Development of a Mass Sensitive Quartz Crystal Microbalance (QCM)-Based DNA Biosensor Using a 50 MHz Electronic Oscillator Circuit

This work deals with the design of a high sensitivity DNA sequence detector using a 50 MHz quartz crystal microbalance (QCM) electronic oscillator circuit. The oscillator circuitry is based on Miller topology, which is able to work in damping media. Calibration and experimental study of frequency noise are carried out, finding that the designed sensor has a resolution of 7.1 ng/cm2 in dynamic conditions (with circulation of liquid). Then the oscillator is proved as DNA biosensor. Results show that the system is able to detect the presence of complementary target DNAs in a solution with high selectivity and sensitivity. DNA target concentrations higher of 50 ng/mL can be detected

Anharmonic acoustic effects during DNA hybridization on an electrochemical quartz crystal resonator

The paper describes a sensor for single-stranded DNA (ssDNA) biomarker based on anharmonic acoustic signals arising during hybridization with complementary thiolated ssDNA functionalised on the gold electrode of an electrochemical quartz crystal resonator. The steps of sensor preparation and hybridization are carried out in an electrochemical microfluidic flowcell. While the electrochemical impedance spectroscopy does not allow a definitive interpretation, the changes in resonance frequency and third Fourier harmonic current of the resonator on actuation at the fundamental mode indicate formation of a flexibly bound layer. The functionalization and hybridization steps monitored by the anharmonic detection technique (ADT) are described with a simple model based on Duffing nonlinear equation

Design and simulation of a low-cost biosensor for point-of-care diagnose devices

The principal aim of the work is to help in the development of a low-cost biosensor capable of providing a reliable diagnosis to inexpert users. Rather than being a precise measurement instrument like mass spectrometers, this biosensor should be able to qualitatively detect and distinguish between the most relevant families of the bacteria causing mycobacterium tuberculosis in human fluids. Other approaches pursue outstanding sensitivity relying on expensive materials and microfabrication processes. Despite other requisites, low-cost imposes certain criteria regarding materials, systems, fabrication techniques and user interfaces to be included in the device.Ingeniería Biomédic

Bulk and Surface Acoustic Wave Biosensors for Milk Analysis

Milk and dairy products are common foods and, therefore, are subject to regular controls. Such controls cover both the identification and quantification of specific components and the determination of physical parameters. Components include the usual milk ingredients, mainly carbohydrates, proteins, and fat, and any impurities that may be present. The latter range from small molecules, such as drug residues, to large molecules, e.g., protein-based toxins, to pathogenic microorganisms. Physical parameters of interest include viscosity as an indicator of milk gelation. Bulk and surface acoustic wave sensors, such as quartz crystal microbalance (QCM) and surface acoustic wave (SAW) devices, can principally be used for both types of analysis, with the actual application mainly depending on the device coating and the test format. This review summarizes the achievements of acoustic sensor devices used for milk analysis applications, including the determination of physical liquid parameters and the detection of low- and high-molecular-weight analytes and microorganisms. It is shown how the various requirements resulting from the respective analytes and the complex sample matrix are addressed, and to what extent the analytical demands, e.g., with regard to legal limits, are met

Probing multivalent particle–surface interactions using a quartz crystal resonator

The rise in market-approved cellular therapies demands for advancements in process analytical technology (PAT) capable of fulfilling the requirements of this new industry. Unlike conventional biopharmaceuticals, cell-based therapies (CBT) are complex “live” products, with a high degree of inherent biological variability. This exacerbates the need for in-process monitoring and control of critical product attributes, in order to guarantee safety, efficacious and continuous supply of this CBT. There are therefore mutual industrial and regulatory motivations for high throughput, non-invasive and non-destructive sensors, amenable to integration in an enclosed automated cell culture system. While a plethora of analytical methods is available for direct characterization of cellular parameters, only a few satisfy the requirements for online quality monitoring of industrial-scale bioprocesses. [Continues.

Acoustic Wave Based MEMS Devices, Development and Applications

Acoustic waves based MEMS devices offer a promising technology platform for a wide range of applications due to their high sensitivity and the capability to operate wirelessly. These devices utilize acoustic waves propagating through or on the surface of a piezoelectric material. An acoustic wave device typically consists of two layers, metal transducers on top of piezoelectric substrate or thin films. The piezoelectric material has inherent capabilities of generating acoustic waves related to the input electrical sinusoidal signals placed on the transducers. Using this characteristic, different transducer designs can be placed on top of the piezoelectric material to create acoustic wave based filters, resonators or sensors. Historically, acoustic wave devices have been and are still widely used in telecommunications industry, primarily in mobile cell phones and base stations. Surface Acoustic Wave (SAW) devices are capable of performing powerful signal processing and have been successfully functioning as filters, resonators and duplexers for the past 60 years. Although SAW devices are technological mature and have served the telecommunication industry for several decades, these devices are typically fabricated on piezoelectric substrates and are packaged as discrete components. Considering the wide flexibility and capabilities of the SAW device to form filters, resonators there has been motivation to integrate such devices on silicon substrates as demonstrated in (Nordin et al., 2007; M. J. Vellekoop et al., 1987; Visser et al., 1989). One such example is illustrated in (Nordin et al., 2007) where a CMOS SAW resonator was fabricated using 0.6 m AMIs CMOS technology process with additional MEMS post-processing. The traditional SAW structure of having the piezoelectric at the bottom was inverted. Instead, the IDTs were cleverly manufactured using standard complementary-metal-oxide-semiconductor (CMOS) process and the piezoelectric layer was placed on the top. Active circuitry can be placed adjacent to the CMOS resonator and can be connected using the integrated metal layers. A SAW device can also be designed to have a long propagation path between the input and output transducer. The propagating acoustic waves will then be very sensitive to ambient changes, allowing the device to act as a sensor. Any variations to the characteristics of the propagation path affect the velocity or amplitude of the wave. Important application for acoustic wave devices as sensors include torque and tire pressure sensors (Cullen et al., 1980; Cullen et al., 1975; Pohl et al., 1997), gas sensors (Levit et al., 2002; Nakamoto et al., 1996; Staples, 1999; Wohltjen et al., 1979), biosensors for medical applications (Andle et al., 1995; Ballantine et al., 1996; Cavic et al., 1999; Janshoff et al., 2000), and industrial and commercial applications (vapor, humidity, temperature, and mass sensors) (Bowers et al., 1991; Cheeke et al., 1996; Smith, 2001; N. J. Vellekoop et al., 1999; Vetelino et al., 1996; Weld et al., 1999). In recent years, the interest in the development of highly sensitive acoustic wave devices as biosensor platforms has grown. For biological applications the acoustic wave device is integrated in a microfluidic system and the sensing area is coated with a biospecific layer. When a bioanalyte interacts with this sensing layer, physical, chemical, and/or biochemical changes are produced. Typically, mass and viscosity changes of the biospecific layer can be detected by analyzing changes in the acoustic wave properties such as velocity, attenuation and resonant frequency of the sensor. An important advantage of the acoustic wave biosensors is simple electronic readout that characterizes these sensors. The measurement of the resonant frequency or time delay can be performed with high degree of precision using conventional electronics. This chapter is focused on two important applications of the acoustic-wave based MEMS devices; (1) biosensors and (2) telecommunications. For biological applications these devices are integrated in a microfluidic system and the sensing area is coated with a biospecific layer. When a bioanalyte interacts with this sensing layer, physical, chemical, and/or biochemical changes are produced. Typically, mass and viscosity changes of the biospecific layer can be detected by analyzing changes in the acoustic wave properties such as velocity, attenuation and resonant frequency of the sensor. An important advantage of the acoustic wave biosensors is simple electronic readout that characterizes these sensors. The measurement of the resonant frequency and time delay can be performed with high degree of precision using conventional electronics. Only few types of acoustic wave devices could be integrated in microfluidic systems without significant degradation of the quality factor. The acoustic wave based MEMS devices reported in the literature as biosensors are film bulk acoustic wave resonators (FBAR) and surface acoustic waves (SAW) resonators and SAW delay lines. Different approaches to the realization of FBARs and SAW resonators and SAW delay lines used for various biochemical applications are presented. Next, acoustic wave MEMS devices used in telecommunications applications are presented. Telecommunication devices have different requirements compared to sensors, where acoustic wave devices operating as a filter or resonator are expected to operate at high frequencies (GHz), have high quality factors and low insertion losses. Traditionally, SAW devices have been widely used in the telecommunications industry, however with advancement in lithographic techniques, FBARs are rapidly gaining popularity. FBARs have the advantage of meeting the stringent requirement of telecommunication industry of having Qs in the 10,000 range and silicon compatibility

Development of a novel high resolution and high throughput biosensing technology based on a Monolithic High Fundamental Frequency Quartz Crystal Microbalance (MHFF-QCM). Validation in food control

Tesis por compendio[ES] La sociedad actual demanda un mayor control en la seguridad y calidad de los alimentos que se consumen. Esta preocupación se ve reflejada en los diferentes planes estatales y europeos de investigación científica, los cuales, plantean la necesidad de innovar y desarrollar nuevas técnicas analíticas que cubran los requerimientos actuales. En el presente documento se aborda el problema de la presencia de residuos químicos en la miel. El origen de los mismos se debe, fundamentalmente, a los tramientos veterinarios con los que se tratan enfermedades y parásitos en las abejas, y a los tratamientos agrícolas con los que las abejas se ponen en contacto cuando recolectan el néctar en cultivos próximos a las colmenas. La Agencia Europea de Seguridad Alimentaria (EFSA) confirma esta realidad al notificar numerosas alertas sanitarias en la miel. En los últimos años, los métodos de análisis basados en inmunosensores piezoeléctricos se han posicionado como la base de una técnica de cribado muy prometedora, la cual puede ser empleada como técnica complementaria a las clásicas de cromatografía, gracias a su sencillez, rapidez y bajo coste. La tecnología de resonadores High-Fundamental Frequency Quartz Crystal Microbalance with Dissipation (HFF-QCMD) combina la detección directa en tiempo real, alta sensibilidad y selectividad con un fácil manejo y coste reducido en comparación con otras técnicas. Además, está tecnología permite aumentar el rendimiento del análisis mediante el diseño de arrays de resonadores en un mismo sustrato (Monolithic HFF-QCMD). En este documento se presenta el diseño de un array de 24 sensores HFF-QCMD, junto con un cartucho de micro-fluídica que traza diversos microcanales sobre los diferentes elementos sensores, a los que hace llegar la muestra de miel diluida a analizar. El cartucho actúa también como interfaz para realizar la conexión entre el array de resonadores y el instrumento de caracterización de los mismos. Para obtener el máximo partido del array diseñado, se desarrolla un método de medida robusto y fiable que permite elevar la tasa de adquisición de datos para facilitar la toma de registros eléctricos de un elevado número de resonadores de forma simultánea, e incluso en varios armónicos del modo fundamental de resonancia. La gran sensibilidad de la tecnología HFF-QCMD a los eventos bioquímicos a caracterizar se extiende también a otro tipo eventos externos, como son los cambios de temperatura o presión, lo que es necesario minimizar con el fin de reducir el impacto que estas perturbaciones no deseadas provocan en la estabilidad y fiabilidad de la medida. Con este fin, se desarrolla un algoritmo de procesado de señal basado en la Discrete Transform Wavelet (DTW). Finalmente, todos los desarrollos tecnológicos realizados se validan mediante la implementación de un inmunoensayo para la detección simultánea, en muestras de mieles reales, de residuos químicos de naturaleza química muy diferente, a saber, el fungicida tiabendazol y el antibiótico sulfatiazol.[CA] La societat actual demanda un major control en la seguretat i qualitat dels aliments que es consumeixen. Aquesta preocupació es veu reflectida en els diferents plans estatals i europeus d'investigació científica, els quals, plantegen la necessitat d'innovar i desenvolupar noves tècniques analítiques que cobrisquen els requeriments actuals. En el present document s'aborda el problema de la presència de residus químics en la mel. L'origen dels mateixos es deu, fonamentalment, als tractaments veterinaris amb els quals es tracten malalties i paràsits en les abelles, i als tractaments agrícoles amb els quals les abelles es posen en contacte quan recol·lecten el nèctar en cultius pròxims als ruscos. L'Agència Europea de Seguretat Alimentària (EFSA) confirma aquesta realitat notificant nombroses alertes sanitàries en la mel. En els últims anys, els mètodes d'anàlisis basades en immunosensors piezoelèctrics s'han posicionat com la base d'una tècnica de garbellat molt prometedora, la qual pot ser emprada com a tècnica complementària a les clàssiques de cromatografia, gràcies a la seua senzillesa, rapidesa i baix cost. La tecnologia de ressonadors High-Fundamental Frequency Quartz Crystal Microbalance with Dissipation (HFF-QCMD) combina la detecció directa en temps real, alta sensibilitat i selectivitat amb un fàcil maneig i cost reduït en comparació amb altres tècniques. A més, està tecnologia permet augmentar el rendiment del anàlisi mitjançant el disseny d'arrays de ressonadors en un mateix substrat (Monolithic HFF-QCMD). En aquest document es presenta el disseny d'un array de 24 sensors HFF-QCMD, juntament amb un cartutx de microfluídica que estableix diversos microcanals sobre els diferents elements sensors, als quals fa arribar la mostra de mel diluïda a analitzar. El cartutx actua també com a interfície per a realitzar la connexió entre l'array de ressonadors i l'instrument de caracterització d'aquests. Per a traure el màxim partit a l'array dissenyat, es desenvolupa un mètode de mesura robust i fiable que permet elevar la taxa d'adquisició de dades per a facilitar la presa de registres elèctrics d'un elevat nombre de ressonadors de manera simultània, i fins i tot en diversos harmònics del mode fonamental de ressonància. La gran sensibilitat de la tecnologia HFF-QCMD als esdeveniments bioquímics a caracteritzar s'estén també a un altre tipus esdeveniments externs, com són els canvis de temperatura o pressió, la qual cosa és necessari minimitzar amb la finalitat de reduir l'impacte que aquestes pertorbacions no desitjades provoquen en l'estabilitat i fiabilitat de la mesura. A aquest efecte, es desenvolupa un algorisme de processament de senyal basat en la Discrete Transform Wavelet (DTW). Finalment, tots els desenvolupaments tecnològics realitzats es validen mitjançant la implementació d'un immunoassaig per a la detecció simultània, en mostres de mel reals, de residus químics de naturalesa química molt diferent, a saber, el fungicida tiabendazol i l'antibiòtic sulfatiazol.[EN] Currently, society demands greater control over the safety and quality of the food consumed. This concern is reflected in the different states and European plans for scientific research, which establish the necessity to innovate and develop new analytical techniques that meet current requirements. This document addresses the problem of the presence of chemical residues in honey. Its origin is fundamentally due to the veterinary treatments against diseases and parasites in bees, and also to the agricultural treatments with which the bees come into contact when they collect the nectar in crops close to the hives. The European Food Safety Agency (EFSA) confirms this reality by notifying numerous health alerts in honey. In recent years, analysis methods based on piezoelectric immunosensors have been positioned as the basis of a very promising screening technique, which can be used as a complementary technique to the classic chromatography, thanks to its simplicity, speed and low cost. High-Fundamental Frequency Quartz Crystal Microbalance with Dissipation (HFF-QCMD) resonator technology combines direct real-time detection, high sensitivity and selectivity with easy handling and low cost compared to other techniques. In addition, this technology allows increasing the performance of the analysis through the design of resonator arrays on the same substrate (Monolithic HFF-QCMD). This document presents the design of an array of 24 HFF-QCMD sensors, together with a microfluidic cartridge that establish various microchannels on the different sensor elements, to provide them the diluted honey sample to be analyzed. The cartridge also acts as an interface to make the connection between the array of resonators and the characterization instrument. To get the most out of the designed array, a robust and reliable measurement method has been developed that allows increasing the data acquisition rate to facilitate electrical parameters readout from a high number of resonators simultaneously, and even in several harmonics of the fundamental resonance mode. The great sensitivity of the HFF-QCMD technology to the biochemical events to be characterized also is extended to other types of external events, such as changes in temperature or pressure, which must be minimized in order to reduce the impact that these unwanted disturbances cause in the stability and reliability of the measurement. To this end, a signal processing algorithm based on the Discrete Transform Wavelet (DTW) is developed. Finally, all the technological developments carried out are validated through the implementation of an immunoassay for the simultaneous detection, in real honey samples, of chemical residues of very different chemical nature, namely, the fungicide thiabendazole and the antibiotic sulfathiazole.The authors would also like to thank Jorge Martínez from the Laboratory of High Frequency Circuits (LCAF) of the Universitat Politècnica de València (UPV) for assistance with profilometry, and Manuel Planes, José Luis Moya, Mercedes Tabernero, Alicia Nuez and Joaquin Fayos from the Electron Microscopy Services of the UPV for helping with the AFM, and SEM measurements. M.Calero is the recipient of the doctoral fellowship BES-2017-080246 from the Spanish Ministry of Economy, Industry and Competitiveness (Madrid, Spain). This research was funded by Spanish Ministry of Economy and Competitiveness with FEDER funds (AGL 2016-77702-R) and European Commission Horizon 2020 Programme (Grant Agreement number H2020-FETOPEN-2016-2017/737212-CATCH-U-DNA - Capturing non-Amplified Tumor Circulating DNA with Ultrasound Hydrodynamics) for which the authors are grateful. Román Fernández is with the Center for Research and Innovation in Bioengineering (Ci2B), Universitat Politècnica de València, València, Spain and with Advanced Wave Sensors S.L., Paterna, València, Spain. (e-mail: [email protected]); Yolanda Jiménez, Antonio Arnau and María Calero are with the Center for Research and Innovation in Bioengineering (Ci2B), Universitat Politècnica de València, València, Spain; Ilya Reiviakine is with Advanced Wave Sensors S.L., Paterna, Valencia, Spain and with the Department of Bioengineering, University of Washington, Seattle, WA, 98150 USA; María Isabel Rocha-Gaso and José Vicente García are with Advanced Wave Sensors S.L., Paterna, València, Spain.Calero Alcarria, MDS. (2022). Development of a novel high resolution and high throughput biosensing technology based on a Monolithic High Fundamental Frequency Quartz Crystal Microbalance (MHFF-QCM). Validation in food control [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/182652TESISCompendi