Shear Horizontal Surface Acoustic Wave Biosensor Research Articles

New Advances in Rapid Pretreatment for Small Dense LDL Cholesterol Measurement Using Shear Horizontal Surface Acoustic Wave (SH-SAW) Technology.

Atherosclerosis is an inflammatory disease of the arteries associated with alterations in lipid and other metabolism and is a major cause of cardiovascular disease (CVD). LDL consists of several subclasses with different sizes, densities, and physicochemical compositions. Small dense LDL (sd-LDL) is a subclass of LDL. There is growing evidence that sd-LDL-C is associated with CVD risk, metabolic dysregulation, and several pathophysiological processes. In this study, we present a straightforward membrane device filtration method that can be performed with simple laboratory methods to directly determine sd-LDL in serum without the need for specialized equipment. The method consists of three steps: first, the precipitation of lipoproteins with magnesium harpin; second, the collection of effluent from a 100 nm filter; and third, the quantification of sd-LDL-ApoB in the effluent with an SH-SAW biosensor. There was a good correlation between ApoB values obtained using the centrifugation (y = 1.0411x + 12.96, r = 0.82, n = 20) and filtration (y = 1.0633x + 15.13, r = 0.88, n = 20) methods and commercially available sd-LDL-C assay values. In addition to the filtrate method, there was also a close correlation between sd-LDL-C and ELISA assay values (y = 1.0483x - 4489, r = 0.88, n = 20). The filtration treatment method also showed a high correlation with LDL subfractions and NMR spectra ApoB measurements (y = 2.4846x + 4.637, r = 0.89, n = 20). The presence of sd-LDL-ApoB in the effluent was also confirmed by ELISA assay. These results suggest that this filtration method is a simple and promising pretreatment for use with the SH-SAW biosensor as a rapid in vitro diagnostic (IVD) method for predicting sd-LDL concentrations. Overall, we propose a very sensitive and specific SH-SAW biosensor with the ApoB antibody in its sensitive region to monitor sd-LDL levels by employing a simple delay-time phase shifted SH-SAW device. In conclusion, based on the demonstration of our study, the SH-SAW biosensor could be a strong candidate for the future measurement of sd-LDL.

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 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.

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.

Frequency Shift of a SH-SAW Biosensor with Glutaraldehyde and 3-Aminopropyltriethoxysilane Functionalized Films for Detection of Epidermal Growth Factor.

The frequency shift of a shear-horizontal surface-acoustic-wave (SH-SAW) biosensor in which the concentration of biomolecule is determined by the amount of its adsorption on the sensing film was studied. Simulation results were compared with experimental results to investigate its sensitivity and to develop a model to estimate the concentration of a cancer-related biomarker antigen epidermal growth factor (EGF) in the sample, with two types of sensing films, 3-aminopropyltriethoxysilane (APTES) and glutaraldehyde. With the concentration of the targeted biomarker varying from 0.2 to 5 ng/mL, a typical exponential relationship was found between the concentration and the frequency shift of the SH-SAW sensor. Measurement results showed a clear response of this immunosensor to the mass-loading effects of the antibody–antigen. The sensitivity of the glutaraldehyde film is greater than that of the APTES film owing to the chemisorption of the antibody. In the simulation, a shift of the SH-SAW resonant frequency due to added mass occurred on applying an incremental surface mass density on the sensing film, while in real applications, the concentration of the targeted biomarker to be absorbed in the sensing film is demanded. An empirical model was proposed to calculate the frequency shift in the simulation of the SH-SAW biosensor, corresponding to the concentration of specific biomolecules absorbed on a specific film. From the semi-empirical model, the sensitivity level is found to be 0.641 and 1.709 kHz/(ng/mL) for APTES and glutaraldehyde sensing films, respectively, at a biomarker concentration of less than 1 ng/mL. The developed method is useful for quickly estimating the frequency shift with respect to the concentration of the target molecules in the simulation for SH-SAW sensors.

An aptamer-based shear horizontal surface acoustic wave biosensor with a CVD-grown single-layered graphene film for high-sensitivity detection of a label-free endotoxin

The thickness of the sensitive layer has an important influence on the sensitivity of a shear horizontal surface acoustic wave (SH-SAW) biosensor with a delay-line structure and lower layer numbers of graphene produce better sensitivity for biological detection. Therefore, a label-free and highly sensitive SH-SAW biosensor with chemical vapor deposition (CVD-)-grown single-layered graphene (SLG) for endotoxin detection was developed in this study. With this methodology, SH-SAW biosensors were fabricated on a 36° Y-90° X quartz substrate with a base frequency of 246.2 MHz, and an effective detection cell was fabricated using acrylic material. To increase the surface hydrophilicity, chitosan was applied to the surface of the SLG film. Additionally, the aptamer was immobilized on the surface of the SLG film by cross-linking with glutaraldehyde. Finally, the sensitivity was verified by endotoxin detection with a linear detection ranging from 0 to 100 ng/mL, and the detection limit (LOD) was as low as 3.53 ng/mL. In addition, the stability of this type of SH-SAW biosensor from the air phase to the liquid phase proved to be excellent and the specificity was tested and verified by detecting the endotoxin obtained from Escherichia coli (E. coli), the endotoxin obtained from Pseudomonas aeruginosa (P. aeruginosa), and aflatoxin. Therefore, this type of SH-SAW biosensor with a CVD-grown SLG film may offer a promising approach to endotoxin detection, and it may have great potential in clinical applications.

Highly Sensitive Escherichia coli Shear Horizontal Surface Acoustic Wave Biosensor with Silicon Dioxide Nanostructures

In this study, a shear horizontal surface acoustic wave (SHSAW) was used for Escherichia coli (E. coli) detection. E. coli O157:H7 serotype, a dangerous strain among 225 E. coli unique serotypes was chosen as test sample. A few cells of this bacterium are able to cause young children to be most vulnerable to serious complications. Presence of higher than 1 cfu E. coli O157:H7 in 25 g of food has been considered as a dangerous level. The SHSAW biosensor was fabricated on 640 YX LiNbO3 substrate. Five different interdigital transducers (IDT) parameters (Fig. 1(a)-1(e)) SHSAW were fabricated to compare the mass loading sensitivity before advancing to the bio sensing application: four of them were 32 μm pitch size (with the average of synchronous frequency, f0 = 144.303 MHz) and one was 12 μm pitch size (with f0 = 384.948 MHz). All of the four 32 μm pitch sizes were with different combination of delay line length (3.904 mm and 7.296 mm) and aperture size (1.376 mm and 2.464 mm). As for the 12 μm pitch size device, it was with 2.1 mm delay line length and 0.72 mm aperture size. Eventually, 12 μm pitch size device was selected for oligonucleotides detection and its mass loading sensitivity was 1558.04 MHz/ (mg/mm2), 4.8-fold higher than the most sensitivity one in 32 μm pitch size).Its sensitivity was enhanced by depositing 130 nm thin layer of SiO2 nanostructures with particle size less than 70 nm (Fig. 1(f)-1(g)). The nanostructures act both as a waveguide as well as the surface physical modification of the sensor prior to biomolecule immobilization. For this work, a specific DNA sequence from E. coli O157:H7 having 22 mers as an amine-terminated probe ssDNA was immobilized on the thin film sensing area through chemical functionalization [(CHO-(CH2)3-CHO) and (APTES; NH2-(CH2)3-Si(OC2H5)3]. The high-performance of sensor was shown with the oligonucleotide target and attained the sensitivity of 0.6439 nM/ 0.1 kHz and detection limit was down to 1.8 femtomolar (1.8 x 10-15 M). Further evidence was provided by specificity analysis using single mismatched and complementary oligonucleotide sequences.

Highly sensitive Escherichia coli shear horizontal surface acoustic wave biosensor with silicon dioxide nanostructures

Surface acoustic wave mediated transductions have been widely used in the sensors and actuators applications. In this study, a shear horizontal surface acoustic wave (SHSAW) was used for the detection of food pathogenic Escherichia coli O157:H7 (E.coli O157:H7), a dangerous strain among 225 E. coli unique serotypes. A few cells of this bacterium are able to cause young children to be most vulnerable to serious complications. Presence of higher than 1cfu E.coli O157:H7 in 25g of food has been considered as a dangerous level. The SHSAW biosensor was fabricated on 64° YX LiNbO3 substrate. Its sensitivity was enhanced by depositing 130.5nm thin layer of SiO2 nanostructures with particle size lesser than 70nm. The nanostructures act both as a waveguide as well as a physical surface modification of the sensor prior to biomolecular immobilization. A specific DNA sequence from E. coli O157:H7 having 22 mers as an amine-terminated probe ssDNA was immobilized on the thin film sensing area through chemical functionalization [(CHO-(CH2)3-CHO) and APTES; NH2-(CH2)3-Si(OC2H5)3]. The high-performance of sensor was shown with the specific oligonucleotide target and attained the sensitivity of 0.6439nM/0.1kHz and detection limit was down to 1.8femto-molar (1.8×10−15M). Further evidence was provided by specificity analysis using single mismatched and complementary oligonucleotide sequences.

Effect of viscoelastic film for shear horizontal surface acoustic wave on quartz

A numerical analysis for the mass loading sensitivity of shear horizontal surface acoustic wave (SH-SAW) immunoassay biosensors on quartz has already been studied. However, the mass loading analysis is insufficient to explain the actual biosensor performance. To understand the SH-SAW biosensor performance, we analyze the effect of a viscoelastic film on SH-SAW biosensors. In this paper, a numerical analysis using a simple viscoelastic model for the SH-SAW biosensors is presented. In the theoretical model, the bioreaction layer on the SH-SAW biosensors can be treated as a viscoelastic film. The velocity changes of the 250 MHz SH-SAWs on quartz substrates, which are covered with bovine serum albumin (BSA) layers of different thicknesses, were measured and compared with the theoretical results obtained using the proposed viscoelastic model. Good agreement of the velocity changes of SH-SAWs versus changes in the viscoelastic film thickness between theoretical and experimental results was obtained.

Analysis of Mass Loading Effect on Guided Shear Horizontal Surface Acoustic Wave on Liquid/Au/Quartz Structure for Biosensor Application

A numerical analysis of a shear horizontal surface acoustic wave (SH-SAW) immunoassay biosensor has been investigated. The numerical calculation method proposed by Campbell and Jones has generally been used to calculate SAW propagation characteristics. In this paper, a numerical calculation method for SH-SAW propagation characteristics, which is a modified method of Campbell and Jones involving the effect of liquid viscosity is applied to quartz. A mass loading sensitivity of a 250 MHz SH-SAW delay-line biosensor was calculated and compared with the experimental results. It can be concluded from the results that viscosity sensitivity is dominant over mass loading sensitivity for 250 MHz SH-SAW biosensors.