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Comparative Analysis of Static and Non-Static Assays for Biochemical Sensing Using On-Chip Integrated Field Effect Transistors and Solidly Mounted Resonators
Abstract
The advancement in micro/nanotechnologies has been continuously providing possibilities for inventing novel biochemical sensors. However, variations in the transducer type can cause different sensing results due to the differences in their mechanisms of analyzing biomolecular interactions. In this work, we focused on the comparative analysis of static and non-static assays for molecular interactions using on-chip integrated extended-gate field effect transistor (EGFET) as a static sensing interface and solidly mounted resonator (SMR) as a non-static sensing interface. Analysis of polyelectrolytes (PETs) surface assembly and antigen-antibody interaction using the two types of biochemical sensors presented consistent and complementary sets of information. Meanwhile, due to the difference in their operating mechanisms, variations on the detection efficiency, kinetics and thermodynamics were observed. Our results highlighted the critical dependence of signal detection on biochemical sensors’ operating mechanisms and provided a valuable guidance for static and non-static assays for biomolecular detections.
... The integrated device was fabricated according to a previously published process (see ESI, † Fig. S1). 43 Before the fabrication of the resonator, a Bragg reflector was mounted on the silicon wafer by alternately depositing three pairs of silicon dioxide (SiO 2 ) and molybdenum (Mo) thin films. 44 The first layer of the Bragg reflector was used as the bottom electrode followed by the sputtering of highly c-axis oriented aluminum nitride (AlN) film as a piezoelectric layer and another layer of Mo as the top electrode. ...
... The embedded gold electrode was connected with the gate electrode of a back-end field effect transistor (FET) and was used as the charge-sensitive interface for current measurements. 43 Since an SLB is highly resistive and impermeable to hydrated ions, 49 the electrical potential of the underlying gold electrode can remain constant in a pure electrolyte solution by the isolation from the intact SLB. However, when hypersound is applied, the transduction of the motion of the resonator to and through the SLB may deform the membrane structure. ...
Hypersound (ultrasound of gigahertz (GHz) frequency) has been recently introduced as a new type of membrane-disruption method for cells, vesicles and supported lipid bilayers (SLBs), with the potential to improve the efficiency of drug and gene delivery for biomedical applications. Here, we fabricated an integrated microchip, composed of a nano-electromechanical system (NEMS) resonator and a gold electrode as the extended gate of a field effect transistor (EGFET), to study the effects of hypersonic poration on an SLB in real time. The current recordings revealed that hypersound enabled ion conduction through the SLB by inducing transient nanopores in the membrane, which act as the equivalent of ion channels and show gating behavior. The mechanism of pore formation was studied by cyclic voltammetry (CV), atomic force microscopy (AFM) and laser scanning microscopy (LSM), which support the causality between hypersound-triggered deformation and the reversible membrane disruption of the SLB. This finding contributes to the development of an approach to reversibly control membrane permeability by hypersound.
... Another study on integrated systems that deserves to be mentioned was the one developed by Liu et al. [59], who performed a static and dynamic analysis. The sensor had, in addition to the EGFET, solidly mounted resonators (SMRs) and allowed for a multiplexed analysis to detect molecular interactions and obtain different physical-chemical parameters involved in the interactions. ...
Devices that integrate field effect transistors into microfluidic channels are becoming increasingly promising in the medical, environmental, and food realms, among other applications. The uniqueness of this type of sensor lies in its ability to reduce the background signals existing in the measurements, which interfere in obtaining good limits of detection for the target analyte. This and other advantages intensify the development of selective new sensors and biosensors with coupling configuration. This review work focused on the main advances in the fabrication and application of field effect transistors integrated into microfluidic devices as a way of identifying the potentialities that exist in these systems when used in chemical and biochemical analyses. The emergence of research on integrated sensors is not a recent study, although more recently the progress of these devices is more accentuated. Among the studies that used integrated sensors with electrical and microfluidic parts, those that investigated protein binding interactions seem to be the ones that expanded the most due, among other things, to the possibility of obtaining several physicochemical parameters involved in protein–protein interactions. Studies in this area have a great possibility of advancing innovations in sensors with electrical and microfluidic interfaces in new designs and applications.
... Here, a miniaturized virtual impactor (VI) with a 50% cutoff diameter of 2 μm is used to separate mixed particles based on inertial classification. The separated PM2 is collected and analyzed by an extended-gate field-effect transistor (EGFET) in which a gold auxiliary electrode is connected to the gate terminal of a conventional metal-oxide-semiconductor field-effect transistor (MOS-FET), which can directly measure the charges of the deposited PM. 19 Both the VI and EGFET can be mass-producible and are replaceable. Our results suggest that the proposed microsystem has potential for accurate real-time analysis of PM2 containing mixed particles in complex environments. ...
HIGHLIGHTS
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A proposed microsystem system potentially provides accurate and calibration-free measurements based on a combination of front-end particulate matter (PM) separation and back-end PM detection.
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Silicon dioxide particles are used to verify the performance of the system’s virtual impactor, and an accurate cutoff diameter of 2 μm in low air flow rates can be obtained.
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Experimental results suggest that the proposed microsystem has potential for accurate real-time analysis of PM pollution under complex conditions.
... Micropumps can be used for drug delivery with the desired amount of medicines at the appropriate time, which improves the therapeutic effect. With an increasing interest in the development of biochemical microsystems (Wenpeng et al. 2017), there have been numerous reports on the design and fabrication of a variety of micropumps (Partha and Toufique, 2017). These micropumps have recently emerged as a critical area of research because of their extensive potential applications in the fields of chemical analysis (Zolotov. ...
This paper reviews the development of a new microvalve with a cone-shaped tube, inspired by venous valves in the human body. Our microvalves allow fluid flow in one direction while restricting the flow in the opposite direction. When a microvalve is used to control the amount of drug delivery, the efficiency between inlet and outlet flow rate is the key control parameter for regulating and controlling the micro channel (opening/closing). This paper is devoted to the numerical study of flow rate changes in different microvalve geometries (3D) using a Fluid Structure Interaction (FSI) method with an Arbitrary Lagrangian Eulerian (ALE) approach. Numerical simulations were carried out in comsol Multiphysics. In addition, the macrovalve performance was analysed for several pressures where the effect of different geometrical parameters such as the length of the anchor, the diameter at the base and the angle of the cone were studied. An efficiency parameter was employed to compare the different structures. For the best design obtained, it was found that the cone angle was the parameter having the most effect on the microvalves' characteristics, and the forward flow rate was more than doubled compared to the reverse leakage rate.
... Thus, glucose concentration can be successfully monitored with a sensitivity of 0.422 mΩ/mM in logarithmic form and a detection limit of 0.05 mM. Fig. 8 (a) Schematic of the chip from an oblique view in which the electrochemical sensor and SMR share the same film stack with good compatibility; (b) schematic of the electrochemicalpiezoelectric integrated sensing system Liu et al. used the same integrated dual sensor device for static and non-static assays for detection of polyelectrolyte (PETs) surface assembly and antibody-antigen interactions (Liu et al. 2017). Due to the different sensing mechanism, complementary and inconsistence information can be extracted, which emphasize the critical dependence of the sensing results on the sensing mechanisms and offer valuable guidance for better understanding of bioreactions. ...
... FBAR usually operates at two orders of magnitude higher frequency than QCM and is cable of detecting small mass changes at even pictogram order. Hence, it has been extensively employed for fundamental science and practical engineering [13,14]. It features a greatly reduced dimension of the sensing element, which is beneficial to integrate into miniaturized platform and obtain lab-on-a-chip. ...
We developed a highly sensitive humidity sensor based on the combination of ultrahigh-frequency film bulk acoustic resonator (FBAR) and nano-assembled polyelectrolyte (PET) thin films. The water molecule absorption efficiency was optimized by forming loosely-packed PET nanostructures. Then, the humidity sensing characteristics were analyzed in terms of sensitivity, linearity, reversibility, stability and detection limit. As a result, PET-coated FBAR exhibits excellent humidity sensitivity of 2202.20 Hz/ppm, which is five orders of magnitude higher than quartz crystal microbalance (QCM). Additionally, temperature dependence was investigated with the result that PET-coated FBAR possessed a higher sensitivity at low temperature. Furthermore, we realized the selective detection of water vapor from volatile organic compounds (VOCs) with respect to the polarity property. Owing to the high sensitivity, miniaturized size and ultrahigh operating frequency, PET-coated FBAR is uniquely favorable as a wireless humidity sensor node to integrate into wireless sensor networks (WSNs).
This paper demonstrates design and simulation of an acoustic wave resonator-based ammonia gas sensor which is resonating at * 3.95 GHz resonant frequency. The bulk acoustic wave-based solidly mounted resonators (SMRs) are robust, simple to manufacture, miniaturized and most suitable for high sensitivity gas sensors. The designed sensor consists of multilayer stack of (Si/SiO 2 /BR/Mo/ZnO/Mo/rGO) thin films. Here, reduced graphene oxide (rGO) is utilized as an ammonia gas-sensing layer and zinc oxide (ZnO) as piezoelectric layer. Bragg reflector (BR) is applied to confine the acoustic wave. The simulated sensor is based on the resonant frequency shift due to mass loading effect by adsorbed ammonia gas molecules. The amount of frequency shift depends on the concentration of ammonia gas. The ammonia gas sensor results in sensitivity (S) of 7.113 MHz pg-1 with outstanding selectivity over other exposed gases to the sensor. The SMR-based ammonia gas sensor is designed using air tight gas chamber and gases are exposed to the sensor in controlled ambient. In addition, the number of BR pairs (Mo/SiO 2) is also optimized for optimal performance of sensor, using transfer matrix model. The simulated ammonia gas sensor is small in size, highly sensitive and can find applications in safety and environmental monitoring in various industries, including chemical plants, refrigeration systems and agricultural applications.
This work reports a novel acoustic resonator system integrated dual functions of biological samples capture and amount monitoring on a single chip. The system could capture samples from nano-sized proteins to micro-sized cells on micro-sized chip precisely with controllable concentration, meanwhile the high sensitivity mass sensing was achieved during the capture process. The devices were further applied to study the cell growth and cytotoxicity. Results indicated that it was possible to capture and monitor the physiological changes in a single cell level. This work explores a new opportunity on the development of miniaturized multiplex biosensing devices on a single chip.
This paper reports the effect of the design parameters of a solidly mounted film bulk acoustic resonator (SMFBAR) for better gas sensing performance. The electrical equivalent circuit of the proposed device has been developed with the help of a Butterworth Van-Dyke (BVD) circuit. The electro-mechanical response of the SMFBAR has been obtained with the help of 3-D finite element method (FEM) analysis. The analytical modeling and FEM simulation results are compared. The physical parameters of the proposed design such as piezoelectric layer material, its thickness, active area of the
device and sensing layer thickness affecting the characteristics of the SMFBAR have been investigated in detail. To achieve enhanced sensitivity, the variation of square active area has been analysed with one side dimension ranging from 700 lm to 300 lm. Gas sensing performance of the proposed sensor is tested by exposing toluene gas concentration ranging from 0 ppm to 500 ppm and enhanced sensitivity of 20 kHz/ppm has been achieved and reported. Also, it is reported that the variation in ratio of electrode layer
thickness to piezoelectric layer thickness results in improvement of coupling coefficient (keff 2 ) up to 7.46%.
We present a nanoscale acoustofluidic trap (AFT) which manipulates nanoparticles in a microfluidic system actuated by a gigahertz acoustic resonator. The AFT generates independent standing closed vortices with high-speed rotation. By carefully designing and optimizing the geometric confinements, the AFT is able to effectively capture and enrich sub-100 nm nanoparticles with low power consumption (0.25~5 μW/μm2) and rapid trapping (within 30 s), showing greatly enhanced particle operating ability towards its acoustic and optical counterparts. Using specifically functionalized nanoparticles (SFNPs) to selectively capture target molecules from the sample, the AFT produces a molecular concentration enhancement of ~200 times. We investigated the feasibility of the SFNPs-assisted AFT preconcentration method for biosensing applications, and successfully demonstrated its capability for serum prostate specific antigen (PSA) detection. The AFT is prepared with a fully CMOS-compatible process, and thus can be conveniently integrated on a single chip, with potential for “lab-on-a-chip” or point-of-care (POC) nanoparticle-based biosensing applications.
In designing bioassay systems for low-abundance biomolecule detection, most research focuses on improving transduction mechanisms while ignoring the intrinsically fundamental limitations in solution: mass transfer and binding affinity. We demonstrate enhanced biomolecular surface binding using an acoustic nano-electromechanical system (NEMS) resonator, as an on-chip biomolecular concentrator which breaks both mass transfer and binding affinity limitations. As a result, a concentration factor of 10⁵ has been obtained for various biomolecules. The resultantly enhanced surface binding between probes on the absorption surface and analytes in solution enables us to lower the limit of detection for representative proteins. We also integrated the biomolecular concentrator into an optoelectronic bioassay platform to demonstrate delivery of proteins from buffer/serum to the absorption surface. Since the manufacture of the resonator is CMOS-compatible, we expect it to be readily applied to further analysis of biomolecular interactions in molecular diagnostics.
Quantitative biomarker detection methods featured with rapidity, high accuracy, and label-free are demonstrated for the development of point-of-care (POC) technologies or "beside" diagnostics. Microbead aggregation via protein-specific linkage provides an effective approach for selective capture of biomarkers from the samples, and can directly readout the presence and amount of the targets. However, sensors or microfluidic analyzers that can accurately quantify the microbead aggrega-tion are scared. In this work, we demonstrate a microwell-based microbeads analyzing system, by which online manipula-tions of microbeads including trapping, arraying, and rotations can be realized, providing a series of microfluidic approaches to layout the aggregated microbeads for further convenient characterizations. Prostate specific antigen is detected using the proposed system, demonstrating the limit of detection as low as 0.125 ng/mL (3.67 pM). A two-step reaction kinetics model is proposed for the first time to explain the dynamic process of microbeads aggregation. The developed microbeads aggrega-tion analysis system has the advantages of label-free detection, high throughput, and low cost, showing great potential for portable biomarker detection.
This chapter reports on the state of the art of piezoelectric micro-/nano-mechanical devices in frequency control and sensing applications. Recent studies on bulk acoustic wave (BAW) devices are introduced, including investigation of high-coupling materials and filter and oscillator designs. A novel class of frequency devices based on Lamb waves is also reviewed. Micro- and nano-mechanical sensors for various sensing applications and integrated module are outlined.
In this paper, we described a novel fabrication technique for Silicon nanobelt field-effect transistors (SiNB FETs) based on the Local Oxidation of Silicon (LOCOS) process widely utilized in the manufacturing of microelectronics components and explore it to screened alpha-fetoprotein (AFP). LOCOS process is used as it enables thorough compatibility with complementary metal oxide semiconductor (CMOS) technology. Thus, the need for expensive lithography tools to define the nanoscale pattern is avoided. The SiNB FETs emerges as a powerful biosensor for ultrasensitive, label-free for biological/chemical detection and direct electrical readout. The analytical sensing results of the fabricated SiNB FETs employed as a biomolecular sensor for the early, real-time, and label-free screening of AFP can be understood in term of the change in charge density at the silicon nanobelt surface after functionalization. It is observed that, by tuning the gate voltage, the electrical linearity response of the system towards AFP extends its concentration range from 3 ng/mL to 100 ng/mL, allowing the screening of hepatocellular carcinoma (HCC). These results indicate that the detection of AFP under our direct, label-free, and ultrasensitive biosensor in a microfluidic channel could be one of the promising state-of-the-art techniques applicable as an AFP detector in real samples.
Nonspecific binding (NSB) is a general issue for surface based biosensors. Various approaches have been developed to prevent or remove the NSBs. However, these approaches either increased the background signals of the sensors or limited to specific transducers interface. In this work, we developed a hydrodynamic approach to selectively remove the NSBs using a microfabricated hypersonic resonator with 2.5 gigahertz (GHz) resonant frequency. The high frequency device facilitates to generate multiple controlled micro-vortices which then create cleaning forces at the solid-liquid interfaces. The competitive adhesive and cleaning forces have been investigated using the finite element method (FEM) simulation, identifying the feasibility of the vortices induced NSB removal. NSB proteins have been selectively removed experimentally both on the surface of the resonator and on other substrates which contact the vortices. Thus, the developed hydrodynamic approach is believed to be a simple and versatile tool for NSB removal and compatible to many sensor systems. The unique feature of the hypersonic resonator is that it can be used as a gravimetric sensor as well, thus a combined NSB removal and protein detection dual functional biosensor system is developed.
Zinc oxide (ZnO) nanostructures are promising candidates as electronic components for biological and chemical applications. In this study, ZnO ultra-fine nanowire (NW) and nanoflake (NF) hybrid structures have been prepared by Au-assisted chemical vapor deposition (CVD) under ambient pressure. Their surface morphology, lattice structures, and crystal orientation were investigated by scanning electron microscopy (SEM), x-ray diffraction (XRD), and transmission electron microscopy (TEM). Two types of ZnO nanostructures were successfully integrated as gate electrodes in extended-gate field-effect transistors (EGFETs). Due to the amphoteric properties of ZnO, such devices function as pH sensors. We found that the ultra-fine NWs, which were more than 50 μm in length and less than 100 nm in diameter, performed better in the pH sensing process than NW–NF hybrid structures because of their higher surface-to-volume ratio, considering the Nernst equation and the Gouy–Chapman–Stern model. Furthermore, the surface coating of (3-Aminopropyl)triethoxysilane (APTES) protects ZnO nanostructures in both acidic and alkaline environments, thus enhancing the device stability and extending its pH sensing dynamic range.
Live-cell imaging and particle tracking provide rich information on mechanisms of intracellular transport. However, trajectory analysis procedures to infer complex transport dynamics involving stochastic switching between active transport and diffusive motion are lacking. We applied Bayesian model selection to hidden Markov modeling to infer transient transport states from trajectories of mRNA-protein complexes in live mouse hippocampal neurons and metaphase kinetochores in dividing human cells. The software is available at http://hmm-bayes.org/.
Blood contains a range of protein biomarkers that could be used in the early detection of disease. To achieve this, however, requires sensors capable of detecting (with high reproducibility) biomarkers at concentrations one million times lower than the concentration of the other blood proteins. Here, we show that a sandwich assay that combines mechanical and optoplasmonic transduction can detect cancer biomarkers in serum at ultralow concentrations. A biomarker is first recognized by a surface-anchored antibody and then by an antibody in solution that identifies a free region of the captured biomarker. This second antibody is tethered to a gold nanoparticle that acts as a mass and plasmonic label; the two signatures are detected by means of a silicon cantilever that serves as a mechanical resonator for 'weighing' the mass of the captured nanoparticles and as an optical cavity that boosts the plasmonic signal from the nanoparticles. The capabilities of the approach are illustrated with two cancer biomarkers: the carcinoembryonic antigen and the prostate specific antigen, which are currently in clinical use for the diagnosis, monitoring and prognosis of colon and prostate cancer, respectively. A detection limit of 1 × 10(-16) g ml(-1) in serum is achieved with both biomarkers, which is at least seven orders of magnitude lower than that achieved in routine clinical practice. Moreover, the rate of false positives and false negatives at this concentration is extremely low, ∼10(-4).
Affinity biosensors use biorecognition elements and transducers to convert a biochemical event into a recordable signal. They provides the molecule binding information, which includes the dynamics of biomolecular association and dissociation, and the equilibrium association constant. Complementary metal oxide semiconductor-compatible silicon (Si) nanowires configured as a field-effect transistor (NW FET) have shown significant advantages for real-time, label-free and highly sensitive detection of a wide range of biomolecules. Most research has focused on reducing the detection limit of Si-NW FETs but has provided less information about the real binding parameters of the biomolecular interactions. Recently, Si-NW FETs have been demonstrated as affinity biosensors to quantify biomolecular binding affinities and kinetics. They open new applications for NW FETs in the nanomedicine field and will bring such sensor technology a step closer to commercial point-of-care applications. This article summarizes the recent advances in bioaffinity measurement using Si-NW FETs, with an emphasis on the different approaches used to address the issues of sensor calibration, regeneration, binding kinetic measurements, limit of detection, sensor surface modification, biomolecule charge screening, reference electrode integration and nonspecific molecular binding.
Field-effect transistor-based biosensors (bioFETs) have shown great promise in the field of fast, ultra-sensitive, label-free detection of biomolecules. Reliability and accuracy, when trying to measure small concentrations, is of paramount importance for the translation of these research devices into the clinical setting. Our knowledge and experience with these sensors has reached a stage where we are able to identify three main aspects of bioFET sensing that currently limit their applications. By considering the intrinsic device noise as a limitation to the smallest measurable signal, we show how various parameters, processing steps and surface modifications, affect the limit of detection. We also introduce the signal-to-noise ratio of bioFETs as a universal performance metric, which allows us to gain better insight into the design of more sensitive devices. Another aspect that places a limit on the performance of bioFETs is screening by the electrolyte environment, which reduces the signal that could be potentially measured. Alternative functionalization and detection schemes that could enable the use of these charge-based sensors in physiological conditions are highlighted. Finally, the binding kinetics of the receptor–analyte system are considered, both in the context of extracting information about molecular interactions using the bioFET sensor platform and as a fundamental limitation to the number of molecules that bind to the sensor surface at steady-state conditions and to the signal that is generated. Some strategies to overcome these limitations are also proposed. Taken together, these performance-limiting issues, if solved, would bring bioFET sensors closer to clinical applications. WIREs Nanomed Nanobiotechnol 2013, 5:629–645. doi: 10.1002/wnan.1227
Conflict of interest: The authors have declared no conflicts of interest for this article.
For further resources related to this article, please visit the WIREs website.
The ability to correctly track objects in time-lapse sequences is important in many applications of microscopy. Individual object motions typically display a level of dynamic regularity reflecting the existence of an underlying physics or biology. Best results are obtained when this local information is exploited. Additionally, if the particle number is known to be approximately constant, a large number of tracking scenarios may be rejected on the basis that they are not compatible with a known maximum particle velocity. This represents information of a global nature, which should ideally be exploited too. Some time ago, we devised an efficient algorithm that exploited both types of information. The tracking task was reduced to a max-flow min-cost problem instance through a novel graph structure that comprised vertices representing objects from three consecutive image frames. The algorithm is explained here for the first time. A user-friendly implementation is provided, and the specific relaxation mechanism responsible for the method's effectiveness is uncovered. The software is particularly competitive for complex dynamics such as dense antiparallel flows, or in situations where object displacements are considerable. As an application, we characterize a remarkable vortex structure formed by bacteria engaged in interstitial motility.
Surface plasmon resonance (SPR) plays a dominant role in real-time interaction sensing of biomolecular binding events.The book is intended as a hands-on guide for all interested in SPR
In part 14 of the tutorial series "Acoustofluidics--exploiting ultrasonic standing wave forces and acoustic streaming in microfluidic systems for cell and particle manipulation", we provide a qualitative description of acoustic streaming and review its applications in lab-on-a-chip devices. The paper covers boundary layer driven streaming, including Schlichting and Rayleigh streaming, Eckart streaming in the bulk fluid, cavitation microstreaming and surface-acoustic-wave-driven streaming.
Monitoring the binding affinities and kinetics of protein interactions is important in clinical diagnostics and drug development because such information is used to identify new therapeutic candidates. Surface plasmon resonance is at present the standard method used for such analysis, but this is limited by low sensitivity and low-throughput analysis. Here, we show that silicon nanowire field-effect transistors can be used as biosensors to measure protein-ligand binding affinities and kinetics with sensitivities down to femtomolar concentrations. Based on this sensing mechanism, we develop an analytical model to calibrate the sensor response and quantify the molecular binding affinities of two representative protein-ligand binding pairs. The rate constant of the association and dissociation of the protein-ligand pair is determined by monitoring the reaction kinetics, demonstrating that silicon nanowire field-effect transistors can be readily used as high-throughput biosensors to quantify protein interactions.
A droplet-based micro-total-analysis system involving biosensor performance enhancement by integrated surface-acoustic-wave (SAW) microstreaming is shown. The bioreactor consists of an encapsulated droplet with a biosensor on its periphery, with in situ streaming induced by SAW. This paper highlights the characterization by particle image tracking of the speed distribution inside the droplet. The analyte-biosensor interaction is then evaluated by finite element simulation with different streaming conditions. Calculation of the biosensing enhancement shows an optimum in the biosensor response. These results confirm that the evaluation of the Damköhler and Peclet numbers is of primary importance when designing biosensors enhanced by streaming.
The past decade has seen researchers develop and apply novel technologies for biomolecular detection, at times approaching hard limits imposed by physics and chemistry. In nearly all sensors, the transport of target molecules to the sensor can play as critical a role as the chemical reaction itself in governing binding kinetics, and ultimately performance. Yet rarely does an analysis of the interplay between diffusion, convection and reaction motivate experimental design or interpretation. Here we develop a physically intuitive and practical understanding of analyte transport for researchers who develop and employ biosensors based on surface capture. We explore the qualitatively distinct behaviors that result, develop rules of thumb to quickly determine how a given system will behave, and derive order-of-magnitude estimates for fundamental quantities of interest, such as fluxes, collection rates and equilibration times. We pay particular attention to collection limits for micro- and nanoscale sensors, and highlight unexplained discrepancies between reported values and theoretical limits.
Selective gas sensing is of immense importance for industrial as well as safety purposes. Here it is shown that metallic 1T phase transition metal dichalcogenides, such as tungsten sulfide (WS2), provide sensitive and selective platform for gas sensing. Using impedance spectroscopy distinguishable alterations can be detected on the impedance phase spectrum of interdigitated gold electrode modified with chemically exfoliated 1T-WS2 caused by different vapors. In particular, it is found that the impedance phase spectra of 1T-WS2 device present different resonant frequencies with maximum around 1 Hz in the presence of methanol vapor and around 1 kHz in the presence of water vapor. Such a well-distinguished signal allows their selective detection also when they are present in a mixture. The impedance phase spectra allow the selective methanol and water vapor sensing with an impedimetric device based on 1T-WS2. This system utilizing 1T phase of WS2 for selective gas sensing based on impedance spectroscopy opens new avenues for gas sensing and shall find wide spectra of applications.
In this work, a new method to tune the resonant frequency of micro-fabricated resonator using molecular layer-by-layer (LbL) self-assembly approach is demonstrated. By simply controlling the polymer concentration and the number of layers deposited, precisely tuning the frequency of micro-fabricated resonators is realized. Due to its selective deposition through specific molecular recognitions, such technique avoids the high-cost and complex steps of conventional semiconductor fabrications and is able to tune individual diced device. Briefly, Film Bulk Acoustic Resonator (FBAR) is used to demonstrate the tuning process and two types of LbL deposition methods are compared. The film thickness and morphology have been characterized by UV-vis reflection spectra, ellipsometer and AFM. As a result, the maximum resonant frequency shift of FBAR reaches more than 20 MHz, meaning 1.4% tunability at least. The minimum frequency shift is nearly 10 kHZ per bilayer, indicating 7 ppm tuning resolution. Besides, Pressure Cooker Test (PCT) is performed to evaluate the reliability of LbL coated FBAR. Furthermore, applications for wireless broadband communication and chemical sensors of LbL coated FBAR have been demonstrated.
We demonstrate the versatility of Al2O3-passivated Si nanowire devices ("nanoribbons") in the analysis of enzyme-substrate interactions via the monitoring of pH change. Our approach is shown to be effective through the detection of urea in phosphate buffered saline (PBS), and penicillinase in PBS and urine, at limits of detection of <200 μM and 0.02 units/mL, respectively. The ability to extract accurate enzyme kinetics and the Michaelis-Menten constant (Km) from the acetylcholine-acetylcholinesterase reaction is also demonstrated.
Phase separation of polyelectrolyte complexes (PECs) between the polyacid (sodium salt) and polybase (hydrochloride) of poly(acrylic acid) (PAA) and poly(allylamine) (PAH), respectively, has been investigated in aqueous solution. Chain length of the PAA was varied (25 < Pw < 700) holding Pw of the PAH constant at 765. The polyacid/polybase mixing ratio (10–90 wt %) and the ionic strength as salt concentration (0–3,000 mM) were systematically varied. Sample turbidity was utilized as an indicator of PEC formation, complemented by optical microscopy for discrimination between precipitate and coacervate. Salt-free systems always resulted in PEC precipitates; however, coacervates or polyelectrolyte solutions, respectively, were formed upon exceeding critical salt concentrations, the PEC formation also depending on the employed PAA/PAH ratio. The lower the PAA molecular weight, the lower were the critical salt concentrations required for both the precipitate/coacervate and coacervate/solution transitions. The experimental phase behavior established here is explained by molecular models of coacervate complexation, addressing effects of polyelectrolyte molecular weight and salt screening.
Strategies to detect human DNA methyltransferases are needed, given that aberrant methylation by these enzymes is associated with cancer initiation and progression. Here we describe a nonradioactive, antibody-free, electrochemical assay in which methyltransferase activity on DNA-modified electrodes confers protection from restriction for signal-on detection. We implement this assay with a multiplexed chip platform and show robust detection of both bacterial (SssI) and human (Dnmt1) methyltransferase activity. Essential to work with human methyltransferases, our unique assay design allows activity measurements on both unmethylated and hemimethylated DNA substrates. We validate this assay by comparison with a conventional radioactive method. The advantages of electrochemistry over radioactivity and fluorescence make this assay an accessible and promising new approach for the sensitive, label-free detection of human methyltransferase activity.
A potentiometric non-enzymatic sensor using off-chip extended-gate field effect transistor (EGFET) with a ferrocenyl-alkanethiol modified gold electrode is demonstrated for determining the uric acid concentration in human serum and urine. Hexacyanoferrate (II) and (III) ions are used as redox reagent. This potentiometric sensor measures the interface potential on the ferrocene immobilized gold electrode, which is modulated by the redox reaction between uric acid and hexacyanoferrate ions. The device shows a near Nernstian response to uric acid and is highly specific. The interference that comes from glucose, bilirubin, ascorbic acid and hemoglobin is negligible in normal concentration range of these interferents. The sensor also exhibits excellent long term reliability. This extended gate field effect transistor based sensors can be used as a point of care UA testing tool, due to the small size, low cost, and low sample volume consumption.
The intrinsic charging status at the dielectric-electrolyte interface (DEI) plays a critical role for electrofluidic gating in microfluidics and nanofluidics, which offers opportunities for integration of wet ionics with dry electronics. A convenient approach to quantitatively probe the surface charges at the DEI for material pre-selection purpose has been lacking so far. We report here a low-cost, off-chip extended gate field effect transistor configuration for direct electrostatic probing the charging status at the DEI. Capacitive coupling between the surface charges and the floating extended gate is utilized for signal transducing. The relationship between the surface charge density and the experimentally accessible quantities is given by device modeling. The multiplexing ability makes measuring a local instead of a globally averaged surface charge possible.
Multilayer films of organic compounds on solid surfaces have been studied for more than 60 years because they allow fabrication
of multicomposite molecular assemblies of tailored architecture. However, both the Langmuir-Blodgett technique and chemisorption
from solution can be used only with certain classes of molecules. An alternative approach—fabrication of multilayers by consecutive
adsorption of polyanions and polycations—is far more general and has been extended to other materials such as proteins or
colloids. Because polymers are typically flexible molecules, the resulting superlattice architectures are somewhat fuzzy structures,
but the absence of crystallinity in these films is expected to be beneficial for many potential applications.
Floating multilayers of polyallylamine/polystyrenesulfonate (PAH/PSS) adsorbed to a dimethyldioctadecylammonium bromide (DODAB) monolayer at the air/water interface (at pH 5.5) are analyzed quantitatively with respect to the internal charge stoichiometry of the polyelectrolytes. To this end the adsorbed amount of polymer is translated into charge per area. It is postulated that the strong acid PSS is fully charged (fully dissociated). The weak polyelectrolyte PAH is also assumed to be fully dissociated. This is vindicated by theoretical considerations and deduced from pH-dependent studies of the isotherms of Langmuir monolayers of negatively charged dimyristoylphosphatidic acid (DMPA) with PAH adsorbed from the subphase. With both polyelectrolytes fully charged the PAH charges significantly overcompensate the PSS charges. The polyelectrolyte charge stoichiometry is not 1:1, and the multilayer must contain some small counterions (Cl-) for charge compensation. It is suggested that this “extrinsic compensation” occurs when the charge density of the polyelectrolytes exceeds a certain threshold. If the distance between the charges is less than the Bjerrum length, the strong binding of the counterions (“Manning condensation”) prevents their release upon multilayer formation. This explains why multilayers with PAH with its unusually high charge density contain counterions whereas other polyelectrolyte films are ion-free. Literature data support this hypothesis.
We describe the highly sensitive detection of the nonspecific adsorption of proteins onto a 1-undecanethiol self-assembled monolayer (SAM)-formed gold electrode by parallel analysis using field effect transistor (FET), surface plasmon resonance (SPR), and quartz crystal microbalance (QCM) sensors. The FET sensor detects the innate electric charges of the adsorbed protein at the electrode/solution interface, transforming the change in charge density into a potentiometric signal in real time, without the requirement for labels. In particular, using the Debye-Huckel model, the degree of potential shift was proportional to the dry mass of adsorbed albumin and β-casein. A comparison of the FET signal with SPR and QCM data provided information on the conformation and orientation of the surface-bound protein by observing characteristic break points in the correlation slopes between the signals. These slope transitions reflect a multistage process that occurs upon protein adsorption as a function of protein concentration, including interim coverage, film dehydration, and monolayer condensation. The FET biosensor, in combination with SPR and QCM, represents a new technology for interrogating protein-material interactions both quantitatively and qualitatively.
We developed an integrated device comprising a quartz crystal microbalance (QCM) and a field-effect transistor (FET) with a single common gold electrode in a flow chamber. An alternating current inducing oscillations in the piezoelectric quartz of the QCM sensor is electrically independent of the circuit for the FET output so that the two sensors in different detection mechanisms simultaneously record binding kinetics from a single protein solution on the same electrode. A conjunction of adsorbed mass from QCM with electric nature of bound protein from FET provided deeper understanding on a complex process of nonspecific protein adsorption and subsequent conformational changes at a solid/liquid interface. Lower apparent k(on) values obtained by FET than those obtained by QCM on hydrophobic surfaces are interpreted as preferred binding of protein molecules facing uncharged domains to the electrode surface, whereas higher k(off) values by FET than those by QCM imply active macromolecular rearrangements on the surfaces mainly driven by hydrophobic association in an aqueous medium. The advanced features of the combined sensor including in situ, label-free, and real-time monitoring provide information on structural dynamics, beyond measurements of affinities and kinetics in biological binding reactions.
Very recently, electric-field-induced superconductivity in an insulator was realized by tuning charge carrier to a high density level (1 × 1014 cm−2). To increase the maximum attainable carrier density for electrostatic tuning of electronic states in semiconductor field-effect transistors is a hot issue but a big challenge. Here, ultrahigh density carrier accumulation is reported, in particular at low temperature, in a ZnO field-effect transistor gated by electric double layers of ionic liquid (IL). This transistor, called an electric double layer transistor (EDLT), is found to exhibit very high transconductance and an ultrahigh carrier density in a fast, reversible, and reproducible manner. The room temperature capacitance of EDLTs is found to be as large as 34 µF cm−2, deduced from Hall-effect measurements, and is mainly responsible for the carrier density modulation in a very wide range. Importantly, the IL dielectric, with a supercooling property, is found to have charge-accumulation capability even at low temperatures, reaching an ultrahigh carrier density of 8×1014 cm−2 at 220 K and maintaining a density of 5.5×1014 cm−2 at 1.8 K. This high carrier density of EDLTs is of great importance not only in practical device applications but also in fundamental research; for example, in the search for novel electronic phenomena, such as superconductivity, in oxide systems.
Cellular production of such cytokines as interferon (IFN)-γ and tumor necrosis factor (TNF)-α is used to determine disease-specific immune responses and may be used to diagnose infectious diseases such as tuberculosis. In this paper, we describe the development of micropatterned electrodes functionalized with electroactive aptamers for multiplexed detection of immune-cell-produced cytokines. A sequence of electrode deprotection and aptamer incubation steps were used to assemble anti-IFN-γ DNA aptamers and anti-TNF-α RNA aptamers on individually addressable half-ring electrodes. Aptamer molecules were thiolated for assembly on gold and were functionalized with methylene blue redox reporter for electrochemical signal transduction. Specificity of individual sensors to the correct cytokine species was confirmed by exposure to recombinant cytokines. For cell detection experiments, electrode arrays were integrated into microfluidic devices and incubated with immune cells. Design of the surface was such that a small group of ~400 cells attached in the circular adhesion sites surrounded by half-ring electrodes sensing IFN-γ and TNF-α. The microdevice consisted of two parallel microfluidic channels, each channel containing four cell capture/sensing sites. Upon mitogenic activation, secreted IFN-γ and TNF-α molecules were monitored by performing square wave voltammetry (SWV) at different time points at individually addressable electrodes. This biosensing platform was used to analyze the quantity and rate of cytokine release from primary T cells and a monocyte cell line. Upon further development of this platform may be enhanced to enable detection of larger number of cytokines and used to correlate the levels and dynamics of cytokine release in immune cells to diagnosis and treatment of infectious diseases.
Unlike the quartz crystal microbalance, which has been used extensively for the analysis of biochemical interactions, only few measurements with biochemical adsorbent have been done with film bulk acoustic resonators (FBAR). In this paper, the FBAR behaviour on exposure to a lipid vesicle solution and the formation of a polyelectrolyte multilayer structure is investigated and compared with the results obtained with the quartz crystal microbalance. Differences in the resonator response were found between the two techniques and depending on the resonators resonance frequency ranging from the MHz to the GHz regime. As an explanation, we suggest that the penetration depth and the influence on viscoelastic properties, which are both known to be frequency dependent, cause the variations in the results. As a consequence, the higher operating resonance frequencies of the FBAR increase the sensitivity to changes in the viscoelasticity of the adsorbent and also decrease the sensing length of the device.
Alkanethiols are commonly used self-assembly reagents in the preparation of modified electrode. In this study, self-assembled monolayers (SAMs) of alkanethiols with different chain lengths formed on gold electrode were mainly investigated by electrochemical impedance spectroscopy (EIS) in addition to cyclic voltammetry (CV). Two types of immobilization strategies applied to the electrode were also examined. The interfacial properties of all self-assembled electrodes were evaluated in the presence of Fe(CN)63−/4− redox probe. A simple equivalent circuit model with a constant phase element was used to interpret the obtained impedance spectra. The results of cyclic voltammetry revealed that the voltammetric behavior of the redox probe was influenced by the electrode surface modification. The EIS data indicated that the formation of saturated SAMs on gold required at least 30 min and more apparent differences occurred with immobilization time compared to that obtained by CV. Depending on the alkyl chain length of the SAMs, the electron-transfer resistance increased and the capacitance decreased. It was found a good linear relationship between inverse capacitance and chain length for carboxyl-terminated SAMs. To achieve a defect-free structure and/or close packing, the direct application method may provide a more effective, rapid, and facile self-assembly immobilization than immersion method from the viewpoint of the electron-transfer resistance.
This paper presents a review of acoustic-wave based MEMS devices that offer a promising technology platform for the development of sensitive, portable, real-time biosensors. MEMS fabrication of acoustic wave based biosensors enables device miniaturization, power consumption reduction and integration with electronic circuits. For biological applications, the biosensors are integrated in a microfluidic system and the sensing area is coated with a biospecific layer. When a bioanalyte interacts with the sensing layer, mass and viscosity variations of the biospecific layer can be detected by monitoring changes in the acoustic wave properties such as velocity, attenuation, resonant frequency and delay time. Few types of acoustic wave devices could be integrated in microfluidic systems without significant degradation of the quality factor. The acoustic wave based MEMS devices reported in the literature as biosensors and presented in this review are film bulk acoustic wave resonators (FBAR), surface acoustic waves (SAW) resonators and SAW delay lines. Different approaches to the realization of FBARs, SAW resonators and SAW delay lines for various biochemical applications are presented. Methods of integration of the acoustic wave MEMS devices in the microfluidic systems and functionalization strategies will be also discussed.
Piezoelectric microelectromechanical systems (MEMS) resonant sensors, known for their excellent mass resolution, have been studied for many applications, including DNA hybridization, protein-ligand interactions, and immunosensor development. They have also been explored for detecting antigens, organic gas, toxic ions, and explosives. Most piezoelectric MEMS resonant sensors are acoustic sensors (with specific coating layers) that enable selective and label-free detection of biological events in real time. These label-free technologies have recently garnered significant attention for their sensitive and quantitative multi-parameter analysis of biological systems. Since piezoelectric MEMS resonant sensors do more than transform analyte mass or thickness into an electrical signal (e.g., frequency and impedance), special attention must be paid to their potential beyond microweighing, such as measuring elastic and viscous properties, and several types of sensors currently under development operate at different resonant modes (i.e., thickness extensional mode, thickness shear mode, lateral extensional mode, flexural mode, etc.). In this review, we provide an overview of recent developments in micromachined resonant sensors and activities relating to biochemical interfaces for acoustic sensors.
Silicon nanowire field effect transistors (FETs) have emerged as ultrasensitive, label-free biodetectors that operate by sensing bound surface charge. However, the ionic strength of the environment (i.e., the Debye length of the solution) dictates the effective magnitude of the surface charge. Here, we show that control of the Debye length determines the spatial extent of sensed bound surface charge on the sensor. We apply this technique to different methods of antibody immobilization, demonstrating different effective distances of induced charge from the sensor surface.
In this study, we have successfully demonstrated that a GaN nanowire (GaNNW) based extended-gate field-effect-transistor (EGFET) biosensor is capable of specific DNA sequence identification under label-free in situ conditions. Our approach shows excellent integration of the wide bandgap semiconducting nature of GaN, surface-sensitivity of the NW-structure, and high transducing performance of the EGFET-design. The simple sensor-architecture, by direct assembly of as-synthesized GaNNWs with a commercial FET device, can achieve an ultrahigh detection limit below attomolar level concentrations: about 3 orders of magnitude higher in resolution than that of other FET-based DNA-sensors. Comparative in situ studies on mismatches ("hotspot" mutations related to human p53 tumor-suppressor gene) and complementary targets reveal excellent selectivity and specificity of the sensor, even in the presence of noncomplementary DNA strands, suggesting the potential pragmatic application in complex clinical samples. In comparison with GaN thin film, NW-based EGFET exhibits excellent performance with about 2 orders higher sensitivity, over a wide detection range, 10(-19)-10(-6) M, reaching about a 6-orders lower detection limit. Investigations illustrate the unique and distinguished feature of nanomaterials. Detailed studies indicate a positive effect of energy band alignment at the biomaterials-semiconductor hybrid interface influencing the effective capacitance and carrier-mobility of the system.
In this study, a highly sensitive and label-free multianalyte capacitive immunosensor was developed based on gold interdigitated electrodes (GID) capacitor arrays to detect a panel of disease biomarkers. C-reactive protein (CRP), TNFalpha, and IL6 have strong and consistent relationships between markers of inflammation and future cardiovascular risk (CVR) events. Early detection of a panel of biomarkers for a disease could enable accurate prediction of a disease risk. The detection of protein biomarkers was based on relative change in capacitive/dielectric properties. Two different lab-on-a-chip formats were employed for multiple biomarker detection on GID-capacitors. In format I, capacitor arrays were immobilized with pure forms of anti-CRP, -TNFalpha, and -IL6 antibodies in which each capacitor array contained a different immobilized antibody. Here, the CRP and IL6 were detected in the range 25 pg/ml to 25 ng/ml and 25 pg/ml to 1 ng/ml for TNFalpha in format I. Sensitive detection was achieved with chips co-immobilized (diluted) with equimolar mixtures of anti-CRP, -IL6, and -TNFalpha antibodies (format II) in which all capacitors in an array were identical and tested for biomarkers with sequential incubation. The resulting response to CRP, IL6, and TNFalpha in format II for all biomarkers was found to be within 25 pg/ml to 25 ng/ml range. The capacitive biosensor for panels of inflammation and CVR markers show significant clinical value and provide great potential for detection of biomarker panel in suspected subjects for early diagnosis.
SPR imaging (SPRi) is at the forefront of optical label-free and real-time detection. It offers the possibility of monitoring hundreds of biological interactions simultaneously and from the binding profiles, allows the estimation of the kinetic parameters of the interactions between the immobilised probes and the ligands in solution. We review the current state of development of SPRi technology and its application including commercially available SPRi instruments. Attention is also given to surface chemistries for biochip functionalisation and suitable approaches to improve sensitivity.
Electrochemical processes at the electrode-electrolyte (body fluid) interface are of ultimate importance for stimulating/sensing electrode function. A high electrode surface area is desirable for safe stimulation through double-layer charging and discharging. Pt and Pt-Ir alloys have been the most common electrode materials. The use of TiN coating as the surface layer on the electrode has found increasing interest because of its metal-like conductivity, excellent mechanical and chemical properties, and the fact that it can be deposited with a high surface area. In this work, electrochemical impedance spectroscopy (EIS), which is a sensitive and non-destructive technique and widely used for characterization of electrical properties of electrode-electrolyte interfaces, was applied to investigate pure Pt and Ti, and TiN coated electrodes exposed to a phosphate-buffered-saline (PBS) solution. Platinized Pt and Ti were also studied for comparison. The capacitance value of the electrodes in PBS was obtained through quantitative analysis of the EIS spectra. The results reveal that the capacitance of the TiN coated electrodes with a rough surface is several hundreds times higher than that of a smooth Pt surface. Platinization of Ti can also increase the capacitance to the same extent as platina. EIS has been shown to be a powerful technique for characterization of stimulating/sensing electrodes.
The labelfree detection of nucleic acid sequences is one of the modern attempts to develop quick, cheap and miniaturised hand-held devices for the future genetic testing in biotechnology and medical diagnostics. We present an approach to detect the hybridisation of DNA sequences using electrolyte-oxide-semiconductor field-effect transistors (EOSFETs) with micrometer dimensions. These semiconductor devices are sensitive to electrical charge variations that occur at the surface/electrolyte interface, i.e. upon hybridisation of oligonucleotides with complementary single-stranded (ss) oligonucleotides, which are immobilised on the oxide surface of the transistor gate. This method allows direct, time-resolved and in situ detection of specific nucleic acid binding events without any labelling. We focus on the detection mechanism of our sensors by using oppositely charged polyelectrolytes (PAH and PSS) subsequently attached to the transistor structures. Our results indicate that the sensor output is charge sensitive and distance dependent from the gate surface, which pinpoints the need for very defined surface chemistry at the device surface. The hybridisation of natural 19 base-pair sequences has been successfully detected with the sensors. In combination with nano-transistors a PCR free detection system might be feasible in future.
Electrostatic layer-by-layer film assembly is an attractive way to non-covalently incorporate proteins and bioactive moieties into the surface of conventional biomaterials. Selection of polycationic and polyanionic components and deposition conditions can be used to control the interfacial properties, and through them protein adsorption, cell adhesion, and tissue development. In this study the polycation was poly(allylamine hydrochloride) (PAH), which is a weak base and consequently adsorbs at interfaces in a pH-dependent manner, and the polyanion was heparin, which is capable of interacting with many adhesion ligands and growth factors. PAH/heparin multilayer films were formed using PAH solutions of pH 6.4, 7.4, 8.4, and 9.4. Film thickness increased both with the number of PAH/heparin bilayers and the pH of the PAH solution. Films consisting of 10 bilayers with heparin topmost exhibited similar bulk atomic compositions and penetration of PAH into the heparin top layer. Finally, fibronectin adsorption and cell adhesion were maximal at an intermediate pH (pH 8.4>pH 9.4>pH 7.4). These results demonstrate that heparin-containing electrostatic films support cell adhesion and protein adsorption in a manner sensitive to film deposition conditions.
The dispersion of nanotubes by pH-responsive polymers (i.e., weak polyelectrolytes) enables the macroscopic properties of aqueous suspensions to be tuned. Microstructural changes were achieved as a function of pH in aqueous suspensions containing single-walled carbon nanotubes and imaged by cryogenic-TEM. Clear evidence of pH-sensitive nanotube dispersion is shown. We expect that many useful properties of these nanotube-polymer systems could be sensitive to microstructure, making this technique important for aqueous processing of carbon nanotubes and macroscopic tailoring of solid polymer nanocomposite behavior.
Nanowire field effect transistors (NW-FETs) can serve as ultrasensitive detectors for label-free reagents. The NW-FET sensing mechanism assumes a controlled modification in the local channel electric field created by the binding of charged molecules to the nanowire surface. Careful control of the solution Debye length is critical for unambiguous selective detection of macromolecules. Here we show the appropriate conditions under which the selective binding of macromolecules is accurately sensed with NW-FET sensors.
- L Mu
- I A Droujinine
- N K Rajan
- S D Sawtelle
- M A Reed
L. Mu, I.A. Droujinine, N.K. Rajan, S.D. Sawtelle, M.A. Reed, Direct, Rapid, and Label-Free
Detection of Enzyme–Substrate Interactions in Physiological Buffers Using CMOS-Compatible
Nanoribbon Sensors, Nano letters, 14(2014) 5315-22.
Buildup of ultrathin multilayer films by a self-assembly process
- G Decher
- J Hong
- J Schmitt
G. Decher, J. Hong, J. Schmitt, Buildup of ultrathin multilayer films by a self-assembly process: