Properties and Applications of Love Surface Waves in Seismology and Biosensors

Shear horizontal (SH) surface waves of the Love type are elastic surface waves propagating in layered waveguides, in which surface layer is “slower” than the substrate. Love surface waves are of primary importance in geophysics and seismology, since most structural damages in the wake of earthquakes are attributed to the devastating SH motion inherent to the Love surface waves. On the other hand, Love surface waves found benign applications in biosensors used in biology, medicine, and chemistry. In this chapter, we briefly sketch a mathematical model for Love surface waves and present examples of the resulting dispersion curves for phase and group velocities, attenuation as well as the amplitude distribution as a function of the depth. We illustrate damages due to Love surface waves generated by earthquakes on real-life examples. In the following of this chapter, we present a number of representative examples for Love wave biosensors, which have been already used to DNA characterization, bacteria and virus detection, measurements of toxic substances, etc. We hope that the reader, after studying this chapter, will have a clear idea that deadly earthquakes and a beneficiary biosensor technology share the same physical phenomenon, which is the basis of a fascinating interdisciplinary research

Love Wave Biosensors: A Review

In the fields of analytical and physical chemistry, medical diagnostics and biotechnology there is an increasing demand of highly selective and sensitive analytical techniques which, optimally, allow an in real-time label-free monitoring with easy to use, reliable, miniaturized and low cost devices. Biosensors meet many of the above features which have led them to gain a place in the analytical bench top as alternative or complementary methods for routine classical analysis. Different sensing technologies are being used for biosensors. Categorized by the transducer mechanism, optical and acoustic wave sensing technologies have emerged as very promising biosensors technologies. Optical sensing represents the most often technology currently used in biosensors applications. Among others, Surface Plasmon Resonance (SPR) is probably one of the better known label-free optical techniques, being the main shortcoming of this method its high cost. Acoustic wave devices represent a cost-effective alternative to these advanced optical approaches [1], since they combine their direct detection, simplicity in handling, real-time monitoring, good sensitivity and selectivity capabilities with a more reduced cost. The main challenges of the acoustic techniques remain on the improvement of the sensitivity with the objective to reduce the limit of detection (LOD), multi-analysis and multi-analyte detection (High-Throughput Screening systems-HTS), and integration capabilities. Acoustic sensing has taken advantage of the progress made in the last decades in piezoelectric resonators for radio-frequency (rf) telecommunication technologies. The so-called gravimetric technique [2], which is based on the change in the resonance frequency experimented by the resonator due to a mass attached on the sensor surface, has opened a great deal of applications in bio-chemical sensing in both gas and liquid media. Traditionally, the most commonly used acoustic wave biosensors were based on QCM devices. This was primarily due to the fact that the QCM has been studied in detail for over 50 years and has become a mature, commercially available, robust and affordable technology [3, 4]. LW acoustic sensors have attracted a great deal of attention in the scientific community during the last two decades, due to its reported high sensitivity in liquid media compared to traditional QCM-based sensors. Nevertheless, there are still some issues to be further understood, clarified and/or improved about this technology; mostly for biosensor applications. LW devices are able to operate at higher frequencies than traditional QCMs [5]; typical operation frequencies are between 80-300 MHz. Higher frequencies lead, in principle, to higher sensitivity because the acoustic wave penetration depth into the adjacent media is reduced [6]. However, the increase in the operation frequency also results in an increased noise level, thus restricting the LOD. The LOD determines the minimum surface mass that can be detected. In this sense, the optimization of the read out and characterization system for these high frequency devices is a key aspect for improving the LOD [7]. Another important aspect of LW technology is the optimization of the fluidics, specially the flow cell. This is of extreme importance for reducing the noise and increasing the biosensor system stability; aspects that will contribute to improve the LOD. The analysis and interpretation of the results obtained with LW biosensors must be deeper understood, since the acoustic signal presents a mixed contribution of changes in the mass and the viscoelasticity of the adsorbed layers due to interactions of the biomolecules. A better understanding of the transduction mechanism in LW sensors is a first step to advance in this issue; however its inherent complexity leads, in many cases, to frustration [8]. The fabrication process of the transducer, unlike in traditional QCM sensors, is another aspect under investigation in LW technology, where features such as: substrate materials, sizes, structures and packaging must be still optimized. This chapter aims to provide an updated insight in the mentioned topics focused on biosensors applications

Based acoustic waves microsensor for the detection of bacteria in complex liquid media

Cette thèse s’inscrit dans le cadre d’une cotutelle internationale entre l’Université de Bourgogne Franche-Comté en France et l’Université de Sherbrooke au Canada. Elle porte sur le développement d'un biocapteur miniature pour la détection et la quantification de bactéries dans des milieux liquides complexes. La bactérie visée est l’Escherichia coli (E. coli), régulièrement mise en cause dans des épidémies d'infections alimentaires, et parfois meurtrière. La géométrie du biocapteur consiste en une membrane en arséniure de gallium (GaAs) sur laquelle est déposé un film mince piézoélectrique d’oxyde de zinc (ZnO). L'apport du ZnO structuré en couche mince constitue un réel atout pour atteindre de meilleures performances du transducteur piézoélectrique et consécutivement une meilleure sensibilité de détection. Une paire d'électrodes déposée sur le film de ZnO permet de générer, sous une tension sinusoïdale, des ondes acoustiques se propageant dans le GaAs, à une fréquence donnée. La face arrière de la membrane, quant à elle, est fonctionnalisée avec une monocouche auto-assemblée (SAM) d'alkanethiols et des anticorps contre l’E. coli, conférant la spécificité de la détection. Ainsi, le biocapteur bénéficie à la fois des technologies de microfabrication et de bio-fonctionnalisation du GaAs, déjà validées au sein de l’équipe de recherche, et des propriétés piézoélectriques prometteuses du ZnO, afin d’atteindre potentiellement une détection hautement sensible et spécifique de la bactérie d’intérêt. Le défi consiste à pouvoir détecter et quantifier cette bactérie à de très faibles concentrations dans un échantillon liquide et/ou biologique complexe. Les travaux de recherche ont en partie porté sur les dépôts et caractérisations de couches minces piézoélectriques de ZnO sur des substrats de GaAs. L’effet de l’orientation cristalline du GaAs ainsi que l’utilisation d’une couche intermédiaire de Platine entre le ZnO et le GaAs ont été étudiés par différentes techniques de caractérisation structurale (diffraction des rayons X, spectroscopie Raman, spectrométrie de masse à ionisation secondaire), topographique (microscopie à force atomique), optique (ellipsométrie) et électrique. Après la réalisation des contacts électriques, la membrane en GaAs a été usinée par gravure humide. Une fois fabriqué, le transducteur a été testé en air et en milieu liquide par des mesures électriques, afin de déterminer les fréquences de résonance pour les modes de cisaillement d’épaisseur. Un protocole de bio-fonctionnalisation de surface, validé au sein du laboratoire, a été appliqué à la face arrière du biocapteur pour l’ancrage des SAMs et des anticorps, tout en protégeant la face avant. De plus, les conditions de greffage d’anticorps en termes de concentration utilisée, pH et durée d’incubation, ont été étudiées, afin d’optimiser la capture de bactérie. Par ailleurs, l’impact du pH et de la conductivité de l’échantillon à tester sur la réponse du biocapteur a été déterminé. Les performances du biocapteur ont été évaluées par des tests de détection de la bactérie cible, E. coli, tout en corrélant les mesures électriques avec celles de fluorescence. Des tests de détection ont été réalisés en variant la concentration d’E. coli dans des milieux de complexité croissante. Différents types de contrôles ont été réalisés pour valider les critères de spécificité. En raison de sa petite taille, de son faible coût de fabrication et de sa réponse rapide, le biocapteur proposé pourrait être potentiellement utilisé dans les laboratoires de diagnostic clinique pour la détection d’E. coli.Abstract: This thesis was conducted in the frame of an international collaboration between Université de Bourgogne Franche-Comté in France and Université de Sherbrooke in Canada. It addresses the development of a miniaturized biosensor for the detection and quantification of bacteria in complex liquid media. The targeted bacteria is Escherichia coli (E. coli), regularly implicated in outbreaks of foodborne infections, and sometimes fatal. The adopted geometry of the biosensor consists of a gallium arsenide (GaAs) membrane with a thin layer of piezoelectric zinc oxide (ZnO) on its front side. The contribution of ZnO structured in a thin film is a real asset to achieve better performances of the piezoelectric transducer and consecutively a better sensitivity of the detection. A pair of electrodes deposited on the ZnO film allows the generation of acoustic waves propagating in GaAs under a sinusoidal voltage, at a given frequency. The backside of the membrane is functionalized with a self-assembled monolayer (SAM) of alkanethiols and antibodies against E. coli, providing the specificity of the detection. Thus, the biosensor benefits from the microfabrication and bio-functionalization technologies of GaAs, validated within the research team, and the promising piezoelectric properties of ZnO, to potentially achieve a highly sensitive and specific detection of the bacteria of interest. The challenge is to be able to detect and quantify these bacteria at very low concentrations in a complex liquid and/or biological sample. The research work was partly focused on the deposition and characterization of piezoelectric ZnO thin films on GaAs substrates. The effect of the crystalline orientation of GaAs and the use of a titanium/platinum buffer layer between ZnO and GaAs were studied using different structural (X-ray diffraction, Raman spectroscopy, secondary ionization mass spectrometry), topographic (atomic force microscopy), optical (ellipsometry) and electrical characterizations. After the realization of the electrical contacts on top of the ZnO film, the GaAs membrane was micromachined using chemical wet etching. Once fabricated, the transducer was tested in air and liquid medium by electrical measurements, in order to determine the resonance frequencies for thickness shear mode. A protocol for surface bio-functionalization, validated in the laboratory, was applied to back side of the biosensor for anchoring SAMs and antibodies, while protecting the top side. Furthermore, different conditions of antibody immobilization such as the concentration, pH and incubation time, were tested to optimize the immunocapture of bacteria. In addition, the impact of the pH and the conductivity of the solution to be tested on the response of the biosensor has been determined. The performance of the biosensor was evaluated by detection tests of the targeted bacteria, E. coli, while correlating electrical measurements with fluorescence microscopy. Detection tests were completed by varying the concentration of E. coli in environments of increasing complexity. Various types of controls were performed to validate the specificity criteria. Thanks to its small size, low cost of fabrication and rapid response, the proposed biosensor has the potential of being applied in clinical diagnostic laboratories for the detection of E. coli

Combined surface acoustic wave and surface plasmon resonance measurement of collagen and fibrinogen layers

We use an instrument combining optical (surface plasmon resonance) and acoustic (Love mode acoustic wave device) real-time measurements on a same surface for the identification of water content in collagen and fibrinogen protein layers. After calibration of the surface acoustic wave device sensitivity by copper electrodeposition, the bound mass and its physical properties -- density and optical index -- are extracted from the complementary measurement techniques and lead to thickness and water ratio values compatible with the observed signal shifts. Such results are especially usefully for protein layers with a high water content as shown here for collagen on an hydrophobic surface. We obtain the following results: collagen layers include 70+/-20 % water and are 16+/-3 to 19+/-3 nm thick for bulk concentrations ranging from 30 to 300 ug/ml. Fibrinogen layers include 50+/-10 % water for layer thicknesses in the 6+/-1.5 to 13+/-2 nm range when the bulk concentration is in the 46 to 460 ug/ml range.Comment: 50 pages, 8 figures, 1 tabl

Acoustic Wave Biosensors for Biomechanical and Biological Characterization of Cells

During past decades, interest in development of cell-based biosensors has increased considerably. In this study, two kinds of acoustic wave sensors are adopted as the cell-based biosensors to investigate the biomechanical and biological behaviors of cells, the quartz thickness shear mode (TSM) resonator and Love wave sensor. For the first part, the quartz TSM resonator is applied to detect the structural and mechanical properties of tendon stem/progenitor cells (TSCs), which are one kind of newly discovered adult cells in tendons, and the platelets from blood. Through the TSM resonator, the related viscoelastic properties of cells are extracted, which could indicate the state of cells in different physiological conditions. The TSM resonator sensor is utilized to characterize the aging-related viscoelasticity differences between the aging and young TSCs, and also to monitor the dynamic activation process of platelets. For the second part, a 36˚ YX-LiTaO3 Love wave sensor with a parylene-C wave guiding layer is proposed as a cell-based biosensor. A theoretical model is derived, to describe the Love wave propagation in the wave guiding layer, the adherent cell layer, and penetration into the liquid medium. The Love wave sensor is used to monitor the adhesion process of cells. Compared with TSM resonator, the response of Love wave sensor to the cell adhesion is primarily induced by the formation of bonds between cells and the substrate. The numerical results indicate that the adherent cell layer of various storage or loss shear modulus in certain range can cause evident, characteristic variations in propagation velocity and propagation loss, revealing the potential of Love wave sensors in providing useful quantitative measures on cellular mechanical properties. In addition, a Love wave sensor with a phononic wave guiding layer is introduced for non-operation signal filtering and sensor sensitivity improvement. Both two kinds of acoustic wave sensors present their own advantages as the cell-based biosensors, indicating advisable techniques for investigating cell biology in general and certain physiological processes in particular

SENSORS: Detecting Microbial Pathogens with Novel Surface Acoustic Wave Devices in Liquid Environments

This SENSORS proposal integrates research and education to exploit the sensitivity of a new family of LGX crystal devices operated in novel Shear Horizontal Surface Acoustic Wave (SH-SAW) propagation directions by combining them with highly selective molecular padlock probes to detect specific nucleic acid sequences associated with bacteria such as Escherichia coli O157:H7, Salmonella typhi, and Vibrio cholerae in aqueous solutions. The anticipated fundamental advances in sensor science and engineering will be relevant to numerous applications, including rapid response to bioterrorism, healthcare, epidemiology, agriculture, food safety, and pollution avoidance and mitigation. This SENSORS program builds upon the initial proof-of-concept results provided by an NSF SGER project funded by the divisions of Electrical and Communication Systems, and Bioengineering and Environmental Systems. The intellectual merit of this proposal rests in the creative, integrated research and education activities related to combining the recently identified LGX SH-SAW devices with molecular padlock probe technology to permit the design, fabrication, testing, and optimization of prototype biosensors. The specific research objectives of this SENSORS program are to: (i) Identify the surface density chemistry for increased sensitivity; (ii) Investigate and identify the optimal LGX SH-SAW orientation and device design for operation with the padlock technology; (iii) Study and develop the molecular padlock probe system to operate effectively in conjunction with the LGX SH-SAW device; (iv) Fabricate and test the prototype SH-SAW liquid biosensors; (v) Identify and optimize a procedure for sensor regeneration; and (vi) Characterize and optimize the sensor\u27s dynamic range and cross-effects due to temperature and other physical and chemical factors. The educational objective of this SENSORS program is to provide a multidisciplinary learning experience to students ranging from high school to graduate student level in the area of sensors in general, and biosensors in particular. Broader impacts will be achieved through the following programs and activities to: (i) Train and interact with high school audiences through two major ongoing programs at University of Maine (UMaine), NSF Research Experiences for Teachers (RET) and the GK-12 Sensors; (ii) Involve undergraduates from Maine and other institutions directly into the research project under the umbrella of the ongoing NSF Research Experience for Undergraduates (REU) program at the UMaine; (iii) Expand existing undergraduate Sensor Technology and Instrumentation and Biochemical Engineering Engineering courses at the UMaine by adding modules relating to biosensors devices and systems; (iv) Identify appropriate Capstone projects for undergraduates involving cross-disciplinary research and design projects; (v) Enhance existing graduate level courses Microscale Bioengineering and Design and Fabrication of Acoustic Wave Devices by incorporating research results into the course; (vi) Contribute to the new interdisciplinary multi-institutional NSF Integrative Graduate Education and Research Traineeship (IGERT) program in functional genomics, which involves UMaine, the Jackson Laboratory, and the Maine Medical Center Research Institute; (vi) Provide a experimental and/or theoretical thesis topics for Masters and Ph.D. students; (vii) Disseminate the research and educational material on a project website, and through conferences and printed literature. The SENSORS project proposed here is designed to result in tangible research and educational benefits. It will provide a knowledge base critical to creation of the next generation of biosensors for single unit production and future integration into arrays. It also seeks to establish a model program whereby cross-disciplinary education is integrated with a state-of-the-art research program, providing a rich learning experience for students ranging from high school to graduate student level. Finally, the project will help to strengthen U.S. research and educational capabilities in an area of high technology that currently is in need of highly trained industry and academic professionals

SURFACE ACOUSTIC WAVE (SAW)-ENHANCED SURFACE PLASMON RESONANCE (SPR) BIOSENSOR

The aim of this master thesis work was the realization of a biosensor based on surface plasmon resonance (SPR) which was integrated with surface acoustic wave (SAW)-driven microfluidics. Over the last 15 years SAW-induced mixers for microfluidic devices have been developed owing to their fast mixing capabilities. Meanwhile, SPR sensors have also been developed because of their high reliability and quantitative real-time measurements. Following the Drude model of electrical conduction, surface plasmons (SPs) can be considered as propagating electron density waves occurring at the interface between a metal and a dielectric and can alternatively be viewed as electromag- netic waves that are strongly bound to this interface. The resonance condition for SP excitation varies with the refractive index of the dielectric in the proximity (about 200 nm for visible light) of the surface of the metal film supporting the SP. A change in the resonance condition measured with an optical setup can be used to detect changes in the refractive index. SPR sensing is particularly useful for biological applications. By functionalizing the SPR sensor surface it is pos- sible to detect binding events in real-time and quantify the concentration of the analyte to be studied with high reliability. SPR biosensors have applications in numerous important fields including medical diagnostics, environmental mon- itoring, and food safety and security with resolution as low as 10 − 7 refractive index units (RIU) (§1). Lab-on-a-chip (LOC) devices are typically being developed for use in the life sciences and diagnostics and represent a fast moving field in which efforts are being made in order to increase portability and efficiency. Microfluidic systems are characterized by small Reynolds numbers which indicates that fluid flow is in general laminar. Efficient mixing is a challenge at these scales that can, how- ever, be overcome with the use of SAW-induced streaming. SAWs are mechanical oscillations which propagate along the surface of a given crystal. In piezoelectric materials they can be generated using interdigitated transducers (IDT), which are fabricated using thin-film metal deposition. When a SAW comes into con- tact with the edge of a liquid in its path, the acoustic energy diffracts into the fluid due to the mismatch between the sound velocity in the substrate and the liquid, causing a longitudinal pressure wave front that gives rise to the acoustic streaming. This phenomenon can be exploited to efficiently mix solutions with 1times that are significantly shorter than without SAWs (§2). By using micro- and nano- fabrication techniques (§3.1) a biosensor was de- veloped where SAW-driven active mixing and SPR sensing were integrated onto a common substrate. The optical setup (§3.4) was based on wavelength modula- tion and Kretschmann geometry where a polychromatic light is totally reflected through a high refractive index prism (on which the chip is placed). A spectrom- eter was used to analyze the reflected spectra. The SPR surface was functional- ized in order to study a biotin-streptavidin system (§3.3). SPR was first character- ized in droplets and then in polydimethylsiloxan (PDMS) microchannels (§3.2). By adding an IDT onto the chip it was possible to induce acoustic streaming in the channel while the biotin functionalization or the biotin-streptavidin event occurred. SAW was characterized by using a laser doppler vibrometer and a vector network analyzer. Owing to the chip design it was possible to decouple the two effects induced by SAW in the microchannel: streaming and heating. A thermocamera was used to study the second effect. The effect of the SAWs was studied both on the functionalization process and the streptavidin-biotin binding (§4). SAW streaming resulted in better surface functionalization than in the case without SAW. The signal due to the functionalization of gold with biotin was about 4.4 times higher than the signal detected without SAW-assisted functionalization. It is possible, then, to conclude that SAW streaming increases the probability that the biotin will be in contact with the gold surface and attach to it. Preliminary data also suggest better streptavidin biosensing than control device. The biosensor made for this master thesis work was the first SAW-driven mi- crofluidic device with SPR integrated on the same substrate. It showed promis- ing results that might be exploited for improving sensitivity and limit of detec- tion of SPR biosensors

Surface Generated Acoustic Wave Biosensors for the Detection of Pathogens: A Review

This review presents a deep insight into the Surface Generated Acoustic Wave (SGAW) technology for biosensing applications, based on more than 40 years of technological and scientific developments. In the last 20 years, SGAWs have been attracting the attention of the biochemical scientific community, due to the fact that some of these devices - Shear Horizontal Surface Acoustic Wave (SH-SAW), Surface Transverse Wave (STW), Love Wave (LW), Flexural Plate Wave (FPW), Shear Horizontal Acoustic Plate Mode (SH-APM) and Layered Guided Acoustic Plate Mode (LG-APM) - have demonstrated a high sensitivity in the detection of biorelevant molecules in liquid media. In addition, complementary efforts to improve the sensing films have been done during these years. All these developments have been made with the aim of achieving, in a future, a highly sensitive, low cost, small size, multi-channel, portable, reliable and commercially established SGAW biosensor. A setup with these features could significantly contribute to future developments in the health, food and environmental industries. The second purpose of this work is to describe the state-of-the-art of SGAW biosensors for the detection of pathogens, being this topic an issue of extremely importance for the human health. Finally, the review discuses the commercial availability, trends and future challenges of the SGAW biosensors for such applications