Electroacoustic Biosensor Systems for Evaluating Antibiotic Action on Microbial Cells.

Antibiotics are widely used to treat infectious diseases. This leads to the presence of antibiotics and their metabolic products in the ecosystem, especially in aquatic environments. In many countries, the growth of pathogen resistance to antibiotics is considered a threat to national security. Therefore, methods for determining the sensitivity/resistance of bacteria to antimicrobial drugs are important. This review discusses the mechanisms of the formation of antibacterial resistance and the various methods and sensor systems available for analyzing antibiotic effects on bacteria. Particular attention is paid to acoustic biosensors with active immobilized layers and to sensors that analyze antibiotics directly in liquids. It is shown that sensors of the second type allow analysis to be done within a short period, which is important for timely treatment.

Microwave resonator-based sensor system for specific antibody detection.

An antibody-detecting sensor is described that is based on a microwave electrodynamic resonator. A polystyrene film with immobilized bacteria deposited on a lithium niobate plate was placed at one end of the resonator and was used as the sensing element. The second end was electrically shorted. The frequency and depth of the reflection coefficient S11 for three resonances in the range 6.5-8.5 GHz were used as an analytical signal to examine antibody interactions with bacteria and determine the time required for cell immobilization. The sensor distinguished between situations in which bacteria interacted with specific antibodies and those in which no such interaction occurred (control). Although the cell-antibody interaction changed the frequency and depth of the second and third resonance peaks, the parameters of the first resonance peak did not change. The interaction of cells with nonspecific antibodies did not change the parameters of any of the peaks. These results are promising for use in the design of methods to detect specific antibodies, which can supplement the existing methods of antibody analysis.

Sensor System Based on a Piezoelectric Resonator with a Lateral Electric Field for Virus Diagnostics.

A sensor system based on a piezoelectric resonator with a lateral electric field in the frequency range 6-7 MHz of the electric field for virus detection is described. Through use of the transmissible virus causing gastroenteritis in pigs and specific antibodies, the possibility of detecting the virus in suspension in real time was determined. It was found that the frequency dependence of the real and imaginary parts of the electrical impedance of such a resonator loaded with a virus suspension changes significantly after the addition of specific antibodies to the suspension. No changes are observed if the antibodies are not specific. Thus, the results obtained illustrate the possibility of detecting viruses in situ, directly in the liquid phase, if the change in the real or imaginary parts of the electrical impedance after the addition of antibodies is used as an analytical signal. The possibility of virus detection in the presence of foreign viral particles has been illustrated.

A new liquid sensor based on a piezoelectric resonator with a radial electric field.

The possibility of development a liquid sensor based on a piezoelectric resonator with radial concentric electrodes is shown. The specified resonator has a large number of resonance peaks corresponding to different vibrational modes. The influence of two types of liquid container with distilled water on the resonance characteristics of these vibrational modes is experimentally investigated. As a result, the optimal type of container and two vibrational modes with frequencies of 0.128 and 0.317 MHz were selected, which retain acceptable Q-factors in the presence of distilled water. The dependences of the resonance frequency and the maximum value of the real part of the electrical impedance of these resonance peaks on the conductivity of the liquid were measured. It has been found that with an increase in the conductivity of the liquid, the resonance frequency of the parallel resonance initially remains practically unchanged, but after reaching a certain value of the conductivity of water, it decreases for both resonances. In this case, the maximum value of the real part of the electrical impedance first decreases, reaches a minimum, and then increases in all cases. It is shown that using these dependences as calibration curves, one can unambiguously determine the conductivity of a liquid in the range of 45-5000 µS/cm.

Prospects for the Use of Gold Nanoparticles to Increase the Sensitivity of an Acoustic Sensor in the Detection of Microbial Cells.

The interaction of microbial cells with antibody-gold nanoparticle conjugates in conductive suspensions was experimentally studied by using an acoustic slot-mode sensor. The sensor consisted of a piezoelectric plate with a propagating acoustic wave and a liquid container located above this plate with a given gap. An analysis of the measured parameters of the sensor revealed that the specific interaction of bacterial cells with the conjugates led to a stronger change in the sensor output signal than the specific interaction of bacterial cells with antibodies. The measurements were made for Azospirillum brasilense Sp7 cells in buffer with an initial conductivity of 5-30 µS/cm. The limit of cell detection with the conjugates was 103 cells/mL, and the analysis took about 4 min. The advantage of the sensor is the possibility of repeated use and cleaning of the liquid container without damaging the sensor's elements. These results are promising for use in rapid test systems for the direct detection of microbial cells in actual samples of liquids in medical diagnostics.

Evaluation of Elastic Properties and Conductivity of Chitosan Acetate Films in Ammonia and Water Vapors Using Acoustic Resonators.

Novel bio-materials, like chitosan and its derivatives, appeal to finding a new niche in room temperature gas sensors, demonstrating not only a chemoresistive response, but also changes in mechanical impedance due to vapor adsorption. We determined the coefficients of elasticity and viscosity of chitosan acetate films in air, ammonia, and water vapors by acoustic spectroscopy. The measurements were carried out while using a resonator with a longitudinal electric field at the different concentrations of ammonia (100-1600 ppm) and air humidity (20-60%). It was established that, in the presence of ammonia, the longitudinal and shear elastic modules significantly decreased, whereas, in water vapor, they changed slightly. At that, the viscosity of the films increased greatly upon exposure to both vapors. We found that the film's conductivity increased by two and one orders of magnitude, respectively, in ammonia and water vapors. The effect of analyzed vapors on the resonance properties of a piezoelectric resonator with a lateral electric field that was loaded by a chitosan film on its free side was also experimentally studied. In these vapors, the parallel resonance frequency and maximum value of the real part of the electrical impedance decreased, especially in ammonia. The results of a theoretical analysis of the resonance properties of such a sensor in the presence of vapors turned out to be in a good agreement with the experimental data. It has been also found that with a growth in the concentration of the studied vapors, a decrease in the elastic constants, and an increase in the viscosity factor and conductivity lead to reducing the parallel resonance frequency and the maximum value of the real part of the electric impedance of the piezoelectric resonator with a lateral electric field that was loaded with a chitosan film. This leads to an increase in the sensitivity of such a sensor during exposure to these gas vapors.

Analysis of Microbial Cell Viability in a Liquid Using an Acoustic Sensor.

A method was developed for the rapid analysis and evaluation of the viability of bacteriophage-infected Escherichia coli (E.coli) XL-1 directly in a conducting suspension by using a slot-mode sensor. The method is based on recording the changes in the depth and frequency of resonant absorption peaks in the frequency dependence of the insertion loss of the sensor before and after the biologic interaction of E. coli with specific bacteriophages. The possibility was shown of recording the infection of E. coli with specific bacteriophages and assessing its viability in suspensions with a conductivity of 4.5-30 µS/cm. Сontrol experiments were carried out with non-specific interactions of E. coli cells with bacteriophages, in which no changes in the sensor variables were observed. The optimal informational variable for estimating the number of viable cells was the degree of change in the depth of the resonant peaks in the frequency dependence of the insertion loss of the sensor. The limit of cell detection was ∼102-103 cells mL-1, with an analysis time of about 5 min. An additional advantage of the sensor was the availability of a removable liquid container, which allows one to use it repeatedly and to facilitate the cleaning of the container from spent samples. The results are promising for the detection of bacteria and assessment of their viability in solutions with conductivity in the range 4.5-30 µS/cm.

Super high sensitive plate acoustic wave humidity sensor based on graphene oxide film.

The changes of density and elastic modules due to water vapor adsorption are measured for graphene oxide film at room temperature. Dominant mechanism for acoustic wave humidity sensing by the film is shown to be related with variation of its electric conductivity. Basing on the data, super high sensitive humidity sensor employing high-order Lamb wave with large coupling constant, standard lithium niobate plate, and graphene oxide sorbent film is developed. The minimal detectable level of the sensor is as low 0.03% RH, response times are 60/120s, and reproducibility is ±2.5%. The sensor is completely selective towards H2, CO, CH4, NO, O2.

Gasoline identifier based on SH0 plate acoustic waves.

The present paper is devoted to the development of gasoline identifier based on zero order shear-horizontal (SH0) acoustic wave propagating in piezoelectric plate. It has been found that the permittivity of gasoline is increased when its octane number rises. The development of such identifier is experimentally demonstrated to be possible.

Liquid sensor based on a piezoelectric lateral electric field-excited resonator.

The influence of viscous and conducting liquid on the characteristics of a piezoelectric lateral electric field-excited resonator based on the X-cut lithium niobate plate has been investigated. It has been found that the contact of a free surface of such resonator with conducting or viscous liquid leads to the substantial variation of its electrical impedance/admittance. The analysis has shown the modulus of electrical impedance or admittance at any frequency near the parallel or series resonance to be a parameter unambiguously associated with the conductivity or the viscosity. This parameter is more sensitive to the variation of the liquid conductivity or viscosity as compared to the widely used for this purpose resonant frequency whose variation area is essentially smaller. By this means the liquid conductivity and viscosity affects unambiguously on the change of electrical impedance and admittance modulus whose measurement at a fixed frequency should present no problem in practice. Consequently, the lateral field excited resonator we have described may be employed as a liquid conductivity and viscosity meter with an appropriate graduation.

Acoustic waves in a structure containing two piezoelectric plates separated by an air (vacuum) gap.

This paper presents experimental results for the characteristics of acoustic waves propagating in a structure containing two parallel piezoelectric plates (I and II) separated by an air gap. Plate I, made of Y-X lithium niobate, contained two interdigital transducers that excited and received an acoustic wave with shear-horizontal polarization. Piezoelectric plate II, made of lithium niobate, was placed above and between the transducers, separated by a fixed gap. For its certain orientation, the amplitude-frequency characteristic showed sharply defined resonant attenuation peaks, which were situated at an equidistant separation from each other. The depth of the peaks was observed to decrease with a wider gap between the plates. It has been stated that these peaks are associated with the resonant reflections of a slot acoustic wave across the width of plate II. Experimentally determined phase velocities and electromechanical coupling coefficient for the slot wave in the structure under study are in a good agreement with theoretical values for various crystallographic orientations of plate II. A comparison between the experimental and theoretical results has allowed us to state two conditions for the slot wave to exist. The structures described may be employed for noncontact excitation of acoustic waves in the plates and for the development of various liquid, gas, and temperature sensors.

Biological sensor based on a lateral electric field-excited resonator.

This paper describes a biological sensor based on a lateral electric field-excited resonator using an X-cut lithium niobate plate. Its potential was shown through the example of biological interaction between bacterial cells and specific bacteriophages. The detection was based on the analysis of the measured real and imaginary parts of electrical impedance for a resonator loaded by the biological suspension under study. It has been shown that the sensor is sensitive to specific interactions between bacterial cells and specific bacteriophages in a pure state as well as in the presence of extraneous microflora. The degree of electrical impedance variation resulting from the biological interaction depends on the numbers of phage particles and bacteria cells. The sensor may be used not only for the qualitative analysis of bacteria but also for their quantitative detection.

Elastic and viscous properties of nanocomposite films based on low-density polyethylene.

The physical properties of nanocomposite materials which contain nanoparticles of metals and their compounds stabilized within a polymeric dielectric matrix are currently being studied. These materials show a low acoustic impedance, resulting in their use as low-perturbing substrates for thin piezoelectric plates, as well as matching and damping layers for bulk acoustic wave devices. Mechanical properties of the materials which contain various Fe and Fe2O3 nanoparticle concentrations are experimentally defined in the paper. It has been shown that the acoustic impedance of the materials under study may be varied by more than 30% and 40% for longitudinal and shear acoustic waves, respectively, within the same technological procedure. The velocities of bulk longitudinal and shear acoustic waves can be purposefully changed by more than 40% and 29%, respectively.

The power flow angle of acoustic waves in thin piezoelectric plates.

The curves of slowness and power flow angle (PFA) of quasi-antisymmetric (A(0)) and quasi-symmetric (S(0)) Lamb waves as well as quasi-shear-horizontal (SH(0)) acoustic waves in thin plates of lithium niobate and potassium niobate of X-,Y-, and Z-cuts for various propagation directions and the influence of electrical shorting of one plate surface on these curves and PFA have been theoretically investigated. It has been found that the group velocity of such waves does not coincide with the phase velocity for the most directions of propagation. It has been also shown that S(0) and SH(0) wave are characterized by record high values of PFA and its change due to electrical shorting of the plate surface in comparison with surface and bulk acoustic waves in the same material. The most interesting results have been verified by experiment. As a whole, the results obtained may be useful for development of various devices for signal processing, for example, electrically controlled acoustic switchers.

The peculiarities of energy characteristics of acoustic waves in piezoelectric materials and structures.

This paper is devoted to detailed theoretical investigation of energy density and power flow of homogeneous (bulk) and inhomogeneous (surface and plate) plane acoustic waves in piezoelectric materials and structures. The analysis of these waves in different materials of various crystallographic orientations allowed us to establish some energy regularities. These regularities are the same for instantaneous energy characteristics of homogeneous waves and for time-average energy characteristics on unit of aperture of inhomogeneous waves if the electrical energy and power flow in vacuum are taken into account. It has been shown that, for strong piezoactive waves, the electric energy density may exceed the mechanical energy density more than three times.

New method of change in temperature coefficient delay of acoustic waves in thin piezoelectric plates.

As is well-known, the development of high-effective and thermostable acoustic devices assumes using the acoustic waves with high coefficient of electromechanical coupling (K2) and low temperature coefficient of delay (TCD). At present, it also is well-known that fundamental shear horizontal (SH0) acoustic waves in thin piezoelectric plates possess by significantly more electromechanical coupling compared to surface acoustic waves (SAW) in the same material. However, although the value of TCD of SH0 waves is insignificantly less than for SAW, this is not enough for development of thermostable devices. This paper suggests a new way of decreasing TCD of SH0 waves in piezoelectric plates at a high level of electromechanical coupling. This way assumes to use the structure containing the piezoelectric plate and liquid with the special dependence of permittivity on temperature. Theoretical and experimental investigation showed that, for SH0 wave in YX LiNbO3 plate at hf = 700 m/s (h = plate thickness, f = wave frequency) the presence of butyl acetate can decrease the value of TCD by six times at K2 = 30%. In a whole the obtained results open the wide prospect of using SH0 wave in thin piezoelectric plate for development of high effective and thermo-stable acoustic devices.

Transmission of surface acoustic waves through the interface based on discontinuity of electrical boundary conditions on surface of potassium niobate.

This paper presents theoretical investigation of the propagation of surface acoustic waves (SAWs) across the boundary between metallized (electrically shorted) and unmetallized (electrically open) regions on the surface of potassium niobate crystals. Potassium niobate is a very strong piezoelectric material and has the interesting property that only one type of SAW, namely a Rayleigh wave, can exist on unmetallized surface, where as two types of SAWs, namely Rayleigh and Bleustein-Gulyaev (BG), can exist on a metallized surface. Analysis shows that the Rayleigh wave propagates through the interface with very little change in amplitude or polarization. On the other hand, almost total reflection of the BG wave is expected. Details of the theoretical analysis and calculated results will be presented.

The energy density and power flow of acoustic waves propagating in piezoelectric materials.

It is shown that, for plane bulk acoustic wave propagating in arbitrary piezoelectric media, the densities of mechanoelectrical and electromechanical energies are always equal in absolute value and have opposite signs. However, in general, the mechanoelectrical and electromechanical power flows of such wave calculated by traditional expression for Poynting vector do not compensate each other, although the total density of these energies is always equal to zero. Discovered discrepancy based on the dissymmetry of piezoelectric constants with respect to the electrical and mechanical indexes may cause difficulties for calculation of important parameters for practical applications such as energy transport velocity of acoustic waves in piezoelectric materials.

Reflection of plate acoustic waves produced by a periodic array of mechanical load strips or grooves.

The reflection of fundamental acoustic waves propagating in a thin piezoelectric plate by a periodic array of conducting strips of finite thickness or grooves has been theoretically and experimentally investigated. The analysis has shown that electrical shorting and mass loading affect the relationship of neighboring region impedances in a contrary manner. In some cases, these effects are comparable, and there exists a certain strip thickness for each piezoactive fundamental plate mode at which the reflection coefficient can become zero. A high efficiency of grooved reflector for plate acoustic waves was theoretically revealed. Experimental results for mass loading and grooved reflections, which have been obtained for an SH0 wave propagating in the Y-X lithium niobate plate, are in a good agreement with the theory. They show a high efficiency of such reflectors and confirm an opportunity of using a model based on equivalent circuit for the analysis of their operation. Investigations indicate that nearly 100% reflection of the SH0 wave in the lithium niobate plate can be obtained with the use of a mass loading reflector containing 10 silver strips of thickness d/h = 0.08 or a grooved reflector containing eight grooves of depth d/h = 0.25. Here h is the plate thickness and d is the reflector thickness or depth.