Lab-on-a-chip based integrated hybrid technologies for biofluids manipulation and characterization

[EN] The goal of this work is to develop an original and high performance hybrid lab-on-a-chip, coupling actuators and biosensors for the active control and broad range characterization of biofluids samples. These biofluids will be controlled (moved and mixed) actively by Rayleigh-Surface Acoustic Wave (R-SAW) and analysed in real-time by combining Love-SAW (L-SAW) and Surface Plasmon Resonance (SPR) technologies. A microfluidic chamber and specific transducer were designed for this application to interact efficiently with the liquid sample entrapped in the chamber and to detect modifications of its physical properties such as viscosity. AT-cut quartz and LiNbO3 36Y-X substrates were used to generate both Rayleigh and Shear-Horizontal waves and ZnO material as guiding layer. The whole system has proven is full efficiency to interact with the fluid and to detect the signal perturbations with no significant loss compared with the measurements carried out with a probe station.The authors wish to thank the ANR for its financial support through the AWESOM project (ANR-12-BS09-021); the operators Laurent Bouvot, Jean Georges Mussot and Emmanuel Vatoux, for their support and assistance in the fabrication of the cell and the SAW devices; and the engineering students, Olivier Bettoni and Bastien Lafont, for their contribution to the cell design.Rocha Gaso, MI.; Renaudin, A.; Sarry, F.; Beyssen, D. (2015). Lab-on-a-chip based integrated hybrid technologies for biofluids manipulation and characterization. Procedia Engineering. 120:687-690. https://doi.org/10.1016/j.proeng.2015.08.75068769012

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

Biosensors to diagnose Chagas disease: A review

International audienceChagas disease (CD), which mostly affects underprivileged people, has turned into one 9 of Latin America's main public health problems. Prevention of the disease requires early diagnosis, 10 initiation of therapy, and regular blood monitoring of the infected individual. However, the majority 11 of the infections go undiagnosed because of general mild symptoms and lack of access to medical 12 care. Therefore, more affordable and accessible detection technologies capable of providing early 13 diagnosis and parasite load measurements in settings where CD is prevalent are needed to enable 14 enhanced intervention strategies. This review discusses currently available detection technologies 15 and emerging biosensing technologies for a future application to CD. Even if biosensing 16 technologies still require further research efforts to develop portable systems, we arrive to the 17 conclusion that biosensors could improve diagnosis and the patients' treatment follow-up, in terms 18 of rapidity, small sample volume, high integration, ease of use, real-time and low cost detection 19 compared to current conventional technologies. 2

High Fundamental Frequency (HFF) Monolithic Resonator Arrays for Biosensing Applications: Design, Simulations, Experimental, Characterization

© 2020 IEEE. Personal use of this material is permitted. Permissíon from IEEE must be obtained for all other uses, in any current or future media, including reprinting/republishing this material for advertisíng or promotional purposes, creating new collective works, for resale or redistribution to servers or lists, or reuse of any copyrighted component of this work in other works.[EN] Miniaturized, high-throughput, cost-effective sensing devices are needed to advance lab-on-a-chip technologies for healthcare, security, environmental monitoring, food safety, and research applications. Quartz crystal microbalance with dissipation (QCMD) is a promising technology for the design of such sensing devices, but its applications have been limited, until now, by low throughput and significant costs. In this work, we present the design and characterization of 24-element monolithic QCMD arrays for high-throughput and low-volume sensing applications in liquid. Physical properties such as geometry and roughness, and electrical properties such as resonance frequency, quality factor, spurious mode suppression, and interactions between array elements (crosstalk), are investigated in detail. In particular, we show that the scattering parameter, S 21 , commonly measured experimentally to investigate crosstalk, contains contributions from the parasitic grounding effects associated with the acquisition circuitry. Finite element method simulations do not take grounding effects into account explicitly. However, these effects can be effectively modelled with appropriate equivalent circuit models, providing clear physical interpretation of the different contributions. We show that our array design avoids unwanted interactions between elements and discuss in detail aspects of measuring these interactions that are often-overlooked.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.Fernández Díaz, R.; Calero-Alcarria, MDS.; Reviakine, I.; García, JV.; Rocha-Gaso, MI.; Arnau Vives, A.; Jiménez Jiménez, Y. (2021). High Fundamental Frequency (HFF) Monolithic Resonator Arrays for Biosensing Applications: Design, Simulations, Experimental, Characterization. IEEE Sensors Journal. 21(1):284-295. https://doi.org/10.1109/JSEN.2020.3015011S28429521

Love Wave Immunosensor for the Detection of Carbaryl Pesticide

A Love Wave (LW) immunosensor was developed for the detection of carbaryl pesticide. The experimental setup consisted on: a compact electronic characterization circuit based on phase and amplitude detection at constant frequency; an automated flow injection system; a thermal control unit; a custom-made flow-through cell; and Quartz /SiO2 LW sensors with a 40 μm wavelength and 120 MHz center frequency. The carbaryl detection was based on a competitive immunoassay format using LIB-CNH45 monoclonal antibody (MAb). Bovine Serum Albumin-CNH (BSA-CNH) carbaryl hapten-conjugate was covalently immobilized, via mercaptohexadecanoic acid self-assembled monolayer (SAM), onto the gold sensing area of the LW sensors. This immobilization allowed the reusability of the sensor for at least 70 assays without significant signal losses. The LW immunosensor showed a limit of detection (LOD) of 0.09 μg/L, a sensitivity of 0.31 μg/L and a linear working range of 0.14–1.63 μg/L. In comparison to other carbaryl immunosensors, the LW immunosensor achieved a high sensitivity and a low LOD. These features turn the LW immunosensor into a promising tool for applications that demand a high resolution, such as for the detection of pesticides in drinking water at European regulatory levels.The author also would like to acknowledge the Spanish Ministry of Economy and Competitiveness and the European Regional Development Fund (ERDF) for their financing support through the grant of the INNPACTO 2012 project (DETECTA IPT-2012-0154-300000), J.V. Garcia's Fellowship, AP2007-03745, of FPU (Formacion de Profesorado Universitario) program, M. I. Rocha-Gaso's PhD CONACyT Fellowship and AWSensors Inc. for its collaboration.Rocha-Gaso, M.; García Narbón, JV.; García, P.; March Iborra, MDC.; Jiménez Jiménez, Y.; Francis, L.; Montoya Baides, Á.... (2014). Love Wave Immunosensor for the Detection of Carbaryl Pesticide. Sensors. 14(9):16434-16453. https://doi.org/10.3390/s140916434S1643416453149March, C., Manclús, J. J., Jiménez, Y., Arnau, A., & Montoya, A. (2009). A piezoelectric immunosensor for the determination of pesticide residues and metabolites in fruit juices. Talanta, 78(3), 827-833. doi:10.1016/j.talanta.2008.12.058Janshoff, A., Galla, H.-J., & Steinem, C. (2000). Piezoelectric Mass-Sensing Devices as Biosensors—An Alternative to Optical Biosensors? Angewandte Chemie, 39(22), 4004-4032. doi:10.1002/1521-3773(20001117)39:223.0.co;2-2Rocha-Gaso, M.-I., March-Iborra, C., Montoya-Baides, Á., & Arnau-Vives, A. (2009). Surface Generated Acoustic Wave Biosensors for the Detection of Pathogens: A Review. Sensors, 9(7), 5740-5769. doi:10.3390/s90705740Gronewold, T. M. A. (2007). Surface acoustic wave sensors in the bioanalytical field: Recent trends and challenges. Analytica Chimica Acta, 603(2), 119-128. doi:10.1016/j.aca.2007.09.056Länge, K., Rapp, B. E., & Rapp, M. (2008). Surface acoustic wave biosensors: a review. Analytical and Bioanalytical Chemistry, 391(5), 1509-1519. doi:10.1007/s00216-008-1911-5Arnau, A., Montagut, Y., García, J. V., & Jiménez, Y. (2009). A different point of view on the sensitivity of quartz crystal microbalance sensors. Measurement Science and Technology, 20(12), 124004. doi:10.1088/0957-0233/20/12/124004Montagut, Y. J., García, J. V., Jiménez, Y., March, C., Montoya, A., & Arnau, A. (2011). Frequency-shift vs phase-shift characterization of in-liquid quartz crystal microbalance applications. Review of Scientific Instruments, 82(6), 064702. doi:10.1063/1.3598340Abad, A., Primo, J., & Montoya, A. (1997). Development of an Enzyme-Linked Immunosorbent Assay to Carbaryl. 1. Antibody Production from Several Haptens and Characterization in Different Immunoassay Formats. Journal of Agricultural and Food Chemistry, 45(4), 1486-1494. doi:10.1021/jf9506904Manclús, J. J., Abad, A., Lebedev, M. Y., Mojarrad, F., Micková, B., Mercader, J. V., … Montoya, A. (2004). Development of a Monoclonal Immunoassay Selective for Chlorinated Cyclodiene Insecticides. Journal of Agricultural and Food Chemistry, 52(10), 2776-2784. doi:10.1021/jf035382hFrancis, L. A., Friedt, J.-M., Zhou, C., & Bertrand, P. (2006). In Situ Evaluation of Density, Viscosity, and Thickness of Adsorbed Soft Layers by Combined Surface Acoustic Wave and Surface Plasmon Resonance. Analytical Chemistry, 78(12), 4200-4209. doi:10.1021/ac051233hMauriz, E., Calle, A., Abad, A., Montoya, A., Hildebrandt, A., Barceló, D., & Lechuga, L. M. (2006). Determination of carbaryl in natural water samples by a surface plasmon resonance flow-through immunosensor. Biosensors and Bioelectronics, 21(11), 2129-2136. doi:10.1016/j.bios.2005.10.013Abad, A., & Montoya, A. (1997). Development of an Enzyme-Linked Immunosorbent Assay to Carbaryl. 2. Assay Optimization and Application to the Analysis of Water Samples. Journal of Agricultural and Food Chemistry, 45(4), 1495-1501. doi:10.1021/jf950691wMontagut, Y., García, J. V., Jiménez, Y., March, C., Montoya, Á., & Arnau, A. (2011). Validation of a Phase-Mass Characterization Concept and Interface for Acoustic Biosensors. Sensors, 11(5), 4702-4720. doi:10.3390/s11050470

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

Analysis, implementation and validation of a Love mode surface acoustic wave device for its application as sensor of biological processes in liquid media

En las últimas dos décadas, han surgido diferentes tecnologías acústicas para aplicaciones biosensoras como alternativas a tecnologías de detección bien establecidas ¿acústicas o ópticas¿ como son la Microbalanza de Cuarzo (QCM, por sus siglas en inglés) y la Resonancia de Plasmón de Superficie (SPR, de sus siglas en inglés). En la primera parte de este documento se revisan dichas tecnologías alternativas para aplicaciones en medio líquido. Como resultado de esta revisión, se determina que los dispositivos de onda acústica superficial Love (LW, de sus siglas en inglés) son los más prometedores y viables para conseguir el principal objetivo de esta Tesis, que es establecer una comparativa en las mismas condiciones entre inmnosensores desarrollados con la tecnología seleccionada en esta tesis y los inmunosensores desarrollados con QCMs de Alta Frecuencia Fundamental (HFF-QCM, por sus siglas en inglés). Después de esta revisión se presenta el estado del arte de los dispositivos LW en su aplicación como biosensores, así como una discusión de las tendencias y retos actuales de este tipo de sensores. Posteriormente se reúne la información más actualizada sobre aspectos de diseño, principios de operación y modelado de estos sensores. Algunos aspectos de diseño son estudiados y probados para establecer el diseño final de los dispositivos LW. Previamente a su fabricación, también se realizan simulaciones para modelar el comportamiento del dispositivo elegido previamente a su fabricación. Posteriormente, se describe la fabricación del dispositivo así como la celda de flujo diseñada para trabajar con el dispositivo en medios líquidos. Adicionalmente, un sistema electrónico de caracterización, previamente validado para sensores QCM, se adapta para sensores LW. Como resultados, se valida el sistema electrónico para caracterizar los sensores LW fabricados y montados en la celda de flujo y, finalmente, se desarrolla un inmunosensor para la detección del pesticida carbaril que se compara con otras tecnologías inmunosensoras.In the last two decades, different acoustic technologies for biosensors applications have emerged as promising alternatives to other better established detection technologies ¿ acoustic or optic ones- such as traditional Quartz Crystal Microbalance (QCM) and Surface Plasmon Resonance (SPR). The alternative acoustic technologies for in liquid measurements are reviewed in this manuscript. Surface Acoustic Wave (SAW) Love Mode or Love Wave (LW) sensors are determined to be the most promising and viable option to work with for achieving the main aim of this Thesis. Such aim is the development of a LW immunosensor for its comparison with the same application based on High Fundamental Frequency-QCM (HFF-QCM) sensors and under the same conditions. Consequently, the state-of-the-art of LW devices for biosensing is provided and a discussion about the current trends and future challenges of these sensors is presented. In order to start working with suitable LW devices, upto- date information regarding the design aspects, operation principles and modeling of such devices is gathered. Some design aspects are explored and tested to establish the design of the final LW device. Different simulations for modeling the chosen device behavior are carried out before its fabrication. Later, the device fabrication is described. Next, to start working with the fabricated device in liquid media, a flow cell is designed and implemented. In addition, an electronic characterization system, previously validated for QCM sensors, is adapted and tested for the fabricated LW device. As results, the adapted electronic characterization system is validated for LW devices mounted in the fabricated flow cell and, finally, a LW-based immunosensor for the determination of carbaryl pesticide was developed and compared with other immunosensor technologies.Rocha Gaso, MI. (2013). Analysis, implementation and validation of a Love mode surface acoustic wave device for its application as sensor of biological processes in liquid media [Tesis doctoral]. Editorial Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/32492TESI

Lab-on-a-chip based integrated hybrid technologies for biofluids manipulation and characterization

[EN] The goal of this work is to develop an original and high performance hybrid lab-on-a-chip, coupling actuators and biosensors for the active control and broad range characterization of biofluids samples. These biofluids will be controlled (moved and mixed) actively by Rayleigh-Surface Acoustic Wave (R-SAW) and analysed in real-time by combining Love-SAW (L-SAW) and Surface Plasmon Resonance (SPR) technologies. A microfluidic chamber and specific transducer were designed for this application to interact efficiently with the liquid sample entrapped in the chamber and to detect modifications of its physical properties such as viscosity. AT-cut quartz and LiNbO3 36Y-X substrates were used to generate both Rayleigh and Shear-Horizontal waves and ZnO material as guiding layer. The whole system has proven is full efficiency to interact with the fluid and to detect the signal perturbations with no significant loss compared with the measurements carried out with a probe station.The authors wish to thank the ANR for its financial support through the AWESOM project (ANR-12-BS09-021); the operators Laurent Bouvot, Jean Georges Mussot and Emmanuel Vatoux, for their support and assistance in the fabrication of the cell and the SAW devices; and the engineering students, Olivier Bettoni and Bastien Lafont, for their contribution to the cell design.Rocha Gaso, MI.; Renaudin, A.; Sarry, F.; Beyssen, D. (2015). Lab-on-a-chip based integrated hybrid technologies for biofluids manipulation and characterization. Procedia Engineering. 120:687-690. https://doi.org/10.1016/j.proeng.2015.08.75068769012

High Frequency (100, 150 MHz) Quartz Crystal Microbalance (QCM) Piezoelectric Genosensor for the Determination of the Escherichia coli O157 rfbE Gene

[EN] Escherichia coli O157 (E. coli O157) is responsible for outbreaks of high morbidity in food-borne infections. The development of sensitive, reliable, and selective detection systems is of great importance in food safety. In this work, two high fundamental frequency (HFF) piezoelectric genosensors (100 and 150 MHz) were designed and validated for the rfbE gene detection, which encodes O-antigen in E. coli O157. HFF resonators offer improved sensitivity, small sample volumes, and the possibility of integration into lab-on-a-chip devices, but their sensing capabilities have not yet been fully explored. This HFF-QCM genosensor uses the method of physisorption based on the union between the streptavidin and the biotin to immobilize the genetic bioreceptor on the surface and detect its hybridization with the target sequence. Parameters such as molecular coating, specificity, and variability were tested to enhance its performance. Although both genosensors evaluated are able to determine the target, the 100 MHz device has a higher response to the analyte than the 150 MHz platform. This is the first step in the development of an HFF-QCM genosensor that may be used as a trial test of E. coli O157 in large batches of samples.This work was supported by the EIA University and the National University of Colombia under internal call in 2016.Barrientos, K.; Rocha Gaso, MI.; Jaramillo, M.; Aldrín Vásquez, N. (2022). High Frequency (100, 150 MHz) Quartz Crystal Microbalance (QCM) Piezoelectric Genosensor for the Determination of the Escherichia coli O157 rfbE Gene. Analytical Letters. 55(17):2697-2709. https://doi.org/10.1080/00032719.2022.206856626972709551