Acoustic Wave Biosensor Research Articles

Isolation and Characterization of Exosomes from Cancer Cells Using Antibody-Functionalized Paddle Screw-Type Devices and Detection of Exosomal miRNA Using Piezoelectric Biosensor.

Exosomes are small extracellular vesicles produced by almost all cell types in the human body, and exosomal microRNAs (miRNAs) are small non-coding RNA molecules that are known to serve as important biomarkers for diseases such as cancer. Given that the upregulation of miR-106b is closely associated with several types of malignancies, the sensitive and accurate detection of miR-106b is important but difficult. In this study, a surface acoustic wave (SAW) biosensor was developed to detect miR-106b isolated from cancer cells based on immunoaffinity separation technique using our unique paddle screw device. Our novel SAW biosensor could detect a miR-106b concentration as low as 0.0034 pM in a linear range from 0.1 pM to 1.0 μM with a correlation coefficient of 0.997. Additionally, we were able to successfully detect miR-106b in total RNA extracted from the exosomes isolated from the MCF-7 cancer cell line, a model system for human breast cancer, with performance comparable to commercial RT-qPCR methods. Therefore, the exosome isolation by the paddle screw method and the miRNA detection using the SAW biosensor has the potential to be used in basic biological research and clinical diagnosis as an alternative to RT-qPCR.

Design and Simulation of MEMS Surface Acoustic Wave Biosensor Based on PVDF Thin Film

Abstract: Surface acoustic wave sensors, as a class of MEMS, are widely used recently. The sensor can transform an input electrical signal into a mechanical wave which can be easily influenced by physical phenomena. Then, the changed mechanical wave is transduced back into an electrical signal. The presence of the desired phenomenon can be detected through the difference between the input and output electrical signal (amplitude, phase, frequency, or time delay). The basic surface acoustic wave device consists of a piezoelectric substrate, an input interdigitated transducer (IDT) on one side of the surface of the substrate, and a second output interdigitated transducer on the other side of the substrate.

Interaction of Pseudomonas aeruginosa with surface-modified silica studied by ultra-high frequency acoustic wave biosensor

Aim: This study aimed to examine the amount of surface non-specific adsorption, or fouling, observed by Pseudomonas aeruginosa (P. aeruginosa) on a quartz crystal based acoustic wave biosensor under different flow conditions with and without an anti-fouling layer. Methods: An electromagnetic piezoelectric acoustic sensor (EMPAS) based on electrode free quartz crystals was used to perform the analysis. Phosphate buffered saline (PBS) was flowed over the crystal surface at various flow rates from 50 μL/min to 200 μL/min, with measurements being taken at the 43rd harmonic (~864 MHz). The crystal was either unmodified, or modified with a monoethylene glycol [2-(3-silylpropyloxy)-hydroxy-ethyl (MEG-OH)] anti-fouling layer. Overnight culture of P. aeruginosa PAO1 (PAO1) in lysogeny broth (LB) was injected into the system, and flow maintained for 30 min. Results: The frequency change of the EMPAS crystal after injection of bacteria into the system was found to change based on the flow rate of buffer, suggesting the flow rate has a strong effect on the level of non-specific adsorption. The MEG-OH layer drastically reduced the level of fouling observed under all flow conditions, as well as reduced the amount of variation between experiments. Flow rates of 150 μL/min or higher were found to best reduce the level of fouling observed as well as experimental variation. Conclusions: The MEG-OH anti-fouling layer is important for accurate and reproducible biosensing measurements due to the reduced fouling and variation during experiments. Additionally, a flow rate of 150 μL/min may prove better for measurement compared to the current standard of 50 μL/min for this type of instrument.

Interaction of Staphylococcus aureus and Candida albicans with surface-modified silica studied by ultra-high frequency acoustic wave biosensor.

In this work the bacteria S. aureus and fungi C. albicans were allowed to interact with quartz-based biosensor devices under different flow rates, with and without an anti-fouling coating. These experiments were conducted in order to determine if the level of fouling observed was affected by the flow rate. The biosensor used was an ultra-high frequency acoustic wave device (EMPAS) for investigation of device surface initial interaction of S. aureus or C. albicans under flow of PBS buffer at flow rates between 50 and 200 μL min-1. Surface-bound microbes were also visualized by fluorescence microscopy following these experiments. S. aureus bacteria was able to foul the bare quartz sensors at each flow rate tested, with the greatest degree of fouling observed at a flow rate of 100 μL min-1. C. albicans showed far less fouling of bare devices with the maximum fouling observed at a flow rate of 75 μL min-1. Antifouling MEG-OH coated sensors showed greatly reduced fouling for S. aureus, with between a 90 and 99% reduction in observed frequency change depending on the flow rate used, and between 22 and 90% for C. albicans. Fluorescence images of the microbes following the experiments correlated well with the frequency data, showing a marked decrease in the amount of bacteria seen on MEG-OH-coated surfaces compared to controls.

Integrated Rayleigh wave streaming-enhanced sensitivity of shear horizontal surface acoustic wave biosensors

Shear horizontal surface acoustic wave (SH-SAW) sensors are regarded as a promising alternative for label-free, sensitive, real time and low-cost detection. Nevertheless, achieving high sensitivity with SH-SAW has approached its limit imposed by the mass transport and probe-target affinity. We present here an SH-SAW biosensor accompanied by a unique Rayleigh wave-based actuator. The platform assembled on an ST-quartz substrate consists of dual-channel SH-SAW delay lines fabricated along a 90°-rotated direction, whilst another interdigital electrode (IDT) is orthogonally placed to generate Rayleigh waves so as to induce favourable streaming in the bio-chamber, enhancing the binding efficiency of the bio-target. Theoretical foundation and simulation have shown that Rayleigh acoustic streaming generates a level of agitation that accelerates the mass transport of the biomolecules to the surface. A fourfold improvement in sensitivity is achieved compared with conventional SH-SAW biosensors by means of complementary DNA hybridization with the aid of the Rayleigh wave device, giving a sensitivity level up to 6.15 Hz/(ng/mL) and a limit of detection of 0.617 ng/mL. This suggests that the proposed scheme could improve the sensitivity of SAW biosensors in real-time detection.

Sensitivity measurements for a 250 MHz quartz shear-horizontal surface acoustic wave biosensor under liquid viscous loading

Surface acoustic wave (SAW) devices have been used in biochemical assays due to their high sensitivity. The device sensitivity is a function of changes in the density and viscosity of the liquid. Here, we studied the effect of fluid viscosity using a 250 MHz quartz shear-horizontal (SH)-SAW biosensor by monitoring different concentrations of binary aqueous/glycerol solutions. In this study, the sensitivity of the biosensor was determined by fitting the data to models derived from perturbation theory. Measurements in water were used as the reference. For a 0% to 50% glycerol solution, an 87°–204° separation in the phase shift was observed. The slope of the plot of the phase shift vs (ηρ)0.5 was used to indicate the sensor’s sensitivity. The sensitivity for our 250 MHz quartz SH-SAW sensors was calculated to be 3.7×10−3m2sKg. The corresponding mass sensitivity was determined to be 9.25 × 105m2Kg. The limit of detection was calculated to be 36 picograms (pg), while the limit of quantification or LOQ was calculated to be 109 pg. Traditionally, liquid phase measurements have been challenging for SAW devices because liquids dampen the vibrating sensors severely. This problem has been largely solved using a transverse (shear) wave instead of the more popular longitudinal or Rayleigh waves. Liquid measurements are now possible using transverse waves, also known as shear waves, because transverse waves are only minimally attenuated by liquids. Shear-horizontal SAW sensors (SH-SAW) show great promise as label-free biosensors because of their ability to handle liquid samples. However, the viscosity of the liquid still induces loading effects and can be measured when the liquid is loaded onto the SH-SAW propagating surface (delay line). When the liquid above the delay line is perturbed by physical or chemical changes, such as binding to a receptor, it alters the propagating acoustic wave. The SH-SAW device can measure these changes in liquid properties as a change in the wave’s phase compared to the original wave. The device’s phase shift was recorded as a function of the changes in the density and viscosity of the binary glycerol solution and used to determine the sensitivity in the linear dynamic range of responses.

Application of Shear Horizontal Surface Acoustic Wave (SH-SAW) Immunosensor in Point-of-Care Diagnosis.

Point-of-care testing (POCT), also known as on-site or near-patient testing, has been exploding in the last 20 years. A favorable POCT device requires minimal sample handling (e.g., finger-prick samples, but plasma for analysis), minimal sample volume (e.g., one drop of blood), and very fast results. Shear horizontal surface acoustic wave (SH-SAW) biosensors have attracted a lot of attention as one of the effective solutions to complete whole blood measurements in less than 3 min, while providing a low-cost and small-sized device. This review provides an overview of the SH-SAW biosensor system that has been successfully commercialized for medical use. Three unique features of the system are a disposable test cartridge with an SH-SAW sensor chip, a mass-produced bio-coating, and a palm-sized reader. This paper first discusses the characteristics and performance of the SH-SAW sensor system. Subsequently, the method of cross-linking biomaterials and the analysis of SH-SAW real-time signals are investigated, and the detection range and detection limit are presented.

A novel leaky surface acoustic wave biosensor for detection of PKA activity in cell lysates based on peptide biomineralized metal nanoclusters

A novel leaky surface acoustic wave biosensor for detection of PKA activity in cell lysates based on peptide biomineralized metal nanoclusters

Highly sensitive surface acoustic wave biosensor for the detection of Hg2+ based on the thymine-Hg2+-thymine structure.

The detection of mercury ions (Hg2+) with an ultralow concentration is of great significance for the treatment of heavy metal pollution in industrial waste water and the monitoring of drinking water quality. In this study, a simple, sensitive, and portable surface acoustic wave (SAW) biosensor for the determination of Hg2+ concentration was developed. First, a DNA fragment with a specific binding capacity for Hg2+ was chemically adsorbed on to the sensitive region through forming Au-S bonds with the SAW biosensor. The DNA probe could then form a thymine-Hg2+-thymine (T-Hg2+-T) complex with strong affinity for Hg2+ after immersion in a test solution containing Hg2+, resulting in a significant change in the response frequency of the device. The linear detection range of the device was 10 pM to 1 nM, while the limit of detection (LOD) was as low as 6.3 pM. Furthermore, the SAW biosensor exhibited excellent selectivity to Hg2+ compared with that of interfering ions, e.g., Ag+, Ba2+, Cu2+, Zn2+, Mn2+, Fe3+, Ca2+, and Na+. The results provide a new strategy for the preparation of portable devices that can monitor toxic heavy metal ions with high sensitivity and selectivity.

A Surface Acoustic Wave (SAW)-Based Lab-on-Chip for the Detection of Active α-Glycosidase

Enzyme detection in liquid samples is a complex laboratory procedure, based on assays that are generally time- and cost-consuming, and require specialized personnel. Surface acoustic wave sensors can be used for this application, overcoming the cited limitations. To give our contribution, in this work we present the bottom-up development of a surface acoustic wave biosensor to detect active α-glycosidase in aqueous solutions. Our device, optimized to work at an ultra-high frequency (around 740 MHz), is functionalized with a newly synthesized probe 7-mercapto-1-eptyl-D-maltoside, bringing one maltoside terminal moiety. The probe is designed ad hoc for this application and tested in-cuvette to analyze the enzymatic conversion kinetics at different times, temperatures and enzyme concentrations. Preliminary data are used to optimize the detection protocol with the SAW device. In around 60 min, the SAW device is able to detect the enzymatic conversion of the maltoside unit into glucose in the presence of the active enzyme. We obtained successful α-glycosidase detection in the concentration range 0.15-150 U/mL, with an increasing signal in the range up to 15 U/mL. We also checked the sensor performance in the presence of an enzyme inhibitor as a control test, with a signal decrease of 80% in the presence of the inhibitor. The results demonstrate the synergic effect of our SAW Lab-on-a-Chip and probe design as a valid alternative to conventional laboratory tests.

Development of nanobiosensors for human immunodeficiency virus detection—a mini review

Acquired immunodeficiency syndrome (AIDS) is a global disease caused by human immunodeficiency virus (HIV). About 50 million people have died worldwide due to HIV-1 infection alone. HIV is a primary sexually transmitted infection but can also spread via breastfeeding, blood transfer, organ transfer, etc. Early detection with the maintenance of the disease is the only way to reduce the spread and severity of the disease. There are many conventional techniques for the detection of the virus. Still, recently nano-based diagnostic method remains a little ahead of these techniques due to advancements in nanotechnology. Nanomaterial-based biosensors constitute a significant part of the discussion because of their high sensitivity and accuracy. Nanobiosensors like electronic nano biosensors, quantum dot (QD)-based biosensors, optical biosensors, electronic biosensors, electrochemiluminescence nanosensors, field-effect transistor (FET) biosensors, surface acoustic wave (SAW) biosensors, graphene-based biosensors, etc. have been widely used for detecting HIV in human blood samples. All these biosensors offer promising results in the detection of the virus. In this article, different types of nanobiosensors and their application in the field of diagnosis and maintenance of HIV was reviewed.

Glycocalyx Sensing with a Mathematical Model of Acoustic Shear Wave Biosensor.

The article deals with an idea of exploiting an acoustic shear wave biosensor for investigating the glycocalyx, a polysaccharide polymer molecule layer on the endothelium of blood vessels that, according to recent studies, plays an important role in protecting against diseases. To test this idea, a mathematical model of an acoustic shear wave sensor and corresponding software developed earlier for proteomic applications are used. In this case, the glycocalyx is treated as a layer homogenized over the thin polymer “villi”. Its material characteristics depend on the density, thickness, and length of the villi and on the viscous properties of the surrounding liquid (blood plasma). It is proved that the model used has a good sensitivity to the above parameters of the villi and blood plasma. Numerical experiments performed using real data collected retrospectively from premature infants show that the use of acoustic shear wave sensors may be a promising approach to investigate properties of glycocalyx-like structures and their role in prematurity.

Silica Inverse Opal Nanostructured Sensors for Enhanced Immunodetection of Extracellular Vesicles by Quartz Crystal Microbalance with Dissipation Monitoring.

Extracellular vesicles (EVs) are nanosized circulating assemblies that contain biomarkers considered promising for early diagnosis within neurology, cardiology, and oncology. Recently, acoustic wave biosensors, in particular based on quartz crystal microbalance with dissipation monitoring (QCM-D), have emerged as a sensitive, label-free, and selective EV characterization platform. A rational approach to further improving sensing detection limits relies on the nanostructuration of the sensor surfaces. To this end, inorganic inverse opals (IOs) derived from colloidal self-assembly present a highly tunable and scalable nanoarchitecture of suitable feature sizes and surface chemistry. This work systematically investigates their use in two-dimensional (2D) and three-dimensional (3D) for enhanced QCM-D EV detection. Precise tuning of the architecture parameters delivered improvements in detection performance to sensitivities as low as 6.24 × 107 particles/mL. Our findings emphasize that attempts to enhance acoustic immunosensing via increasing the surface area by 3D nanostructuration need to be carefully analyzed in order to exclude solvent and artifact entrapment effects. Moreover, the use of 2D nanostructured electrodes to compartmentalize analyte anchoring presents a particularly promising design principle.

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.

A Novel Rapid Test to Detect Anti-SARS-CoV-2 N Protein IgG Based on Shear Horizontal Surface Acoustic Wave (SH-SAW).

Since the Coronavirus disease 2019 (COVID-19) pandemic outbreak, many methods have been used to detect antigens or antibodies to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), including viral culture, nucleic acid test, and immunoassay. The shear-horizontal surface acoustic wave (SH-SAW) biosensor is a novel pathogen detection platform with the advantages of high sensitivity and short detection time. The objective of this study is to develop a SH-SAW biosensor to detect the anti-SARS-CoV-2 nucleocapsid antibody. The rabbit sera collected from rabbits on different days after SARS-CoV-2 N protein injection were evaluated by SH-SAW biosensor and enzyme-linked immunosorbent assay (ELISA). The results showed that the SH-SAW biosensor achieved a high correlation coefficient (R = 0.9997) with different concentrations (34.375–1100 ng/mL) of the “spike-in” anti-N protein antibodies. Compared to ELISA, the SH-SAW biosensor has better sensitivity and can detect anti-N protein IgG signals earlier than ELISA on day 6 (p < 0.05). Overall, in this study, we demonstrated that the SH-SAW biosensor is a promising platform for rapid in vitro diagnostic (IVD) testing, especially for antigen or antibody testing.

Surface Adsorption of the Cancer Biomarker Lysophosphatidic Acid in Serum Studied by Acoustic Wave Biosensor.

The thickness shear mode acoustic wave device is of interest for the sensing of biomarkers for diseases in various biological fluids, but suffers from the issue of non-specific adsorption of compounds other than those of interest to the electrode surface, thus affecting the device’s output. The aim of this present study was to determine the level of non-specific adsorption on gold electrodes from serum samples with added ovarian cancer biomarker lysophosphatidic acid in the presence of a surface anti-fouling layer. The latter was an oligoethylene molecule with thiol group for attachment to the electrode surface. It was found that the anti-fouling layer had a minimal effect on the level of both adsorption of components from serum and the marker. This result stands in sharp contrast to the analogous monolayer employed for anti-fouling reduction on silica.

Evaluation of Shear Horizontal Surface Acoustic Wave Biosensors Using "Layer Parameter" Obtained from Sensor Responses during Immunoreaction.

Shear horizontal surface acoustic wave (SH-SAW) biosensors measure the reaction of capture antibodies immobilized on the sensing surface to capture test molecules (antigens) by using the change in SH-SAW propagation characteristics. SH-SAW displacement exists not only on the SH-SAW propagating surface, but also partially penetrates the specimen liquid to a certain depth, which is determined by the liquid properties of the specimen and the operating frequency of the SH-SAW. This phenomenon is called viscosity penetration. In previous studies, the effect of viscosity penetration was not considered in the measurement of SH-SAW biosensors, and the mass or viscosity change caused by the specific binding of capture antibodies to the target antigen was mainly used for the measurement. However, by considering the effect of viscosity penetration, it was found that the antigen–antibody reaction could be measured and the detection characteristics of the biosensor could be improved. Therefore, this study aims to evaluate the detection properties of SH-SAW biosensors in the surface height direction by investigating the relationship between molecular dimensions and SH-SAW propagation characteristics, which are pseudo-changed by varying the diameter of gold nanoparticles. For the evaluation, we introduced a layer parameter defined by the ratio of the SH-SAW amplitude change to the SH-SAW velocity change caused by the antigen–antibody reaction. We found a correlation between the layer parameter and pseudo-varied molecular dimensions. The results suggest that SH-SAW does not only measure the mass and viscosity but can also measure the size of the molecule to be detected. This shows that SH-SAW biosensors can be used for advanced functionality.

Surface Acoustic Wave Biosensor with Laser-Deposited Gold Layer Having Controlled Porosity

Laser-deposited gold immobilization layers having different porosities were incorporated into love wave surface acoustic wave sensors (LW-SAWs). Variation of pulsed laser deposition parameters allows good control of the gold film morphology. Biosensors with various gold film porosities were tested using the biotin–avidin reaction. Control of the Au layer morphology is important since the biotin and avidin layer morphologies closely follow that of the gold. The response of the sensors to biotin/avidin, which is a good indicator of biosensor performance, is improved when the gold layer has increased porosity. Given the sizes of the proteins, the laser-deposited porous gold interfaces have optimal pore dimensions to ensure protein stability.

Regenerable ZnO/GaAs Bulk Acoustic Wave Biosensor for Detection of Escherichia coli in "Complex" Biological Medium.

A regenerable bulk acoustic wave (BAW) biosensor is developed for the rapid, label-free and selective detection of Escherichia coli in liquid media. The geometry of the biosensor consists of a GaAs membrane coated with a thin film of piezoelectric ZnO on its top surface. A pair of electrodes deposited on the ZnO film allows the generation of BAWs by lateral field excitation. The back surface of the membrane is functionalized with alkanethiol self-assembled monolayers and antibodies against E. coli. The antibody immobilization was investigated as a function of the concentration of antibody suspensions, their pH and incubation time, designed to optimize the immunocapture of bacteria. The performance of the biosensor was evaluated by detection tests in different environments for bacterial suspensions ranging between 103 and 108 CFU/mL. A linear dependence between the frequency response and the logarithm of E. coli concentration was observed for suspensions ranging between 103 and 107 CFU/mL, with the limit of detection of the biosensor estimated at 103 CFU/mL. The 5-fold regeneration and excellent selectivity towards E. coli detected at 104 CFU/mL in a suspension tinted with Bacillus subtilis at 106 CFU/mL illustrate the biosensor potential for the attractive operation in complex biological media.

Shear Horizontal Phononic Metasurface for In-Liquid Gravimetric Biosensing

In-liquid wave attenuation is the major factor that limits the resolution of acoustic wave biosensors. Existing acoustic wave devices rely on shear mode operation to reduce the mechanical motion transferred to the liquid. However, the energy leakage still exists through acoustic radiation. In this article, we combined phononic metasurface based on local mechanical resonators with shear horizontal surface acoustic wave (SH-SAW) to address the above issue. The shear-horizontal phononic metasurface (SH-PM) resonator is made of periodic high-aspect-ratio electrode (HARE). It produces a slow phononic wave that cannot propagate in water due to its low velocity, which suppresses the acoustic radiation. Furthermore, as the particle motion of the HARE is polarized in shear mode, the SH-PM also reduces the mechanical motion transfer to water. The minimization of the total acoustic energy leakage in the SH-PM results in high quality factor (Q) resonance in water. Besides, the increased surface-to-volume ratio of the HAREs also enhances the mass sensitivity of the device. Experimental results show that the SH-PM resonator achieved an in-liquid Q factor of 230 and a normalized mass sensitivity of - 192.4 <i xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">cm</i> <sup xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">2</sup> /g. The high in-liquid Q factor together with the high mass sensitivity makes the SH-PM a promising candidate for high-resolution gravimetric detection of biomolecules.