This paper demonstrates a highly sensitive surface-scanning detector used for magnetoelastic (ME) biosensors for the detection of Salmonella on the surface of a polyethylene (PE) food preparation surface. The design and fabrication methods of the new planar spiral coil are introduced. Different concentrations of Salmonella were measured on the surface of a PE board. The efficacy of Salmonella capture and detection is discussed.
This paper investigates the effects of surface-scanning detector position on the resonant frequency and signal amplitude of a wireless magnetoelastic (ME) biosensor for direct pathogen detection on solid surfaces. The experiments were conducted on the surface of a flat polyethylene (PE) plate as a model study. An ME biosensor (1 mm × 0.2 mm × 30 μm) was placed on the PE surface, and a surface-scanning detector was brought close and aligned to the sensor for wireless resonant frequency measurement. The position of the detector was accurately controlled by using a motorized three-axis translation system (i.e., controlled X, Y, and Z positions). The results showed that the resonant frequency variations of the sensor were -125 to +150 Hz for X and Y detector displacements of ±600 μm and Z displacements of +100 to +500 μm. These resonant frequency variations were small compared to the sensor's initial resonant frequency (< 0.007% of 2.2 MHz initial resonant frequency) measured at the detector home position, indicating high accuracy of the measurement. In addition, the signal amplitude was, as anticipated, found to decrease exponentially with increasing detection distance (i.e., Z distance). Finally, additional experiments were conducted on the surface of cucumbers. Similar results were obtained.
This paper investigates a phage-based biomolecular filter that enables the evaluation of large volumes of liquids for the presence of small quantities of bacterial pathogens. The filter is a planar arrangement of phage-coated, strip-shaped magnetoelastic (ME) biosensors (4 mm × 0.8 mm × 0.03 mm), magnetically coupled to a filter frame structure, through which a liquid of interest flows. This "phage filter" is designed to capture specific bacterial pathogens and allow non-specific debris to pass, eliminating the common clogging issue in conventional bead filters. ANSYS Maxwell was used to simulate the magnetic field pattern required to hold ME biosensors densely and to optimize the frame design. Based on the simulation results, a phage filter structure was constructed, and a proof-in-concept experiment was conducted where a Salmonella solution of known concentration were passed through the filter, and the number of captured Salmonella was quantified by plate counting.
Phage based magneto-elastic (ME) biosensors have been shown to be able to rapidly detect Salmonella in various food systems to serve food pathogen monitoring purposes. In this ME biosensor platform, the free-standing strip-shaped magneto-elastic sensor is the transducer and the phage probe that recognizes Salmonella in food serves as the bio-recognition element. According to Sorokulova et al. at 2005, a developed oligonucleotide probe E2 was reported to have high specificity to Salmonella enterica Typhimurium. In the report, the specificity tests were focused in most of Enterobacterace groups outside of Salmonella family. Here, to understand the specificity of phage E2 to different Salmonella enterica serotypes within Salmonella Family, we further tested the specificity of the phage probe to thirty-two Salmonella serotypes that were present in the major foodborne outbreaks during the past ten years (according to Centers for Disease Control and Prevention). The tests were conducted through an Enzyme linked Immunosorbent Assay (ELISA) format. This assay can mimic probe immobilized conditions on the magnetoelastic biosensor platform and also enable to study the binding specificity of oligonucleotide probes toward different Salmonella while avoiding phage/ sensor lot variations. Test results confirmed that this oligonucleotide probe E2 was high specific to Salmonella Typhimurium cells but showed cross reactivity to Salmonella Tennessee and four other serotypes among the thirty-two tested Salmonella serotypes.
This paper presents a method for detection of a few pathogenic bacteria and determination of live versus dead cells. The method combines wireless phage-coated magnetoelastic (ME) biosensors and a surface-scanning dectector, enabling real-time monitoring of the growth of specific bacteria in a nutrient broth. The ME biosensor used in this investigation is composed of a strip-shaped ME resonator upon which an engineered bacteriophage is coated to capture a pathogen of interest. E2 phage with high binding affinity for Salmonella Typhimurium was used as a model study. The specificity of E2 phage has been reported to be 1 in 105 background bacteria. The phage-coated ME biosensors were first exposed to a low-concentration Salmonella suspension to capture roughly 300 cells on the sensor surface. When the growth of Salmonella in the broth occurs, the mass of the biosensor increases, which results in a decrease in the biosensor's resonant frequency. Monitoring of this mass- induced resonant frequency change allows for real-time detection of the presence of Salmonella. Detection of a few bacteria is also possible by growing them to a sufficient number. The surface-scanning detector was used to measure resonant frequency changes of 25 biosensors sequentially in an automated manner as a function of time. This methodology offers direct, real-time detection, quantification, and viability determination of specific bacteria. The rate of the sensor's resonant frequency change was found to be largely dependent on the number of initially bound cells and the efficiency of cell growth.
The wireless phage-based magnetoelastic (ME) biosensor has proven to be promising for real-time detection of pathogenic bacteria on fresh produces. The ME biosensor consists of a freestanding ME resonator as the signal transducer and filamentous phage as the biomolecular-recognition element, which can specifically bind to a pathogen of interest. Due to the Joule magnetostriction effect, the biosensors can be placed into mechanical resonance when subjected to a time-varying magnetic field alternating at the sensor’s resonant frequency. Upon the attachment of the target pathogen, the mass of the biosensor increases, thereby decreasing its resonant frequency. This paper presents an investigation of blocking reagents immobilization for detecting Salmonella Typhimurium on fresh food surfaces. Three different blocking reagents (BSA, SuperBlock blocking buffer, and blocker BLOTTO) were used and compared. The optical microscope was used for bacterial cells binding observation. Student t-test was used to statistically analysis the experiment results. The results shows that SuperBlock blocking buffer and blocker BLOTTO have much better blocking performance than usually used BSA.
This paper describes the results of a research project to investigate magnetoelastic (ME) biosensors actuated with a pulse excitation to measure the concentration of Salmonella Typhimurium of globe fruits. The ME biosensors are based on an acoustic wave resonator platform that is a freestanding (free-free) thin ribbon of magnetostrictive material with a lengthto- width ratio of 5:1. A biorecognition probe coated on the surface of the resonator platform binds with a targeted pathogen, i.e. E2 phage that binds with S. Typhimurium. The biosensor was actuated to vibrate longitudinally such that the resonant frequency depended primarily on the length of sensor and its overall mass. A pulsed excitation and measurement system was used to actuate micron scale ME biosensors to vibrate. The biosensor responds in a ring-down manner, a damped decay of the resonance amplitude, from which the resonant frequency was measured. An increase in mass due to the binding of the target pathogen resulted in a decrease in the resonant frequency. The pulsed excitation and measurement system that was developed under this effort and the characterization of its performance on the measurement of Salmonella concentrations on globe fruits is described.
This paper presents an investigation into magnetoelastic (ME) biosentinels that capture and detect low-concentration pathogenic bacteria in stagnant liquids. The ME biosentinels are designed to mimic a variety of white blood cell types, known as the main defensive mechanism in the human body against different pathogenic invaders. The ME biosentinels are composed of a freestanding ME resonator coated with an engineered phage that specifically binds with the pathogens of interest. These biosentinels are ferromagnetic and thus can be moved through a liquid by externally applied magnetic fields. In addition, when a time-varying magnetic field is applied, the ME biosentinels can be placed into mechanical resonance by magnetostriction. As soon as the biosentinels bind with the target pathogen through the phage-based biomolecular recognition, a change in the biosentinel’s resonant frequency occurs, and thereby the presence of the target pathogen can be detected. Detection of Bacillus anthracis spores under stagnant flow conditions was demonstrated.
This paper presents the concept of self-propelled magnetoelastic (ME) biosentinels that seek out and capture pathogenic
bacteria in stagnant liquids. These biosentinels are composed of a free-standing, asymmetric-shaped ME resonator coated
with a filamentous landscape phage that specifically binds with a pathogen of interest. When a time-varying magnetic pulse
is applied, the ME biosentinels can be placed into mechanical resonance by magnetostriction. The resultant asymmetric
vibration then generates a net force on the surroundings and hence generates autonomous motion in the liquid. As soon
as the biosentinels find and bind with the target pathogen through the phage-based biomolecular recognition, a change
in the biosentinel’s resonant frequency occurs, and thereby the presence of the target pathogen can be detected. In order
to actuate the ME biosentinels into mechanical resonance of a desired mode, modal analysis using the three-dimensional
finite element method was performed. In addition, the design of a magnetic chamber that can control the orientation and/or
translation of a biosentinel is discussed.
An electrical circuit was designed and tested to measure the resonant frequency of micron-scale magnetoelastic (ME)
biosensors using a pulsed wave excitation technique. In this circuit, a square pulse current is applied to an excitation coil
to excite the vibration of ME biosensors and a pick-up coil is used to sense the ME biosensor’s mechanical vibration and
convert it to an electrical output signal. The output signal is filtered and amplified by a custom designed circuit to allow
the measurement of the resonant frequency of the ME biosensor from which the detection of specific pathogens can be
made. As a proof-in-concept experiment, JRB7 phage-coated ME biosensors were used to detect different concentrations
of Bacillus anthracis Sterne strain spores. A statistically significant difference was observed for concentrations of 5 ×
102 spore/ml and above.
Phage-based magnetoelastic (ME) biosensors have proven useful in rapidly and inexpensively detecting food surface con- tamination. These biosensors are wireless, mass-sensitive biosensors and can be placed directly on food surfaces to detect the presence of target pathogens. Previously, millimeter-scale strip-shaped ME biosensors have been used to demonstrate direct detection of Salmonella Typhimurium on various fresh produce surfaces, including tomatoes, shell eggs, watermel- ons, and spinach leaves. Since the topography of these produce surfaces are different, and the biosensor must come into direct contact with Salmonella bacteria, food surfaces with large roughness and curvatures (e.g., spinach leaf surfaces) may allow the bacteria to avoid direct contact, thereby avoiding detection. The primary objective of this paper is, hence, to investigate the effects of food surface topography on the detection capabilities of the biosensors. Spinach leaf surfaces were selected as model surfaces, and detection experiments were conducted with differently sized biosensors (2 mm, 0.5 mm, and 150 μm in length). Spinach leaf roughness and curvatures of both adaxial (top) and abaxial (underside) surfaces were measured using a confocal laser scanning microscope. The experimental results showed that in spinach as the sen- sor was made smaller, the physical contact between the biosensors and bacteria were improved. Smaller sensors thereby enhance detection capabilities. When proper numbers of biosensors are used, micron-scale biosensors are anticipated to yield improved limits of detection over previously investigated millimeter-scale biosensors.
This paper presents a new device and method for the in-situ detection of Salmonella Typhimurium on tomato surfaces. This real-time in-situ detection was accomplished with phage-based magnetoelastic (ME) biosensors on fresh food surfaces. The E2 phage from a landscape phage library serves as the bio-recognition element that has the capability of binding specifically with S. Typhimurium. This mass-sensitive ME biosensor is wirelessly actuated into mechanical resonance by an externally applied time-varying magnetic field. When the biosensor binds with S. Typhimurium, the mass of the sensor increases, resulting in a decrease in the sensor's resonant frequency. Until now, ME sensors had to be collected from the tomato surface where they are exposed to S. Typhimurium and inserted into a measurement coil for the detection of the bacterium. In contrast, the newly designed test device allows the whole detection process to take place directly on the tomato. Changes in resonant frequency over time due to the accumulation of S. Typhimurium on the sensor were measured and are presented. Real-time in-situ detection of 20 minutes was achieved. In addition, this new methodology effectively decreases the measurement error and enables the simultaneous detection of multiple pathogens.
Hydrazine is mostly used as a propellant in the control/propulsion system of missiles, spacecraft and satellites. However with its highly toxic and strong reducing nature, hydrazine is very dangerous to humans and the environment. In this research, a low cost, passive, and highly sensitive micro-sensor has been developed as an alarm device for real-time monitoring for the accidental release of hydrazine, and to insure the safety of personnel and the readiness of the system before lift-off. The micro-sensor is fabricated using standard microelectronic manufacturing techniques and is composed of interdigitated electrodes and a hydrazine-sensitive poly (3-hexylthiophene) (P3HT) thin film. When exposed to 1ppm of hydrazine gas, the compensation interaction between the reducing hydrazine gas and p-type doped P3HT leads to a five order magnitude increase in the resistance of the device. The sensor is capable of detecting hydrazine leaks from tens of ppb to tens of ppm concentration. The sensitivity of sensor increases with the increasing of hydrazine concentration and the decreasing of the polymer film thickness. A numerical simulation result based on the possible theoretical model is compared with the experimental data, which shows a good agreement.
Infrared thermography was employed to identify changes in plate thickness during the arc welding of steel plates. The step changes in plate thickness were found to uniquely affect the surface temperature distributions of the plates during the welding process. A technique was developed to identify the thermal profile irregularities ahead of the weld that were indicative of changes in plate thickness. The results of this investigation suggest that the calculation of the area under the linescan can be successfully used to identify thickness variations in a plate as it is being welded. The gradient of the area beneath the linescan was further found to be indicative of changes in plate thickness.
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