding conversion matrix.

G-force measurements at Mount Nyiragongo

To evaluate the mechanical stress on the volcanic edifice that results from lava lake level variations, we deployed a self-recording, differential capacitance (MEMS Inertial Sensor STMicroelectronics LIS3LV02DQ), 3-axis X6-1A accelerometer (Gulf Coast Data Concepts, LLC) at a distance of ~100m from the center of the Nyiragongo lava lake on freshly erupted lava flows. The device range was used in high (12-bit) resolution mode, which corresponds to a sensitivity of about 1 mg. The device was set to high-sensitivity mode with four additional bits to improve resolution, yet with a much lower signal-noise ratio. Once in position, the accelerometer continuously recorded data for three-day periods in June 2010. The system was oriented so that the X- and Y-axes form a plain parallel to the lava lake. During data collection, we did not attempt to calibrate the precision of the angle because relative G-force measurements were required instead of absolute G-force measurements. To distinguish the tiny accelerations caused by temperature differentials of the atmosphere, from the forces caused by magma movements, the temperature of the X6-1A device was continuously recorded. Temperature variations were corrected for by applying a de-correlation method to the recorded signal. Data was collected at 20 Hz, regrouped into batches that cover 1 hour per observation and associated with one averaged temperature measurement. This method was reproducible because diurnal temperature variations were the main cause for heating and cooling.

Measurement of the intrinsic damping constant in individual nanodisks of Y3Fe5O12 and Y3Fe5O12|Pt

We report on an experimental study on the spin-waves relaxation rate in two series of nanodisks of diameter ϕ=300 , 500, and 700 nm, patterned out of two systems: a 20 nm thick yttrium iron garnet (YIG) film grown by pulsed laser deposition either bare or covered by 13 nm of Pt. Using a magnetic resonance force microscope, we measure precisely the ferromagnetic resonance linewidth of each individual YIG and YIG|Pt nanodisks. We find that the linewidth in the nanostructure is sensibly smaller than the one measured in the extended film. Analysis of the frequency dependence of the spectral linewidth indicates that the improvement is principally due to the suppression of the inhomogeneous part of the broadening due to geometrical confinement, suggesting that only the homogeneous broadening contributes to the linewidth of the nanostructure. For the bare YIG nano-disks, the broadening is associated to a damping constant α=4 × 10−4 . A threefold increase of the linewidth is observed for the series with Pt cap layer, attributed to the spin pumping effect. The measured enhancement allows to extract the spin mixing conductance found to be G↑↓=1.55 × 1014 Ω−1 m−2 for our YIG(20nm)|Pt interface, thus opening large opportunities for the design of YIG based nanostructures with optimized magnetic losses.

Laboratory measurement of vehicle characteristics for predicting highway behavior. Final report.

Federal Highway Administration, Washington, D.C.

Investigation of a method for the measurement of subsonic oscillatory aerodynamic influence coefficients,

"ASTIA document no. AD-142201."