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1.
Real time device for biosensing: design of a bacteriophage model using love acoustic waves 总被引:2,自引:0,他引:2
Tamarin O Comeau S Déjous C Moynet D Rebière D Bezian J Pistré J 《Biosensors & bioelectronics》2003,18(5-6):755-763
Love wave sensors (ST-cut quartz substrate with interdigital transducers, SiO(2) guiding layer and sensitive coating) have been receiving a great deal of attention for a few years. Indeed, the wave coupled in a guiding layer confers a high gravimetric sensitivity and the shear horizontal (SH) polarization allows to work in liquid media. In this paper, an analytical method is proposed to calculate the Love wave phase velocity and the gravimetric sensitivity for a complete multilayer structure. This allows us to optimize the Love wave devices design in order to improve their gravimetric sensitivity in liquid media. As a model for virus or bacteria detection in liquids (drinking or bathing water, food em leader ) we design a model using M13 bacteriophage. The first step is the anti-M13 (AM13) monoclonal antibody grafting, on the device surface (SiO(2)). The second step is an immunoreaction in between the M13 bacteriophage and the AM13 antibody. The Love wave device allows to detect in real time the graft of the AM13 sensitive coating, as well as the immobilization of the M13 bacteriophages. With a pH change, the M13 bacteriophages can be removed from the sensor surface, in order to be numerated as plaque forming unit (pfu). Results on the sensitivity of Love waves are compared with similar immunological works with bulk acoustic wave devices, and demonstrate the high potentialities of Love waves sensors. 相似文献
2.
P.W. Walton M.R. O''Flaherty M.E. Butler P. Compton 《Biosensors & bioelectronics》1993,8(9-10):401-407
Most gravimetric biosensors use thin piezoelectric quartz crystals, either as resonating crystals (quartz crystal microbalance, QCM), or as bulk/surface acoustic wave (SAW) devices. In the majority of these the mass response is inversely proportional to the crystal thickness which, at a limit of about 150 microns, gives inadequate sensitivity. A new system is described in which acoustic waves are launched in very thin (10 microns) tensioned polymer films to produce an oscillatory device. A theoretical equation for this system is almost identical to the well-known Sauerbrey equation used in the QCM method. Because the polymer films are so thin, a 30-fold increase in sensitivity is predicted and verified by adding known surface masses. Temperature sensitivity is a problem so a separate control sensor and careful temperature regulation are necessary. Preliminary results showing the real time binding of protein (IgG), a step towards immunosensor development, and the use of mass enhancing particles are presented. Inexpensive materials are used so disposable gravimetric biosensors may become feasible. 相似文献
3.
Stevenson AC Araya-Kleinsteuber B Sethi RS Mehta HM Lowe CR 《Journal of molecular recognition : JMR》2004,17(3):174-179
A tunable acoustic biosensor for investigating the properties of biomolecules at the solid-liquid interfaces is described. In its current, format the device can be tuned to frequencies between 6.5 MHz and 1.1 GHz in order to provide a unique detection feature: a variable evanescent wave thickness at the sensor surface. The key to its successful implementation required the careful selection of antennae designs that could induce shear acoustic waves at the solid-liquid interface. This non-contact format makes it possible to recover resonant shear acoustic waves over 100 different harmonic frequencies as a result of the electrical characteristics of the spiral coil. For testing this multifrequency sensing concept the surface of a quartz disc was exposed to solutions of immunoglobulin G (IgG) to form an adsorbed monolayer, whence protein A and IgG were added again in order to form multilayers. Spectra at frequencies between 6 and 600 MHz were generated for each successive layer and revealed two characteristic phases: an initial phase at the low megahertz frequencies consistent with the conventional Sauerbrey relation, and a possible additional phase towards the high megahertz to gigahertz frequencies, that we believe relates to the structure of the biomolecular film. This two-phase behaviour evident from differences between high and low frequencies, rather than from any distinct frequency transition, was anticipated from the reduction in evanescent wave thickness down to nanometre dimensions, and thin film resonance phenomena that are known to occur for film and fluid systems. These measurements suggested that the single element acoustic biosensor we present here may form the basis from which to generate acoustic molecular spectra, or "acoustic fingerprints", in a manner akin to optical spectroscopy. 相似文献
4.
Stevenson AC Araya-Kleinsteuber B Sethi RS Metha HM Lowe CR 《Biosensors & bioelectronics》2005,20(7):1298-1304
A transducer format that replaces the electrode of an acoustic resonator with a planar spiral coil is used to extract multifrequency spectral information from adsorbed protein films. Both amorphous silica and crystalline piezoelectric resonators are driven to resonance by forces induced across an air gap by magnetic direct generation and piezoelectric excitation induced by the electromagnetic field of the coil. Inspection of the harmonic frequencies between 6 MHz and 0.6 GHz indicates that the response of these two resonator types is described by different families of shear acoustic standing waves, with similar acoustic features to the quartz crystal microbalance. Exposure of the devices to protein solutions results in reproducible shifts of their harmonic frequencies, up to a maximum of 15 kHz and increasing linearly with frequency and operating mode. The gradient, determined from the ratio of the frequency change to the operating frequency was determined as 21.5 x 10(-6) for the quartz device and 60.9 x 10(-6) for the silica device. Consistency with the Sauerbrey equation for the piezoelectric linear shear mode was comparable at a predicted value of 22.5 x 10(-6), but not for the radial shear mode of the silica device at 12.7 x 10(-6). Opportunities resulting from the wide bandwidth of the planar coil excitation and choice of acoustic mode are discussed with respect to acoustic fingerprinting of adsorbed proteins. 相似文献
5.
Ultrasensitive quartz crystal microbalance sensors for detection of M13-Phages in liquids 总被引:4,自引:0,他引:4
Erich Uttenthaler Michael Schr ml Johannes Mandel Stephan Drost 《Biosensors & bioelectronics》2001,16(9-12):735-743
Quartz crystal microbalance (QCM) sensors are widely used for determining liquid properties or probing interfacial processes. For some applications the sensitivity of the QCM sensors typically used (5–20 MHz) is limited compared with other biosensor methods. In this study ultrasensitive QCM sensors with resonant frequencies from 39 to 110 MHz for measurements in the liquid phase are presented. The fundamental sensor effect of a QCM is the decrease of the resonant frequency of an oscillating quartz crystal due to the binding of mass on a coated surface during the measurement. The sensitivity of QCM sensors increases strongly with an increasing resonant frequency and, therefore, with a decreasing thickness of the sensitive area. The new kind of ultrasensitive QCM sensors used in this study is based on chemically milled shear mode quartz crystals which are etched only in the center of the blank, forming a thin quartz membrane with a thick, mechanically stable outer ring. An immunoassay using a virus specific monoclonal antibody and a M13-Phage showed an increase in the signal to noise ratio by a factor of more than 6 for 56 MHz quartz crystals compared with standard 19 MHz quartz crystals, the detection limit was improved by a factor of 200. Probing of acoustic properties of glycerol/water mixtures resulted in an increase in sensitivity, which is in very good agreement with theory. Chemically milled QCM sensors strongly improve the sensitivity in biosensing and probing of acoustic properties and, therefore, offer interesting new application fields for QCM sensors. 相似文献
6.
A novel high sensitivity ZnO/SiO(2)/Si Love mode surface acoustic wave (SAW) biosensor for the detection of interleukin-6 (IL-6), is reported. The biosensors operating at 747.7MHz and 1.586GHz were functionalized by immobilizing the monoclonal IL-6 antibody onto the ZnO biosensor surface both through direct surface adsorption and through covalent binding on gluteraldehyde. The morphology of the IL-6 antibody-protein complex was studied using scanning electron microscopy (SEM), and the mass of the IL-6 protein immobilized on the surface was measured from the frequency shift of the SAW resonator biosensor. The biosensor was shown to have extended linearity, which was observed to improve with higher sensor frequency and for IL-6 immobilization through the monoclonal antibody. Preliminary results of biosensor measurements of low levels of IL-6 in normal human serum are reported. The biosensor can be fully integrated with CMOS Si chips and developed as a portable real time detection system for the interleukin family of proteins in human serum. 相似文献
7.
The thickness shear mode (TSM) acoustic wave sensor attached with living cells has been shown to be an effective functional biosensing device to monitor the process of cell adhesion to a surface in real time. In this study, a multilayer sensor model that includes a quartz substrate, a cell-substrate interfacial layer and a cell layer was constructed based on the state of cell adhesion to the substrate. The dynamic process of cell adhesions as a function of cell seeding densities was monitored using the acoustic wave sensor. The mechanisms that are responsible for the frequency and resistance change are discussed according to the predictions of the acoustic wave sensor model. In addition, knowing that the actin cytoskeleton is important for cell adhesion, we investigated the motional resistance change caused by the disruption of actin cytoskeleton induced by fungal toxin cytochalasin D in the human skin fibroblasts. The results indicate that resistance changes are related to the disruption of actin cytoskeleton and dosage-dependent. 相似文献
8.
Novel metal clad leaky waveguide (MCLW) sensor devices have been developed for sensing applications. These chips are designed to confine the light in a low refractive index waveguide that encompasses the chemically-selective layer, maximising the overlap between the optical mode and the chemistry, thus improving the sensitivity. In this work, a thin metal layer was inserted between the substrate and the thick waveguide layer, increasing the reflectivity of the waveguide/metal interface and decreasing the light lost at each of reflection in the leaky mode, which in turn increases the propagation distance. The device has been used for a range of biosensing applications, including the detection of organophosphoros pesticides. The limit of detection for paraoxon, based on absorbance detection, was calculated to be 6 nM. Refractive index detection was demonstrated by monitoring the change in the out-coupled angle resulting from the binding of protein A to anti-protein A immobilized on agarose. The sensor was also used for detecting the quenching of the fluorescence of an acid-base sensitive ruthenium complex immobilized within the sol-gel and with glucose oxidase enzyme. The limit of detection for glucose was 3 microM. The advantage of using the metal layer in the MCLW was that an electrical potential could be applied to accelerate the diffusion of the analyte to the immobilised antibody, which resulted in a shortened analysis time and a reduction in non-specific binding. 相似文献
9.
In this work, we describe a novel pulse mode shear horizontal-surface acoustic wave (SH-SAW) polymer coated biosensor that monitors rapid changes in both amplitude and phase. The SH-SAW sensors were fabricated on 36 degrees rotated Y-cut X propagating lithium tantalate (36 YX.LT). The sensitivity of the device to both mass loading and visco-elastic effects may be increased by using a thin guiding layer of cross-linked polymer. Two acoustic modes are excited by the electrodes in this crystalline direction. Metallisation of the propagation path of the 36 YX.LT devices allows the two modes to be discriminated. Successive polymer coatings resulted in the observation of resonant conditions in both modes as the layer thickness was increased. Using the 36 YX.LT devices, we have investigated the application of a novel pulse mode system by sensing a sequence of deposition and removal of a biological layer consisting of vesicles of the phospholipid POPC. A continuous wave system was used to verify the accuracy of the pulse mode system by sensing a series of poly(ethylene glycol) (PEG) solutions. The data clearly demonstrates the ability of the 36 YX.LT pulse mode system to provide rapid measurements of both amplitude and phase for biosensing applications. 相似文献
10.
J Freudenberg S Schelle K Beck M von Schickfus S Hunklinger 《Biosensors & bioelectronics》1999,14(4):423-425
Surface acoustic wave sensors operating in liquid generally cause problems resulting from wire bonding. The authors present an approach for a biosensor where the need for bonding wires is eliminated by utilizing inductive coupling of the sensor device to the RF circuitry. Protection of the electrodes from the liquid is achieved by coating the device surface with a SiO2 layer, resulting in a simplified handling of the devices. The first measurements with a sensor operating at 420 MHz are presented, demonstrating the potential of this operating principle for biosensing. 相似文献
11.
Gabl R Feucht HD Zeininger H Eckstein G Schreiter M Primig R Pitzer D Wersing W 《Biosensors & bioelectronics》2004,19(6):615-620
A novel integrated bio-sensor technology based on thin-film bulk acoustic wave resonators on silicon is presented and the feasibility of detecting DNA and protein molecules proofed. The detection principle of these sensors is label-free and relies on a resonance frequency shift caused by mass loading of an acoustic resonator, a principle very well known from quartz crystal micro balances. Integrated ZnO bulk acoustic wave resonators with resonance frequencies around 2 GHz have been fabricated, employing an acoustic mirror for isolation from the silicon substrate. DNA oligos have been thiol-coupled to the gold electrode by on-wafer dispensing. In a further step, samples have either been hybridised or alternatively a protein has been coupled to the receptor. The measurement results show the new bio-sensor being capable of both, detecting proteins as well as the DNA hybridisation without using a label. Due to the substantially higher oscillation frequency, these sensors already show much higher sensitivity and resolution comparable to quartz crystal micro balances. The potential for these sensors and sensors arrays as well as technological challenges will be discussed in detail. 相似文献
12.
Background
Nucleic acid based recognition of viral sequences can be used together with label-free biosensors to provide rapid, accurate confirmation of viral infection. To enhance detection sensitivity, gold nanoparticles can be employed with mass-sensitive acoustic biosensors (such as a quartz crystal microbalance) by either hybridising nanoparticle-oligonucleotide conjugates to complimentary surface-immobilised ssDNA probes on the sensor, or by using biotin-tagged target oligonucleotides bound to avidin-modified nanoparticles on the sensor. We have evaluated and refined these signal amplification assays for the detection from specific DNA sequences of Herpes Simplex Virus (HSV) type 1 and defined detection limits with a 16.5 MHz fundamental frequency thickness shear mode acoustic biosensor. 相似文献13.
A thickness shear-mode acoustic wave device, operated in a flow-through format, was used to detect the binding of ions or peptides to surface-attached calmodulin. On-line surface attachment of the protein was achieved by immobilisation of the biotinylated molecule via a neutravidin-biotin linkage onto the surface of the gold electrode of the detector. The interaction between calmodulin, and calcium and magnesium ions induced an increase in resonant frequency and a decrease in motional resistance, which were reversible on washing with buffer. Interestingly, the changes in resonant frequency and motional resistance induced by the binding were opposite to the normal operation of the detector. The response was interpreted as a decrease in surface coupling (partial slip at the liquid/solid interface) instigated by exposure of hydrophobic domains on the protein, and an increase in the thickness, and hence effective wavelength, of the acoustic device, corresponding to an increase in the length of calmodulin by 1.5 A. This result is consistent with the literature value of 4 A. In addition, the interaction of the protein with peptide together with calcium ions was detected successfully, despite the relatively low molecular mass of the 2-kDa peptide. These results confirm the potential of acoustic wave physics for the detection of changes in the conformational chemistry of monolayer of biochemical macromolecules at the solid/liquid interface. 相似文献
14.
Quantification of the effect of glycocalyx condition on membrane receptor interactions using an acoustic wave sensor 总被引:1,自引:0,他引:1
The effect of the cell glycocalyx on the binding of a membrane receptor, class I major histocompatibility complex (MHC) human
leukocyte antigen (HLA)-A2, to an immobilized anti-HLA antibody was investigated using an acoustic sensor based on a Love
wave geometry. The enzyme neuraminidase was used to remove sialic acid residues from the cell glycocalyx. Real-time measurements
of the amplitude of the acoustic wave showed that treatment with neuraminidase facilitates HLA/anti-HLA-mediated cell attachment
via a 3.6-fold increase of the two-dimensional (2D) binding constant of the interaction. This could be attributed to better
approach of binding partners due to favorable condition of the desialylated glycocalyx. The results underline the importance
of microtopological factors in membrane receptor binding and reveal the potential of the Love wave sensor and 2D binding parameters
for studying cell–substrate binding events. 相似文献
15.
Rapid and sensitive biosensor for Salmonella 总被引:2,自引:0,他引:2
Pathirana ST Barbaree J Chin BA Hartell MG Neely WC Vodyanoy V 《Biosensors & bioelectronics》2000,15(3-4):135-141
The rapid and sensitive detection of Salmonella typhymurium based on the use of a polyclonal antibody immobilized by the Langmuir-Blodgett method on the surface of a quartz crystal acoustic wave device was demonstrated. The binding of bacteria to the surface changed the crystal resonance parameters; these were quantified by the output voltage of the sensor instrumentation. The sensor had a lower detection limit of a few hundred cells/ml, and a response time of < 100 s over the range of 10(2)-10(10) cells/ml. The sensor response was linear between bacterial concentrations of 10(2)-10(7) cells/ml, with a sensitivity of 18 mV/decade. The binding of bacteria was specific with two binding sites needed to bind a single cell. The sensors preserve approximately 75% of their sensitivity over a period of 32 days. 相似文献
16.
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. 相似文献
17.
A new technique has been developed that combines evanescent-field optical sensing with electrochemical control of surface adsorption processes. This new technique, termed "electrochemical optical waveguide lightmode spectroscopy" (EC-OWLS), proved efficient in monitoring molecular surface adsorption and layer thickness changes of an adsorbed polymer layer examined in situ as a function of potential applied to a waveguide in a pilot study. For optical sensing, a layer of indium tin oxide (ITO) served as both a high-refractive-index waveguide and a conductive electrode. In addition, an electrochemical flow-through fluid cell was provided, which incorporated working, reference, and counter electrodes, and was compatible with the constraints of optical sensing. Poly(L-lysine)-grafted-poly(ethylene glycol) (PLL-g-PEG) served as a model, polycation adsorbate. Adsorption of PLL-g-PEG from aqueous buffer solution increased from 125 to 475 ng/cm(2 )along a sigmoidal path as a function of increasing potential between 0 and 1.5 V versus the Ag reference electrode. Upon buffer rinse, adsorption was partially reversible when a potential of >/=0.93 V was maintained on the ITO waveguide. However, reducing the applied potential back to 0 V before rinsing resulted in irreversible polymer adsorption. PLL-g-PEG modified with biotin demonstrated similar adsorption characteristics, but subsequent streptavidin binding was independent of biotin concentration. Applying positive potentials resulted in increased adsorbed mass, presumably due to polymer chain extension and reorganization in the molecular adlayer. 相似文献
18.
Naima Maouche Nadia Ktari Idriss Bakas Najla Fourati Chouki Zerrouki Mahamadou Seydou François Maurel Mohammed Mehdi Chehimi 《Journal of molecular recognition : JMR》2015,28(11):667-678
A surface acoustic wave sensor operating at 104 MHz and functionalized with a polypyrrole molecularly imprinted polymer has been designed for selective detection of dopamine (DA). Optimization of pyrrole/DA ratio, polymerization and immersion times permitted to obtain a highly selective sensor, which has a sensitivity of 0.55°/mM (≈550 Hz/mM) and a detection limit of ≈ 10 nM. Morphology and related roughness parameters of molecularly imprinted polymer surfaces, before and after extraction of DA, as well as that of the non imprinted polymer were characterized by atomic force microscopy. The developed chemosensor selectively recognized dopamine over the structurally similar compound 4‐hydroxyphenethylamine (referred as tyramine), or ascorbic acid,which co‐exists with DA in body fluids at a much higher concentration. Selectivity tests were also carried out with dihydroxybenzene, for which an unexpected phase variation of order of 75% of the DA one was observed. Quantum chemical calculations, based on the density functional theory, were carried out to determine the nature of interactions between each analyte and the PPy matrix and the DA imprinted PPy polypyrrole sensing layer in order to account for the important phase variation observed during dihydroxybenzene injection. Copyright © 2015 John Wiley & Sons, Ltd. 相似文献
19.
We present an acoustic Love-wave biosensor for detection of the Bacillus anthracis simulant, Bacillus thuringiensis at or below inhalational infectious levels. The present work is an experimental study of 36 degrees YX cut LiTaO3 based Love-wave devices for detection of pathogenic spores in aqueous conditions. Given that the detection limit (D1) of Love-wave-based sensors is a strong function of the overlying waveguide, two waveguide materials have been investigated, which are polyimide and polystyrene. To determine the mass sensitivity of Love-wave sensor, bovine serum albumin (BSA) protein was injected into the Love-wave test cell while recording the magnitude and phase shift across each sensor. Polyimide had the lowest mass detection limit with an estimated value of 1.0-2.0 ng/cm2, as compared to polystyrene where D1 = 2.0 ng/cm2. Suitable chemistries were used to orient antibodies on the Love-wave sensor using protein G. The thickness of each biofilm was measured using ellipsometry from which the surface concentrations were calculated. The monoclonal antibody BD8 with a high degree of selectivity for anthrax spores was used to capture the non-pathogenic simulant B. thuringiensis B8 spores. Bacillus subtilis spores were used as a negative control to determine whether significant non-specific binding would occur. Spore aliquots were prepared using an optical counting method, which permitted removal of background particles for consistent sample preparation. This work demonstrates that Love-wave biosensors are promising for low-level detection for whole-cell biological pathogens. 相似文献
20.
Cell property determination from the acoustic microscope generated voltage versus frequency curves
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Among the methods for the determination of mechanical properties of living cells acoustic microscopy provides some extraordinary advantages. It is relatively fast, of excellent spatial resolution and of minimal invasiveness. Sound velocity is a measure of the stiffness or Young's modulus of the cell. Attenuation of cytoplasm is a measure of supramolecular interactions. These parameters are of crucial interest for studies of cell motility, volume regulations and to establish the functional role of the various elements of the cytoskeleton. Using a phase and amplitude sensitive modulation of a scanning acoustic microscope (Hillman et al., 1994, J. Alloys Compounds. 211/212:625-627) longitudinal wave speed, attenuation and thickness profile of a biological cell are obtained from the voltage versus frequency or V(f) curves. A series of pictures, for instance in the frequency range 980-1100 MHz with an increment of 20 MHz, allows the experimental generation of V(f) curves for each pixel while keeping the lens-specimen distance unchanged. Both amplitude and phase values of the V(f) curves are used for obtaining the cell properties and the cell thickness profile. The theoretical analysis shows that the thin liquid layer, between the cell and the substrate, has a strong influence on the reflection coefficient and should not be ignored during the analysis. Cell properties, cell profile and the thickness of the thin liquid layer are obtained from the V(f) curves by the simplex inversion algorithm. The main advantages of this new method are that imaging can be done near the focal plane, therefore an optimal signal to noise ratio is achieved, no interference with Rayleigh waves occurs, and the method requires only an approximate estimate of the material properties of the solid substratum where the cells are growing on. 相似文献