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Sample records for achieve signal amplification

  1. A Simple Structure for Signal Amplification

    NASA Astrophysics Data System (ADS)

    Ding, Wan-Xiang; Gu, Chang-Gui; Liang, Xiao-Ming

    2016-02-01

    It has been found that a triple-node feed-forward motif has a function of signal amplification, where two input nodes receive the external weak signal and jointly modulate the response of the third output node [Liang et al., Phys. Rev. E 88 (2013) 012910]. We here show that the signal amplification can be further enhanced by adding a link between the two input nodes in the feed-forward motif. We further reveal that the coupling strength of the link regulates the enhancement of signal amplification in the modified feed-forward motif. We finally analyze the mechanism of signal amplification of such simple structure. Supported by the Program for Professor of Special Appointment (Eastern Scholar) at Shanghai Institutions of Higher Learning under Grant No. QD2015016, the National Natural Science Foundation of China under Grant Nos. 11505114 and 11305078

  2. Tyramide Signal Amplification for Immunofluorescent Enhancement.

    PubMed

    Faget, Lauren; Hnasko, Thomas S

    2015-01-01

    Enzyme-linked signal amplification is a key technique used to enhance the immunohistochemical detection of protein, mRNA, and other molecular species. Tyramide signal amplification (TSA) is based on a catalytic reporter deposit in close vicinity to the epitope of interest. The advantages of this technique are its simplicity, enhanced sensitivity, high specificity, and compatibility with modern multi-label fluorescent microscopy. Here, we describe the use of a TSA kit to increase the signal of enhanced green fluorescent protein (eGFP) expressed under the control of Slc17a6 regulatory elements in the brain of a transgenic mouse. The labeling procedure consists of 6 basic steps: (1) tissue preparation, (2) blocking of nonspecific epitopes, (3) binding with primary antibody, (4) binding with horseradish peroxidase-conjugated secondary antibody, (5) reacting with fluorescent tyramide substrate, and (6) imaging of the signal. The procedures described herein detail these steps and provide additional guidance and background to assist novice users.

  3. Amplification of signaling events in bacteria.

    PubMed

    Dahlquist, Frederick W

    2002-05-14

    Bacteria respond to extremely shallow chemical gradients by modifying their motility in a process called chemotaxis. This chemotactic response is characterized by high sensitivity to small concentration differences, which extends over a large range of concentrations. This combination of high signal gain and large dynamic range results from both a memory of past events and the ability to amplify small differences in signal between the memory and the current environment. Dahlquist describes the signaling mechanism used by bacteria to regulate the flagellar motor and the places in this pathway where signal amplification may occur.

  4. Signal Amplification of Bioassay Using Zinc Nanomaterials

    NASA Astrophysics Data System (ADS)

    Cowles, Chad L.

    An emerging trend in the analytical detection sciences is the employment of nanomaterials for bioassay signal transduction to identify analytes critical to public health. These nanomaterials have been specifically investigated for applications which require identification of trace levels of cells, proteins, or other molecules that can have broad ranging impacts to human health in fields such as clinical diagnostics, environmental monitoring, food and drink control, and the prevention of bioterrorism. Oftentimes these nanoparticle-based signal transduction or amplification approaches offer distinct advantages over conventional methods such as increased sensitivity, rapidity, or stability. The biological application of nanoparticles however, does suffer from drawbacks that have limited more widespread adoption of these techniques. Some of these drawbacks are, high cost and toxicity, arduous synthesis methods, functionalization and bioconjugation challenges, and laboratory disposal and environmental hazard issues, all of which have impeded the progression of this technology in some way or another. This work aims at developing novel techniques that offer solutions to a number of these hurdles through the development of new nanoparticle-based signal transduction approaches and the description of a previously undescribed nanomaterial. Zinc-based nanomaterials offer the opportunity to overcome some of the limitations that are encountered when other nanomaterials are employed for bioassay signal transduction. On the other hand, the biological application of zinc nanomaterials has been difficult because in general their fluorescence is in the blue range and the reported quantum yields are usually too low for highly sensitive applications. The advantages of using zinc nanomaterials for biological applications, such as reduced toxicity, simple synthesis, low cost, and straightforward functionalization strategies contribute to the research interest in their application as

  5. Method Of Signal Amplification In Multi-Chromophore Luminescence Sensors

    DOEpatents

    Levitsky, Igor A.; Krivoshlykov, Sergei G.

    2004-02-03

    A fluorescence-based method for highly sensitive and selective detection of analyte molecules is proposed. The method employs the energy transfer between two or more fluorescent chromophores in a carefully selected polymer matrix. In one preferred embodiment, signal amplification has been achieved in the fluorescent sensing of dimethyl methylphosphonate (DMMP) using two dyes, 3-aminofluoranthene (AM) and Nile Red (NR), in a hydrogen bond acidic polymer matrix. The selected polymer matrix quenches the fluorescence of both dyes and shifts dye emission and absorption spectra relative to more inert matrices. Upon DMMP sorption, the AM fluorescence shifts to the red at the same time the NR absorption shifts to the blue, resulting in better band overlap and increased energy transfer between chromophores. In another preferred embodiment, the sensitive material is incorporated into an optical fiber system enabling efficient excitation of the dye and collecting the fluorescent signal form the sensitive material on the remote end of the system. The proposed method can be applied to multichromophore luminescence sensor systems incorporating N-chromophores leading to N-fold signal amplification and improved selectivity. The method can be used in all applications where highly sensitive detection of basic gases, such as dimethyl methylphosphonate (DMMP), Sarin, Soman and other chemical warfare agents having basic properties, is required, including environmental monitoring, chemical industry and medicine.

  6. First demonstration of high-order QAM signal amplification in PPLN-based phase sensitive amplifier.

    PubMed

    Umeki, T; Tadanaga, O; Asobe, M; Miyamoto, Y; Takenouchi, H

    2014-02-10

    We demonstrate the phase sensitive amplification of a high-order quadrature amplitude modulation (QAM) signal using non-degenerate parametric amplification in a periodically poled lithium niobate (PPLN) waveguide. The interaction between the pump, signal, and phase-conjugated idler enables us to amplify arbitrary phase components of the signal. The 16QAM signals are amplified without distortion because of the high gain linearity of the PPLN-based phase sensitive amplifier (PSA). Both the phase and amplitude noise reduction capabilities of the PSA are ensured. Phase noise cancellation is achieved by using the interaction with the phase-conjugated idler. A degraded signal-to-noise ratio (SNR) is restored by using the gain difference between a phase-correlated signal-idler pair and uncorrelated excess noise. The applicability of the simultaneous amplification of multi-carrier signals and the amplification of two independent polarization signals are also confirmed with a view to realizing ultra-high spectrally efficient signal amplification.

  7. Signal amplification in a qubit-resonator system

    NASA Astrophysics Data System (ADS)

    Karpov, D. S.; Oelsner, G.; Shevchenko, S. N.; Greenberg, Ya. S.; Il'ichev, E.

    2016-03-01

    We study the dynamics of a qubit-resonator system, when the resonator is driven by two signals. The interaction of the qubit with the high-amplitude driving we consider in terms of the qubit dressed states. Interaction of the dressed qubit with the second probing signal can essentially change the amplitude of this signal. We calculate the transmission amplitude of the probe signal through the resonator as a function of the qubit's energy and the driving frequency detuning. The regions of increase and attenuation of the transmitted signal are calculated and demonstrated graphically. We present the influence of the signal parameters on the value of the amplification, and discuss the values of the qubit-resonator system parameters for an optimal amplification and attenuation of the weak probe signal.

  8. Signal bi-amplification in networks of unidirectionally coupled MEMS

    NASA Astrophysics Data System (ADS)

    Tchakui, Murielle Vanessa; Woafo, Paul; Colet, Pere

    2016-01-01

    The purpose of this paper is to analyze the propagation and the amplification of an input signal in networks of unidirectionally coupled micro-electro-mechanical systems (MEMS). Two types of external excitations are considered: sinusoidal and stochastic signals. We show that sinusoidal signals are amplified up to a saturation level which depends on the transmission rate and despite MEMS being nonlinear the sinusoidal shape is well preserved if the number of MEMS is not too large. However, increasing the number of MEMS, there is an instability that leads to chaotic behavior and which is triggered by the amplification of the harmonics generated by the nonlinearities. We also show that for stochastic input signals, the MEMS array acts as a band-pass filter and after just a few elements the signal has a narrow power spectra.

  9. Parametric signal amplification to create a stiff optical bar

    NASA Astrophysics Data System (ADS)

    Somiya, K.; Kataoka, Y.; Kato, J.; Saito, N.; Yano, K.

    2016-02-01

    An optical cavity consisting of optically trapped mirrors makes a resonant bar that can be stiffer than diamond. A limitation of the stiffness arises in the length of the optical bar as a consequence of the finite light speed. High laser power and light mass mirrors are essential for realization of a long and stiff optical bar that can be useful for example in the gravitational-wave detector aiming at the observation of a signal from neutron-star collisions, supernovae, etc. In this letter, we introduce a parametric signal amplification scheme that realizes the long and stiff optical bar with a non-linear crystal inside the signal-recycling cavity.

  10. Utilization of nanoparticle labels for signal amplification in ultrasensitive electrochemical affinity biosensors: a review.

    PubMed

    Ding, Liang; Bond, Alan M; Zhai, Jianping; Zhang, Jie

    2013-10-03

    Nanoparticles with desirable properties not exhibited by the bulk material can be readily synthesized because of rapid technological developments in the fields of materials science and nanotechnology. In particular their highly attractive electrochemical properties and electrocatalytic activity have facilitated achievement of the high level of signal amplification needed for the development of ultrasensitive electrochemical affinity biosensors for the detection of proteins and DNA. This review article explains the basic principles of nanoparticle based electrochemical biosensors, highlights the recent advances in the development of nanoparticle based signal amplification strategies, and provides a critical assessment of the likely drawbacks associated with each strategy. Finally, future perspectives for achieving advanced signal simplification in nanoparticles based biosensors are considered.

  11. Preferential amplification of rising versus falling frequency whistler mode signals

    NASA Astrophysics Data System (ADS)

    Li, J. D.; Harid, V.; Spasojevic, M.; Gołkowski, M.; Inan, U. S.

    2015-01-01

    Analysis of ground-based ELF/VLF observations of injected whistler mode waves from the 1986 Siple Station experiment demonstrates the preferential magnetospheric amplification of rising over descending frequency-time ramps. From examining conjugate region receptions of ±1 kHz/s frequency-time ramps, we find that rising ramps generate an average total power 1.9 times higher than that of falling frequency ramps when both are observed during a transmission. And in 17% of receptions, only rising ramps are observed above the noise floor. Furthermore, the amplification ratio inversely correlates with the noise and total signal power. Using a narrowband Vlasov-Maxwell numerical simulation, we explore the preferential amplification due to differences in linear growth rate as a function of frequency, relative to the frequency which maximizes the linear growth rate for a given anisotropy, and in nonlinear phase trapping. These results contribute to the understanding of magnetospheric wave amplification and the preference for structured rising elements in chorus.

  12. Ultrasensitive electrochemical aptasensor for ochratoxin A based on two-level cascaded signal amplification strategy.

    PubMed

    Yang, Xingwang; Qian, Jing; Jiang, Ling; Yan, Yuting; Wang, Kan; Liu, Qian; Wang, Kun

    2014-04-01

    Ochratoxin A (OTA) has a number of toxic effects to both humans and animals, so developing sensitive detection method is of great importance. Herein, we describe an ultrasensitive electrochemical aptasensor for OTA based on the two-level cascaded signal amplification strategy with methylene blue (MB) as a redox indicator. In this method, capture DNA, aptamers, and reporter DNA functionalized-gold nanoparticles (GNPs) were immobilized on the electrode accordingly, where GNPs were used as the first-level signal enhancer. To receive the more sensitive response, a larger number of guanine (G)-rich DNA was bound to the GNPs' surface to provide abundant anchoring sites for MB to achieve the second-level signal amplification. By employing this novel strategy, an ~8.5 (±0.3) fold amplification in signal intensity was obtained. Afterward, OTA was added to force partial GNPs/G-rich DNA to release from the sensing interface and thus decreased the electrochemical response. An effective sensing range from 2.5pM to 2.5nM was received with an extremely low detection limit of 0.75 (±0.12) pM. This amplification strategy has the potential to be the main technology for aptamer-based electrochemical biosensor in a variety of fields.

  13. Coherent signal amplification in bistable nanomechanical oscillators by stochastic resonance

    NASA Astrophysics Data System (ADS)

    Badzey, Robert L.; Mohanty, Pritiraj

    2005-10-01

    Stochastic resonance is a counterintuitive concept: the addition of noise to a noisy system induces coherent amplification of its response. First suggested as a mechanism for the cyclic recurrence of ice ages, stochastic resonance has been seen in a wide variety of macroscopic physical systems: bistable ring lasers, superconducting quantum interference devices (SQUIDs), magnetoelastic ribbons and neurophysiological systems such as the receptors in crickets and crayfish. Although fundamentally important as a mechanism of coherent signal amplification, stochastic resonance has yet to be observed in nanoscale systems. Here we report the observation of stochastic resonance in bistable nanomechanical silicon oscillators. Our nanomechanical systems consist of beams that are clamped at each end and driven into transverse oscillation with the use of a radiofrequency source. Modulation of the source induces controllable switching of the beams between two stable, distinct states. We observe that the addition of white noise causes a marked amplification of the signal strength. Stochastic resonance in nanomechanical systems could have a function in the realization of controllable high-speed nanomechanical memory cells, and paves the way for exploring macroscopic quantum coherence and tunnelling.

  14. Extended amplification of acoustic signals by amphibian burrows.

    PubMed

    Muñoz, Matías I; Penna, Mario

    2016-07-01

    Animals relying on acoustic signals for communication must cope with the constraints imposed by the environment for sound propagation. A resource to improve signal broadcast is the use of structures that favor the emission or the reception of sounds. We conducted playback experiments to assess the effect of the burrows occupied by the frogs Eupsophus emiliopugini and E. calcaratus on the amplitude of outgoing vocalizations. In addition, we evaluated the influence of these cavities on the reception of externally generated sounds potentially interfering with conspecific communication, namely, the vocalizations emitted by four syntopic species of anurans (E. emiliopugini, E. calcaratus, Batrachyla antartandica, and Pleurodema thaul) and the nocturnal owls Strix rufipes and Glaucidium nanum. Eupsophus advertisement calls emitted from within the burrows experienced average amplitude gains of 3-6 dB at 100 cm from the burrow openings. Likewise, the incoming vocalizations of amphibians and birds were amplified on average above 6 dB inside the cavities. The amplification of internally broadcast Eupsophus vocalizations favors signal detection by nearby conspecifics. Reciprocally, the amplification of incoming conspecific and heterospecific signals facilitates the detection of neighboring males and the monitoring of the levels of potentially interfering biotic noise by resident frogs, respectively.

  15. General Signal Amplification Strategy for Nonfaradic Impedimetric Sensing: Trastuzumab Detection Employing a Peptide Immunosensor.

    PubMed

    Liu, Juan; Chisti, Mohammad Muhsin; Zeng, Xiangqun

    2017-04-04

    A label-free and reagent-free peptide mimotope capacitive biosensor has been developed for cancer drug (trastuzumab) quantification based on nonfaradic readout. The low sensitivity issue of capacitive biosensors was overcome with two innovations: peptide mimotope mixed self-assembled monolayer (SAM) biointerface and dilution of the analysis buffer. Signal amplification was achieved through dilution of phosphate-buffered saline (PBS) to tune Cdl to dominate the overall capacitance change upon target binding, which contribution is often negligible without dilution. After 1000× dilution, the limit of detection was lowered 500-fold (0.22 μg/mL) and the sensitivity was increased 20-fold [0.04192 (μg/mL)(-1)] in comparison with undiluted PBS. The proposed signal amplification strategy is more straightforward and practical compared to biorecognition element engineering and other strategies. The proposed method was further applied to planar electrodes for optimizing sensing response time to less than 1 min.

  16. Colocalization recognition-activated cascade signal amplification strategy for ultrasensitive detection of transcription factors.

    PubMed

    Zhu, Desong; Wang, Lei; Xu, Xiaowen; Jiang, Wei

    2017-03-15

    Transcription factors (TFs) bind to specific double-stranded DNA (dsDNA) sequences in the regulatory regions of genes to regulate the process of gene transcription. Their expression levels sensitively reflect cell developmental situation and disease state. TFs have become potential diagnostic markers and therapeutic targets of cancers and some other diseases. Hence, high sensitive detection of TFs is of vital importance for early diagnosis of diseases and drugs development. The traditional exonucleases-assisted signal amplification methods suffered from the false positives caused by incomplete digestion of excess recognition probes. Herein, based on a new recognition way-colocalization recognition (CR)-activated dual signal amplification, an ultrasensitive fluorescent detection strategy for TFs was developed. TFs-induced the colocalization of three split recognition components resulted in noticeable increases of local effective concentrations and hybridization of three split components, which activated the subsequent cascade signal amplification including strand displacement amplification (SDA) and exponential rolling circle amplification (ERCA). This strategy eliminated the false positive influence and achieved ultra-high sensitivity towards the purified NF-κB p50 with detection limit of 2.0×10(-13)M. Moreover, NF-κB p50 can be detected in as low as 0.21ngμL(-1) HeLa cell nuclear extracts. In addition, this proposed strategy could be used for the screening of NF-κB p50 activity inhibitors and potential anti-NF-κB p50 drugs. Finally, our proposed strategy offered a potential method for reliable detection of TFs in medical diagnosis and treatment research of cancers and other related diseases.

  17. NLA-QAM - A method for generating high-power QAM signals through nonlinear amplification

    NASA Astrophysics Data System (ADS)

    Morais, D. H.; Feher, K.

    1982-03-01

    A generalized technique for generating quadrature amplitude modulated (QAM) signals that permits nonlinear amplification is presented. With this technique a high-power (2 to the 2n power)-state QAM signal is generated by combining n unfiltered, nonlinear amplified, QPSK signals, n being a positive integer. The specific methods of generating 16- and 64-state signals using this technique are presented. An attractive feature of the technique is that despite significant difference with the conventional method of generating QAM signals, the same straightforward demodulation techniques apply to both, and the Pe versus S/N performances are essentially identical. In the 16-state version employing traveling wave tubes (TWT's) or GaAs FET amplifiers, the technique is shown to result in a transmitter output power advantage that is on the order of 5 dB compared to the conventional method. This advantage is achieved, however, at the expense of an additional output power amplifier.

  18. A sensitive SERS assay for detecting proteins and nucleic acids using a triple-helix molecular switch for cascade signal amplification.

    PubMed

    Ye, Sujuan; Wu, Yanying; Zhang, Wen; Li, Na; Tang, Bo

    2014-08-25

    A novel surface-enhanced Raman scattering (SERS) detection system is developed for proteins and nucleic acids based on a triple-helix molecular switch for multiple cycle signal amplification, achieving high sensitivity, universality, rapid analysis, and high selectivity.

  19. Use of Peltier effect for small signal amplification and conversion

    NASA Astrophysics Data System (ADS)

    Ageyev, Y. I.; Akperov, M. M.; Kobakhidze, K. Z.; Nebuchinov, M. V.

    1984-04-01

    It is possible to use thermocouples operating as heat pumps with small temperature gradients to effect the control of elements whose properties are temperature dependent. This enables the construction of a number of electrical and optical signal transducers. The cooling or heating gain of a thermocouple used as a heat pump is proportional to the ratio of the cold or hot junction temperature to the temperature drop across the thermocouple. As this temperature gradient becomes quite small, the efficiency of such converters theoretically rises without limit. Under these conditions, the thermocouple can control any device whose properties change sharply in a narrow temperature range. Simple circuits for small signal amplification, frequency conversion, and detection were discussed. The gain of one such amplifier was plotted as a function of the input signal using various metal-semiconductor phase transition devices; the detection gain was plotted as a function of the input signal for a posistor and a metal-semiconductor phase transition device. Gains on the order of 100 and more were obtained with the latter. While such devices have the advantage of electrically isolating the input from the output, the speed is governed primarily by the rate of the thermal processes and is approximately inversely proportional to the square of thermocouple branch length. The speed is presently limited to tens of milliseconds, though with the transition to film technology, it may increase by a few orders of magnitude.

  20. Polydimethylsiloxane microfluidic chemiluminescence immunodevice with the signal amplification strategy for sensitive detection of human immunoglobin G.

    PubMed

    Li, Huifang; Zhao, Mei; Liu, Wei; Chu, Weiru; Guo, Yumei

    2016-01-15

    A polydimethylsiloxane (PDMS) microfluidic chemiluminescence (CL) immunodevice for sensitive detection of human immunoglobin G (IgG) with the signal amplification strategy was developed in this work. The immunodevice was prepared by covalently immobilizing capture antibodies (Abs) on the silanized microchannel of microfluidic chip. Gold nanoparticles (AuNPs) functionalized with a high molar ratio of horseradish peroxidase (HRP) were used as an Ab label for signal amplification. Using a sandwich immunoassay, the multi-HRP conjugated AuNPs can catalyze the luminol-H2O2 CL system to achieve the high sensitivity. In addition, the double spiral flow-channel was adopted here, which can still contribute to the high sensitivity. Based on signal amplification strategy, the performance of human IgG tests revealed a lower detection limit (DL) of 0.03ng/mL and showed an increase of 7.4-fold in detection sensitivity compared to a commercial Ab-HRP conjugation. This microfluidic immunodevice can provide an alternative approach for sensitive detection of human IgG in the field of clinic diagnostic and therapeutic.

  1. Trichotomous noise controlled signal amplification in a generalized Verhulst model

    NASA Astrophysics Data System (ADS)

    Mankin, Romi; Soika, Erkki; Lumi, Neeme

    2014-10-01

    The long-time limit of the probability distribution and statistical moments for a population size are studied by means of a stochastic growth model with generalized Verhulst self-regulation. The effect of variable environment on the carrying capacity of a population is modeled by a multiplicative three-level Markovian noise and by a time periodic deterministic component. Exact expressions for the moments of the population size have been calculated. It is shown that an interplay of a small periodic forcing and colored noise can cause large oscillations of the mean population size. The conditions for the appearance of such a phenomenon are found and illustrated by graphs. Implications of the results on models of symbiotic metapopulations are also discussed. Particularly, it is demonstrated that the effect of noise-generated amplification of an input signal gets more pronounced as the intensity of symbiotic interaction increases.

  2. Chemical signal amplification in two-dimensional paper networks

    PubMed Central

    Fu, Elain; Kauffman, Peter; Lutz, Barry; Yager, Paul

    2010-01-01

    Two-dimensional paper networks (2DPNs) hold great potential for extending the utility of paper-based chemical and biochemical diagnostics at a cost and ease-of-use that is comparable to conventional lateral flow strips. 2DPNs enable the automated sequential delivery of multiple reagents to a detection region with a single user activation step, and therefore have the potential to extend the processing capabilities of inexpensive paper-based assays with comparable ease of use to conventional lateral flow tests. In this communication, we used a simple 3 inlet 2DPN to perform signal amplification of a colloidal gold label using a gold enhancement solution, thus demonstrating the capability of 2DPNs to perform processes that can improve limits of detection. PMID:20706615

  3. Signal amplification strategies for DNA and protein detection based on polymeric nanocomposites and polymerization: A review.

    PubMed

    Zhou, Shaohong; Yuan, Liang; Hua, Xin; Xu, Lingling; Liu, Songqin

    2015-06-02

    Demand is increasing for ultrasensitive bioassays for disease diagnosis, environmental monitoring and other research areas. This requires novel signal amplification strategies to maximize the signal output. In this review, we focus on a series of significant signal amplification strategies based on polymeric nanocomposites and polymerization. Some common polymers are used as carriers to increase the local concentration of signal probes and/or biomolecules on their surfaces or in their interiors. Some polymers with special fluorescence and optical properties can efficiently transfer the excitation energy from a single site to the whole polymer backbone. This results in superior fluorescence signal amplification due to the resulting collective effort (integration of signal). Recent polymerization-based signal amplification strategies that employ atom transfer radical polymerization (ATRP) and photo-initiated polymerization are also summarized. Several distinctive applications of polymers in ultrasensitive bioanalysis are highlighted.

  4. Wingless/Wnt signal transduction requires distinct initiation and amplification steps that both depend on Arrow/LRP

    PubMed Central

    Baig-Lewis, Shahana; Peterson-Nedry, Wynne; Wehrli, Marcel

    2007-01-01

    Members of the Wg/Wnt family provide key intercellular signals during embryonic development and in the maintenance of homeostatic processes, but critical aspects of their signal transduction pathways remain controversial. We have found that canonical Wg signaling in Drosophila involves distinct initiation and amplification steps, both of which require Arrow/LRP. Expressing a chimeric Frizzled2-Arrow protein in flies that lack endogenous Wg or Arrow showed that this construct functions as an activated Wg receptor but is deficient in signal amplification. In contrast, a chimeric Arrow protein containing the dimerization domain of Torso acted as a potent amplifier of Wg signaling but could not initiate Wg signaling on its own. The two chimeric proteins synergized, so that their co-expression largely reconstituted the signaling levels achieved by expressing Wg itself. The amplification function of Arrow/LRP appears to be particularly important for long-range signaling, and may reflect a general mechanism for potentiating signals in the shallow part of a morphogen gradient. PMID:17433287

  5. Dual Electrophoresis Detection System for Rapid and Sensitive Immunoassays with Nanoparticle Signal Amplification

    NASA Astrophysics Data System (ADS)

    Zhang, Fangfang; Ma, Junjie; Watanabe, Junji; Tang, Jinlong; Liu, Huiyu; Shen, Heyun

    2017-02-01

    An electrophoretic technique was combined with an enzyme-linked immunosorbent assay (ELISA) system to achieve a rapid and sensitive immunoassay. A cellulose acetate filter modified with polyelectrolyte multilayer (PEM) was used as a solid substrate for three-dimensional antigen-antibody reactions. A dual electrophoresis process was used to induce directional migration and local condensation of antigens and antibodies at the solid substrate, avoiding the long diffusion times associated with antigen-antibody reactions in conventional ELISAs. The electrophoretic forces drove two steps in the ELISA process, namely the adsorption of antigen, and secondary antibody-labelled polystyrene nanoparticles (NP-Ab). The total time needed for dual electrophoresis-driven detection was just 4 min, nearly 2 h faster than a conventional ELISA system. Moreover, the rapid NP-Ab electrophoresis system simultaneously achieved amplification of the specific signal and a reduction in noise, leading to a more sensitive NP-Ab immunoassay with a limit of detection (LOD) of 130 fM, and wide range of detectable concentrations from 0.13 to 130 pM. These results suggest that the combination of dual electrophoresis detection and NP-Ab signal amplification has great potential for future immunoassay systems.

  6. Dual Electrophoresis Detection System for Rapid and Sensitive Immunoassays with Nanoparticle Signal Amplification.

    PubMed

    Zhang, Fangfang; Ma, Junjie; Watanabe, Junji; Tang, Jinlong; Liu, Huiyu; Shen, Heyun

    2017-02-15

    An electrophoretic technique was combined with an enzyme-linked immunosorbent assay (ELISA) system to achieve a rapid and sensitive immunoassay. A cellulose acetate filter modified with polyelectrolyte multilayer (PEM) was used as a solid substrate for three-dimensional antigen-antibody reactions. A dual electrophoresis process was used to induce directional migration and local condensation of antigens and antibodies at the solid substrate, avoiding the long diffusion times associated with antigen-antibody reactions in conventional ELISAs. The electrophoretic forces drove two steps in the ELISA process, namely the adsorption of antigen, and secondary antibody-labelled polystyrene nanoparticles (NP-Ab). The total time needed for dual electrophoresis-driven detection was just 4 min, nearly 2 h faster than a conventional ELISA system. Moreover, the rapid NP-Ab electrophoresis system simultaneously achieved amplification of the specific signal and a reduction in noise, leading to a more sensitive NP-Ab immunoassay with a limit of detection (LOD) of 130 fM, and wide range of detectable concentrations from 0.13 to 130 pM. These results suggest that the combination of dual electrophoresis detection and NP-Ab signal amplification has great potential for future immunoassay systems.

  7. Dual Electrophoresis Detection System for Rapid and Sensitive Immunoassays with Nanoparticle Signal Amplification

    PubMed Central

    Zhang, Fangfang; Ma, Junjie; Watanabe, Junji; Tang, Jinlong; Liu, Huiyu; Shen, Heyun

    2017-01-01

    An electrophoretic technique was combined with an enzyme-linked immunosorbent assay (ELISA) system to achieve a rapid and sensitive immunoassay. A cellulose acetate filter modified with polyelectrolyte multilayer (PEM) was used as a solid substrate for three-dimensional antigen-antibody reactions. A dual electrophoresis process was used to induce directional migration and local condensation of antigens and antibodies at the solid substrate, avoiding the long diffusion times associated with antigen-antibody reactions in conventional ELISAs. The electrophoretic forces drove two steps in the ELISA process, namely the adsorption of antigen, and secondary antibody-labelled polystyrene nanoparticles (NP-Ab). The total time needed for dual electrophoresis-driven detection was just 4 min, nearly 2 h faster than a conventional ELISA system. Moreover, the rapid NP-Ab electrophoresis system simultaneously achieved amplification of the specific signal and a reduction in noise, leading to a more sensitive NP-Ab immunoassay with a limit of detection (LOD) of 130 fM, and wide range of detectable concentrations from 0.13 to 130 pM. These results suggest that the combination of dual electrophoresis detection and NP-Ab signal amplification has great potential for future immunoassay systems. PMID:28198385

  8. Quantum dot layer-by-layer assemblies as signal amplification labels for ultrasensitive electronic detection of uropathogens.

    PubMed

    Xiang, Yun; Zhang, Haixia; Jiang, Bingying; Chai, Yaqin; Yuan, Ruo

    2011-06-01

    The preparation and use of a new class of signal amplification label, quantum dot (QD) layer-by-layer (LBL) assembled polystyrene microsphere composite, for amplified ultrasensitive electronic detection of uropathogen-specific DNA sequences is described. The target DNA is sandwiched between the capture probes immobilized on the magnetic beads and the signaling probes conjugated to the QD LBL assembled polystyrene beads. Because of the dramatic signal amplification by the numerous QDs involved in each single DNA binding event, subfemtomolar level detection of uropathogen-specific DNA sequences is achieved, which makes our strategy among the most sensitive electronic approach for nucleic acid-based monitoring of pathogens. Our signal amplified detection scheme could be readily expanded to monitor other important biomolecules (e.g., proteins, peptides, amino acids, cells, etc.) in ultralow levels and thus holds great potential for early diagnosis of disease biomarkers.

  9. Signal amplification in biological and electrical engineering systems: universal role of cascades.

    PubMed

    Grubelnik, Vladimir; Dugonik, Bogdan; Osebik, Davorin; Marhl, Marko

    2009-08-01

    In this paper we compare the cascade mechanisms of signal amplification in biological and electrical engineering systems, and show that they share the capacity to considerably amplify signals, and respond to signal changes both quickly and completely, which effectively preserves the form of the input signal. For biological systems, these characteristics are crucial for efficient and reliable cellular signaling. We show that this highly-efficient biological mechanism of signal amplification that has naturally evolved is mathematically fully equivalent with some man-developed amplifiers, which indicates parallels between biological evolution and successful technology development.

  10. Irreversible Catalyst Activation Enables Hyperpolarization and Water Solubility for NMR Signal Amplification by Reversible Exchange

    DTIC Science & Technology

    2016-09-12

    Irreversible Catalyst Activation Enables Hyperpolarization and Water Solubility for NMR Signal Amplification by Reversible Exchange Milton L. Truong...Supporting Information ABSTRACT: Activation of a catalyst [IrCl(COD)(IMes)] (IMes = 1,3-bis(2,4,6-trimethylphenyl)imidazol-2-ylidene; COD = cyclooctadiene...for signal amplification by reversible exchange (SABRE) was monitored by in situ hyperpolarized proton NMR at 9.4 T. During the catalyst -activation

  11. Microwave signal amplification and Pierce instability on radial electron flows in cylindrical and spherical diodes

    SciTech Connect

    Gnavi, G.; Gratton, F.T. )

    1994-11-01

    Linear space charge perturbations of focused electron beams flowing between cylindrical and spherical electrodes on convergent or divergent trajectories are studied, and the amplification of high-frequency signals when the flow is modulated at one electrode is computed. It is shown that divergent beams give the largest amplification effect. The instability of electron beams drifting through grounded grids (Pierce instability in cylindrical or spherical diodes) is also considered. The instability threshold occurs at higher critical currents when the curvature of the electrodes is large. Results for planar electrodes are recovered in the limit of zero curvature devices. Spherical configurations have better signal amplification and stability properties than similar planar or cylindrical systems.

  12. In situ amplification signaling-based autonomous aptameric machine for the sensitive fluorescence detection of cocaine.

    PubMed

    Xie, Su-Jin; Zhou, Hui; Liu, Dengyou; Shen, Guo-Li; Yu, Ruqin; Wu, Zai-Sheng

    2013-06-15

    The development of autonomous DNA machines and their use for specific sensing purpose have recently attracted considerable research attention. In existing autonomous machines, the target recognition process and signal transduction are separated from each other. This results in misunderstanding of the operation behavior, and the assay capability is compromised when serving as a sensing tool. In this communication, the integrated signal transduction-based autonomous aptameric machine, in which the recognition element and signal reporters are integrated into a DNA strand, is developed. This new machine can execute the in situ amplification of target binding-induced signal. The authentic operation behavior of autonomous DNA machine is discovered: the machine's products directly hybridize to the "track" rather than to the signaling probes. Along this line, the machine is employed to detect the cocaine in a more straightforward fashion, and improved assay characteristics (for example, the dynamic response range is widened by more than 500-fold) are achieved. Our efforts not only clarify the concept described in traditional autonomous DNA machines but also have made technological advancements that are expected to be especially valuable in designing nucleic acid-based machines employed in basic research and medical diagnosis.

  13. Label-free electrochemical aptasensor for adenosine detection based on cascade signal amplification strategy.

    PubMed

    Shen, Jing; Wang, Hongyang; Li, Chunxiang; Zhao, Yanyan; Yu, Xijuan; Luo, Xiliang

    2017-04-15

    In this work, a simple and highly sensitive label-free electrochemical aptasensor for adenosine detection was developed based on target-aptamer binding triggered nicking endonuclease-assisted strand-replacement DNA polymerization and rolling circle amplification (RCA) strategy. The magnetic beads (MB) probe, which was attached the aptamer of adenosine and mDNA, was firstly fabricated. In the presence of adenosine, mDNA was released from MB upon recognition of the aptamer to target adenosine. The released mDNA as the primer activated autonomous DNA polymerization/nicking process and accompanied by the continuous release of replicated DNA fragments. Subsequently, numerous released DNA fragments were captured on the working electrode, and then as initiators to trigger the downstream RCA process leading to the formation of a long ssDNA concatemer for loading large amounts of Ru(NH3)6(3+). Therefore, a conspicuously amplified electrochemical signal through the developed dual-amplification strategy could be achieved. This method exhibited a high sensitivity toward adenosine with a detection limit of 0.032nM. Also, it exhibited high selectivity to different nucleoside families and good reproducibility. This design opens new horizons for integrating different disciplines, presenting a versatile tool for ultrasensitive detecting organic small molecules in medical research and clinical diagnosis.

  14. Engineering self-contained DNA circuit for proximity recognition and localized signal amplification of target biomolecules.

    PubMed

    Ang, Yan Shan; Yung, Lin-Yue Lanry

    2014-08-01

    Biomolecular interactions have important cellular implications, however, a simple method for the sensing of such proximal events is lacking in the current molecular toolbox. We designed a dynamic DNA circuit capable of recognizing targets in close proximity to initiate a pre-programmed signal transduction process resulting in localized signal amplification. The entire circuit was engineered to be self-contained, i.e. it can self-assemble onto individual target molecules autonomously and form localized signal with minimal cross-talk. α-thrombin was used as a model protein to evaluate the performance of the individual modules and the overall circuit for proximity interaction under physiologically relevant buffer condition. The circuit achieved good selectivity in presence of non-specific protein and interfering serum matrix and successfully detected for physiologically relevant α-thrombin concentration (50 nM-5 μM) in a single mixing step without any further washing. The formation of localized signal at the interaction site can be enhanced kinetically through the control of temperature and probe concentration. This work provides a basic general framework from which other circuit modules can be adapted for the sensing of other biomolecular or cellular interaction of interest.

  15. An ultrasensitive and universal photoelectrochemical immunoassay based on enzyme mimetics enhanced signal amplification.

    PubMed

    Wang, Guang-Li; Shu, Jun-Xian; Dong, Yu-Ming; Wu, Xiu-Ming; Li, Zai-Jun

    2015-04-15

    An ultrasensitive photoelectrochemical (PEC) immunoassay based on signal amplification by enzyme mimetics was fabricated for the detection of mouse IgG (as a model protein). The PEC immunosensor was constructed by a layer-by-layer assembly of poly (diallyldimethylammonium chloride) (PDDA), CdS quantum dots (QDs), primary antibody (Ab1, polyclonal goat antimouse IgG), and the antigen (Ag, mouse IgG) on an indium-tin oxide (ITO) electrode. Then, the secondary antibody (Ab2, polyclonal goat antimouse IgG) combined to a bio-bar-coded Pt nanoparticle(NP)-G-quadruplex/hemin probe was used for signal amplification. The bio-bar-coded Pt NP-G-quadruplex/hemin probe could catalyze the oxidation of hydroquinone (HQ) using H2O2 as an oxidant, demonstrating its intrinsic enzyme-like activity. High sensitivity for the target Ag was achieved by using the bio-bar-coded probe as signal amplifier due to its high catalytic activity, a competitive nonproductive absorption of hemin and the steric hindrance caused by the polymeric oxidation products of HQ. For most important, the oxidation product of HQ acted as an efficient electron acceptor of the illuminated CdS QDs. The target Ag could be detected from 0.01pg/mL to 1.0ng/mL with a low detection limit of 6.0fg/mL. The as-obtained immunosensor exhibited high sensitivity, good stability and acceptable reproducibility. This method might be attractive for clinical and biomedical applications.

  16. Drastic disorder-induced reduction of signal amplification in scale-free networks.

    PubMed

    Chacón, Ricardo; Martínez, Pedro J

    2015-07-01

    Understanding information transmission across a network is a fundamental task for controlling and manipulating both biological and manmade information-processing systems. Here we show how topological resonant-like amplification effects in scale-free networks of signaling devices are drastically reduced when phase disorder in the external signals is considered. This is demonstrated theoretically by means of a starlike network of overdamped bistable systems, and confirmed numerically by simulations of scale-free networks of such systems. The taming effect of the phase disorder is found to be sensitive to the amplification's strength, while the topology-induced amplification mechanism is robust against this kind of quenched disorder in the sense that it does not significantly change the values of the coupling strength where amplification is maximum in its absence.

  17. Tiny grains give huge gains: nanocrystal-based signal amplification for biomolecule detection.

    PubMed

    Tong, Sheng; Ren, Binbin; Zheng, Zhilan; Shen, Han; Bao, Gang

    2013-06-25

    Nanocrystals, despite their tiny sizes, contain thousands to millions of atoms. Here we show that the large number of atoms packed in each metallic nanocrystal can provide a huge gain in signal amplification for biomolecule detection. We have devised a highly sensitive, linear amplification scheme by integrating the dissolution of bound nanocrystals and metal-induced stoichiometric chromogenesis, and demonstrated that signal amplification is fully defined by the size and atom density of nanocrystals, which can be optimized through well-controlled nanocrystal synthesis. Further, the rich library of chromogenic reactions allows implementation of this scheme in various assay formats, as demonstrated by the iron oxide nanoparticle linked immunosorbent assay (ILISA) and blotting assay developed in this study. Our results indicate that, owing to the inherent simplicity, high sensitivity and repeatability, the nanocrystal based amplification scheme can significantly improve biomolecule quantification in both laboratory research and clinical diagnostics. This novel method adds a new dimension to current nanoparticle-based bioassays.

  18. Noiseless intensity amplification of repetitive signals by coherent addition using the temporal Talbot effect

    PubMed Central

    Maram, Reza; Van Howe, James; Li, Ming; Azaña, José

    2014-01-01

    Amplification of signal intensity is essential for initiating physical processes, diagnostics, sensing, communications and measurement. During traditional amplification, the signal is amplified by multiplying the signal carriers through an active gain process, requiring the use of an external power source. In addition, the signal is degraded by noise and distortions that typically accompany active gain processes. We show noiseless intensity amplification of repetitive optical pulse waveforms with gain from 2 to ~20 without using active gain. The proposed method uses a dispersion-induced temporal self-imaging (Talbot) effect to redistribute and coherently accumulate energy of the original repetitive waveforms into fewer replica waveforms. In addition, we show how our passive amplifier performs a real-time average of the wave-train to reduce its original noise fluctuation, as well as enhances the extinction ratio of pulses to stand above the noise floor. Our technique is applicable to repetitive waveforms in any spectral region or wave system. PMID:25319207

  19. Optimization of noise in non-integrated instrumentation amplifier for the amplification of very low electrophysiological [corrected] signals. Case of electro cardio graphic signals (ECG).

    PubMed

    Ngounou, Guy Merlin; Kom, Martin

    2014-12-01

    In this paper we present an instrumentation amplifier with discrete elements and optimized noise for the amplification of very low signals. In amplifying signals of very weak amplitude, the noise can completely absorb these signals if the used amplifier does not present the optimal guarantee to minimize the noise. Based on related research and re-viewing of recent patents Journal of Medical Systems, 30:205-209, 2006, we suggest an approach of noise reduction in amplification much more thoroughly than re-viewing of recent patents and we deduce from it the general criteria necessary and essential to achieve this optimization. The comparison of these criteria with the provisions adopted in practice leads to the inadequacy of conventional amplifiers for effective noise reduction. The amplifier we propose is an instrumentation amplifier with active negative feedback and optimized noise for the amplification of signals with very low amplitude. The application of this method in the case of electro cardio graphic signals (ECG) provides simulation results fully in line with forecasts.

  20. Experimental investigation of chirp properties induced by signal amplification in quantum-dot semiconductor optical amplifiers.

    PubMed

    Matsuura, Motoharu; Ohta, Hiroaki; Seki, Ryota

    2015-03-15

    We experimentally show the dynamic frequency chirp properties induced by signal amplification in a quantum-dot semiconductor optical amplifier (QD-SOA) for the first time. We also compare the red and blue chirp peak values and temporal chirp changes while changing the gain and injected signal powers of the QD-SOA with those of a common SOA.

  1. Battery-triggered ultrasensitive electrochemiluminescence detection on microfluidic paper-based immunodevice based on dual-signal amplification strategy.

    PubMed

    Li, Weiping; Li, Meng; Ge, Shenguang; Yan, Mei; Huang, Jiadong; Yu, Jinghua

    2013-03-12

    Dual-signal amplification strategy for ultrasensitive electrochemiluminescence (ECL) multiplexed immunoassay on microfluidic paper-based analytical devices (μ-PADs) was demonstrated. This dual-signal amplification technique was achieved by employing graphene oxide-chitosan/gold nanoparticles (GCA) immunosensing platform and [4,4'-(2,5-dimethoxy-1,4-phenylene)bis(ethyne-2,1-diyl) dibenzoic acid] (P-acid) functionalized nanoporous silver (P-acid/NPS) signal amplification label. For further low-cost and disposable applications, battery-triggered constant-potential ECL (+1.0V for P-acid label (vs. Ag/AgCl auxiliary electrode)) was applied on this paper-based immunodevice with the aid of a home-made voltage-tunable power device, allowing the traditional electrochemical workstation to be abandoned. We found that two tumor markers could be sequentially detected in the linear ranges of 0.003-20 and 0.001-10ngmL(-1) with the detection limits down to 1.0 and 0.8pgmL(-1), respectively, by simply reversing the connection mode on two working electrodes. The results exhibited excellent precision and high sensitivity of such immunoassay, and it also demonstrated that this battery-triggered ECL paper-based immunodevice could provide a rapid, simple and simultaneous multiplex immunoassay with high throughput, low-cost and low detection limits for point-of-care testing.

  2. Nonlinear photoacoustic signal amplification from single targets in absorption background☆

    PubMed Central

    Sarimollaoglu, Mustafa; Nedosekin, Dmitry A.; Menyaev, Yulian A.; Juratli, Mazen A.; Zharov, Vladimir P.

    2013-01-01

    Photoacoustic (PA) detection of single absorbing targets such as nanoparticles or cells can be limited by absorption background. We show here that this problem can be overcome by using the nonlinear photoacoustics based on the differences in PA signal dependences on the laser energy from targets and background. Among different nonlinear phenomena, we focused on laser generation of nanobubbles as more efficient PA signal amplifiers from strongly absorbing, highly localized targets in the presence of spatially homogenous absorption background generating linear signals only. This approach was demonstrated by using nonlinear PA flow cytometry platform for label-free detection of circulating melanoma cells in blood background in vitro and in vivo. Nonlinearly amplified PA signals from overheated melanin nanoclusters in melanoma cells became detectable above still linear blood background. Nonlinear nanobubble-based photoacoustics provide new opportunities to significantly (5–20-fold) increase PA contrast of single nanoparticles, cells, viruses and bacteria in complex biological environments. PMID:24921062

  3. Novel signal amplification approach for HRP-based colorimetric genosensors using DNA binding protein tags.

    PubMed

    Aktas, Gülsen Betül; Skouridou, Vasso; Masip, Lluis

    2015-12-15

    The need for sensitive detection of DNA is growing as more specific DNA sequences are being correlated to gene markers for disease diagnosis, food safety and other security related applications. Detection in hybridization-based assays is usually achieved with target-specific ssDNA probes conjugated directly to enzyme labels like HRP that provide signal amplification or with nanoparticles functionalized with DNA and multiple HRP molecules. In order to overcome some of the drawbacks presented by these approaches, we developed a unique DNA sensing platform based on an HRP-DNA binding protein tag conjugate and a hybrid ssDNA-dsDNA detection probe. Specifically, in this work we describe the preparation and characterization of an HRP conjugate with scCro DNA binding protein tag and its application for the detection of a model ssDNA target sequence. By using the HRP-scCro conjugate together with a hybrid detection probe containing three scCro-specific dsDNA binding sites, we demonstrate an improvement by over 3-fold in both sensitivity and limit of detection of high-risk human papillomavirus (HPV16), compared to the standard ssDNA-HRP conjugate. These results show that the HRP-DNA binding protein tag conjugate can be used as an alternative and universal tool for signal enhancement in enzyme-linked assays suitable for integration in point-of-care systems.

  4. A new ultrasonic signal amplification method for detection of bacteria

    NASA Astrophysics Data System (ADS)

    Kant Shukla, Shiva; Resa López, Pablo; Sierra Sánchez, Carlos; Urréjola, José; Segura, Luis Elvira

    2012-10-01

    A new method is presented that increases the sensitivity of ultrasound-based techniques for detection of bacteria. The technique was developed for the detection of catalase-positive microorganisms. It uses a bubble trapping medium containing hydrogen peroxide that is mixed with the sample for microbiological evaluation. The enzyme catalase is present in catalase-positive bacteria, which induces a rapid hydrolysis of hydrogen peroxide, forming bubbles which remain in the medium. This reaction results in the amplification of the mechanical changes that the microorganisms produce in the medium. The effect can be detected by means of ultrasonic wave amplitude continuous measurement since the bubbles increase the ultrasonic attenuation significantly. It is shown that microorganism concentrations of the order of 105 cells ml-1 can be detected using this method. This allows an improvement of three orders of magnitude in the ultrasonic detection threshold of microorganisms in conventional culture media, and is competitive with modern rapid microbiological methods. It can also be used for the characterization of the enzymatic activity.

  5. Impulse-induced optimum signal amplification in scale-free networks.

    PubMed

    Martínez, Pedro J; Chacón, Ricardo

    2016-04-01

    Optimizing information transmission across a network is an essential task for controlling and manipulating generic information-processing systems. Here, we show how topological amplification effects in scale-free networks of signaling devices are optimally enhanced when the impulse transmitted by periodic external signals (time integral over two consecutive zeros) is maximum. This is demonstrated theoretically by means of a star-like network of overdamped bistable systems subjected to generic zero-mean periodic signals and confirmed numerically by simulations of scale-free networks of such systems. Our results show that the enhancer effect of increasing values of the signal's impulse is due to a correlative increase of the energy transmitted by the periodic signals, while it is found to be resonant-like with respect to the topology-induced amplification mechanism.

  6. Nonenzymatic catalytic signal amplification for nucleic acid hybridization assays

    NASA Technical Reports Server (NTRS)

    Fan, Wenhong (Inventor); Cassell, Alan M. (Inventor); Han, Jie (Inventor)

    2006-01-01

    Devices, methods, and kits for amplifying the signal from hybridization reactions between nucleic acid probes and their cognate targets are presented. The devices provide partially-duplexed, immobilized probe complexes, spatially separate from and separately addressable from immobilized docking strands. Cognate target acts catalytically to transfer probe from the site of probe complex immobilization to the site of immobilized docking strand, generating a detectable signal. The methods and kits of the present invention may be used to identify the presence of cognate target in a fluid sample.

  7. A fast and sensitive quantitative lateral flow immunoassay for Cry1Ab based on a novel signal amplification conjugate.

    PubMed

    Chen, Chunxiang; Wu, Jian

    2012-01-01

    A novel lateral flow immunoassay (LFIA) signal amplification strategy for the detection of Cry1Ab based on amplification via a polylysine (PL) chain and biotin-streptavidin system (BSAS) is described. In this system, multiple fluorescence dyes (FL) were directly coated on the surface of PL and conjugated with antibody via the BSAS for construction of novel signal amplification (FLPL-BSAS-mAb1) conjugates, in which FL, PL and BSAS were employed to improve the sensitivity of LFIA. Compared with conventional LFIA, the sensitivity of FLPL-BSAS-mAb1-based LFIA was increased by approximately 100-fold. Quantified linearity was achieved in the value range of 0-1,000 pg/mL. The limit of detection (LOD) was reached 10 pg/mL after optimization of reaction conditions. To our knowledge, this represents one of the most sensitive LFIA for Cry1Ab yet reported. Furthermore, the detection time for this method was about 10 min. Therefore, it should be an attractive alternative compared to conventional immunoassays in routine control for Cry1Ab.

  8. Thousand-fold fluorescent signal amplification for mHealth diagnostics

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The low sensitivity of Mobile Health (mHealth) optical detectors, such as those found on mobile phones, is a limiting factor for many mHealth clinical applications. To improve sensitivity, we have combined two approaches for optical signal amplification: (1) a computational approach based on an imag...

  9. Ultrasensitive fluorescence polarization DNA detection by target assisted exonuclease III-catalyzed signal amplification.

    PubMed

    Zhang, Min; Guan, Yi-Meng; Ye, Bang-Ce

    2011-03-28

    Single stranded DNA sequences can be detected by target assisted exonuclease III-catalyzed signal amplification fluorescence polarization (TAECA-FP). The method offers an impressive detection limit of 83 aM within one hour for DNA detection and exhibits high discrimination ability even against a single base mismatch.

  10. Nucleic acid tool enzymes-aided signal amplification strategy for biochemical analysis: status and challenges.

    PubMed

    Qing, Taiping; He, Dinggeng; He, Xiaoxiao; Wang, Kemin; Xu, Fengzhou; Wen, Li; Shangguan, Jingfang; Mao, Zhengui; Lei, Yanli

    2016-04-01

    Owing to their highly efficient catalytic effects and substrate specificity, the nucleic acid tool enzymes are applied as 'nano-tools' for manipulating different nucleic acid substrates both in the test-tube and in living organisms. In addition to the function as molecular scissors and molecular glue in genetic engineering, the application of nucleic acid tool enzymes in biochemical analysis has also been extensively developed in the past few decades. Used as amplifying labels for biorecognition events, the nucleic acid tool enzymes are mainly applied in nucleic acids amplification sensing, as well as the amplification sensing of biorelated variations of nucleic acids. With the introduction of aptamers, which can bind different target molecules, the nucleic acid tool enzymes-aided signal amplification strategies can also be used to sense non-nucleic targets (e.g., ions, small molecules, proteins, and cells). This review describes and discusses the amplification strategies of nucleic acid tool enzymes-aided biosensors for biochemical analysis applications. Various analytes, including nucleic acids, ions, small molecules, proteins, and cells, are reviewed briefly. This work also addresses the future trends and outlooks for signal amplification in nucleic acid tool enzymes-aided biosensors.

  11. Comparison of sensor structures for the signal amplification of surface plasmon resonance immunoassay using enzyme precipitation

    NASA Astrophysics Data System (ADS)

    Yang, Chih-Tsung; Thierry, Benjamin

    2015-12-01

    Surface plasmon resonance (SPR) biosensing has been successfully applied for the label-free detection of a broad range of bioanalytes ranging from bacteria, cell, exosome, protein and nucleic acids. When it comes to the detection of small molecules or analytes found at low concentration, amplification schemes are desirable to enhance binding signals and in turn increase sensitivity. A number of SPR signal amplification schemes have been developed and validated; however, little effort has been devoted to understanding the effect of the SPR sensor structures on the amplification of binding signals and therefore on the overall sensing performance. The physical phenomenon of long-range SPR (LRSPR) relies on the propagation of coupled surface plasmonic waves on the opposite sides of a nanoscale-thick noble metal film suspended between two dielectrics with similar refractive indices. Importantly, as compared with commonly used conventional SPR (cSPR), LRSPR is not only characterized by a longer penetration depth of the plasmonic waves in the analyzed medium but also by a greater sensitivity to bulk refractive index changes. In this work, an immunoassay signal amplification platform based on horseradish peroxidase (HRP) catalyzed precipitation was utilized to investigate the sensing performance with regards to cSPR and LRSPR. The enzymatic precipitation of 3, 3'-diaminobenzidine tetrahydrochloride (DAB)/H2O2 was used to amplify SPR signals. The structure-function relationship of cSPR and LRSPR sensors is presented for both standard refractometric measurements and the enzymatic precipitation scheme. Experimental data shows that despite its inherent higher sensitivity to bulk refractive index changes and higher figure of merit, LRSPR was characterized by a lower angular sensitivity in the model enzymatic amplification scheme used here.

  12. Cycling excitation process: An ultra efficient and quiet signal amplification mechanism in semiconductor

    SciTech Connect

    Liu, Yu-Hsin; Yan, Lujiang; Zhang, Alex Ce; Hall, David; Niaz, Iftikhar Ahmad; Zhou, Yuchun; Sham, L. J.; Lo, Yu-Hwa

    2015-08-03

    Signal amplification, performed by transistor amplifiers with its merit rated by the efficiency and noise characteristics, is ubiquitous in all electronic systems. Because of transistor thermal noise, an intrinsic signal amplification mechanism, impact ionization was sought after to complement the limits of transistor amplifiers. However, due to the high operation voltage (30-200 V typically), low power efficiency, limited scalability, and, above all, rapidly increasing excess noise with amplification factor, impact ionization has been out of favor for most electronic systems except for a few applications such as avalanche photodetectors and single-photon Geiger detectors. Here, we report an internal signal amplification mechanism based on the principle of the phonon-assisted cycling excitation process (CEP). Si devices using this concept show ultrahigh gain, low operation voltage, CMOS compatibility, and, above all, quantum limit noise performance that is 30 times lower than devices using impact ionization. Established on a unique physical effect of attractive properties, CEP-based devices can potentially revolutionize the fields of semiconductor electronics.

  13. Signal-to-Noise Enhancement of a Nanospring Redox-Based Sensor by Lock-in Amplification

    PubMed Central

    Bakharev, Pavel V.; McIlroy, David N.

    2015-01-01

    A significant improvement of the response characteristics of a redox chemical gas sensor (chemiresistor) constructed with a single ZnO coated silica nanospring has been achieved with the technique of lock-in signal amplification. The comparison of DC and analog lock-in amplifier (LIA) AC measurements of the electrical sensor response to toluene vapor, at the ppm level, has been conducted. When operated in the DC detection mode, the sensor exhibits a relatively high sensitivity to the analyte vapor, as well as a low detection limit at the 10 ppm level. However, at 10 ppm the signal-to-noise ratio is 5 dB, which is less than desirable. When operated in the analog LIA mode, the signal-to-noise ratio at 10 ppm increases by 30 dB and extends the detection limit to the ppb range. PMID:26053754

  14. Efficient Sub-Bandgap Light Absorption and Signal Amplification in Silicon Photodetectors

    NASA Astrophysics Data System (ADS)

    Liu, Yu-Hsin

    This thesis focuses on two areas in silicon photodetectors, the first being enhancing the sub-bandgap light absorption of IR wavelenghts in silicon, and the second being intrinsic signal amplification in silicon photodetectors. Both of these are achieved using heavily doped p-n junction devices which create localized states that relax the k-selection rule of indirect bandgap material. The probability of transitions between impurity band and the conduction/valence band would be much more efficient than the one between band-to-band transition. The waveguide-coupled epitaxial p-n photodetector was demonstrated for 1310 nm wavelength detection. Incorporated with the Franz-Keldysh effect and the quasi-confined epitaxial layer design, an absorption coefficient around 10 cm-1 has been measured and internal quantum efficiency nearly 100% at -2.5V. The absorption coefficient is calculated from the wave function of the electron and hole in p-n diode. The heavily doped impurity wave function can be formulated as a delta function, and the quasi-confined conduction band energy states, and the wave function on each level can be obtained from the Silvaco software. The calculated theoretical absorption coefficient increases with the increasing applied bias and the doping concentration, which matches the experimental results. To solve the issues of large excess noise and high operation bias for avalanche photodiodes based on impact ionization, I presented a detector using the Cycling Excitation Process (CEP) for signal amplification. This can be realized in a heavily doped and highly compensated Si p-n junction, showing ultra high gain about 3000 at very low bias (<4 V), and possessing an intrinsic, phonon-mediated regulation process to keep the device stable without any quenching device required in today's Geiger-mode avalanche detectors. The CEP can be formulated with the rate equations in conduction bands and impurity states. The gain expression, which is a function of the

  15. Novel Immunohistochemical Techniques Using Discrete Signal Amplification Systems for Human Cutaneous Peripheral Nerve Fiber Imaging

    PubMed Central

    Wang, Ningshan; Gibbons, Christopher H.; Freeman, Roy

    2011-01-01

    Confocal imaging uses immunohistochemical binding of specific antibodies to visualize tissues, but technical obstacles limit more widespread use of this technique in the imaging of peripheral nerve tissue. These obstacles include same-species antibody cross-reactivity and weak fluorescent signals of individual and co-localized antigens. The aims of this study were to develop new immunohistochemical techniques for imaging of peripheral nerve fibers. Three-millimeter punch skin biopsies of healthy individuals were fixed, frozen, and cut into 50-µm sections. Tissues were stained with a variety of antibody combinations with two signal amplification systems, streptavidin-biotin-fluorochrome (sABC) and tyramide-horseradish peroxidase-fluorochrome (TSA), used simultaneously to augment immunohistochemical signals. The combination of the TSA and sABC amplification systems provided the first successful co-localization of sympathetic adrenergic and sympathetic cholinergic nerve fibers in cutaneous human sweat glands and vasomotor and pilomotor systems. Primary antibodies from the same species were amplified individually without cross-reactivity or elevated background interference. The confocal fluorescent signal-to-noise ratio increased, and image clarity improved. These modifications to signal amplification systems have the potential for widespread use in the study of human neural tissues. PMID:21411809

  16. Signal amplification in immunohistochemistry: loose-jointed deformable heteropolymeric HRP conjugates vs. linear polymer backbone HRP conjugates.

    PubMed

    Buchwalow, Igor; Boecker, Werner; Wolf, Eduard; Samoilova, Vera; Tiemann, Markus

    2013-07-01

    Improvements in reagents and protocols for immunohistochemistry have led to increased sensitivity of detection systems. A significant level of signal amplification was achieved by the chain-polymer conjugate technology utilizing enzyme-labeled inert "backbone" molecule of dextran (Dako). However, the relatively large size of the dextran molecule in aqueous phase appears to create spatial hindrance compromising the penetrative ability of the detection reagent. Novel AmpliStain™ detection systems (SDT GmbH, Baesweiler, Germany) seem to overcome these constraints offering a more compact and deformable conjugate design that facilitates agile penetration through the narrowest diffusion pathways in tissue sections. Here, we compared the level of signal amplification achievable with AmpliStain™-HRP (SDT) and EnVision™+-HRP (Dako). Our results show that the AmpliStain™-HRP systems allow higher dilutions of primary antibodies in both immunohistochemistry and ELISA. Compared with EnVision™+, anti-mouse AmpliStain™ enables at least three times more sensitive detection of mouse antibodies, whereas anti-rabbit AmpliStain™ is ten times more sensitive than anti-rabbit EnVision™+.

  17. Signal amplification by 1/f noise in silicon-based nanomechanical resonators.

    PubMed

    Guerra, Diego N; Dunn, Tyler; Mohanty, Pritiraj

    2009-09-01

    We report signal amplification by 1/f(alpha) noise with stochastic resonance in a nonlinear nanomechanical resonator. The addition of 1/f(alpha) noise to a subthreshold modulation signal enhances the probability of an electrostatically driven resonator switching between its two vibrational states in the hysteretic region. Considering the prevalence of 1/f noise in the materials in integrated circuits, signal enhancement demonstrated here, using a fully on-chip electronic actuation/detection scheme, suggests beneficial use of the otherwise detrimental noise.

  18. Sensitive detection of Escherichia coli O157:H7 based on cascade signal amplification in ELISA.

    PubMed

    Shan, Shan; Liu, Daofeng; Guo, Qi; Wu, Songsong; Chen, Rui; Luo, Kai; Hu, Liming; Xiong, Yonghua; Lai, Weihua

    2016-09-01

    In this study, cascade signal amplification in ELISA involving double-antibody sandwich ELISA and indirectly competitive ELISA was established to sensitively detect Escherichia coli O157:H7. In the double-antibody sandwich ELISA, a complex was formed comprising anti-E. coli O157:H7 polyclonal antibody, E. coli O157:H7, biotinylated anti-E. coli O157:H7 monoclonal antibody, streptavidin, and biotinylated β-lactamase. Penicillin solution was then added into the ELISA well and hydrolyzed by β-lactamase. Afterward, the penicillin solution was transferred to indirectly competitive ELISA. The concentration of penicillin can be sensitively detected in indirectly competitive ELISA. In the cascade signal amplification system, increasing the amount of added E. coli O157:H7 resulted in more β-lactamase and less penicillin. The detection sensitivity of E. coli O157:H7, which was 20cfu/mL with the cascade signal amplification in ELISA, was 1,000-fold higher than that of traditional ELISA. Furthermore, the novel method can be used to detect E. coli O157:H7 in milk (2cfu/g). Therefore, this new signaling strategy will facilitate analyses of highly sensitive foodborne pathogens.

  19. Signal-beam amplification by two-wave mixing in a liquid-crystal light valve.

    PubMed

    Brignon, A; Bongrand, I; Loiseaux, B; Huignard, J P

    1997-12-15

    A new two-wave-mixing interaction with gain through a Bi(12)SiO(20) liquid-crystal light valve is presented. We show that the diffraction of a pump beam in the direction of a weak signal leads to a net amplification of the signal beam, with no need for a phase shift between the interference pattern and the induced index grating. A two-wave-mixing gain of 10 and a 150-ms response time are obtained with a 8.8-microm -thick liquid-crystal layer, a total intensity of the interacting beams of only 200 muW/cm(2), and an ac external voltage of +/-6 V . Image amplification is also demonstrated.

  20. Highly enhanced electrochemiluminescent strategy for tumor biomarkers detection with in situ generation of L-homocysteine for signal amplification.

    PubMed

    Wang, Haijun; Chai, Yaqin; Yuan, Ruo; Cao, Yaling; Bai, Lijuan

    2014-03-07

    In this work, an ultrasensitive peroxydisulfate electrochemiluminescence (ECL) immunosensor using in situ generation of L-homocysteine (L-Hcys) for signal amplification was successfully constructed for detection of carcinoembryonic antigen (CEA). In the reaction of biological methylation, S-adenosyl-L-homocysteine hydrolase (SAHH) catalyzed the reversible hydrolysis of S-adenosyl-L-homocysteine (SAH) to produce L-Hcys, which was inducted into ECL system to construct the immunosensor for signal amplification in this work. Simultaneously, Gold and palladium nanoparticles functionalized multi-walled carbon nanotubes (Au-PdNPs@MWCNTs) were prepared, which were introduced to immobilize the secondary antibody (Ab2) and SAHH with high loading amount and good biological activity due to their improved surface area and excellent biocompatibility. Then the proposed ECL immunosensor was developed by a sandwich-type format using Au-PdNPs@MWCNTs-SAHH-Ab2 as tracer and graphene together with AuNPs as substrate. Besides the enhancement of Au-PdNPs, the enzymatic catalysis reaction also amplified the ECL signal dramatically, which was achieved by efficient catalysis of the SAHH towards the hydrolysis of SAH to generate improved amount of L-Hcys in situ. Furthermore, due to the special interaction between Au-PdNPs and -SH or -NH2 in L-Hcys, L-Hcys would gradually accumulate on the surface of the immunosensor, which greatly enhanced the concentration of L-Hcys on the immunosensor surface and further improved the ECL intensity. With the amplification factors above, a wide linear ranged from 0.1 pg mL(-1) to 80 ng mL(-1) was acquired with a relatively low detection limit of 33 fg mL(-1) for CEA.

  1. mRNA-targeted Fluorescent in Situ Hybridization (FISH) of Gram-negative Bacteria Without Template Amplification or Tyramide Signal Amplification

    SciTech Connect

    Coleman, James R.; Culley, David E.; Chrisler, William B.; Brockman, Fred J.

    2007-12-01

    Technologies are needed to study gene expression at the level of individual cells within a population or microbial community. Fluorescent in situ hybridization (FISH) supplies high-resolution spatial information and has been widely applied to study microbial communities at the rRNA level. While mRNA-targeted FISH has been popular for studying gene expression in eukaryotic cells, very little success has been achieved with prokaryotes. At present, detection of specific mRNAs in individual prokaryotic cells requires the use of in situ-RT-PCR or tyramide signal amplification (TSA). In this study we used DNA oligonucleotide probes labeled with a single near-infrared dye in FISH assays to detect multicopy plasmid-based and endogenous mRNA molecules in Escherichia coli and Shewanella oneidensis MR-1. We took advantage of the fact there is much less background signal produced by biological materials and support matrices in the near-infrared spectrum and thus long camera exposure times could be used. In addition, we demonstrate that a combination of probes targeting both rRNA and mRNA could be successfully employed within the same FISH assay. These results, as well as ongoing R&D improvements in NIR and infrared dyes, indicate the FISH approach we demonstrated could be applied in certain environmental settings to monitor gene expression in mixed populations.

  2. New Signal Amplification Strategy Using Semicarbazide as Co-reaction Accelerator for Highly Sensitive Electrochemiluminescent Aptasensor Construction.

    PubMed

    Ma, Meng-Nan; Zhuo, Ying; Yuan, Ruo; Chai, Ya-Qin

    2015-11-17

    A highly sensitive electrochemiluminescent (ECL) aptasensor was constructed using semicarbazide (Sem) as co-reaction accelerator to promote the ECL reaction rate of CdTe quantum dots (CdTe QDs) and the co-reactant of peroxydisulfate (S2O8(2-)) for boosting signal amplification. The co-reaction accelerator is a species that when it is introduced into the ECL system containing luminophore and co-reactant, it can interact with co-reactant rather than luminophore to promote the ECL reaction rate of luminophore and co-reactant; thus the ECL signal is significantly amplified in comparison with that in which only luminophore and co-reactant are present. In this work, the ECL signal probes were first fabricated by alternately assembling the Sem and Au nanoparticles (AuNPs) onto the surfaces of hollow Au nanocages (AuNCs) via Au-N bond to obtain the multilayered nanomaterials of (AuNPs-Sem)n-AuNCs for immobilizing amino-terminated detection aptamer of thrombin (TBA2). Notably, the Sem with two -NH2 terminal groups could not only serve as cross-linking reagent to assemble AuNPs and AuNCs but also act as co-reaction accelerator to enhance the ECL reaction rate of CdTe QDs and S2O8(2-) for signal amplification. With the sandwich-type format, TBA2 signal probes could be trapped on the CdTe QD-based sensing interface in the presence of thrombin (TB) to achieve a considerably enhanced ECL signal in S2O8(2-) solution. As a result, the Sem in the TBA2 signal probes could accelerate the reduction of S2O8(2-) to produce the more oxidant mediators of SO4(•-), which further boosted the production of excited states of CdTe QDs to emit light. With the employment of the novel co-reaction accelerator Sem, the proposed ECL biosensor exhibited ultrahigh sensitivity to quantify the concentration of TB from 1 × 10(-7) to 1 nM with a detection limit of 0.03 fM, which demonstrated that the co-reaction accelerator could provide a simple, efficient, and low-cost approach for signal

  3. Signal Amplification Technique (SAT): an approach for improving resolution and reducing image noise in computed tomography

    SciTech Connect

    Phelps, M.E.; Huang, S.C.; Hoffman, E.J.; Plummer, D.; Carson, R.

    1981-01-01

    Spatial resolution improvements in computed tomography (CT) have been limited by the large and unique error propagation properties of this technique. The desire to provide maximum image resolution has resulted in the use of reconstruction filter functions designed to produce tomographic images with resolution as close as possible to the intrinsic detector resolution. Thus, many CT systems produce images with excessive noise with the system resolution determined by the detector resolution rather than the reconstruction algorithm. CT is a rigorous mathematical technique which applies an increasing amplification to increasing spatial frequencies in the measured data. This mathematical approach to spatial frequency amplification cannot distinguish between signal and noise and therefore both are amplified equally. We report here a method in which tomographic resolution is improved by using very small detectors to selectively amplify the signal and not noise. Thus, this approach is referred to as the signal amplification technique (SAT). SAT can provide dramatic improvements in image resolution without increases in statistical noise or dose because increases in the cutoff frequency of the reconstruction algorithm are not required to improve image resolution. Alternatively, in cases where image counts are low, such as in rapid dynamic or receptor studies, statistical noise can be reduced by lowering the cutoff frequency while still maintaining the best possible image resolution. A possible system design for a positron CT system with SAT is described.

  4. Rapid and visual detection of Listeria monocytogenes based on nanoparticle cluster catalyzed signal amplification.

    PubMed

    Zhang, Lisha; Huang, Ru; Liu, Weipeng; Liu, Hongxing; Zhou, Xiaoming; Xing, Da

    2016-12-15

    Foodborne pathogens pose a significant threat to human health worldwide. The identification of foodborne pathogens needs to be rapid, accurate and convenient. Here, we constructed a nanoparticle cluster (NPC) catalyzed signal amplification biosensor for foodborne pathogens visual detection. In this work, vancomycin (Van), a glycopeptide antibiotic for Gram-positive bacteria, was used as the first molecular recognition agent to capture Listeria monocytogenes (L. monocytogenes). Fe3O4 NPC modified aptamer, was used as the signal amplification nanoprobe, specifically recognize to the cell wall of L. monocytogenes. As vancomycin and aptamer recognize L. monocytogenes at different sites, the sandwich recognition showed satisfied specificity. Compared to individual Fe3O4 nanoparticle (NP), NPC exhibit collective effect-enhanced catalytic activity for the color reaction of chromogenic substrate. The change in absorbance or color could represent the concentration of target. Using the Fe3O4 NPC-based signal amplification method, L. monocytogenes whole cells could be directly assayed within a linear range of 5.4×10(3)-10(8) cfu/mL and a visual limit of detection of 5.4×10(3) cfu/mL. Fe3O4 NPC-based method was more sensitive than the Fe3O4 NP-based method. All these attractive characteristics of highly sensitivity, visual and labor-saving, make the biosensor possess a potential application for foodborne pathogenic bacteria detection.

  5. Sensitive Method for Biomolecule Detection Utilizing Signal Amplification with Porphyrin Nanoparticles.

    PubMed

    Gibson, Lauren E; Wright, David W

    2016-06-07

    Disease diagnosis requires identification of biomarkers that occur in small quantities, making detection a difficult task. Effective diagnosis is an even greater challenge in low-resource areas of the world. Methods must be simple, stable, and sensitive so that tests can be easily administered and withstand uncontrolled environmental conditions. One approach to this issue is development of stable signal amplification strategies. In this work, we applied the nanocrystal-based signal amplification method to tetra(4-carboxyphenyl)porphyrin nanoparticles (TCPP NPs). The dissolution of the nanoparticle into thousands of porphyrin molecules results in amplified detection of the biomarker. By using nanoparticles as the signal-generating moiety, stability of the detection method is increased relative to commonly used enzyme-based assays. Additionally, the inherent fluorescent signal of TCPP molecules can be measured after nanoparticle dissolution. The ability to directly read the TCPP fluorescent signal increases assay simplicity by reducing the steps required for the test. This detection method was optimized by detecting rabbit IgG and then was applied to the detection of the malarial biomarker Plasmodium falciparum histidine-rich protein II (pfHRPII) from a complex matrix. The results for both biomarkers were assays with low picomolar limits of detection.

  6. Enzyme- and affinity biomolecule-mediated polymerization systems for biological signal amplification and cell screening.

    PubMed

    Malinowska, Klara H; Nash, Michael A

    2016-06-01

    Enzyme-mediated polymerization and polymerization-based signal amplification have emerged as two closely related techniques that are broadly applicable in the nanobio sciences. We review recent progress on polymerization systems mediated by biological molecules (e.g., affinity molecules and enzymes), and highlight newly developed formats and configurations of these systems to perform such tasks as non-instrumented biodetection, synthesis of core-shell nanomaterials, isolation of rare cells, and high-throughput screening. We discuss useful features of biologically mediated polymerization systems, such as multiple mechanisms of amplification (e.g., enzymatic, radical chain propagation), and the ability to localize structures at interfaces and at cell surfaces with microscopic spatial confinement. We close with a perspective on desirable improvements that need to be addressed to adapt these molecular systems to future applications.

  7. Magnetic micro/nanoparticle flocculation-based signal amplification for biosensing

    PubMed Central

    Mzava, Omary; Taş, Zehra; İçöz, Kutay

    2016-01-01

    We report a time and cost efficient signal amplification method for biosensors employing magnetic particles. In this method, magnetic particles in an applied external magnetic field form magnetic dipoles, interact with each other, and accumulate along the magnetic field lines. This magnetic interaction does not need any biomolecular coating for binding and can be controlled with the strength of the applied magnetic field. The accumulation can be used to amplify the corresponding pixel area that is obtained from an image of a single magnetic particle. An application of the method to the Escherichia coli 0157:H7 bacteria samples is demonstrated in order to show the potential of the approach. A minimum of threefold to a maximum of 60-fold amplification is reached from a single bacteria cell under a magnetic field of 20 mT. PMID:27354793

  8. Thousand-fold fluorescent signal amplification for mHealth diagnostics

    PubMed Central

    Balsam, Joshua; Rasooly, Reuven; Bruck, Hugh Alan; Rasooly, Avraham

    2013-01-01

    The low sensitivity of Mobile Health (mHealth) optical detectors, such as those found on mobile phones, is a limiting factor for many mHealth clinical applications. To improve sensitivity, we have combined two approaches for optical signal amplification: (1) a computational approach based on an image stacking algorithm to decrease the image noise and enhance weak signals, and (2) an optical signal amplifier utilizing a capillary tube array. These approaches were used in a detection system which includes a multi-wavelength LEDs capable of exciting many fluorophores in multiple wavelengths, a mobile phone or a webcam as a detector, and capillary tube array configured with 36 capillary tubes for signal enhancement. The capillary array enables a ~100X increase in signal sensitivity for fluorescein, reducing the limit of detection (LOD) for mobile phones and webcams from 1000 nM to 10 nM. Computational image stacking enables another ~10X increase in signal sensitivity, further reducing the LOD for webcam from 10 nM to 1 nM. To demonstrate the feasibility of the device for the detection of disease-related biomarkers, Adenovirus DNA labeled with SYBR Green or fluorescein was analyzed by both our capillary array and a commercial plate reader. The LOD for the capillary array was 5ug/mL, and that of the plate reader was 1 ug/mL. Similar results were obtained using DNA stained with fluorescein. The combination of the two signal amplification approaches enables a ~1000X increase in LOD for the webcam platform. This brings it into the range of a conventional plate reader while using a smaller sample volume (10ul) than the plate reader requires (100 ul). This suggests that such a device could be suitable for biosensing applications where up to 10 fold smaller sample sizes are needed. The simple optical configuration for mHealth described in this paper employing the combined capillary and image processing signal amplification is capable of measuring weak fluorescent signals

  9. Thousand-fold fluorescent signal amplification for mHealth diagnostics.

    PubMed

    Balsam, Joshua; Rasooly, Reuven; Bruck, Hugh Alan; Rasooly, Avraham

    2014-01-15

    The low sensitivity of Mobile Health (mHealth) optical detectors, such as those found on mobile phones, is a limiting factor for many mHealth clinical applications. To improve sensitivity, we have combined two approaches for optical signal amplification: (1) a computational approach based on an image stacking algorithm to decrease the image noise and enhance weak signals, and (2) an optical signal amplifier utilizing a capillary tube array. These approaches were used in a detection system which includes multi-wavelength LEDs capable of exciting many fluorophores in multiple wavelengths, a mobile phone or a webcam as a detector, and capillary tube array configured with 36 capillary tubes for signal enhancement. The capillary array enables a ~100× increase in signal sensitivity for fluorescein, reducing the limit of detection (LOD) for mobile phones and webcams from 1000 nM to 10nM. Computational image stacking enables another ~10× increase in signal sensitivity, further reducing the LOD for webcam from 10nM to 1 nM. To demonstrate the feasibility of the device for the detection of disease-related biomarkers, adenovirus DNA labeled with SYBR green or fluorescein was analyzed by both our capillary array and a commercial plate reader. The LOD for the capillary array was 5 ug/mL, and that of the plate reader was 1 ug/mL. Similar results were obtained using DNA stained with fluorescein. The combination of the two signal amplification approaches enables a ~1000× increase in LOD for the webcam platform. This brings it into the range of a conventional plate reader while using a smaller sample volume (10 ul) than the plate reader requires (100 ul). This suggests that such a device could be suitable for biosensing applications where up to 10 fold smaller sample sizes are needed. The simple optical configuration for mHealth described in this paper employing the combined capillary and image processing signal amplification is capable of measuring weak fluorescent signals

  10. Amplification of optical signals in Bi{sub 12}TiO{sub 20} crystal by photorefractive surface waves

    SciTech Connect

    Khomenko, A.V.; Garcia-Weidner, A.; Kamshilin, A.A.

    1996-07-01

    We have demonstrated experimentally beam amplification by coupling between the signal beam and the photorefractive surfaces wave in Bi{sub 12}TiO{sub 20} crystal. A gain of 16,000 has been measured, with an output signal-to-noise ratio of {approx_gt}20 for weak input signals. {copyright} {ital 1996 Optical Society of America.}

  11. Preferential Amplification of Rising Versus Falling Frequency Whistler-Mode Signals

    NASA Astrophysics Data System (ADS)

    Li, J.; Harid, V.; Spasojevic, M.; Golkowski, M.; Inan, U.

    2014-12-01

    Analysis of ground-based ELF/VLF observations of injected whistler-mode waves from the 1986 Siple Station experiment indicate a distinct preference for the magnetospheric amplification of rising frequency-time ramps over descending frequency-time ramps. In examining conjugate region receptions of ±1 kHz/sec frequency ramps, we find that the rising ramps generate a total power that is on average 1.9 times higher than that of falling frequency ramps when both are observed within a given transmission. Also, in 17% of receptions, only the rising frequency ramps are observed, and the descending ramps are below the detectable signal level. Using a narrowband Vlasov-Maxwell numerical simulation, we explore the preferential amplification of rising ramps due to differences in the linear growth rate as a function of frequency and the onset and duration of nonlinear phase trapping. These results contribute to the understanding of magnetospheric wave amplification and the formation of structured rising elements in chorus.

  12. Electrochemical signal amplification for immunosensor based on 3D interdigitated array electrodes.

    PubMed

    Han, Donghoon; Kim, Yang-Rae; Kang, Chung Mu; Chung, Taek Dong

    2014-06-17

    We devised an electrochemical redox cycling based on three-dimensional interdigitated array (3D IDA) electrodes for signal amplification to enhance the sensitivity of chip-based immunosensors. The 3D IDA consists of two closely spaced parallel indium tin oxide (ITO) electrodes that are positioned not only on the bottom but also the ceiling, facing each other along a microfluidic channel. We investigated the signal intensities from various geometric configurations: Open-2D IDA, Closed-2D IDA, and 3D IDA through electrochemical experiments and finite-element simulations. The 3D IDA among the four different systems exhibited the greatest signal amplification resulting from efficient redox cycling of electroactive species confined in the microchannel so that the faradaic current was augmented by a factor of ∼100. We exploited the enhanced sensitivity of the 3D IDA to build up a chronocoulometric immunosensing platform based on the sandwich enzyme-linked immunosorbent assay (ELISA) protocol. The mouse IgGs on the 3D IDA showed much lower detection limits than on the Closed-2D IDA. The detection limit for mouse IgG measured using the 3D IDA was ∼10 fg/mL, while it was ∼100 fg/mL for the Closed-2D IDA. Moreover, the proposed immunosensor system with the 3D IDA successfully worked for clinical analysis as shown by the sensitive detection of cardiac troponin I in human serum down to 100 fg/mL.

  13. A chronocoulometric aptasensor based on gold nanoparticles as a signal amplification strategy for detection of thrombin.

    PubMed

    Jiao, Xiao Xia; Chen, Jing Rong; Zhang, Xi Yuan; Luo, Hong Qun; Li, Nian Bing

    2013-10-15

    A sensitive chronocoulometric aptasensor for the detection of thrombin has been developed based on gold nanoparticle amplification. The functional gold nanoparticles, loaded with link DNA (LDNA) and report DNA (RDNA), were immobilized on an electrode by thrombin aptamers performing as a recognition element and capture probe. LDNA was complementary to the thrombin aptamers and RDNA was noncomplementary, but could combine with [Ru(NH₃)₆]³⁺ (RuHex) cations. Electrochemical signals obtained by RuHex that bound quantitatively to the negatively charged phosphate backbone of DNA via electrostatic interactions were measured by chronocoulometry. In the presence of thrombin, the combination of thrombin and thrombin aptamers and the release of the functional gold nanoparticles could induce a significant decrease in chronocoulometric signal. The incorporation of gold nanoparticles in the chronocoulometric aptasensor significantly enhanced the sensitivity. The performance of the aptasensor was further increased by the optimization of the surface density of aptamers. Under optimum conditions, the chronocoulometric aptasensor exhibited a wide linear response range of 0.1-18.5 nM with a detection limit of 30 pM. The results demonstrated that this nanoparticle-based amplification strategy offers a simple and effective approach to detect thrombin.

  14. Ternary surface monolayers for ultrasensitive (zeptomole) amperometric detection of nucleic acid hybridization without signal amplification.

    PubMed

    Wu, Jie; Campuzano, Susana; Halford, Colin; Haake, David A; Wang, Joseph

    2010-11-01

    A ternary surface monolayer, consisting of coassembled thiolated capture probe, mercaptohexanol and dithiothreitol, is shown to offer dramatic improvements in the signal-to-noise characteristics of electrochemical DNA hybridization biosensors based on common self-assembled monolayers. Remarkably low detection limits down to 40 zmol (in 4 μL samples) as well as only 1 CFU Escherichia coli per sensor are thus obtained without any additional amplification step in connection to the commonly used horseradish peroxidase/3,3',5,5'-tetramethylbenzidine system. Such dramatic improvements in the detection limits (compared to those of common binary alkanethiol interfaces and to those of most electrochemical DNA sensing strategies without target or signal amplification) are attributed primarily to the remarkably higher resistance to nonspecific adsorption. This reflects the highly compact layer (with lower pinhole density) produced by the coupling of the cyclic- and linear-configuration "backfillers" that leads to a remarkably low background noise even in the presence of complex sample matrixes. A wide range of surface compositions have been investigated, and the ternary mixed monolayer has been systematically optimized. Detailed impedance spectroscopy and cyclic voltammetric studies shed useful insights into the surface coverage. The impressive sensitivity and high specificity of the simple developed methodology indicate great promise for a wide range of nucleic acid testing, including clinical diagnostics, biothreat detection, food safety, and forensic analysis.

  15. Simple and rapid chemiluminescence aptasensor for Hg(2+) in contaminated samples: A new signal amplification mechanism.

    PubMed

    Qi, Yingying; Xiu, Fu-Rong; Yu, Gending; Huang, Lili; Li, Baoxin

    2017-01-15

    Detection of ultralow concentration of heavy metal ion Hg(2+) is important for human health protection and environment monitoring because of the gradual accumulation in environmental and biological fields. Herein, we report a convenient chemiluminescence (CL) biosensing platform for ultrasensitive Hg(2+) detection by signal amplification mechanism from positively charged gold nanoparticles ((+)AuNPs). It is based on (+)AuNPs charge effect and aptamer conformation change induced by target to stimulate the generation of CL in the presence of H2O2 and luminol without high salt medium. Notably particularly, the typical problem of the high salt medium from (-) AuNPs system, like influencing aptamers' bind with target and hindering CL reaction can be effectively addressed through the direct introduction of (+)AuNPs. Therefore, the proposed biosensing exhibits a high sensitivity toward target Hg(2+) with a detection limit of 16 pM, which is far below the limit (10nM) defined by the U.S. Environmental Protection Agency in drinkable water, and is about 10-fold lower than the previously reported aptamer-based assays for Hg(2+). This sensing platform provides a simple, rapid, and cost-effective approach for label-free sensitive detection of Hg(2+). Moreover, it is universal for the detection of other targets. Undoubtedly, such a direct utilizing of (+)AuNPs' charge effect will provide a new signal amplification way for label-free aptamer-based CL analysis.

  16. The dynamics of signal amplification by macromolecular assemblies for the control of chromosome segregation

    PubMed Central

    Lee, Semin; Bolanos-Garcia, Victor M.

    2014-01-01

    The control of chromosome segregation relies on the spindle assembly checkpoint (SAC), a complex regulatory system that ensures the high fidelity of chromosome segregation in higher organisms by delaying the onset of anaphase until each chromosome is properly bi-oriented on the mitotic spindle. Central to this process is the establishment of multiple yet specific protein-protein interactions in a narrow time-space window. Here we discuss the highly dynamic nature of multi-protein complexes that control chromosome segregation in which an intricate network of weak but cooperative interactions modulate signal amplification to ensure a proper SAC response. We also discuss the current structural understanding of the communication between the SAC and the kinetochore; how transient interactions can regulate the assembly and disassembly of the SAC as well as the challenges and opportunities for the definition and the manipulation of the flow of information in SAC signaling. PMID:25324779

  17. Sensitive electrochemical detection of telomerase activity using spherical nucleic acids gold nanoparticles triggered mimic-hybridization chain reaction enzyme-free dual signal amplification.

    PubMed

    Wang, Wen-Jing; Li, Jing-Jing; Rui, Kai; Gai, Pan-Pan; Zhang, Jian-Rong; Zhu, Jun-Jie

    2015-03-03

    We report an electrochemical sensor for telomerase activity detection based on spherical nucleic acids gold nanoparticles (SNAs AuNPs) triggered mimic-hybridization chain reaction (mimic-HCR) enzyme-free dual signal amplification. In the detection strategy, SNAs AuNPs and two hairpin probes were employed. SNAs AuNPs as the primary amplification element, not only hybridized with the telomeric repeats on the electrode to amplify signal but also initiated the subsequent secondary amplification, mimic-hybridization chain reaction of two hairpin probes. If the cells' extracts were positive for telomerase activity, SNAs AuNPs could be captured on the electrode. The carried initiators could trigger an alternative hybridization reaction of two hairpin probes that yielded nicked double helices. The signal was further amplified enzyme-free by numerous hexaammineruthenium(III) chloride ([Ru(NH3)6](3+), RuHex) inserting into double-helix DNA long chain by electrostatic interaction, each of which could generate an electrochemical signal at appropriate potential. With this method, a detection limit of down to 2 HeLa cells and a dynamic range of 10-10,000 cells were achieved. Telomerase activities of different cell lines were also successfully evaluated.

  18. 1010 Amplification and phase conjugation with high efficiency achieved by overcoming noise limitations in Brillouin two-beam coupling

    NASA Astrophysics Data System (ADS)

    Glick, Yaakov; Sternklar, Shmuel

    1995-06-01

    A model incorporating noise and pump depletion in a Brillouin amplifier (BA) predicts a fundamental limitation on attainable pump-to-signal-ratio extraction efficiency. Experimental data supporting this model are also presented. In spite of the limitation, an experimental technique is shown that results in a factor-of-7 increase in extraction efficiency. We accomplish this by noise suppressing and subsequently double passing a BA. We report an overall power efficiency of 37% and phase-conjugate amplification of 3.75 \\times 1010 for a signal input power near the noise level. This performance, which is the best to our knowledge recorded to date, is accomplished without requiring additional input energy.

  19. Toward Biocompatible Nuclear Hyperpolarization Using Signal Amplification by Reversible Exchange: Quantitative in Situ Spectroscopy and High-Field Imaging

    PubMed Central

    2014-01-01

    Signal amplification by reversible exchange (SABRE) of a substrate and parahydrogen at a catalytic center promises to overcome the inherent insensitivity of magnetic resonance. In order to apply the new approach to biomedical applications, there is a need to develop experimental equipment, in situ quantification methods, and a biocompatible solvent. We present results detailing a low-field SABRE polarizer which provides well-controlled experimental conditions, defined spins manipulations, and which allows in situ detection of thermally polarized and hyperpolarized samples. We introduce a method for absolute quantification of hyperpolarization yield in situ by means of a thermally polarized reference. A maximum signal-to-noise ratio of ∼103 for 148 μmol of substance, a signal enhancement of 106 with respect to polarization transfer field of SABRE, or an absolute 1H-polarization level of ≈10–2 is achieved. In an important step toward biomedical application, we demonstrate 1H in situ NMR as well as 1H and 13C high-field MRI using hyperpolarized pyridine (d3) and 13C nicotinamide in pure and 11% ethanol in aqueous solution. Further increase of hyperpolarization yield, implications of in situ detection, and in vivo application are discussed. PMID:24397559

  20. Nearly noiseless amplification of microwave signals with a Josephson parametric amplifier

    NASA Astrophysics Data System (ADS)

    Castellanos-Beltran, Manuel

    2009-03-01

    A degenerate parametric amplifier transforms an incident coherent state by amplifying one of its quadrature components while deamplifying the other. This transformation, when performed by an ideal parametric amplifier, is completely deterministic and reversible; therefore the amplifier in principle can be noiseless. We attempt to realize a noiseless amplifier of this type at microwave frequencies with a Josephson parametric amplifier (JPA). To this end, we have built a superconducting microwave cavity containing many dc-SQUIDs. This arrangement creates a non-linear medium in a cavity and it is closely analogous to an optical parametric amplifier. In my talk, I will describe the current performance of this circuit, where I show I can amplify signals with less added noise than a quantum-limited amplifier that amplifies both quadratures. In addition, the JPA also squeezes the electromagnetic vacuum fluctuations by 10 dB. Finally, I will discuss our effort to put two such amplifiers in series in order to undo the first stage of squeezing with a second stage of amplification, demonstrating that the amplification process is truly reversible.[4pt] M. A. Castellanos-Beltran, K. D. Irwin, G. C. Hilton, L. R. Vale and K. W. Lehnert, Nature Physics, published on line, http://dx.doi.org/10.1038/nphys1090 (2008).

  1. Electrochemical detection of protein kinase activity based on carboxypeptidase Y digestion triggered signal amplification.

    PubMed

    Yin, Huanshun; Wang, Xinxu; Guo, Yunlong; Zhou, Yunlei; Ai, Shiyun

    2015-04-15

    An effective assay method for monitoring protein kinase activity and screening inhibitors is greatly beneficial to kinase-related drug discovery, early diagnosis of diseases, and therapeutic effect evaluation. Herein, we develop a simple electrochemical method for detecting the activity of casein kinase II (CK2) based on phosphorylation against carboxypeptidase Y (CPY) digestion triggered signal amplification, where CK2 catalyzed phosphorylation event protects the substrate peptide from the digestion of CPY, maintains the repulsive force of the substrate peptide towards the redox probe, and results in a weak electrochemical signal. Whereas, without phosphorylation, the substrate peptide is digested by CPY and a strong electrochemical signal is obtained. The detection feasibility is demonstrated for the assay of CK2 activity with low detection limit of 0.047unit/mL. Moreover, the biosensor was used for the analysis of kinase inhibition. Based on the electrochemical signal dependent inhibitor concentration, the IC50 value of ellagic acid was estimated to be 39.77nM. The proposed method is also successfully applied to analyze CK2 activity in cell lysates, proving the applicability in complex biological samples.

  2. Biosensors for breast cancer diagnosis: A review of bioreceptors, biotransducers and signal amplification strategies.

    PubMed

    Mittal, Sunil; Kaur, Hardeep; Gautam, Nandini; Mantha, Anil K

    2017-02-15

    Breast cancer is highly prevalent in females and accounts for second highest number of deaths, worldwide. Cumbersome, expensive and time consuming detection techniques presently available for detection of breast cancer potentiates the need for development of novel, specific and ultrasensitive devices. Biosensors are the promising and selective detection devices which hold immense potential as point of care (POC) tools. Present review comprehensively scrutinizes various breast cancer biosensors developed so far and their technical evaluation with respect to efficiency and potency of selected bioreceptors and biotransducers. Use of glycoproteins, DNA biomarkers, micro-RNA, circulatory tumor cells (CTC) and some potential biomarkers are introduced briefly. The review also discusses various strategies used in signal amplification such as nanomaterials, redox mediators, p19 protein, duplex specific nucleases (DSN) and redox cycling.

  3. ASC filament formation serves as a signal amplification mechanism for inflammasomes

    PubMed Central

    Dick, Mathias S.; Sborgi, Lorenzo; Rühl, Sebastian; Hiller, Sebastian; Broz, Petr

    2016-01-01

    A hallmark of inflammasome activation is the ASC speck, a micrometre-sized structure formed by the inflammasome adaptor protein ASC (apoptosis-associated speck-like protein containing a CARD), which consists of a pyrin domain (PYD) and a caspase recruitment domain (CARD). Here we show that assembly of the ASC speck involves oligomerization of ASCPYD into filaments and cross-linking of these filaments by ASCCARD. ASC mutants with a non-functional CARD only assemble filaments but not specks, and moreover disrupt endogenous specks in primary macrophages. Systematic site-directed mutagenesis of ASCPYD is used to identify oligomerization-deficient ASC mutants and demonstrate that ASC speck formation is required for efficient processing of IL-1β, but dispensable for gasdermin-D cleavage and pyroptosis induction. Our results suggest that the oligomerization of ASC creates a multitude of potential caspase-1 activation sites, thus serving as a signal amplification mechanism for inflammasome-mediated cytokine production. PMID:27329339

  4. Delivering strong (1)H nuclear hyperpolarization levels and long magnetic lifetimes through signal amplification by reversible exchange.

    PubMed

    Rayner, Peter J; Burns, Michael J; Olaru, Alexandra M; Norcott, Philip; Fekete, Marianna; Green, Gary G R; Highton, Louise A R; Mewis, Ryan E; Duckett, Simon B

    2017-04-04

    Hyperpolarization turns typically weak NMR and MRI responses into strong signals so that ordinarily impractical measurements become possible. The potential to revolutionize analytical NMR and clinical diagnosis through this approach reflect this area's most compelling outcomes. Methods to optimize the low-cost parahydrogen-based approach signal amplification by reversible exchange with studies on a series of biologically relevant nicotinamides and methyl nicotinates are detailed. These procedures involve specific (2)H labeling in both the agent and catalyst and achieve polarization lifetimes of ca 2 min with 50% polarization in the case of methyl-4,6-d2 -nicotinate. Because a 1.5-T hospital scanner has an effective (1)H polarization level of just 0.0005% this strategy should result in compressed detection times for chemically discerning measurements that probe disease. To demonstrate this technique's generality, we exemplify further studies on a range of pyridazine, pyrimidine, pyrazine, and isonicotinamide analogs that feature as building blocks in biochemistry and many disease-treating drugs.

  5. Hyperbranched Hybridization Chain Reaction for Triggered Signal Amplification and Concatenated Logic Circuits.

    PubMed

    Bi, Sai; Chen, Min; Jia, Xiaoqiang; Dong, Ying; Wang, Zonghua

    2015-07-06

    A hyper-branched hybridization chain reaction (HB-HCR) is presented herein, which consists of only six species that can metastably coexist until the introduction of an initiator DNA to trigger a cascade of hybridization events, leading to the self-sustained assembly of hyper-branched and nicked double-stranded DNA structures. The system can readily achieve ultrasensitive detection of target DNA. Moreover, the HB-HCR principle is successfully applied to construct three-input concatenated logic circuits with excellent specificity and extended to design a security-mimicking keypad lock system. Significantly, the HB-HCR-based keypad lock can alarm immediately if the "password" is incorrect. Overall, the proposed HB-HCR with high amplification efficiency is simple, homogeneous, fast, robust, and low-cost, and holds great promise in the development of biosensing, in the programmable assembly of DNA architectures, and in molecular logic operations.

  6. Homogeneously ultrasensitive electrochemical detection of adenosine triphosphate based on multiple signal amplification strategy.

    PubMed

    Chen, Xiaojun; Ge, Lingna; Guo, Buhua; Yan, Ming; Hao, Ning; Xu, Lin

    2014-08-15

    An ultrasensitive electrochemical aptasensor was successfully fabricated for the detection of adenosine triphosphate (ATP). For the first time, one detection system combined several elements: magnetic aptamer sequences for target recognition and separation, a DNAzyme assisted cyclic signal amplification strategy, layer-by-layer (LBL) quantum dots (QDs) composites for promoting square wave anodic stripping voltammetric (SWASV) analysis and Bi, Nafion (Nf) and three-dimensional ordered macroporous polyaniline-ionic liquid (Bi/Nf/3DOM PANI-IL) film modified glassy carbon electrode (GCE) for monitoring enhanced SWASV signal. The modification of Nf/3DOM PANI-IL on GCE showed that the preconcentration efficiency was improved by the electrostatic absorption of Cd(2+) with negative Nf layer with the enhanced analytical sensitivity due to a large active surface area of 3DOM structure. The increased SWASV peak current values of the label (CdS)4@SiO2 composites were found to be proportional to the logarithmic value of ATP concentrations in the range of 1pM-10nM and 10nM-1µM, with the detection limit as low as 0.5pM. The proposed aptasensor has shown an excellent performance such as high sensitivity, good selectivity and analytical application in real samples. The results demonstrated that the multiple signal amplified strategy we developed was feasible for clinical ATP assay and would provide a promising model for the detection of other small molecules.

  7. Optical amplification and optical filter based signal processing for cost and energy efficient spatial multiplexing.

    PubMed

    Krummrich, Peter M

    2011-08-15

    Spatial division multiplexing has been proposed as an option for further capacity increase of transmission fibers. Application of this concept is attractive only, if cost and energy efficient implementations can be found. In this work, optical amplification and optical filter based signal processing concepts are investigated. Deployment of multi mode fibers as the waveguide type for erbium doped fiber amplifiers potentially offers cost and energy efficiency advantages compared to using multi core fibers in preamplifier as well as booster stages. Additional advantages can be gained from optimization of the amplifier module design. Together with transponder design optimizations, they can increase the attractiveness of inverse spatial multiplexing, which is proposed as an intermediate step. Signal processing based on adaptive passive optical filters offers an alternative approach for the separation of channels at the receiver which have experienced mode coupling along the link. With this optical filter based approach, fiber capacity can potentially be increased faster and more energy efficiently than with solutions relying solely on electronic signal processing.

  8. Dual signal amplification for highly sensitive electrochemical detection of uropathogens via enzyme-based catalytic target recycling.

    PubMed

    Su, Jiao; Zhang, Haijie; Jiang, Bingying; Zheng, Huzhi; Chai, Yaqin; Yuan, Ruo; Xiang, Yun

    2011-11-15

    We report an ultrasensitive electrochemical approach for the detection of uropathogen sequence-specific DNA target. The sensing strategy involves a dual signal amplification process, which combines the signal enhancement by the enzymatic target recycling technique with the sensitivity improvement by the quantum dot (QD) layer-by-layer (LBL) assembled labels. The enzyme-based catalytic target DNA recycling process results in the use of each target DNA sequence for multiple times and leads to direct amplification of the analytical signal. Moreover, the LBL assembled QD labels can further enhance the sensitivity of the sensing system. The coupling of these two effective signal amplification strategies thus leads to low femtomolar (5fM) detection of the target DNA sequences. The proposed strategy also shows excellent discrimination between the target DNA and the single-base mismatch sequences. The advantageous intrinsic sequence-independent property of exonuclease III over other sequence-dependent enzymes makes our new dual signal amplification system a general sensing platform for monitoring ultralow level of various types of target DNA sequences.

  9. Sensitive Immunosensor for Cancer Biomarker Based on Dual Signal Amplification Strategy of Graphene Sheets and Multi-Enzyme Functionalized Carbon Nanospheres

    SciTech Connect

    Du, Dan; Zou, Zhexiang; Shin, Yongsoon; Wang, Jun; Wu, Hong; Engelhard, Mark H.; Liu, Jun; Aksay, Ilhan A.; Lin, Yuehe

    2010-03-30

    A novel electrochemical immunosensor for sensitive detection of cancer biomarker α fetoprotein (AFP) is described that uses a graphene sheet sensor platform and functionalized carbon nanospheres (CNSs) labeling with horseradish peroxidase-secondary antibodies (HRP-Ab2). Greatly enhanced sensitivity for the cancer biomarker is based on a dual signal amplification strategy: first, the synthesized CNSs yielded a homogeneous and narrow size distribution, which allowed several binding events of HRP-Ab2 on each nanosphere. Enhanced sensitivity was achieved by introducing the multi-bioconjugates of HRP-Ab2-CNSs onto the electrode surface through sandwich immunoreactions. Secondly, functionalized graphene sheets used for the biosensor platform increased the surface area to capture a large amount of primary antibodies (Ab1), thus amplifying the detection response. This amplification strategy is a promising platform for clinical screening of cancer biomarkers and point-of-care diagnostics.

  10. Signal amplification for DNA detection based on the HRP-functionalized Fe3O4 nanoparticles.

    PubMed

    Dong, Xiao-Ya; Mi, Xiao-Na; Wang, Bo; Xu, Jing-Juan; Chen, Hong-Yuan

    2011-04-15

    An electrochemical approach for the sensitive detection of sequence-specific DNA has been developed. Horseradish peroxidase (HRP) assembled on the Fe(3)O(4) nanoparticles (NPs) were utilized as signal amplification sources. High-content HRP was adsorbed on the Fe(3)O(4) NPs via layer-by-layer (LbL) technique to prepare HRP-functionalized Fe(3)O(4) NPs. Signal probe and diluting probe were then immobilized on the HRP-functionalized Fe(3)O(4) NPs through the bridge of Au NPs. Thereafter, the resulting DNA-Au-HRP-Fe(3)O(4) (DAHF) bioconjugates were successfully anchored to the gold nanofilm (GNF) modified electrode surface for the construction of sandwich-type electrochemical DNA biosensor. The electrochemical behaviors of the prepared biosensor had been investigated by the cyclic voltammetry (CV), chronoamperometry (i-t), and electrochemical impedance spectroscopy (EIS). Under optimal conditions, the proposed strategy could detect the target DNA down to the level of 0.7 fmol with a dynamic range spanning 4 orders of magnitude and exhibited excellent discrimination to two-base mismatched DNA and non-complementary DNA sequences.

  11. Development and significance of a fetal electrocardiogram recorded by signal-averaged high-amplification electrocardiography.

    PubMed

    Hayashi, Risa; Nakai, Kenji; Fukushima, Akimune; Itoh, Manabu; Sugiyama, Toru

    2009-03-01

    Although ultrasonic diagnostic imaging and fetal heart monitors have undergone great technological improvements, the development and use of fetal electrocardiograms to evaluate fetal arrhythmias and autonomic nervous activity have not been fully established. We verified the clinical significance of the novel signal-averaged vector-projected high amplification ECG (SAVP-ECG) method in fetuses from 48 gravidas at 32-41 weeks of gestation and in 34 neonates. SAVP-ECGs from fetuses and newborns were recorded using a modified XYZ-leads system. Once noise and maternal QRS waves were removed, the P, QRS, and T wave intervals were measured from the signal-averaged fetal ECGs. We also compared fetal and neonatal heart rates (HRs), coefficients of variation of heart rate variability (CV) as a parasympathetic nervous activity, and the ratio of low to high frequency (LF/HF ratio) as a sympathetic nervous activity. The rate of detection of a fetal ECG by SAVP-ECG was 72.9%, and the fetal and neonatal QRS and QTc intervals were not significantly different. The neonatal CVs and LF/HF ratios were significantly increased compared with those in the fetus. In conclusion, we have developed a fetal ECG recording method using the SAVP-ECG system, which we used to evaluate autonomic nervous system development.

  12. Enhanced signal-to-noise ratios in frog hearing can be achieved through amplitude death

    PubMed Central

    Ahn, Kang-Hun

    2013-01-01

    In the ear, hair cells transform mechanical stimuli into neuronal signals with great sensitivity, relying on certain active processes. Individual hair cell bundles of non-mammals such as frogs and turtles are known to show spontaneous oscillation. However, hair bundles in vivo must be quiet in the absence of stimuli, otherwise the signal is drowned in intrinsic noise. Thus, a certain mechanism is required in order to suppress intrinsic noise. Here, through a model study of elastically coupled hair bundles of bullfrog sacculi, we show that a low stimulus threshold and a high signal-to-noise ratio (SNR) can be achieved through the amplitude death phenomenon (the cessation of spontaneous oscillations by coupling). This phenomenon occurs only when the coupled hair bundles have inhomogeneous distribution, which is likely to be the case in biological systems. We show that the SNR has non-monotonic dependence on the mass of the overlying membrane, and find out that the SNR has maximum value in the region of amplitude death. The low threshold of stimulus through amplitude death may account for the experimentally observed high sensitivity of frog sacculi in detecting vibration. The hair bundles' amplitude death mechanism provides a smart engineering design for low-noise amplification. PMID:23883956

  13. Signal Amplification in Field Effect-Based Sandwich Enzyme-Linked Immunosensing by Tuned Buffer Concentration with Ionic Strength Adjuster.

    PubMed

    Kumar, Satyendra; Kumar, Narendra; Panda, Siddhartha

    2016-04-01

    Miniaturization of the sandwich enzyme-based immunosensor has several advantages but could result in lower signal strength due to lower enzyme loading. Hence, technologies for amplification of the signal are needed. Signal amplification in a field effect-based electrochemical immunosensor utilizing chip-based ELISA is presented in this work. First, the molarities of phosphate buffer saline (PBS) and concentrations of KCl as ionic strength adjuster were optimized to maximize the GOx glucose-based enzymatic reactions in a beaker for signal amplification measured by change in the voltage shift with an EIS device (using 20 μl of solution) and validated with a commercial pH meter (using 3 ml of solution). The PBS molarity of 100 μM with 25 mM KCl provided the maximum voltage shift. These optimized buffer conditions were further verified for GOx immobilized on silicon chips, and similar trends with decreased PBS molarity were obtained; however, the voltage shift values obtained on chip reaction were lower as compared to the reactions occurring in the beaker. The decreased voltage shift with immobilized enzyme on chip could be attributed to the increased Km (Michaelis-Menten constant) values in the immobilized GOx. Finally, a more than sixfold signal enhancement (from 8 to 47 mV) for the chip-based sandwich immunoassay was obtained by altering the PBS molarity from 10 to 100 μM with 25 mM KCl.

  14. Nuclease-aided target recycling signal amplification strategy for ochratoxin A monitoring.

    PubMed

    Lv, Lei; Li, Donghao; Cui, Chengbi; Zhao, Yangyang; Guo, Zhijun

    2017-01-15

    Ochratoxin A (OTA), a toxin produced by Aspergillus ochraceus and Penicillium verrucosum, is one of the most abundant food-contaminating mycotoxins worldwide. OTA mainly exerts nephrotoxicity, immunotoxicity, mutagenicity, carcinogenicity, teratogenicity, and neurotoxicity. This paper describes a simple and sensitive aptamer/single-walled carbon nanohorn (SWCNH)-based assay for OTA detection. SWCNHs can protect DNA from DNase I cleavage. However, aptamers can be detached from the surface of SWCNHs through specific target binding, exposing them to enzymatic cleavage and releases the target for a new cycle. Cycling of targets leads to significant signal amplification and low limit of detection (LOD), resulting in a nearly 20-fold reduction in LOD for OTA assay compared with non-target recycling under the same experimental parameters. This technique responded specifically to OTA without interference from other analogues (Ochratoxin B, Ochratoxin C, warfarin, and N-acetyl-l-phenylalanine). Moreover, the application of this technique in real sample has been verified using red wine samples spiked with a series of OTA concentrations. This aptasensor offers a great practical importance in food safety and can be widely extended for detection of other toxins by replacing the sequence of the recognition aptamer.

  15. Visualization of mitochondrial DNA replication in individual cells by EdU signal amplification.

    PubMed

    Haines, Kristine M; Feldman, Eva L; Lentz, Stephen I

    2010-11-15

    Mitochondria are key regulators of cellular energy and mitochondrial biogenesis is an essential component of regulating mitochondria numbers in healthy cells. One approach for monitoring mitochondrial biogenesis is to measure the rate of mitochondrial DNA (mtDNA) replication. We developed a sensitive technique to label newly synthesized mtDNA in individual cells in order to study mtDNA biogenesis. The technique combines the incorporation of 5-ethynyl-2'-deoxyuridine (EdU) with a tyramide signal amplification (TSA) protocol to visualize mtDNA replication within subcellular compartments of neurons. EdU is superior to other thymidine analogs, such as 5-bromo-2-deoxyuridine (BrdU), because the initial click reaction to label EdU does not require the harsh acid treatments or enzyme digests that are required for exposing the BrdU epitope. The milder labeling of EdU allows for direct comparison of its incorporation with other cellular markers. The ability to visualize and quantify mtDNA biogenesis provides an essential tool for investigating the mechanisms used to regulate mitochondrial biogenesis and would provide insight into the pathogenesis associated with drug toxicity, aging, cancer and neurodegenerative diseases. Our technique is applicable to sensory neurons as well as other cell types. The use of this technique to measure mtDNA biogenesis has significant implications in furthering the understanding of both normal cellular physiology as well as impaired disease states.

  16. Three different signal amplification strategies for the impedimetric sandwich detection of thrombin.

    PubMed

    Ocaña, Cristina; del Valle, Manel

    2016-03-17

    In this work, we report a comparative study on three highly specific amplification strategies for the ultrasensitive detection of thrombin with the use of aptamer sandwich protocol. The protocol consisted on the use of a first thrombin aptamer immobilized on the electrode surface, the recognition of thrombin protein, and the reaction with a second biotinylated thrombin aptamer forming the sandwich. Through the exposed biotin end, three variants have been tested to amplify the electrochemical impedance signal. The strategies included (a) silver enhancement treatment, (b) gold enhancement treatment and (c) insoluble product produced by the combination of the enzyme horseradish peroxidase (HRP) and 3-amino-9-ethylcarbazole (AEC). The properties of the sensing surface were probed by electrochemical impedance measurements in the presence of the ferrocyanide/ferricyanide redox marker. Insoluble product strategy and silver enhancement treatment resulted in the lowest detection limit (0.3 pM), while gold enhancement method resulted in the highest reproducibility, 8.8% RSD at the pM thrombin concentration levels. Results of silver and gold enhancement treatment also permitted direct inspection by scanning electron microscopy (SEM).

  17. Imaging hydrogen peroxide in Alzheimer’s disease via cascade signal amplification

    PubMed Central

    Yang, Jian; Yang, Jing; Liang, Steven H.; Xu, Yungen; Moore, Anna; Ran, Chongzhao

    2016-01-01

    In brains of Alzheimer’s disease (AD), reactive oxygen species (ROS) levels are significantly higher than that of healthy brains. Evidence suggests that, during AD onset and progression, a vicious cycle revolves around amyloid beta (Aβ) production, aggregation, plaque formation, microglia/immunological responses, inflammation, and ROS production. In this cycle, ROS species play a central role, and H2O2 is one of the most important ROS species. In this report, we have designed a fluorescent imaging probe CRANAD-88, which is capable of cascade amplifying near infrared fluorescence (NIRF) signals at three levels upon interacting with H2O2 in AD brains. We demonstrated that the amplification was feasible in vitro and in vivo. Remarkably, we showed that, for the first time, it was feasible to monitor the changes of H2O2 concentrations in AD brains before and after treatment with an H2O2 scavenger. Our method opens new revenues to investigate H2O2 in AD brains and can be very instructive for drug development. PMID:27762326

  18. Double-probe signal enhancing strategy for toxin aptasensing based on rolling circle amplification.

    PubMed

    Tong, Ping; Zhao, Wei-Wei; Zhang, Lan; Xu, Jing-Juan; Chen, Hong-Yuan

    2012-03-15

    On the basis of aptamer-based rolling circle amplification (RCA) and magnetic beads (MBs), a highly sensitive electrochemical method was developed for the determination of Ochratoxin A (OTA). Initially, an amino-modified capture DNA was immobilized onto MBs for the following hybridization with an OTA aptamer and a phosphate labeled padlock DNA. In the presence of OTA, the aptamer would dissociate from the bioconjugate, and the padlock DNA would subsequently hybridize with the capture DNA to form a circular template with the aid of the T4 ligase. Next, capture DNA would act as primer to initiate a linear RCA reaction and hence generate a long tandem repeated sequences by phi29 DNA polymerase and dNTPs. Then, two quantum dots (QDs) labeled DNA probes were tagged on the resulted RCA product to indicate the OTA recognition event by electrochemical readout. This strategy, based on the novel design of OTA-mediated DNA circularization, the combination of RCA and double signal probes introduction, could detect OTA down to the level of 0.2 pg mL(-1) with a dynamic range spanning more than 4 orders of magnitude. The proposed approach is tested to determine OTA in red wines and shows good application potential in real samples.

  19. DNAzyme-functionalized gold-palladium hybrid nanostructures for triple signal amplification of impedimetric immunosensor.

    PubMed

    Hou, Li; Gao, Zhuangqiang; Xu, Mingdi; Cao, Xia; Wu, Xiaoping; Chen, Guonan; Tang, Dianping

    2014-04-15

    A highly sensitive and selective impedimetric immunosensor with triple signal amplification was designed for ultrasensitive detection of prostate-specific antigen (PSA) by using anti-PSA antibody and DNAzyme-functionalized gold-palladium hybrid nanotags (Ab2-AuPd-DNA). The signal was amplified based on the Ab2-AuPd-DNA toward the catalytic precipitation of 4-choloro-1-naphthol (4-CN). DNAzyme (as a kind of peroxidase mimic) could catalyze the oxidation of 4-CN, whilst AuPd hybrid nanostructures could not only provide a large surface coverage for immobilization of biomolecules but also promote 4-CN oxidation to some extent. The produced insoluble benzo-4-chlorohexadienone via 4-CN was coated on the electrode surface, and hindered the electron transfer between the solution and the electrode, thereby increasing the Faradaic impedance of the base electrode. Three labeling strategies including Ab2-AuNP, Ab2-AuPd and Ab2-AuPd-DNA were investigated for determination of PSA, and improved analytical features were obtained with the Ab2-AuPd-DNA strategy. Under optimal conditions, the dynamic concentration range of the impedimetric immunosensor spanned from 1.0 pg mL(-1) to 50 ng mL(-1) PSA with a detection limit of 0.73 pg mL(-1). Intra- and inter-assay coefficients of variation were below 8.5% and 9.5%, respectively. Importantly, no significant differences at the 0.05 significance level were encountered in the analysis of 6 clinical serum specimens and 6 diluted standards between the impedimetric immunosensor and the commercialized electrochemiluminescent method for PSA detection.

  20. Biobar-coded gold nanoparticles and DNAzyme-based dual signal amplification strategy for ultrasensitive detection of protein by electrochemiluminescence.

    PubMed

    Xia, Hui; Li, Lingling; Yin, Zhouyang; Hou, Xiandeng; Zhu, Jun-Jie

    2015-01-14

    A dual signal amplification strategy for electrochemiluminescence (ECL) aptasensor was designed based on biobar-coded gold nanoparticles (Au NPs) and DNAzyme. CdSeTe@ZnS quantum dots (QDs) were chosen as the ECL signal probes. To verify the proposed ultrasensitive ECL aptasensor for biomolecules, we detected thrombin (Tb) as a proof-of-principle analyte. The hairpin DNA designed for the recognition of protein consists of two parts: the sequences of catalytical 8-17 DNAzyme and thrombin aptamer. Only in the presence of thrombin could the hairpin DNA be opened, followed by a recycling cleavage of excess substrates by catalytic core of the DNAzyme to induce the first-step amplification. One part of the fragments was captured to open the capture DNA modified on the Au electrode, which further connected with the prepared biobar-coded Au NPs-CdSeTe@ZnS QDs to get the final dual-amplified ECL signal. The limit of detection for Tb was 0.28 fM with excellent selectivity, and this proposed method possessed good performance in real sample analysis. This design introduces the new concept of dual-signal amplification by a biobar-coded system and DNAzyme recycling into ECL determination, and it is promising to be extended to provide a highly sensitive platform for various target biomolecules.

  1. Coupling Activity-Based Detection, Target Amplification, Colorimetric and Fluorometric Signal Amplification, for Quantitative Chemosensing of Fluoride Generated from Nerve Agents.

    PubMed

    Sun, Xiaolong; Reuther, James F; Phillips, Scott T; Anslyn, Eric V

    2017-03-17

    The G-class nerve agents, which include sarin, soman, and cyclosarin, react readily with nucleophilic reagents to produce fluoride. Thus, a chemosensing protocol has been designed for these agents that pairs the nucleophilic reactivity of oximates for generating fluoride with an autoinductive target amplification reaction to amplify the quantity of fluoride for facile colorimetric and fluorescent optical quantification. The chemosensing protocol was demonstrated by using the nerve agent surrogate diisopropyl fluorophosphate (DFP) and benzaldoxime as the nucleophile. Autoinductive fluoride amplification responds to fluoride released from DFP by amplifying the fluoride concentration and a yellow reporter molecule. The reporter is a conjugated oligomer with a nominal repeating unit that originates from 4-aminobenzaldehyde. Exposure of the amplified fluoride to a fluoride-specific ratiometric fluorescent reporter provides a fluorescent readout, in which three fluorophores are generated per fluoride. Both colorimetric and fluorescent readouts enable quantitative assays with low micromolar limits of detection for fluoride resulting from DFP. More importantly, this work demonstrates the successful merging of multiple complex reactions for achieving selective, sensitive, and quantitative chemosensing.

  2. Laser-enhanced ionization of mercury atoms in an inert atmosphere with avalanche amplification of the signal.

    PubMed

    Clevenger, W L; Matveev, O I; Cabredo, S; Omenetto, N; Smith, B W; Winefordner, J D

    1997-07-01

    A new method for laser-enhanced ionization detection of mercury atoms in an inert gas atmosphere is described. The method, which is based on the avalanche amplification of the signal resulting from the ionization from a selected Rydberg level reached by a three-step laser excitation of mercury vapor in a simple quartz cell, can be applied to the determination of this element in various matrices by the use of conventional cold atomization techniques. The overall (collisional + photo) ionization efficiency is investigated at different temperatures, and the avalanche amplification effect is reported for Ar and P-10 gases at atmospheric pressure. It is shown that the amplified signal is related to the number of charges produced in the laser-irradiated volume. Under amplifier noise-limited conditions, a detection limit of ∼15 Hg atoms/laser pulse in the interaction region is estimated.

  3. Signal-on electrochemical detection of antibiotics at zeptomole level based on target-aptamer binding triggered multiple recycling amplification.

    PubMed

    Wang, Hongzhi; Wang, Yu; Liu, Su; Yu, Jinghua; Guo, Yuna; Xu, Ying; Huang, Jiadong

    2016-06-15

    In the work, a signal-on electrochemical DNA sensor based on multiple amplification for ultrasensitive detection of antibiotics has been reported. In the presence of target, the ingeniously designed hairpin probe (HP1) is opened and the polymerase-assisted target recycling amplification is triggered, resulting in autonomous generation of secondary target. It is worth noting that the produced secondary target could not only hybridize with other HP1, but also displace the Helper from the electrode. Consequently, methylene blue labeled HP2 forms a "close" probe structure, and the increase of signal is monitored. The increasing current provides an ultrasensitive electrochemical detection for antibiotics down to 1.3 fM. To our best knowledge, such work is the first report about multiple recycling amplification combing with signal-on sensing strategy, which has been utilized for quantitative determination of antibiotics. It would be further used as a general strategy associated with more analytical techniques toward the detection of a wide spectrum of analytes. Thus, it holds great potential for the development of ultrasensitive biosensing platform for the applications in bioanalysis, disease diagnostics, and clinical biomedicine.

  4. Layer-by-layer multienzyme assembly for highly sensitive electrochemical immunoassay based on tyramine signal amplification strategy.

    PubMed

    Zhou, Jun; Tang, Juan; Chen, Guonan; Tang, Dianping

    2014-04-15

    A new sandwich-type electrochemical immunosensor based on nanosilver-doped bovine serum albumin microspheres (Ag@BSA) with a high ratio of horseradish peroxidase (HRP) and detection antibody was developed for quantitative monitoring of biomarkers (carcinoembryonic antigen, CEA, used in this case) by coupling enzymatic biocatalytic precipitation with tyramine signal amplification strategy on capture antibody-modified glassy carbon electrode. Two immunosensing protocols (with and without tyramine signal amplification) were also investigated for the detection of CEA and improved analytical features were acquired with tyramine signal amplification strategy. With the labeling method, the performance and factors influencing the electrochemical immunoassay were studied and evaluated in detail. Under the optimal conditions, the electrochemical immunosensor exhibited a wide dynamic range of 0.005-80 ng mL(-1) toward CEA standards with a low detection limit of 5.0 pg mL(-1). Intra- and inter-assay coefficients of variation were below 11%. No significant differences at the 0.05 significance level were encountered in the analysis of 6 clinical serum specimens and 6 spiked new-born cattle serum samples between the electrochemical immunoassay and the commercialized electrochemiluminescent immunoassay method for the detection of CEA.

  5. Electrochemical current rectification-a novel signal amplification strategy for highly sensitive and selective aptamer-based biosensor.

    PubMed

    Feng, Lingyan; Sivanesan, Arumugam; Lyu, Zhaozi; Offenhäusser, Andreas; Mayer, Dirk

    2015-04-15

    Electrochemical aptamer-based (E-AB) sensors represent an emerging class of recently developed sensors. However, numerous of these sensors are limited by a low surface density of electrode-bound redox-oligonucleotides which are used as probe. Here we propose to use the concept of electrochemical current rectification (ECR) for the enhancement of the redox signal of E-AB sensors. Commonly, the probe-DNA performs a change in conformation during target binding and enables a nonrecurring charge transfer between redox-tag and electrode. In our system, the redox-tag of the probe-DNA is continuously replenished by solution-phase redox molecules. A unidirectional electron transfer from electrode via surface-linked redox-tag to the solution-phase redox molecules arises that efficiently amplifies the current response. Using this robust and straight-forward strategy, the developed sensor showed a substantial signal amplification and consequently improved sensitivity with a calculated detection limit of 114nM for ATP, which was improved by one order of magnitude compared with the amplification-free detection and superior to other previous detection results using enzymes or nanomaterials-based signal amplification. To the best of our knowledge, this is the first demonstration of an aptamer-based electrochemical biosensor involving electrochemical rectification, which can be presumably transferred to other biomedical sensor systems.

  6. A SPR biosensor based on signal amplification using antibody-QD conjugates for quantitative determination of multiple tumor markers

    PubMed Central

    Wang, Huan; Wang, Xiaomei; Wang, Jue; Fu, Weiling; Yao, Chunyan

    2016-01-01

    The detection of tumor markers is very important in early cancer diagnosis; however, tumor markers are usually present at very low concentrations, especially in the early stages of tumor development. Surface plasmon resonance (SPR) is widely used to detect biomolecular interactions; it has inherent advantages of being high-throughput, real-time, and label-free technique. However, its sensitivity needs essential improvement for practical applications. In this study, we developed a signal amplification strategy using antibody-quantum dot (QD) conjugates for the sensitive and quantitative detection of α-fetoprotein (AFP), carcinoembryonic antigen (CEA) and cytokeratin fragment 21-1 (CYFRA 21-1) in clinical samples. The use of a dual signal amplification strategy using AuNP-antibody and antibody-QD conjugates increased the signal amplification by 50-folds. The constructed SPR biosensor showed a detection limit as low as 0.1 ng/mL for AFP, CEA, and CYFRA 21-1. Moreover, the results obtained using this SPR biosensor were consistent with those obtained using the electrochemiluminescence method. Thus, the constructed SPR biosensor provides a highly sensitive and specific approach for the detection of tumor markers. This SPR biosensor can be expected to be readily applied for the detection of other tumor markers and can offer a potentially powerful solution for tumor screening. PMID:27615417

  7. Microelectrode miRNA sensors enabled by enzymeless electrochemical signal amplification.

    PubMed

    Wang, Tanyu; Viennois, Emilie; Merlin, Didier; Wang, Gangli

    2015-08-18

    Better detections of circulating microRNAs (miRNAs) as disease biomarkers could advance diseases diagnosis and treatment. Current analysis methods or sensors for research and applications are challenged by the low concentrations and wide dynamic range (from aM to nM) of miRNAs in a physiological sample. Here, we report a one-step label-free electrochemical sensor comprising a triple-stem DNA-redox probe structure on a gold microelectrode. A new signal amplification mechanism without the need of a redox enzyme is introduced. The novel strategy overcomes the fundamental limitations of microelectrode DNA sensors that fail to generate detectable current, which is primarily due to the limited amount of redox probes in response to the target analyte binding. By employing a reductant, tris(2-carboxyethyl) phosphine hydrochloride (TCEP) in the detection buffer solution, each redox molecule on the detection probe is cyclically oxidized at the electrode and reduced by the reductant; thus, the signal is amplified in situ during the detection period. The combined merits in the diagnosis power of cyclic voltammetry and the high sensitivity of pulse voltammetry enable parallel analysis for method validation and optimization previously inaccessible. As such, the detection limit of miRNA-122 was 0.1 fM via direct readout, with a wide detection range from sub fM to nM. The detection time is within minutes, which is a significant improvement over other macroscopic sensors and other relevant techniques such as quantitative reverse transcription polymerase chain reaction (qRT-PCR). The high selectivity of the developed sensors is demonstrated by the discrimination against two most similar family sequences: miR-122-3p present in serum and 2-mismatch synthetic RNA sequence. Interference such as nonspecific adsorption, a common concern in sensor development, is reduced to a negligible amount by adopting a multistep surface modification strategy. Importantly, unlike qRT-PCR, the

  8. Activity-Based Probes linked with Laser-Cleavable Mass Tags for Signal Amplification in Imaging Mass Spectrometry: Analysis of Serine Hydrolase Enzymes in Mammalian Tissue

    PubMed Central

    Yang, Junhai; Chaurand, Pierre; Norris, Jeremy L.; Porter, Ned A.; Caprioli, Richard M.

    2012-01-01

    A novel functional Imaging Mass Spectrometry technology is described that utilizes activity-based probes for imaging enzyme active sites in tissue sections. We demonstrate this technology using an activity-based probe (fluorophosphate) that is specific for serine hydrolases. A dendrimer containing multiple mass tags that is attached to the activity-based probe is used to analyze the binding sites of the probe through release and measurement of the mass tags on laser irradiation. A generation 8 Poly(amido amine) dendrimer with 1024 amino groups was labeled with an azide group and then more than 900 mass tags were attached in order to achieve signal amplification of nearly three orders of magnitude. The experimental protocol first involves binding of the activity-based probe containing an alkyne group to serine hydrolases in the tissue section followed by attachment of the dendrimer labeled with mass tags to the bound probe by Click chemistry. On irradiation of the labeled tissue by the laser beam in a raster pattern, the mass tags are liberated and recorded by the mass analyzer, consequently, the ion image of the mass tag reveals the distribution of serine hydrolases in the tissue. This process was shown using rat brain and mouse embryo sections. Targeted imaging has the advantage of providing high spatial resolution and high sensitivity through the use of signal amplification chemistry with high target specificity through the use of an enzyme activity probe. PMID:22424244

  9. Sub-femtomolar DNA detection based on layered molybdenum disulfide/multi-walled carbon nanotube composites, Au nanoparticle and enzyme multiple signal amplification.

    PubMed

    Huang, Ke-Jing; Liu, Yu-Jie; Wang, Hai-Bo; Wang, Ya-Ya; Liu, Yan-Ming

    2014-05-15

    A novel 2-dimensional graphene analog molybdenum disulfide/multi-walled carbon nanotube (MoS2/MWCNT) was synthesized by a simple hydrothermal method to achieve excellent electrochemical properties. An ultrasensitive electrochemical DNA biosensor was subsequently constructed by assembling a thiol-tagged DNA probe on a MoS2/MWCNT and gold nanoparticle (AuNP)-modified electrode that has already been coupled with glucose oxidase (GOD). In this work, GOD was used as a redox marker. The heteronanostructure formed on the biosensor surface appeared relatively good conductor for accelerating the electron transfer, while the modification of GOD and AuNPs provided multiple signal amplification for electrochemical biosensing. The multiple signal amplification strategy produced an ultrasensitive electrochemical detection of DNA down to 0.79 fM with a linear range from 10 fM to 10(7)fM, and appeared high selectivity to differentiate three-base mismatched DNA and one-base mismatched DNA. The developed approach provided a simple and reliable method for DNA detection with high sensitivity and specificity, and would open new opportunities for sensitive detection of other biorecognition events.

  10. A sensitive electrochemical aptasensor based on palladium nanoparticles decorated graphene-molybdenum disulfide flower-like nanocomposites and enzymatic signal amplification.

    PubMed

    Jing, Pei; Yi, Huayu; Xue, Shuyan; Chai, Yaqin; Yuan, Ruo; Xu, Wenju

    2015-01-01

    In the present study, with the aggregated advantages of graphene and molybdenum disulfide (MoS2), we prepared poly(diallyldimethylammonium chloride)-graphene/molybdenum disulfide (PDDA-G-MoS2) nanocomposites with flower-like structure, large surface area and excellent conductivity. Furthermore, an advanced sandwich-type electrochemical assay for sensitive detection of thrombin (TB) was fabricated using palladium nanoparticles decorated PDDA-G-MoS2 (PdNPs/PDDA-G-MoS2) as nanocarriers, which were functionalized by hemin/G-quadruplex, glucose oxidase (GOD), and toluidine blue (Tb) as redox probes. The signal amplification strategy was achieved as follows: Firstly, the immobilized GOD could effectively catalyze the oxidation of glucose to gluconolactone, coupling with the reduction of the dissolved oxygen to H2O2. Then, both PdNPs and hemin/G-quadruplex acting as hydrogen peroxide (HRP)-mimicking enzyme could further catalyze the reduction of H2O2, resulting in significant electrochemical signal amplification. So the proposed aptasensor showed high sensitivity with a wide dynamic linear range of 0.0001 to 40 nM and a relatively low detection limit of 0.062 pM for TB determination. The strategy showed huge potential of application in protein detection and disease diagnosis.

  11. All-numerical noise filtering of fluorescence signals for achieving ultra-low limit of detection in biomedical applications.

    PubMed

    Dongre, Chaitanya; Pollnau, Markus; Hoekstra, Hugo J W M

    2011-03-21

    We present an all-numerical method for post-processing of the fluorescent signal as obtained from labeled molecules by capillary electrophoresis (CE) in an optofluidic chip, on the basis of data filtering in the Fourier domain. It is shown that the method outperforms the well-known lock-in amplification during experiments in the reduction of noise by a factor of (square root)2. The method is illustrated using experimental data obtained during CE separation of molecules from a commercial DNA ladder with 17 fluorescently labeled molecules having different base-pair sizes. An improvement in signal-to-noise ratio by a factor of ∼10 is achieved, resulting in a record-low limit of detection of 210 fM.

  12. Amplification of FRS2 and activation of FGFR/FRS2 signaling pathway in high-grade liposarcoma.

    PubMed

    Zhang, Keqiang; Chu, Kevin; Wu, Xiwei; Gao, Hanlin; Wang, Jinhui; Yuan, Yate-Ching; Loera, Sofia; Ho, Kimberley; Wang, Yafan; Chow, Warren; Un, Frank; Chu, Peiguo; Yen, Yun

    2013-02-15

    Fibroblast growth factor (FGF) receptor (FGFR) substrate 2 (FRS2) is an adaptor protein that plays a critical role in FGFR signaling. FRS2 is located on chromosome 12q13-15 that is frequently amplified in liposarcomas. The significance of FRS2 and FGFR signaling in high-grade liposarcomas is unknown. Herein, we first comparatively examined the amplification and expression of FRS2 with CDK4 and MDM2 in dedifferentiated liposarcoma (DDLS) and undifferentiated high-grade pleomorphic sarcoma (UHGPS). Amplification and expression of the three genes were identified in 90% to 100% (9-11 of 11) of DDLS, whereas that of FRS2, CDK4, and MDM2 were observed in 55% (41 of 75), 48% (36 of 75), and 44% (33/75) of clinically diagnosed UHGPS, suggesting that these "UHGPS" may represent DDLS despite lacking histologic evidence of lipoblasts. Immunohistochemical analysis of phosphorylated FRS2 protein indicated that the FGFR/FRS2 signaling axis was generally activated in about 75% of FRS2-positive high-grade liposarcomas. Moreover, we found that FRS2 and FGFRs proteins are highly expressed and functional in three high-grade liposarcoma cell lines: FU-DDLS-1, LiSa-2, and SW872. Importantly, the FGFR selective inhibitor NVP-BGJ-398 significantly inhibited the growth of FU-DDLS-1 and LiSa-2 cells with a concomitant suppression of FGFR signal transduction. Attenuation of FRS2 protein in FU-DDLS-1 and LiSa-2 cell lines decreased the phosphorylated extracellular signal-regulated kinase 1/2 and AKT and repressed cell proliferation. These findings indicate that analysis of FRS2 in combination with CDK4 and MDM2 will more accurately characterize pathologic features of high-grade liposarcomas. Activated FGFR/FRS2 signaling may play a functional role in the development of high-grade liposarcomas, therefore, serve as a potential therapeutic target.

  13. Microwave amplification with nanomechanical resonators.

    PubMed

    Massel, F; Heikkilä, T T; Pirkkalainen, J-M; Cho, S U; Saloniemi, H; Hakonen, P J; Sillanpää, M A

    2011-12-14

    The sensitive measurement of electrical signals is at the heart of modern technology. According to the principles of quantum mechanics, any detector or amplifier necessarily adds a certain amount of noise to the signal, equal to at least the noise added by quantum fluctuations. This quantum limit of added noise has nearly been reached in superconducting devices that take advantage of nonlinearities in Josephson junctions. Here we introduce the concept of the amplification of microwave signals using mechanical oscillation, which seems likely to enable quantum-limited operation. We drive a nanomechanical resonator with a radiation pressure force, and provide an experimental demonstration and an analytical description of how a signal input to a microwave cavity induces coherent stimulated emission and, consequently, signal amplification. This generic scheme, which is based on two linear oscillators, has the advantage of being conceptually and practically simpler than the Josephson junction devices. In our device, we achieve signal amplification of 25 decibels with the addition of 20 quanta of noise, which is consistent with the expected amount of added noise. The generality of the model allows for realization in other physical systems as well, and we anticipate that near-quantum-limited mechanical microwave amplification will soon be feasible in various applications involving integrated electrical circuits.

  14. Porous platinum nanotubes labeled with hemin/G-quadruplex based electrochemical aptasensor for sensitive thrombin analysis via the cascade signal amplification.

    PubMed

    Sun, Aili; Qi, Qingan; Wang, Xuannian; Bie, Ping

    2014-07-15

    For the first time, a sensitive electrochemical aptasensor for thrombin (TB) was developed by using porous platinum nanotubes (PtNTs) labeled with hemin/G-quadruplex and glucose dehydrogenase (GDH) as labels. Porous PtNTs with large surface area exhibited the peroxidase-like activity. Coupling with GDH and hemin/G-quadruplex as NADH oxidase and HRP-mimicking DNAzyme, the cascade signal amplification was achieved by the following ways: in the presence of glucose and NAD(+) in the working buffer, GDH electrocatalyzed the oxidation of glucose with the production of NADH. Then, hemin/G-quadruplex as NADH oxidase catalyzed the oxidation of NADH to in situ generate H2O2. Based on the corporate electrocatalysis of PtNTs and hemin/G-quadruplex toward H2O2, the electrochemical signal was significantly amplified, allowing the detection limit of TB down to 0.15 pM level. Moreover, the proposed strategy was simple because the intercalated hemin offered the readout signal, avoiding the adding of additional redox mediator as signal donator. Such an electrochemical aptasensor is highly promising for sensitive detection of other proteins in clinical diagnostics.

  15. Sensitive mRNA detection by fluorescence in situ hybridization using horseradish peroxidase-labeled oligodeoxynucleotides and tyramide signal amplification.

    PubMed

    van de Corput, M P; Dirks, R W; van Gijlswijk, R P; van Binnendijk, E; Hattinger, C M; de Paus, R A; Landegent, J E; Raap, A K

    1998-11-01

    With the ongoing progress in human genome projects, many genes are discovered whose function and/or expression pattern are not known. Most of these genes are expressed in relatively low abundance compared to housekeeping genes such as elongation factor-1alpha and beta-actin. Gene expression is studied by Northern blot assays or by semiquantitative PCR methods. Another method is the visualization of transcripts in tissue or cell cultures by fluorescence in situ hybridization (FISH). However, for low-abundance RNA detection, this method is hampered by its limited detection sensitivity and by the interference of background signals with specific hybridization signals. Background signals are introduced by nonspecific hybridization of probe sequences or nonspecific binding of antibodies used for visualization. To eliminate background signals derived from both sources and to benefit from the peroxidase-driven tyramide signal amplification (TSA), we directly conjugated horseradish peroxidase (HRP) to oligodeoxynucleotides (ODNs) and used these probes to study in the bladder cancer cell line 5637 the expression of various cytokine genes which, according to Northern hybridization and reverse transcriptase-polymerase chain reaction (RT-PCR) assays, are expressed at levels up to 10,000-fold less than abundantly expressed housekeeping genes. The results show that reduction of probe complexity and the limited use of immunocytochemical detection layers strongly reduces noise signals derived from nonspecific binding of nucleic acid probe and antibodies. The use of the HRP-ODNs in combination with TSA allowed detection of low-abundance cytokine mRNAs by FISH.

  16. Towards the use of protein A-tagged gold nanoparticles for signal amplification of electrochemical immunosensors in virus detection

    NASA Astrophysics Data System (ADS)

    Tran, Quang Huy; Mai, Anh Tuan; Thuy Nguyen, Thanh; Chung Pham, Van; Hanh Nguyen, Thi Hong

    2012-06-01

    In this paper we represent a study on the potential use of protein A-tagged gold nanoparticles applied for signal amplification of electrochemical immunosensors. Gold nanoparticles (GNPs) were synthesized by the chemical reduction of tetrachloroauric (III) acid trihydrate using sodium ascorbate, and then tagged with protein A (PrA) via ultracentrifugation. UV-Vis spectroscopy and transmission electron microscopy were used to verify the characteristics of formed GNPs/PrA complex. The analyzed results indicate that GNPs were found spherically, homogeneously, and with an average diameter of about 10 nm. Immunoelectron microscopy was then used to investigate the bioactivity of the GNPs/PrA complex in solution by the effective binding of GNPs to viral particles. Scanning electron and fluorescence microscopies were also used to investigate the distribution and the bioactivity of the GNPs/PrA complex on the surface of the interdigitated sensor. Consequently, this study provided some assumptions of the potential application of protein A-tagged gold nanoparticles for signal amplification of electrochemical immunosensors in virus detection from clinical samples.

  17. Sensitive detection for coralyne and mercury ions based on homo-A/T DNA by exonuclease signal amplification.

    PubMed

    Huang, Hailiang; Shi, Shuo; Zheng, Xuyue; Yao, Tianming

    2015-09-15

    Based on specific homo-A/T DNA binding properties, a strategy for coralyne and mercury ions detection was realised by exonuclease-aided signal amplification. Coralyne could specifically bind homo-A DNA and protect it from the hydrolysis of exonuclease I. The coralyne-protected DNA was subsequently used as a trigger strand to hydrolyze DNA2 in exonuclease-aided signal amplification process. Thiazole orange was used to quantify the remainder DNA2. Under the optimal condition, the fluorescence intensity was linearly proportional to the concentration of coralyne in the range of 0.2-100 nM with a limit of detection (LOD) of 0.31 nM, which presented the lowest LOD for coralyne among all reported. With homo-T and Hg(2+) taking the place of homo-A DNA and coralyne, respectively, the system could also be used for Hg(2+) detection. The experiments in real samples also showed good results. This method was label-free, low-cost, easy-operating and highly repeatable for the detection of coralyne and mercury ions. It could also be extended to detect various analytes, such as other metal ions, proteins and small molecules by using appropriate aptamers.

  18. Dual Signal Amplification Electrochemical Biosensor for Monitoring the Activity and Inhibition of the Alzheimer's Related Protease β-Secretase.

    PubMed

    Qu, Fengli; Yang, Minghui; Rasooly, Avraham

    2016-11-01

    The protease BACE1 (the β-site amyloid precursor protein cleaving enzyme 1) catalyzes the first step in the synthesis of β-amyloids (Aβ), peptides that accumulate in the brain in Alzheimer's disease (AD). Measurement of BACE1 activity is important for the development of BACE1 inhibitors to slow or stop AD. To measure BACE1 cleavage of the electrode-immobilized substrate peptide, we developed a redox-generating hydroxyapatite (HAP) probe which generates electrochemical current by reaction of the nanoparticle with molybdate (MoO4(2-)). The probe combines alkaline phosphatase (ALP) for dual signal amplification and Aβ antibody to bind the probe to the immobilized peptide substrate on the surface of the electrode. We measured the activity of BACE1 at concentrations ranging from 0.25 to 100 U/mL. The use of the dual-signal HAP-ALP probe increased the signal by an order of magnitude compared to HAP-only probe, enabling detection limits as low as 0.1 U/mL. To measure the inhibition of BACE1 activity, the BACE1 inhibitor OM99-2 was added to 25 U/mL of BACE1 in concentrations ranging from 5 to 150 nM. The observed detection limit of inhibition is 10 nM of OM99-2. These results demonstrate the capabilities of this novel biosensor to measure BACE1 activity and inhibitors of BACE1 activity. To the best of our knowledge this is the first report that reaction of HAP nanoparticles with molybdate can generate electrochemical current. This dual signal amplification strategy can be extended to other electrochemical assays and adapted for wide applications.

  19. Electrochemical biosensor based on enzyme substrate as a linker: Application for aldolase activity with pectin-thionine complex as recognization element and signal amplification probe.

    PubMed

    Wang, Xiaonan; Wang, Meiwen; Zhang, Yuanyuan; Miao, Xiaocao; Huang, Yuanyuan; Zhang, Juan; Sun, Lizhou

    2016-09-15

    A new strategy to fabricate electrochemical biosensor is reported based on the linkage of enzyme substrate, thereby an electrochemical method to detect aldolase activity is established using pectin-thionine complex (PTC) as recognization element and signal probe. The linkage effect of fructose-1,6-bisphosphate (FBP), the substrate of aldolase, can be achieved via its strong binding to magnetic nanoparticles (MNPs)/aminophenylboronic acid (APBA) and the formation of phosphoramidate bond derived from its reaction with p-phenylenediamine (PDA) on the surface of electrode. Aldolase can reversibly catalyze the substrates into the products which have no binding capacity with MNPs/APBA, resulting in the exposure of the corresponding binding sites and its subsequent recognization on signal probe. Meanwhile, signal amplification can be accomplished by using the firstly prepared PTC which can bind with MNPs/APBA, and accuracy can be strengthened through magnetic separation. With good precision and accuracy, the established sensor may be extended to other proteins with reversible catalyzed ability.

  20. Signal-enhancer molecules encapsulated liposome as a valuable sensing and amplification platform combining the aptasensor for ultrasensitive ECL immunoassay.

    PubMed

    Mao, Li; Yuan, Ruo; Chai, Yaqin; Zhuo, Ying; Xiang, Yun

    2011-06-15

    An innovatory ECL immunoassay strategy was proposed to detect the newly developing heart failure biomarker N-terminal pro-brain natriuretic peptide (NT-proBNP). Firstly, this strategy used small molecules encapsulated liposome as immune label to construct a sandwich immune sensing platform for NT-proBNP. Then the ECL aptasensor was prepared to collect and detect the small molecules released from the liposome. Finally, based on the ECL signal changes caused by the small molecules, the ECL signal indirectly reflected the level of NT-proBNP antigen. In this experiment, the cocaine was chosen as the proper small molecule that can act as signal-enhancer to enhance the ECL of Ru(bpy)(3)(2+). The cocaine-encapsulated liposomes were successfully characterized by TEM. The quantificational calculation proved the ∼5.3×10(3) cocaine molecules per liposome enough to perform the assignment of signal amplification. The cocaine-binding ECL aptasensor further promoted the work aimed at amplifying signal. The performance of NT-proBNP assay by the proposed strategy exhibited high sensitivity and high specificities with a linear relationship over 0.01-500 ng mL(-1) range, and a detection limit down to 0.77 pg mL(-1).

  1. [Oligonucleotide derivatives in the nucleic acid hybridization analysis. II. Isothermal signal amplification in process of DNA analysis by minisequencing].

    PubMed

    Dmitrienko, E V; Khomiakova, E A; Pyshnaia; Bragin, A G; Vedernikov, V E; Pyshnyĭ, D V

    2010-01-01

    The isothermal amplification of reporter signal via limited probe extension (minisequencing) upon hybridization of nucleic acids has been studied. The intensity of reporter signal has been shown to increase due to enzymatic labeling of multiple probes upon consecutive hybridization with one DNA template both in homophase and heterophase assays using various kinds of detection signal: radioisotope label, fluorescent label, and enzyme-linked assay. The kinetic scheme of the process has been proposed and kinetic parameters for each step have been determined. The signal intensity has been shown to correlate with physicochemical characteristics of both complexes: probe/DNA and product/DNA. The maximum intensity has been observed at minimal difference between the thermodynamic stability of these complexes, provided the reaction temperature has been adjusted near their melting temperature values; rising or lowering the reaction temperature reduces the amount of reporting product. The signal intensity has been shown to decrease significantly upon hybridization with the DNA template containing single-nucleotide mismatches. Limited probe extension assay is useful not only for detection of DNA template but also for its quantitative characterization.

  2. Hetero-enzyme-based two-round signal amplification strategy for trace detection of aflatoxin B1 using an electrochemical aptasensor.

    PubMed

    Zheng, Wanli; Teng, Jun; Cheng, Lin; Ye, Yingwang; Pan, Daodong; Wu, Jingjing; Xue, Feng; Liu, Guodong; Chen, Wei

    2016-06-15

    An electrochemical aptasensor for trace detection of aflatoxin B1 (AFB1) was developed by using an aptamer as the recognition unit while adopting the telomerase and EXO III based two-round signal amplification strategy as the signal enhancement units. The telomerase amplification was used to elongate the ssDNA probes on the surface of gold nanoparticles, by which the signal response range of the signal-off model electrochemical aptasensor could be correspondingly enlarged. Then, the EXO III amplification was used to hydrolyze the 3'-end of the dsDNA after the recognition of target AFB1, which caused the release of bounded AFB1 into the sensing system, where it participated in the next recognition-sensing cycle. With this two-round signal amplified electrochemical aptasensor, target AFB1 was successfully measured at trace concentrations with excellent detection limit of 0.6*10(-4)ppt and satisfied specificity due to the excellent affinity of the aptamer against AFB1. Based on this designed two-round signal amplification strategy, both the sensing range and detection limit were greatly improved. This proposed ultrasensitive electrochemical aptasensor method was also validated by comparison with the classic instrumental methods. Importantly, this hetero-enzyme based two-round signal amplified electrochemical aptasensor offers a great promising protocol for ultrasensitive detection of AFB1 and other mycotoxins by replacing the core recognition sequence of the aptamer.

  3. Enzyme-free detection of sequence-specific microRNAs based on nanoparticle-assisted signal amplification strategy.

    PubMed

    Li, Ru-Dong; Wang, Qian; Yin, Bin-Cheng; Ye, Bang-Ce

    2016-03-15

    Developing direct and convenient methods for microRNAs (miRNAs) analysis is of great significance in understanding biological functions of miRNAs, and early diagnosis of cancers. We have developed a rapid, enzyme-free method for miRNA detection based on nanoparticle-assisted signal amplification coupling fluorescent metal nanoclusters as signal output. The proposed method involves two processes: target miRNA-mediated nanoparticle capture, which consists of magnetic microparticle (MMP) probe and CuO nanoparticle (NP) probe, and nanoparticle-mediated amplification for signal generation, which consists of fluorescent DNA-Cu/Ag nanocluster (NC) and 3-mercaptopropionic acid (MPA). In the presence of target miRNA, MMP probe and NP probe sandwich-capture the target miRNA via their respective complementary sequence. The resultant sandwich complex (MMP probe-miRNA-CuO NP probe) is separated using a magnetic field and further dissolved by acidolysis to turn CuO NP into a great amount of copper (II) ions (Cu(2+)). Cu(2+) could disrupt the interactions between thiol moiety of MPA and the fluorescent Cu/Ag NCs by preferentially reacting with MPA to form a disulfide compound as intermediate. By this way, the fluorescence emission of the DNA-Cu/Ag NCs in the presence of MPA increases upon the increasing concentration of Cu(2+), which is directly proportional to the amount of target miRNA. The proposed method allows quantitative detection of a liver-specific miR-221-5p in the range of 5 pM to 1000 pM with a detection limit of ~0.73 pM, and shows a good ability to discriminate single-base difference. Moreover, the detection assay can be applied to detect miRNA in cancerous cell lysates in excellent agreement with that from a commercial miRNA detection kit.

  4. Electrochemical biosensor for microRNA detection based on poly(U) polymerase mediated isothermal signal amplification.

    PubMed

    Zhou, Yunlei; Yin, Huanshun; Li, Jie; Li, Bingchen; Li, Xue; Ai, Shiyun; Zhang, Xiansheng

    2016-05-15

    MicroRNAs play crucial role in post-transcriptional regulation for gene expression in animals, plants, and viruses. For the better understanding of microRNA and its functions, it is very important to develop effectively analytical method for microRNA detection. Herein, a novel electrochemical biosensor was fabricated for sensitive and selective detection of microRNA based on poly(U) polymerase mediated isothermal signal amplification, where poly(U) polymerase can catalyze the template independent addition of UMP from UTP to the 3' end of RNA. Using this activity, the target microRNA can be successfully labeled with biotin conjugated UMPs at its 3'-end using biotin conjugated UTP (biotin-UTP) as donor. Then, the avidin conjugated alkaline phosphatase can be further captured to the 3'-end of the target microRNA based on the specific interaction between biotin and avidin. Finally, under the catalytic activity of alkaline phosphatase, the substrate of p-nitrophenyl phosphate disodium salt hexahydrate can be hydrolyzed to produce 4-nitrophenol. According to the relationship between the electrochemical signal of p-nitrophenol and the concentration of microRNA-319a, the content of microRNA-319a can be detected. This signal amplification method is simple and sensitive. The developed method can detect as low as 1.7 fM microRNA and produce precise and accurate linear dynamic range from 10 to 1000 fM. The fabricated biosensor was further applied to detect the expression level change of microRNA-319a in rice seedlings after incubation with five kinds of different phytohormones.

  5. Pt NPs and DNAzyme functionalized polymer nanospheres as triple signal amplification strategy for highly sensitive electrochemical immunosensor of tumour marker.

    PubMed

    Chang, Honghong; Zhang, Haochun; Lv, Jia; Zhang, Bing; Wei, Wenlong; Guo, Jingang

    2016-12-15

    Highly sensitive determination of tumour markers is the key for early diagnosis of cancer. Herein, triple signal amplification strategy resulting from polymer nanospheres, Pt NPs, and DNAzyme was proposed in the developed electrochemical immunosensor. First, electroactive polymer nanospheres were synthesized by infinite coordination polymerization of ferrocenedicarboxylic acid, which could generate strong electrochemical signals due to plentiful ferrocene molecules. Further, the polymer nanospheres were functionalized by Pt NPs and DNAzyme (hemin/G-quadruplex) with the ability of catalyzing H2O2, which contributes to enhance the electrochemical signals. The prepared conjugations were characterized by transmission electron microscope (TEM) and energy dispersive X-ray spectroscopy (EDX). And the process of preparation was monitored by zeta potential. Based on the sandwich-type immunoassay, the electrochemical immunosensor was constructed employing the conjugations as signal tags. Under optimal conditions, the DPV peak increased with the increasing of alpha fetal protein (AFP) concentration, and the linear range was from 0.1pgmL(-1) to 100ngmL(-1) with low detection limit of 0.086pgmL(-1). Meanwhile, the designed immunosensor exhibited excellent selectivity and anti-interference property, good reproducibility and stability. More importantly, there were no significant differences in analyzing real clinical samples between designed immunosensor and commercial ELISA.

  6. Imaging resolution signal-to-noise ratio in transverse phase amplification from classical information theory

    NASA Astrophysics Data System (ADS)

    French, Doug; Huang, Zun; Pao, Hsueh-Yuan; Jovanovic, Igor

    2009-03-01

    A quantum phase amplifier operated in the spatial domain can improve the signal-to-noise ratio in imaging beyond the classical limit. The scaling of the signal-to-noise ratio with the gain of the quantum phase amplifier is derived from classical information theory.

  7. Silver nanowires-based signal amplification for CdSe quantum dots electrochemiluminescence immunoassay.

    PubMed

    Huang, Tingyu; Meng, Qingmin; Jie, Guifen

    2015-04-15

    A novel silver-cysteine hybrid nanowires (SCNWs) with many reactive carboxyl and amine groups were prepared, which enable them to be used as idea signal amplifying labels in bioassays. A large number of CdSe quantum dots (QDs) were loaded on the SCNWs to develop amplified SCNWs-QDs electrochemiluminescence (ECL) signal probe. The PAMAM dendrimer-SCNWs nanohybrids covered on the electrode constructed an effective antibody immobilization matrix and made the immobilized biomolecules hold high stability and bioactivity. Based on the specific sandwich immunoreaction strategy, the detection antibody (Ab2)-SCNWs-QDs ECL signal probe was applied to the sensitive signal-on ECL immunoassay of human IgG. The SCNWs-QDs ECL not only opens promising new ECL emitting species, but also promotes the development of novel ECL signal-transition platforms for biosensing devices.

  8. The interplay between discrete noise and nonlinear chemical kinetics in a signal amplification cascade

    NASA Astrophysics Data System (ADS)

    Lan, Yueheng; Papoian, Garegin A.

    2006-10-01

    We used various analytical and numerical techniques to elucidate signal propagation in a small enzymatic cascade which is subjected to external and internal noises. The nonlinear character of catalytic reactions, which underlie protein signal transduction cascades, renders stochastic signaling dynamics in cytosol biochemical networks distinct from the usual description of stochastic dynamics in gene regulatory networks. For a simple two-step enzymatic cascade which underlies many important protein signaling pathways, we demonstrated that the commonly used techniques such as the linear noise approximation and the Langevin equation become inadequate when the number of proteins becomes too low. Consequently, we developed a new analytical approximation, based on mixing the generating function and distribution function approaches, to the solution of the master equation that describes nonlinear chemical signaling kinetics for this important class of biochemical reactions. Our techniques work in a much wider range of protein number fluctuations than the methods used previously. We found that under certain conditions the burst phase noise may be injected into the downstream signaling network dynamics, resulting possibly in unusually large macroscopic fluctuations. In addition to computing first and second moments, which is the goal of commonly used analytical techniques, our new approach provides the full time-dependent probability distributions of the colored non-Gaussian processes in a nonlinear signal transduction cascade.

  9. Hairpin DNA Switch for Ultrasensitive Spectrophotometric Detection of DNA Hybridization Based on Gold Nanoparticles and Enzyme Signal Amplification

    SciTech Connect

    Zhang, Youyu; Tang, Zhiwen; Wang, Jun; Wu, Hong; Maham, Aihui; Lin, Yuehe

    2010-08-01

    A novel DNA detection platform based on a hairpin-DNA switch, nanoparticles, and enzyme signal amplification for ultrasensitive detection of DNA hybridization has been developed in this work. In this DNA assay, a “stem-loop” DNA probe dually labeled with a thiol at its 5’ end and a biotin at its 3’ end, respectively, was used. This probe was immobilized on the gold nanoparticles (AuNPs) anchored by a protein, globulin, on a 96-well microplate. In the absence of target DNA, the immobilized probe with the stem-loop structure shields the biotin from being approached by a bulky horseradish peroxidase linked-avidin (avidin-HRP) conjugate due to the steric hindrance. However, in the presence of target DNA, the hybridization between the hairpin DNA probe and the target DNA causes significant conformational change of the probe, which forces biotin away from the surface of AuNPs. As a result, the biotin becomes accessible by the avidin-HRP, and the target hybridization event can be sensitively detected via the HRP catalyzed substrate 3, 3', 5, 5'-tetramethylbenzidine using spectrophometric method. Some experimental parameters governing the performance of the assay have been optimized. At optimal conditions, this DNA assay can detect DNA at the concentration of femtomolar level by means of a signal amplification strategy based on the combination of enzymes and nanoparticles. This approach also has shown excellent specificity to distinguish single-base mismatches of DNA targets because of the intrinsic high selectivity of the hairpin DNA probe.

  10. Electrochemical magnetoimmunosensor for the ultrasensitive determination of interleukin-6 in saliva and urine using poly-HRP streptavidin conjugates as labels for signal amplification.

    PubMed

    Ojeda, I; Moreno-Guzmán, M; González-Cortés, A; Yáñez-Sedeño, P; Pingarrón, J M

    2014-10-01

    A novel magnetoimmunosensor design for interleukin-6 (IL-6) which involved the covalent immobilization of anti-IL-6 antibodies onto carboxyl-functionalized magnetic microparticles and a sandwich-type immunoassay with signal amplification using poly-HRP-streptavidin conjugates is reported. All the variables concerning the preparation and the electroanalytical performance of the immunosensor were optimized. The use of poly-HRP-strept conjugates as enzymatic labels instead of conventional HRP-strept allowed enhanced signal-to-blank current ratios to be obtained. A linear calibration plot between the measured steady-state current and the log of IL-6 concentration was achieved in the 1.75 to 500 pg/mL range, which was not feasible when using HRP-strep as label. A limit of detection of 0.39 pg/mL IL-6 was obtained. The anti-IL-6-MB conjugates exhibited an excellent storage stability providing amperometric responses with no significant loss during at least 36 days. The magnetoimmunosensor showed also an excellent selectivity against potentially interfering substances. The immunosensor was used to determine IL-6 in urine samples spiked at three different concentration levels with clinical relevance. Moreover, IL-6 was measured in three different saliva samples corresponding to a periodontitis patient, a smoker volunteer, and a non-smoker volunteer. The obtained results were statistically in agreement with those provided by a commercial ELISA kit.

  11. Magnetic Particle-Based Immunoassay of Phosphorylated p53 Using Protein-Cage Templated Lead Phosphate and Carbon Nanospheres for Signal Amplification

    SciTech Connect

    Chen, Aiqiong; Bao, Yuanwu; Ge, Xiaoxiao; Shin, Yongsoon; Du, Dan; Lin, Yuehe

    2012-11-20

    Phosphorylated p53 at serin 15 (phospho-p53-15) is a potential biomarker of Gamma-radiation exposure. In this paper, we described a new magnetic particles (MPs)-based electrochemical immunoassay of human phospho-p53-15 using carbon nanospheres (CNS) and protein-cage templated lead phosphate nanoparticles for signal amplification. Greatly enhanced sensitivity was achieved by three aspects: 1) The protein-cage nanoparticle (PCN) and p53-15 signal antibody (p53-15 Ab2) are linked to CNS (PCNof each apoferritin; 3) MPs capture a large amount of primary antibodies. Using apoferritin templated metallic phosphate instead of enzyme as label has the advantage of eliminating the addition of mediator or immunoreagents and thus makes the immunoassay system simpler. The subsequent stripping voltammetric analysis of the released lead ions were detected on a disposable screen printed electrode. The response current was proportional to the phospho-p53-15 concentration in the range of 0.02 to 20 ng mL-1 with detection limit of 0.01 ng mL-1. This method shows a good stability, reproducibility and recovery.

  12. Label-free electrochemical immunosensor based on enhanced signal amplification between Au@Pd and CoFe2O4/graphene nanohybrid

    PubMed Central

    Zhang, Yong; Li, Jiaojiao; Wang, Zhiling; Ma, Hongmin; Wu, Dan; Cheng, Qianhe; Wei, Qin

    2016-01-01

    The improvement of sensitivity of electrochemical immunosensor can be achieved via two approaches: increasing loading capacities of antibody and enlarging responding electrochemical signals. Based on these, CoFe2O4/graphene nanohybrid (CoFe2O4/rGO) as support was firstly used for preparing electrochemical biosensor, and with the addition of Au@Pd nanorods (NRs) as mimic enzyme, a label-free electrochemical immunosensor was prepared. Due to the high electrical conductivity, open porous structure and large loading capacities of CoFe2O4/rGO, the enhanced signal amplification between Au@Pd NRs and CoFe2O4/rGO was studied. Fabricated as a novel substrate, the prepared immunosensor had a good analytical performance and exhibited a wide linear range from 0.01 to 18.0 ng·mL−1 with a low detection limit of 3.3 pg·mL−1 for estradiol, which was succeeded in applying to detect estradiol in the natural water. PMID:26987503

  13. Sensitive immunosensor for tumor necrosis factor α based on dual signal amplification of ferrocene modified self-assembled peptide nanowire and glucose oxidase functionalized gold nanorod.

    PubMed

    Sun, Zhifang; Deng, Liu; Gan, Hao; Shen, Rujuan; Yang, Minghui; Zhang, Yi

    2013-01-15

    Sensitive electrochemical immunosensor for the detection of protein biomarker tumor necrosis factor α (TNF-α) was reported that uses ferrocene carboxylic acid (Fc) functionalized self-assembled peptide nanowire (Fc-PNW) as sensor platform and glucose oxidase (GOx) modified gold nanorod (GNR) as label. Greatly enhanced sensitivity is achieved based on a dual signal amplification strategy: first, the synthesized Fc-PNW used as the sensor platform increased the loading of primary anti-TNF-α antibody (Ab(1)) onto electrode surface due to its large surface area. At the same time, the Fc moiety on the nanowire is used as a mediator for GOx to catalyze the glucose reaction. Second, multiple GOx and secondary anti-TNF-α antibody (Ab(2)) molecules are bounded onto each GNR to increase the sensitivity of the immunosensor. After the preparation of the immunosensor based on the traditional sandwich protocol, the response of the immunosensor towards glucose was used as a signal to differentiate various concentrations of TNF-α. The resulting immunosensor has high sensitivity, wide linear range (0.005-10ng/mL) and good selectivity. This immunosensor preparation strategy is a promising platform for clinical screening of protein biomarkers.

  14. Rapid pathogen detection by lateral-flow immunochromatographic assay with gold nanoparticle-assisted enzyme signal amplification.

    PubMed

    Cho, Il-Hoon; Bhunia, Arun; Irudayaraj, Joseph

    2015-08-03

    To date most LF-ICA format for pathogen detection is based on generating color signals from gold nanoparticle (AuNP) tracers that are perceivable by naked eye but often these methods exhibit sensitivity lower than those associated with the conventional enzyme-based immunological methods or mandated by the regulatory guidelines. By developing AuNP avidin-biotin constructs in which a number of enzymes can be labeled we report on an enhanced LF-ICA system to detect pathogens at very low levels. With this approach we show that as low as 100 CFU/mL of Escherichia coli O157:H7 can be detected, indicating that the limit of detection can be increased by about 1000-fold due to our signal amplification approach. In addition, extensive cross-reactivity experiments were conducted (19 different organisms were used) to test and successfully validate the specificity of the assay. Semi-quantitative analysis can be performed using signal intensities which were correlated with the target pathogen concentrations for calibration by image processing.

  15. Detection of biological molecules using chemical amplification and optical sensors

    DOEpatents

    Van Antwerp, William Peter; Mastrototaro, John Joseph

    2000-01-01

    Methods are provided for the determination of the concentration of biological levels of polyhydroxylated compounds, particularly glucose. The methods utilize an amplification system that is an analyte transducer immobilized in a polymeric matrix, where the system is implantable and biocompatible. Upon interrogation by an optical system, the amplification system produces a signal capable of detection external to the skin of the patient. Quantitation of the analyte of interest is achieved by measurement of the emitted signal.

  16. Detection of biological molecules using chemical amplification and optical sensors

    DOEpatents

    Van Antwerp, William Peter; Mastrototaro, John Joseph

    2004-10-12

    Methods are provided for the determination of the concentration of biological levels of polyhydroxylated compounds, particularly glucose. The methods utilize an amplification system that is an analyte transducer immobilized in a polymeric matrix, where the system is implantable and biocompatible. Upon interrogation by an optical system, the amplification system produces a signal capable of detection external to the skin of the patient. Quantitation of the analyte of interest is achieved by measurement of the emitted signal.

  17. Detection of biological molecules using chemical amplification and optical sensors

    SciTech Connect

    Antwerp, W.P. van; Mastrototaro, J.J.

    2000-01-04

    Methods are provided for the determination of the concentration of biological levels of polyhydroxylated compounds, particularly glucose. The methods utilize an amplification system that is an analyte transducer immobilized in a polymeric matrix, where the system is implantable and biocompatible. Upon interrogation by an optical system, the amplification system produces a signal capable of detection external to the skin of the patient. Quantitation of the analyte of interest is achieved by measurement of the emitted signal.

  18. Full quadrature regeneration of QPSK signals using sequential phase sensitive amplification and parametric saturation.

    PubMed

    Bottrill, K R H; Hesketh, G; Jones, L; Parmigiani, F; Richardson, D J; Petropoulos, P

    2017-01-23

    We demonstrate all-optical regeneration of both the phase and the amplitude of a 10 GBaud quadrature phase shift keying (QPSK) signal using two nonlinear stages. First we regenerate the phase using a wavelength converting phase sensitive amplifier and then we regenerate the amplitude using a saturated single-pump parametric amplifier, returning the signal to its original wavelength at the same time. We exploit the conjugating nature of the two processing stages to eliminate the intrinsic SPM distortion of the system, further improving performance.

  19. Ping-pong amplification of a retroviral vector achieves high-level gene expression: human growth hormone production.

    PubMed Central

    Kozak, S L; Kabat, D

    1990-01-01

    Retroviral vectors offer major advantages for gene transfer studies but have not been useful for producing proteins in large quantities. This deficiency has resulted in part from interference to superinfection, which limits the numbers of active proviruses in cells. Recently, we found that these vectors amplify when they are added as calcium phosphate precipitates to cocultures of cells that package retroviruses into ecotropic and amphotropic host range envelopes. Helper-free virions from either cell type can infect the other without interference, resulting in theoretically limitless back-and-forth (ping-pong) vector replication. In initial studies, however, amplifications of a vector that contained the human growth hormone gene ceased when the hormone produced was 0.3% or less of cellular protein synthesis. This limit was caused by two factors. First, recombinant shutoff viruses that are replication defective and encode envelope glycoproteins form at a low probability during any round of the vector replication cycle and these spread in cocultures, thereby establishing interference. Single cells in shutoff cocultures therefore synthesize both ecotropic and amphotropic envelope glycoproteins, and they release promiscuous (presumably hybrid) virions. The probability of forming shutoff viruses before the vector had amplified to a high multiplicity was reduced by using small cocultures. Second, cells with large numbers of proviruses are unhealthy and their proviral expression can be unstable. Stable expresser cell clones were obtained by selection. Thereby, cell lines were readily obtained that stably produce human growth hormone as 4 to 6% of the total protein synthesis. A ping-pong retroviral vector can be used for high-level protein production in vertebrate cells. Images PMID:2352330

  20. Collective Signal Processing in Cluster Chemotaxis: Roles of Adaptation, Amplification, and Co-attraction in Collective Guidance

    PubMed Central

    Camley, Brian A.; Zimmermann, Juliane; Levine, Herbert; Rappel, Wouter-Jan

    2016-01-01

    Single eukaryotic cells commonly sense and follow chemical gradients, performing chemotaxis. Recent experiments and theories, however, show that even when single cells do not chemotax, clusters of cells may, if their interactions are regulated by the chemoattractant. We study this general mechanism of “collective guidance” computationally with models that integrate stochastic dynamics for individual cells with biochemical reactions within the cells, and diffusion of chemical signals between the cells. We show that if clusters of cells use the well-known local excitation, global inhibition (LEGI) mechanism to sense chemoattractant gradients, the speed of the cell cluster becomes non-monotonic in the cluster’s size—clusters either larger or smaller than an optimal size will have lower speed. We argue that the cell cluster speed is a crucial readout of how the cluster processes chemotactic signals; both amplification and adaptation will alter the behavior of cluster speed as a function of size. We also show that, contrary to the assumptions of earlier theories, collective guidance does not require persistent cell-cell contacts and strong short range adhesion. If cell-cell adhesion is absent, and the cluster cohesion is instead provided by a co-attraction mechanism, e.g. chemotaxis toward a secreted molecule, collective guidance may still function. However, new behaviors, such as cluster rotation, may also appear in this case. Co-attraction and adaptation allow for collective guidance that is robust to varying chemoattractant concentrations while not requiring strong cell-cell adhesion. PMID:27367541

  1. High-sensitive surface plasmon resonance microRNA biosensor based on streptavidin functionalized gold nanorods-assisted signal amplification.

    PubMed

    Hao, Kaihong; He, Yu; Lu, Huiting; Pu, Shaotao; Zhang, Yingnan; Dong, Haifeng; Zhang, Xueji

    2017-02-15

    Herein, a facile and sensitive microRNA (miRNA) biosensor was designed by using interfacial biotinylated thiolated DNA molecular beacon (MB) as probe and streptavidin functionalized gold nanorods (Stre-GNRs) as tag for the enhanced surface plasmon resonance (SPR) signal. The MB probe with two terminals labeled with biotin and thiol groups, respectively, was modified on the gold film via thiol-gold interaction. Upon hybridization with the target, the biotinylated group became accessible to the Stre-GNRs. The introduction of the Stre-GNRs tag to the gold film produced strong SPR signal for detection. Our work has illustrated that the plasmonic field extension generated from the gold film to GNRs and the mass increase due to the GNRs have led to drastic sensitivity enhancement. Under optimal conditions, this proposed approach allowed detection of miRNA with the limit of detection (LOD) down to 0.045 pM. The results have shown that the MB probe functionalized sensing film, together with streptavidin-conjugated GNRs, was readily served as a plasmonic coupling partner that can be used as a powerful ultrasensitive sandwich assay for miRNA detection, and GNRs were readily served as promising amplification labels in SPR sensing technology.

  2. A Nanoporous Alumina Membrane Based Electrochemical Biosensor for Histamine Determination with Biofunctionalized Magnetic Nanoparticles Concentration and Signal Amplification.

    PubMed

    Ye, Weiwei; Xu, Yifan; Zheng, Lihao; Zhang, Yu; Yang, Mo; Sun, Peilong

    2016-10-22

    Histamine is an indicator of food quality and indispensable in the efficient functioning of various physiological systems. Rapid and sensitive determination of histamine is urgently needed in food analysis and clinical diagnostics. Traditional histamine detection methods require qualified personnel, need complex operation processes, and are time-consuming. In this study, a biofunctionalized nanoporous alumina membrane based electrochemical biosensor with magnetic nanoparticles (MNPs) concentration and signal amplification was developed for histamine determination. Nanoporous alumina membranes were modified by anti-histamine antibody and integrated into polydimethylsiloxane (PDMS) chambers. The specific antibody modified MNPs were used to concentrate histamine from samples and transferred to the antibody modified nanoporous membrane. The MNPs conjugated to histamine were captured in the nanopores via specific reaction between histamine and anti-histamine antibody, resulting in a blocking effect that was amplified by MNPs in the nanopores. The blockage signals could be measured by electrochemical impedance spectroscopy across the nanoporous alumina membrane. The sensing platform had great sensitivity and the limit of detection (LOD) reached as low as 3 nM. This biosensor could be successfully applied for histamine determination in saury that was stored in frozen conditions for different hours, presenting a potentially novel, sensitive, and specific sensing system for food quality assessment and safety support.

  3. A Nanoporous Alumina Membrane Based Electrochemical Biosensor for Histamine Determination with Biofunctionalized Magnetic Nanoparticles Concentration and Signal Amplification

    PubMed Central

    Ye, Weiwei; Xu, Yifan; Zheng, Lihao; Zhang, Yu; Yang, Mo; Sun, Peilong

    2016-01-01

    Histamine is an indicator of food quality and indispensable in the efficient functioning of various physiological systems. Rapid and sensitive determination of histamine is urgently needed in food analysis and clinical diagnostics. Traditional histamine detection methods require qualified personnel, need complex operation processes, and are time-consuming. In this study, a biofunctionalized nanoporous alumina membrane based electrochemical biosensor with magnetic nanoparticles (MNPs) concentration and signal amplification was developed for histamine determination. Nanoporous alumina membranes were modified by anti-histamine antibody and integrated into polydimethylsiloxane (PDMS) chambers. The specific antibody modified MNPs were used to concentrate histamine from samples and transferred to the antibody modified nanoporous membrane. The MNPs conjugated to histamine were captured in the nanopores via specific reaction between histamine and anti-histamine antibody, resulting in a blocking effect that was amplified by MNPs in the nanopores. The blockage signals could be measured by electrochemical impedance spectroscopy across the nanoporous alumina membrane. The sensing platform had great sensitivity and the limit of detection (LOD) reached as low as 3 nM. This biosensor could be successfully applied for histamine determination in saury that was stored in frozen conditions for different hours, presenting a potentially novel, sensitive, and specific sensing system for food quality assessment and safety support. PMID:27782087

  4. A non-perturbative analytic expression of signal amplification factor in stochastic resonance

    NASA Astrophysics Data System (ADS)

    Dhara, Asish Kumar

    2017-04-01

    We put forward a non-perturbative scheme to calculate the response of an overdamped bistable system driven by a Gaussian white noise and perturbed by a weak monochromatic force (signal) analytically. The formalism takes into account infinite number of perturbation terms of a perturbation series with amplitude of the signal as an expansion parameter. The contributions of infinite number of relaxation modes of the stochastic dynamics to the response are also taken into account in this formalism. A closed form analytic expression of the response is obtained. Only the knowledge of the first non-trivial eigenvalue and the lowest eigenfunction of the un-perturbed Fokker-Planck operator are needed to evaluate the response. The response calculated from the derived analytic expression matches fairly well with the numerical results.

  5. Studies on the signal amplification in weighted and unweighted small-world networks

    NASA Astrophysics Data System (ADS)

    Gao, Yang; Wang, Jianjun; Ma, Fuqiu

    2017-02-01

    Weighted and unweighted networks composed of coupled bistable oscillators with small-world topology are investigated under the co-presence of a weak signal and multiplicative Gaussian white noise. As the noise intensity is adjusted to one or two optimal values, the temporal periodicity of the output of the system reaches the maximum, indicating the occurrence of stochastic resonance (SR) or stochastic bi-resonance (SBR). The resonance behavior is strongly-dependent on the coupling strength in both networks. At a weak coupling, SR more likely takes place; whereas at a strong coupling, SBR is prone to occur. Compared with unweighted networks, the span of coupling strength for SBR is narrower in weighted networks. In addition, the weak signal cannot be amplified so effectively in the weighted networks as in the unweighted networks, attributing to the weakening effect of the link weight on the coupling between oscillators and the heterogeneity of the whole network connectivity caused by the weight distribution.

  6. In-line phase-sensitive amplification of QPSK signal using multiple quasi-phase matched LiNbO₃ waveguide.

    PubMed

    Asobe, Masaki; Umeki, Takeshi; Takenouchi, Hirokazu; Miyamoto, Yutaka

    2014-11-03

    Phase-sensitive amplifiers (PSA) using periodically poled (PPLN) LiNbO₃ waveguides are promising as low-noise optical amplifiers. However, it is difficult to realize in-line operation for multi-level phase modulated signals using a PPLN based PSA with the conventional configuration. In this paper, we report a PPLN based in-line PSA that can regenerate quadrature phase shift keying (QPSK) signals. Multi-stage frequency mixing in a multiple quasi-phase matched LiNbO₃waveguide allows carrier phase recovery from a QPSK signal. Non-degenerate parametric amplification enables the phase-sensitive amplification of a QPSK signal. Amplitude and phase regeneration is examined utilizing gain saturation and phase squeezing capability.

  7. pH-Induced aggregated melanin nanoparticles for photoacoustic signal amplification

    NASA Astrophysics Data System (ADS)

    Ju, Kuk-Youn; Kang, Jeeun; Pyo, Jung; Lim, Joohyun; Chang, Jin Ho; Lee, Jin-Kyu

    2016-07-01

    We present a new melanin-like nanoparticle (MelNP) and its performance evaluation results. This particle is proposed as an exogenous contrast agent for photoacoustic (PA) imaging. Conventional PA contrast agents are based on non-biological materials. In contrast, the MelNPs are organic nanoparticles inspired by natural melanin. Melanin is an endogenous chromophore that has the ability to produce a PA signal in vivo. The developed MelNPs are capable of aggregating with one another under mildly acidic conditions after introducing hydrolysis-susceptible citraconic amide on the surface of bare MelNPs. We ascertained that the physical aggregation of the MelNPs resulted in an increased PA signal strength in the near-infrared window of biological tissue (i.e., 700 nm) without absorption tuning. This phenomenon is likely because of the overlapping thermal fields of the developed MelNPs. The PA signal produced from the developed MelNPs, after exposure to mildly acidic conditions (i.e., pH 6), is 8.1 times stronger than under neutral conditions. This unique characteristic found in this study can be utilized in a practical strategy for highly sensitive in vivo cancer target imaging in response to its acidic microenvironment. This approach to amplify the PA response of MelNPs in clusters could accelerate the use of MelNPs as an alternative to non-biological nanoprobes, so that MelNPs may be applicable in PA imaging and functional PA imaging such as stimuli sensitive, multimodal, and theranostic imaging.We present a new melanin-like nanoparticle (MelNP) and its performance evaluation results. This particle is proposed as an exogenous contrast agent for photoacoustic (PA) imaging. Conventional PA contrast agents are based on non-biological materials. In contrast, the MelNPs are organic nanoparticles inspired by natural melanin. Melanin is an endogenous chromophore that has the ability to produce a PA signal in vivo. The developed MelNPs are capable of aggregating with one

  8. Quenching of Unincorporated Amplification Signal Reporters in Reverse-Transcription Loop-Mediated Isothermal Amplification Enabling Bright, Single-Step, Closed-Tube, and Multiplexed Detection of RNA Viruses.

    PubMed

    Ball, Cameron S; Light, Yooli K; Koh, Chung-Yan; Wheeler, Sarah S; Coffey, Lark L; Meagher, Robert J

    2016-04-05

    Reverse-transcription-loop-mediated isothermal amplification (RT-LAMP) has frequently been proposed as an enabling technology for simplified diagnostic tests for RNA viruses. However, common detection techniques used for LAMP and RT-LAMP have drawbacks, including poor discrimination capability, inability to multiplex targets, high rates of false positives, and (in some cases) the requirement of opening reaction tubes postamplification. Here, we present a simple technique that allows closed-tube, target-specific detection, based on inclusion of a dye-labeled primer that is incorporated into a target-specific amplicon if the target is present. A short, complementary quencher hybridizes to unincorporated primer upon cooling down at the end of the reaction, thereby quenching fluorescence of any unincorporated primer. Our technique, which we term QUASR (for quenching of unincorporated amplification signal reporters, read "quasar"), does not significantly reduce the amplification efficiency or sensitivity of RT-LAMP. Equipped with a simple LED excitation source and a colored plastic gel filter, the naked eye or a camera can easily discriminate between positive and negative QUASR reactions, which produce a difference in signal of approximately 10:1 without background subtraction. We demonstrate that QUASR detection is compatible with complex sample matrices such as human blood, using a novel LAMP primer set for bacteriophage MS2 (a model RNA virus particle). Furthermore, we demonstrate single-tube duplex detection of West Nile virus (WNV) and chikungunya virus (CHIKV) RNA.

  9. Optical biosensors based on direct coupling of recognition, signal transduction, and amplification

    NASA Astrophysics Data System (ADS)

    Song, Xuedong; Swanson, Basil I.

    1999-02-01

    Highly sensitive, specific and reagent-free optical signal transduction methods for detection of polyvalent proteins have been developed by directly coupling distance-dependent fluorescence self-quenching and/or resonant energy transfer to the protein receptor binding events. The ganglioside GM1 as recognition unit for cholera toxin (CT) was covalently labeled with fluorophores, and then incorporated into a biomimetic membrane surface. In the case using fluorescence self-quenching as a signal transduction mechanism, the fluorescence intensity drops significantly due to aggregation of the fluorophore-labeled GM1 on a biomimetic surface. By labeling GM1 with a fluorescence energy transfer pair, aggregation of the labeled-GM1 results in a decrease in donor and an increase in acceptor fluorescence, providing a unique signature for specific protein-receptor binding. The detection systems can reliably detect less than 0.05 nM CT with fast response (less than five minutes). This approach can easily be adapted to any biosensor scheme that relies on multiple receptors or coreceptors. The methods can also be applied to investigate the kinetics and thermodynamics of the multivalent interactions.

  10. BRIEF COMMUNICATIONS: Amplification of continuously tunable signals in high-pressure CO2 amplifiers

    NASA Astrophysics Data System (ADS)

    Orlovskiĭ, V. M.; Osipov, V. V.; Solov'ev, V. S.

    1981-02-01

    An investigation was made of the emission of high-power continuously tunable signals in a CO2 laser stage consisting of a master oscillator and an amplifier. The active medium in the oscillator and the amplifier was excited by an electric discharge sustained by an electron beam of 1 μsec duration with a current density of 3 A/cm2. The pressure of the CO2:N2 = 1:1 active medium in the master oscillator was kept at 6.5 atm and that in the amplifier was varied between 1 and 6 atm. It was found that when signals were amplified under similar conditions at frequencies corresponding to the peak of the P18 line and to the dip between the P18 and P16 lines of the 00°1-10°0 transition, the radiation energy changed no less than twofold as a result of a single passage through the amplifier. Moreover, it was found that the pressure in the amplifier could be reduced considerably compared with that of the active medium in the master oscillator whilst retaining continuous frequency tuning, which meant that the requirements on the excitation system could be reduced and the efficiency of the amplifier could be enhanced.

  11. Amplification of single molecule translocation signal using β-strand peptide functionalized nanopores.

    PubMed

    Liebes-Peer, Yael; Rapaport, Hanna; Ashkenasy, Nurit

    2014-07-22

    Changes in ionic current flowing through nanopores due to binding or translocation of single biopolymer molecules enable their detection and characterization. It is, however, much more challenging to detect small molecules due to their rapid and small signal signature. Here we demonstrate the use of de novo designed peptides for functionalization of nanopores that enable the detection of a small analytes at the single molecule level. The detection relies on cooperative peptide conformational change that is induced by the binding of the small molecule to a receptor domain on the peptide. This change results in alteration of the nanopore effective diameter and hence induces current perturbation signal. On the basis of this approach, we demonstrate here the detection of diethyl 4-nitrophenyl phosphate (paraoxon), a poisonous organophosphate molecule. Paraoxon binding is induced by the incorporation of the catalytic triad of acetylcholine esterase in the hydrophilic domain of a short amphiphilic peptide and promotes β-sheet assembly of the peptide both in solution and for peptide molecules immobilized on solid surfaces. Nanopores coated with this peptide allowed the detection of paraoxon at the single molecule level revealing two binding arrangements. This unique approach, hence, provides the ability to study interactions of small molecules with the corresponding engineered receptors at the single molecule level. Furthermore, the suggested versatile platform may be used for the development of highly sensitive small analytes sensors.

  12. A cytometric bead assay for sensitive DNA detection based on enzyme-free signal amplification of hybridization chain reaction.

    PubMed

    Ren, Wei; Liu, Hongmei; Yang, Wenxia; Fan, Yunlong; Yang, Lang; Wang, Yucong; Liu, Chenghui; Li, Zhengping

    2013-11-15

    A versatile flow cytometric bead assay (CBA) is developed for sensitive DNA detection by integrating the advantages of hybridization chain reaction (HCR) for enzyme-free signal amplification, flow cytometry for robust and rapid signal readout as well as magnetic beads (MBs) for facile separation. In this HCR-CBA, a biotinylated hairpin DNA (Bio-H1) is firstly immobilized on streptavidin-functionalized MBs. Upon the addition of target DNA, each target would hybridize with one Bio-H1 to open its hairpin structure and subsequently initiate a cascade of hybridization events between two species of fluorescent DNA hairpin probes (H1*/H2*) to form a nicked double helical DNA structure, resulting in amplified accumulation of numerous fluorophores on the MBs. Finally, the fluorescent MBs are directly analyzed by flow cytometry. This technique enables quantitative analysis of the HCR products anchored on the MBs as a function of target DNA concentration, and analysis of each sample can be completed within few minutes. Therefore, the HCR-CBA approach provides a practical DNA assay with greatly improved sensitivity. The detection limit of a model DNA target is 0.5 pM (3σ), which is about 3 orders of magnitude lower compared with traditional hybridization methods without HCR. Furthermore, the signal of complementary target can be clearly distinguished from that of single-base mismatched sequences, indicating the high specificity of the HCR-CBA. Moreover, this strategy is also successfully applied to the DNA analysis in complex biological samples, showing great potential in gene analysis and disease diagnosis in clinical samples.

  13. Gradient sensing by a bistable regulatory motif enhances signal amplification but decreases accuracy in individual cells

    NASA Astrophysics Data System (ADS)

    Sharma, Rati; Roberts, Elijah

    2016-06-01

    Many vital eukaryotic cellular functions require the cell to respond to a directional gradient of a signaling molecule. The first two steps in any eukaryotic chemotactic/chemotropic pathway are gradient detection and cell polarization. Like many processes, such chemotactic and chemotropic decisions are made using a relatively small number of molecules and are thus susceptible to internal and external fluctuations during signal transduction. Large cell-to-cell variations in the magnitude and direction of a response are therefore possible and do, in fact, occur in natural systems. In this work we use three-dimensional probabilistic modeling of a simple gradient sensing pathway to study the capacity for individual cells to accurately determine the direction of a gradient, despite fluctuations. We include a stochastic external gradient in our simulations using a novel gradient boundary condition modeling a point emitter a short distance away. We compare and contrast three different variants of the pathway, one monostable and two bistable. The simulation data show that an architecture combining bistability with spatial positive feedback permits the cell to both accurately detect and internally amplify an external gradient. We observe strong polarization in all individual cells, but in a distribution of directions centered on the gradient. Polarization accuracy in our study was strongly dependent upon a spatial positive feedback term that allows the pathway to trade accuracy for polarization strength. Finally, we show that additional feedback links providing information about the gradient to multiple levels in the pathway can help the cell to refine initial inaccuracy in the polarization direction.

  14. Ultrasensitive amperometric immunosensor for PSA detection based on Cu2O@CeO2-Au nanocomposites as integrated triple signal amplification strategy.

    PubMed

    Li, Faying; Li, Yueyun; Feng, Jinhui; Dong, Yunhui; Wang, Ping; Chen, Lei; Chen, Zhiwei; Liu, Hui; Wei, Qin

    2017-01-15

    In this work, a novel label-free electrochemical immunosensor was developed for the quantitative detection of prostate specific antigen (PSA). To this end, the amino functionalized cuprous oxide @ ceric dioxide (Cu2O@CeO2-NH2) core-shell nanocomposites were prepared to bond gold nanoparticles (Au NPs) by constructing stable Au-N bond between Au NPs and -NH2. Because the synergetic effect presents in Cu2O@CeO2 core-shell loaded with Au NPs (Cu2O@CeO2-Au), it shows better electrocatalytic activity towards the reduction of hydrogen peroxide (H2O2) than single Cu2O, Au NPs and Cu2O@CeO2. Featured by large specific surface area, good biocompatibility and good electrochemical properties which can greatly improve the electronic transmission rate, Cu2O@CeO2-Au was used as transducing materials to achieve efficiently capture antibodies and triple signal amplification of the proposed immunosensor. Under the optimal conditions, the proposed immunosensor exhibited a wide linear range from 0.1pg/mL to 100ng/mL with a low detection limit of 0.03pg/mL (S/N=3). Furthermore, the proposed label-free immunosensor has been used to determine PSA in human serum with satisfactory results. Meanwhile, it displayed good reproducibility, acceptable selectivity, and long-term stability, which had promising application in bioassay analysis.

  15. A novel signal amplification strategy of an electrochemical aptasensor for kanamycin, based on thionine functionalized graphene and hierarchical nanoporous PtCu.

    PubMed

    Qin, Xiaoli; Yin, Yan; Yu, Huijing; Guo, Wenjuan; Pei, Meishan

    2016-03-15

    An ultrasensitive electrochemical aptasensor for the quantitative detection of kanamycin antibiotic was fabricated based on a novel signal amplification strategy. This aptasensor was developed using thionine functionalized graphene (GR-TH) and hierarchical nanoporous (HNP) PtCu alloy as biosensing substrates for the first time. HNP-PtCu alloy with controllable bimodal ligament/pore distributions was successfully prepared by two-step dealloying of a well-designed PtCuAl precursor alloy combined with an annealing operation. GR-TH composite was synthesized by one-step reduction of graphene oxide (GO) in TH solution. Greatly amplified sensitivity was achieved by using GR-TH/HNP-PtCu composite owing to its large specific surface and good electron-transfer ability. Under the optimized conditions, the proposed aptasensor exhibited a high sensitivity and a wider linearity to kanamycin in the range 5 × 10(-7)-5 × 10(-2) μgmL(-1) with a low detection limit of 0.42 pgmL(-1). This aptasensor also displayed a satisfying electrochemical performance with good stability, selectivity and reproducibility. The as-prepared aptasensor was successfully used for the determination of kanamycin in animal derived food.

  16. The Spatial Pattern of Cochlear Amplification

    PubMed Central

    Fisher, Jonathan A.N.; Nin, Fumiaki; Reichenbach, Tobias; Uthaiah, Revathy C.; Hudspeth, A.J.

    2012-01-01

    SUMMARY Sensorineural hearing loss, which stems primarily from the failure of mechanosensory hair cells, changes the traveling waves that transmit acoustic signals along the cochlea. However, the connection between cochlear mechanics and the amplificatory function of hair cells remains unclear. Using an optical technique that permits the targeted inactivation of prestin, a protein of outer hair cells that generates forces on the basilar membrane, we demonstrate that these forces interact locally with cochlear traveling waves to achieve enormous mechanical amplification. By perturbing amplification in narrow segments of the basilar membrane, we further show that a cochlear traveling wave accumulates gain as it approaches its peak. Analysis of these results indicates that cochlear amplification produces negative damping that counters the viscous drag impeding traveling waves; targeted photoinactivation locally interrupts this compensation. These results reveal the locus of amplification in cochlear traveling waves and connect the characteristics of normal hearing to molecular forces. PMID:23217746

  17. The spatial pattern of cochlear amplification.

    PubMed

    Fisher, Jonathan A N; Nin, Fumiaki; Reichenbach, Tobias; Uthaiah, Revathy C; Hudspeth, A J

    2012-12-06

    Sensorineural hearing loss, which stems primarily from the failure of mechanosensory hair cells, changes the traveling waves that transmit acoustic signals along the cochlea. However, the connection between cochlear mechanics and the amplificatory function of hair cells remains unclear. Using an optical technique that permits the targeted inactivation of prestin, a protein of outer hair cells that generates forces on the basilar membrane, we demonstrate that these forces interact locally with cochlear traveling waves to achieve enormous mechanical amplification. By perturbing amplification in narrow segments of the basilar membrane, we further show that a cochlear traveling wave accumulates gain as it approaches its peak. Analysis of these results indicates that cochlear amplification produces negative damping that counters the viscous drag impeding traveling waves; targeted photoinactivation locally interrupts this compensation. These results reveal the locus of amplification in cochlear traveling waves and connect the characteristics of normal hearing to molecular forces.

  18. The 'window' component of the low threshold Ca2+ current produces input signal amplification and bistability in cat and rat thalamocortical neurones.

    PubMed Central

    Williams, S R; Tóth, T I; Turner, J P; Hughes, S W; Crunelli, V

    1997-01-01

    1. The mechanism underlying a novel form of input signal amplification and bistability was investigated by intracellular recording in rat and cat thalamocortical (TC) neurones maintained in slices and by computer simulation with a biophysical model of these neurones. 2. In a narrow membrane potential range centred around -60 mV, TC neurones challenged with small (10-50 pA), short (50-200 ms) current steps produced a stereotyped, large amplitude hyperpolarization (> 20 mV) terminated by the burst firing of action potentials, leading to amplification of the duration and amplitude of the input signal, that is hereafter referred to as input signal amplification. 3. In the same voltage range centred around -60 mV, single evoked EPSPs and IPSPs also produced input signal amplification, indicating that this behaviour can be triggered by physiologically relevant stimuli. In addition, a novel, intrinsic, low frequency oscillation, characterized by a peculiar voltage dependence of its frequency and by the presence of plateau potentials on the falling phase of low threshold Ca2+ potentials, was recorded. 4. Blockade of pure Na+ and K+ currents by tetrodotoxin (1 microM) and Ba2+ (0.1-2.0 mM), respectively, did not affect input signal amplification, neither did the presence of excitatory or inhibitory amino acid receptor antagonists in the perfusion medium. 5. A decrease in [Ca2+]o (from 2 to 1 mM) and an increase in [Mg2+]o (from 2 to 10 mM), or the addition of Ni2+ (2-3 mM), abolished input signal amplification, while an increase in [Ca2+]o (from 2 to 8 mM) generated this behaviour in neurones where it was absent in control conditions. These results indicate the involvement of the low threshold Ca2+ current (IT) in input signal amplification, since the other Ca2+ currents of TC neurones are activated at potentials more positive than -40 mV. 6. Blockade of the slow inward mixed cationic current (Ih) by 4-(N-ethyl-N-phenylamino)-1,2-dimethyl-6-(methylamino

  19. Glucose oxidase-doped magnetic silica nanostrutures as labels for localized signal amplification of electrochemical immunosensors

    NASA Astrophysics Data System (ADS)

    Ren, Jingjing; Tang, Dianping; Su, Biling; Tang, Juan; Chen, Guonan

    2010-07-01

    Herein, we report a novel glucose oxidase (GOD)-doped magnetic silica nanostructure and its possible application in the clinical immunoassays. The doped nanostructures were initially synthesized using the reverse micelle method, and ferritin antibodies (anti-Ft) were then labeled to the surface of the nanostructures, which were employed as signal antibodies for ultrasensitive detection of ferritin (Ft) in the sandwich-type electrochemical enzyme immunoassays. The doped nanostructures were characterized using transmission electron microscopy (TEM), UV-vis absorption spectrometry and vibrating sample magnetometer (VSM). The advantages of the doped nanostructures as labels were investigated in comparison with the conventional label method. Under the optimal conditions, the nanostructures-based immunoassay toward ferritin standards displays a wide dynamic range from 0.1 to 400 ng mL-1 with a low detection limit of 10 pg mL-1 ferritin (at 3σ), which is three-fold higher in the sensitivity than that of directly using GOD-labeled antibodies. The assay results for clinical serum samples with the developed method received in excellent accordance with results obtained from the referenced standard enzyme-linked immunosorbent assay (ELISA) method.

  20. CB[7]-mediated signal amplification approach for sensitive surface plasmon resonance spectroscopy.

    PubMed

    Gao, Yanmin; Zou, Fei; Wu, Beiping; Wang, Xingxin; Zhang, Jiangjiang; Koh, Kwangnak; Chen, Hongxia

    2016-07-15

    Cucurbit[7]uril (CB[7]) has received increasing attention because of its unique structure and multiple recognition properties. To improve the sensitivity of surface plasmon resonance (SPR) biosensors, we designed a novel strategy in which caspase-3 serves as the model analyte and CB[7]-modified AuNPs (CB[7]-AuNPs) act as the intermedium. The substrate peptides can be cleaved and replaced with a new N-terminal Phe residue in presence of caspase-3. The CB[7]-AuNPs combine with the N-terminal Phe on the gold chip surface through incorporating the side chain within the nonpolar CB[7] cavity and chelating the N-terminal ammonium group with CB[7] carbonyl oxygen. Then CB[7]-AuNPs integrate with short peptide-modified AuNPs containing Phe at the N-terminal of the peptide. SPR signals are significantly improved through the layer-by-layer assembly of AuNPs. The well-designed sensing platform allows the detection of caspase-3 in a linear range from 10fg/mL to 10(3)fg/mL with a detection limit of 2.2 fg/mL. Given its high specificity and desirable sensitivity, this CB[7]-assisted SPR method may be a useful tool for the assay of caspase-3 in the future. This work may also afford a new model to improve the sensitivity and selectivity of SPR biosensors in other protein detection experiments and disease diagnosis.

  1. An electrochemical dopamine aptasensor incorporating silver nanoparticle, functionalized carbon nanotubes and graphene oxide for signal amplification.

    PubMed

    Bahrami, Shokoh; Abbasi, Amir Reza; Roushani, Mahmoud; Derikvand, Zohreh; Azadbakht, Azadeh

    2016-10-01

    In this work, immobilization of a dopamine (DA) aptamer was performed at the surface of an amino functionalized silver nanoparticle-carbon nanotube graphene oxide (AgNPs/CNTs/GO) nanocomposite. A 58-mer DA-aptamer was immobilized through the formation of phosphoramidate bonds between the amino group of chitosan and the phosphate group of the aptamer at the 5' end. An AgNPs/CNTs/GO nanocomposite was employed as a highly catalytic label for electrochemical detection of DA based on electrocatalytic activity of the nanocomposite toward hydrogen peroxide (H2O2). Interaction of DA with the aptamer caused conformational changes of the aptamer which, in turn, decreased H2O2 oxidation and reduction peak currents. On the other hand, the presumed folding of the DA-aptamer complexes on the sensing interface inhibited the electrocatalytic activity of AgNPs/CNTs/GO toward H2O2. Sensitive quantitative detection of DA was carried out by monitoring the decrease of differential pulse voltammetric (DPV) responses of AgNPs/CNTs/GO nanocomposite toward H2O2 oxidation. The DPV signal linearly decreased with increased concentration of DA from 3 to 110nmolL(-1) with a detection limit of 700±19.23pmolL(-1). Simple preparation, low operation cost, speed and validity are the decisive factors of this method motivating its application to biosensing investigation.

  2. Multifunctional reduced graphene oxide trigged chemiluminescence resonance energy transfer: Novel signal amplification strategy for photoelectrochemical immunoassay of squamous cell carcinoma antigen.

    PubMed

    Zhang, Yan; Sun, Guoqiang; Yang, Hongmei; Yu, Jinghua; Yan, Mei; Song, Xianrang

    2016-05-15

    Herein, a photoelectrochemical (PEC) immunoassay is constructed for squamous cell carcinoma antigen (SCCA) detection using zinc oxide nanoflower-bismuth sulfide (Bi2S3) composites as photoactive materials and reduced graphene oxide (rGO) as signal labels. Horseradish peroxidase is used to block sites against nonspecific binding, and then participated in luminol-based chemiluminescence (CL) system. The induced CL emission is acted as an inner light source to excite photoactive materials, simplifying the instrument. A novel signal amplification strategy is stem from rGO because of the rGO acts as an energy acceptor, while luminol serves as a donor to rGO, triggering the CL resonance energy transfer phenomenon between luminol and rGO. Thus, the efficient CL emission to photoactive materials decreases. Furthermore, the signal amplification caused by rGO labeled signal antibodies is related to photogenerated electron-hole pairs: perfect matching of energy levels between rGO and Bi2S3 makes rGO a sink to capture photogenerated electrons from Bi2S3; the increased steric hindrance hinders the electron donor to the surface of Bi2S3 for reaction with the photogenerated holes. On the basis of the novel signal amplification strategy, the proposed immunosensor exhibits excellent analytical performance for PEC detection of SCCA, ranging from 0.8 pg mL(-1) to 80 ng mL(-1) with a low detection limit of 0.21 pg mL(-1). Meanwhile, the designed signal amplification strategy provides a general format for future development of PEC assays.

  3. Electrochemical DNA probe for Hg(2+) detection based on a triple-helix DNA and Multistage Signal Amplification Strategy.

    PubMed

    Wang, Huan; Zhang, Yihe; Ma, Hongmin; Ren, Xiang; Wang, Yaoguang; Zhang, Yong; Wei, Qin

    2016-12-15

    In this work, an ultrasensitive electrochemical sensor was developed for detection of Hg(2+). Gold nanoparticles decorated bovine serum albumin reduction of graphene oxide (AuNP-BSA-rGO) were used as subsurface material for the immobilization of triple-helix DNA. The triple-helix DNA containing a thiol labelled single-stranded DNA (sDNA) and a thymine-rich DNA (T-rich DNA), which could be unwinded in the present of Hg(2+) to form more stable thymine-Hg(2+)-thymine (T-Hg(2+)-T) complex. T-Hg(2+)-T complex was then removed and the sDNA was left on the electrode. At this time, gold nanoparticle carrying thiol labelled cytosine-rich complementary DNA (cDNA-AuNP) could bind with the free sDNA. Meanwhile, the other free cDNA on AuNP could bind with each other in the present of Ag(+) to form the stable cytosine-Ag(+)-cytosine (C-Ag(+)-C) complex and circle amplification. Plenty of C-Ag(+)-C could form silver nanoclusters by electrochemical reduction and the striping signal of Ag could be measured for purpose of the final electrochemical detection of Hg(2+). This sensor could detect Hg(2+) over a wide concentration range from 0.1 to 130nM with a detection limit of 0.03nM.

  4. Detection of single-nucleotide polymorphisms with novel leaky surface acoustic wave biosensors, DNA ligation and enzymatic signal amplification.

    PubMed

    Xu, Qinghua; Chang, Kai; Lu, Weiping; Chen, Wei; Ding, Yi; Jia, Shuangrong; Zhang, Kejun; Li, Fake; Shi, Jianfeng; Cao, Liang; Deng, Shaoli; Chen, Ming

    2012-03-15

    This manuscript describes a new technique for detecting single-nucleotide polymorphisms (SNPs) by integrating a leaky surface acoustic wave (LSAW) biosensor, enzymatic DNA ligation and enzymatic signal amplification. In this technique, the DNA target is hybridized with a capture probe immobilized on the surface of a LSAW biosensor. Then, the hybridized sequence is ligated to biotinylated allele-specific detection probe using Taq DNA ligase. The ligation does not take place if there is a single-nucleotide mismatch between the target and the capture probe. The ligated detection probe is transformed into a streptavidin-horseradish peroxidase (SA-HRP) terminal group via a biotin-streptavidin complex. Then, the SA-HRP group catalyzes the polymerization of 3,3-diaminobenzidine (DAB) to form a surface precipitate, thus effectively increasing the sensitivity of detecting surface mass changes and allowing detection of SNPs. Optimal detection conditions were found to be: 0.3 mol/L sodium ion concentration in PBS, pH 7.6, capture probe concentration 0.5 μmol/L and target sequence concentration 1.0 μmol/L. The detection limit was found to be 1 × 10(-12)mol/L. Using this technique, we were able to detect a single-point mutation at nucleotide A2293G in Japanese encephalitis virus.

  5. Sequence amplification via cell passaging creates spurious signals of positive adaptation in influenza virus H3N2 hemagglutinin

    PubMed Central

    McWhite, Claire D.; Meyer, Austin G.; Wilke, Claus O.

    2016-01-01

    Clinical influenza A virus isolates are frequently not sequenced directly. Instead, a majority of these isolates (~70% in 2015) are first subjected to passaging for amplification, most commonly in non-human cell culture. Here, we find that this passaging leaves distinct signals of adaptation, which can confound evolutionary analyses of the viral sequences. We find distinct patterns of adaptation to Madin-Darby (MDCK) and monkey cell culture absent from unpassaged hemagglutinin sequences. These patterns also dominate pooled datasets not separated by passaging type, and they increase in proportion to the number of passages performed. By contrast, MDCK-SIAT1 passaged sequences seem mostly (but not entirely) free of passaging adaptations. Contrary to previous studies, we find that using only internal branches of influenza virus phylogenetic trees is insufficient to correct for passaging artifacts. These artifacts can only be safely avoided by excluding passaged sequences entirely from subsequent analysis. We conclude that future influenza virus evolutionary analyses should appropriately control for potentially confounding effects of passaging adaptations. PMID:27713835

  6. Ultrasensitive sandwich-type electrochemical immunosensor based on trimetallic nanocomposite signal amplification strategy for the ultrasensitive detection of CEA

    PubMed Central

    Tian, Lihui; Liu, Li; Li, Yueyuan; Wei, Qin; Cao, Wei

    2016-01-01

    A novel and ultrasensitive sandwich-type electrochemical immunosensor was designed for the quantitative detection of carcino-embryonic antigen (CEA). This immunosensor was developed by using the trimetallic NiAuPt nanoparticles on graphene nanosheets (NGs) nanosheets (NiAuPt-NGs) as excellent labels and β-cyclodextrin functionalized reduced graphene oxide nanosheets (CD-NGs) as the platform. The CD-NGs with high specific surface area good biocompatibility and the ideal dispersibility was used to capture the primary antibodies (Ab1) efficiently. The trimetallic NiAuPt-NGs nanocomposites were used as the labels for signal amplification, showing better electrocatalytic activity towards the reduction of hydrogen peroxide (H2O2), which is much better than that the monometallic Pt-NGs, bimetallic NiPt-NGs and AuPt-NGs due to the synergetic effect presented in NiAuPt-NGs. The NiAuPt-NGs nanocomposites consist of tightly coupled nanostructures of Au, Ni and Pt, which have neither an alloy nor a core-shell structure. Under the optimal conditions, a linear range from 0.001–100 ng/mL and a low detection limit of 0.27 pg/mL were obtained for CEA. The proposed electrochemical sandwich-type immunosensor may have a promising application in bioassay and it enriches the electrochemical immunoassays. PMID:27488806

  7. Ultrasensitive sandwich-type photoelectrochemical immunosensor based on CdSe sensitized La-TiO2 matrix and signal amplification of polystyrene@Ab2 composites.

    PubMed

    Fan, Dawei; Ren, Xiang; Wang, Haoyuan; Wu, Dan; Zhao, Di; Chen, Yucheng; Wei, Qin; Du, Bin

    2017-01-15

    A novel and sensitive sandwich-type photoelectrochemical (PEC) sensor was fabricated using signal amplification strategy for the quantitative detection of the prostate specific antigen (PSA). CdSe nanoparticles (NPs) sensitized lanthanum-doped titanium dioxide (La-TiO2) composites were used to bind the primary antibodies (Ab1). The doping of lanthanum promoted the visible light absorption of TiO2 and remarkably enhanced the photocurrent. Moreover, 0.3%La-TiO2 displayed the highest photocurrent in the La-TiO2 composites, which was twice as much as that of undoped TiO2. Carboxyl modified CdSe NPs were assembled onto La-TiO2 composites via the dentate binding between -COOH and Ti atom in TiO2 NPs, which dramatically promoted the photocurrent intensity by approximately 2.1 times. Carboxyl functionalized polystyrene (PS) microspheres were coated with the secondary antibodies (Ab2). Owing to the better insulation property and steric hindrance of the prepared polystyrene@Ab2 (PS@Ab2) composites, the significant reduction of the photocurrent signal was achieved after the specific immune recognition. Under the optimum experimental conditions, the fabricated PEC sensor realized ultrasensitive detection of PSA in the range of 0.05-100pgmL(-1) with a detection limit of 17fgmL(-1). Moreover, this well-designed PEC immunoassay exhibited ideal reproducibility, stability, and selectivity, which is a promising platform for the detection of other important tumor targets.

  8. Enzyme-catalysed deposition of ultrathin silver shells on gold nanorods: a universal and highly efficient signal amplification strategy for translating immunoassay into a litmus-type test.

    PubMed

    Yang, Xinjian; Gao, Zhiqiang

    2015-04-25

    On the basis of enzyme-catalysed reduction of silver ions and consequent deposition of ultrathin silver shells on gold nanorods, a highly efficient signal amplification method for immunoassay is developed. For a model analyte prostate-specific antigen, a 10(4)-fold improvement over conventional enzyme-linked immunosorbent assay is accomplished by leveraging on the cumulative nature of the enzymatic reaction and the sensitive response of plasnomic gold nanorods to the deposition the silver shells.

  9. Label-free and sensitive fluorescence detection of nucleic acid, based on combination of a graphene oxid /SYBR green I dye platform and polymerase assisted signal amplification

    NASA Astrophysics Data System (ADS)

    Zhu, Xiao; Xing, Da

    2012-12-01

    A new label-free isothermal fluorescence amplification detection for nucleic acid has been developed. In this paper, we first developed a novel sensitive and specific detection platform with an unmodified hairpin probe (HP) combination of the graphene oxid (GO)/ SYBR green I dye (SG), which was relied on the selective principle of adsorption and the high quenching efficiency of GO. Then for the application of this new strategy, we used Mirco RNA-21 (Mir-21) as the target to evaluate this working principle of our design. When the target was hybridizing with the HP and inducing its conformation of change, an efficient isothermal circular strand-displacement polymerization reaction was activating to assist the first signal amplification. In this format, the formed complex conformation of DNA would interact with its high affinity dye, then detached from the surface of GO after incubating with the platform of GO/intercalating dye. This reaction would accompany with obvious fluorescence recovery, and accomplish farther signal enhancement by a mass of intercalating dye inserting into the minor groove of the long duplex replication product. By taking advantage of the multiple amplification of signal, this method exerted substantial enhancement in sensitivity and could be used for rapid and selective detection of Mir-21 with attomole range. It is expected that this cost-effective GO based sensor might hold considerable potential to apply in bioanalysis studies.

  10. A signal-on electrochemical aptasensor for ultrasensitive detection of endotoxin using three-way DNA junction-aided enzymatic recycling and graphene nanohybrid for amplification

    NASA Astrophysics Data System (ADS)

    Bai, Lijuan; Chai, Yaqin; Pu, Xiaoyun; Yuan, Ruo

    2014-02-01

    Endotoxin, also known as lipopolysaccharide (LPS), is able to induce a strong immune response on its internalization into mammalian cells. To date, aptamer-based biosensors for LPS detection have been rarely reported. This work describes a new signal-on electrochemical aptasensor for the ultrasensitive detection of LPS by combining the three-way DNA hybridization process and nanotechnology-based amplification. With the help of DNA1 (associated with the concentration of target LPS), the capture probe hybridizes with DNA1 and the assistant probe to open its hairpin structure and form a ternary ``Y'' junction structure. The DNA1 can be released from the structure in the presence of nicking endonuclease to initiate the next hybridization process. Then a great deal of cleaved capture probe produced in the cyclic process can bind with DNA2-nanocomposite, which contains the electroactive toluidine blue (Tb) with the amplification materials graphene (Gra) and gold nanoparticles (AuNPs). Thus, an enhanced electrochemical signal can be easily read out. With the cascade signal amplification, this newly designed protocol provides an ultrasensitive electrochemical detection of LPS down to the femtogram level (8.7 fg mL-1) with a linear range of 6 orders of magnitude (from 10 fg mL-1 to 50 ng mL-1). Moreover, the high sensitivity and specificity make this method versatile for the detection of other biomolecules by changing the corresponding sequences of the capture probe and the assistant probe.

  11. A signal-on electrochemical aptasensor for ultrasensitive detection of endotoxin using three-way DNA junction-aided enzymatic recycling and graphene nanohybrid for amplification.

    PubMed

    Bai, Lijuan; Chai, Yaqin; Pu, Xiaoyun; Yuan, Ruo

    2014-03-07

    Endotoxin, also known as lipopolysaccharide (LPS), is able to induce a strong immune response on its internalization into mammalian cells. To date, aptamer-based biosensors for LPS detection have been rarely reported. This work describes a new signal-on electrochemical aptasensor for the ultrasensitive detection of LPS by combining the three-way DNA hybridization process and nanotechnology-based amplification. With the help of DNA1 (associated with the concentration of target LPS), the capture probe hybridizes with DNA1 and the assistant probe to open its hairpin structure and form a ternary "Y" junction structure. The DNA1 can be released from the structure in the presence of nicking endonuclease to initiate the next hybridization process. Then a great deal of cleaved capture probe produced in the cyclic process can bind with DNA2-nanocomposite, which contains the electroactive toluidine blue (Tb) with the amplification materials graphene (Gra) and gold nanoparticles (AuNPs). Thus, an enhanced electrochemical signal can be easily read out. With the cascade signal amplification, this newly designed protocol provides an ultrasensitive electrochemical detection of LPS down to the femtogram level (8.7 fg mL(-1)) with a linear range of 6 orders of magnitude (from 10 fg mL(-1) to 50 ng mL(-1)). Moreover, the high sensitivity and specificity make this method versatile for the detection of other biomolecules by changing the corresponding sequences of the capture probe and the assistant probe.

  12. Development of an efficient signal amplification strategy for label-free enzyme immunoassay using two site-specific biotinylated recombinant proteins.

    PubMed

    Tang, Jin-Bao; Tang, Ying; Yang, Hong-Ming

    2015-02-15

    Constructing a recombinant protein between a reporter enzyme and a detector protein to produce a homogeneous immunological reagent is advantageous over random chemical conjugation. However, the approach hardly recombines multiple enzymes in a difunctional fusion protein, which results in insufficient amplification of the enzymatic signal, thereby limiting its application in further enhancement of analytical signal. In this study, two site-specific biotinylated recombinant proteins, namely, divalent biotinylated alkaline phosphatase (AP) and monovalent biotinylated ZZ domain, were produced by employing the Avitag-BirA system. Through the high streptavidin (SA)-biotin interaction, the divalent biotinylated APs were clustered in the SA-biotin complex and then incorporated with the biotinylated ZZ. This incorporation results in the formation of a functional macromolecule that involves numerous APs, thereby enhancing the enzymatic signal, and in the production of several ZZ molecules for the interaction with immunoglobulin G (IgG) antibody. The advantage of this signal amplification strategy is demonstrated through ELISA, in which the analytical signal was substantially enhanced, with a 32-fold increase in the detection sensitivity compared with the ZZ-AP fusion protein approach. The proposed immunoassay without chemical modification can be an alternative strategy to enhance the analytical signals in various applications involving immunosensors and diagnostic chips, given that the label-free IgG antibody is suitable for the ZZ protein.

  13. A sensitive magnetic nanoparticle-based immunoassay of phosphorylated acetylcholinesterase using protein cage templated lead phosphate for signal amplification with graphite furnace atomic absorption spectrometry detection.

    PubMed

    Liang, Pei; Kang, Caiyan; Yang, Enjian; Ge, Xiaoxiao; Du, Dan; Lin, Yuehe

    2016-04-07

    We developed a new magnetic nanoparticle sandwich-like immunoassay using protein cage nanoparticles (PCN) for signal amplification together with graphite furnace atomic absorption spectrometry (GFAAS) for the quantification of an organophosphorylated acetylcholinesterase adduct (OP-AChE), the biomarker of exposure to organophosphate pesticides (OPs) and nerve agents. OP-AChE adducts were firstly captured by titanium dioxide coated magnetic nanoparticles (TiO2-MNPs) from the sample matrixes through metal chelation with phospho-moieties, and then selectively recognized by anti-AChE antibody labeled on PCN which was packed with lead phosphate in its cavity (PCN-anti-AChE). The sandwich-like immunoreaction was performed among TiO2-MNPs, OP-AChE and PCN-anti-AChE to form a TiO2-MNP/OP-AChE/PCN-anti-AChE immunocomplex. The complex could be easily isolated from the sample solution with the help of magnet, and the released lead ions from PCN were detected by GFAAS for the quantification of OP-AChE. Greatly enhanced sensitivity was achieved because PCN increased the amount of metal ions in the cavity of each apoferritin. The proposed immunoassay yielded a linear response over a broad range of OP-AChE concentrations from 0.01 nM to 2 nM, with a detection limit of 2 pM, which has enough sensitivity for monitoring of low-dose exposure to OPs. This new method showed an acceptable stability and reproducibility and was validated with OP-AChE spiked human plasma.

  14. Wettability Switching of Electrode for Signal Amplification: Conversion of Conformational Change of Stimuli-Responsive Polymer into Enhanced Electrochemical Chiral Analysis.

    PubMed

    Ding, Shushu; Cao, Sumei; Zhu, Anwei; Shi, Guoyue

    2016-12-20

    Signal amplification of chiral interaction is a much needed task for sensing of enantiomers due to nearly identical chemical and physical properties of the chiral isomers. In this article, we established an electrochemical chiral sensing method with high sensitivity and selectivity for monosacharrides based on the stimuli-responsive copolymer/graphene hybrid-modified screen-printed carbon electrodes. The hybrid synthesized by the "grafting from" atom transfer radical polymerization (ATRP) process not only acted as a chiral recognition element but also provided a chiral signal amplification strategy. This occurs due to high sensitivity of conformational transition of copolymer on graphene to the weak chiral interactions that greatly facilitating the diffusion of electroactive probes and monosaccharides to the electrode surface. The described method can quantify monosaccharides, even the concentration of one enantiomer is as low as 1 nM. Apart from the demonstrated chiral distinguish ability, good selectivity toward monosaccharides in comparison to potential interference molecules was also observed. The electrodes with significant analytical performance were successfully applied for discriminating glucose enantiomers in live cells and studying their different transport mechanism. Together, the results show that the coupling of amplification-by-wettability switching concept with electrochemical method offers great promises in providing a sensitive, facile, and cost-effective solution for chiral recognition of molecules in biological process.

  15. Novel signal amplification strategy for ultrasensitive sandwich-type electrochemical immunosensor employing Pd-Fe3O4-GS as the matrix and SiO2 as the label.

    PubMed

    Wang, Yulan; Ma, Hongmin; Wang, Xiaodong; Pang, Xuehui; Wu, Dan; Du, Bin; Wei, Qin

    2015-12-15

    An ultrasensitive sandwich-type electrochemical immunosensor based on a novel signal amplification strategy was developed for the quantitative determination of human immunoglobulin G (IgG). Pd nanocubes functionalized magnetic graphene sheet (Pd-Fe3O4-GS) was employed as the matrix to immobilize the primary antibodies (Ab1). Owing to the synergetic effect between Pd nanocubes and magnetic graphene sheet (Fe3O4-GS), Pd-Fe3O4-GS can provide an obviously increasing electrochemical signal by electrochemical catalysis towards hydrogen peroxide (H2O2). Silicon dioxide (SiO2) was functionalized as the label to conjugate with the secondary antibodies (Ab2). Due to the larger steric hindrance of the obtained conjugate (SiO2@Ab2), the sensitive decrease of the electrochemical signal can be achieved after the specific recognition between antibodies and antigens. In this sense, this proposed immunosensor can achieve a high sensitivity, especially in the presence of low concentrations of IgG. Under optimum conditions, the proposed immunosensor offered an ultrasensitive and specific determination of IgG down to 3.2 fg/mL. This immunoassay method would open up a new promising platform to detect various tumor markers at ultralow levels for early diagnoses of different cancers.

  16. Kerr nonlinearity mitigation in 5 × 28-GBd PDM 16-QAM signal transmission over a dispersion-uncompensated link with backward-pumped distributed Raman amplification.

    PubMed

    Sackey, Isaac; Da Ros, Francesco; Jazayerifar, Mahmoud; Richter, Thomas; Meuer, Christian; Nölle, Markus; Molle, Lutz; Peucheret, Christophe; Petermann, Klaus; Schubert, Colja

    2014-11-03

    We present experimental and numerical investigations of Kerr nonlinearity compensation in a 400-km standard single-mode fiber link with distributed Raman amplification with backward pumping. A dual-pump polarization-independent fiber-based optical parametric amplifier is used for mid-link spectral inversion of 5 × 28-GBd polarization-multiplexed 16-QAM signals. Signal quality factor (Q-factor) improvements of 1.1 dB and 0.8 dB were obtained in the cases of a single-channel and a five-channel wavelength-division multiplexing (WDM) system, respectively. The experimental results are compared to numerical simulations with good agreement. It is also shown with simulations that a maximum transmission reach of 2400 km enabled by the optical phase conjugator is possible for the WDM signal.

  17. A label-free signal amplification assay for DNA detection based on exonuclease III and nucleic acid dye SYBR Green I.

    PubMed

    Zheng, Aihua; Luo, Ming; Xiang, Dongshan; Xiang, Xia; Ji, Xinghu; He, Zhike

    2013-09-30

    We have developed a new fluorescence method for specific single-stranded DNA sequences with exonuclease III (Exo III) and nucleic acid dye SYBR Green I. It is demonstrated by a reverse transcription oligonucleotide sequence (target DNA, 27 bases) of RNA fragment of human immunodeficiency virus (HIV) as a model system. In the absence of the target DNA, the hairpin-probe is in the stem-closed structure, the fluorescence of SYBR Green I is very strong. In the presence of the target DNA, the hairpin-probe hybridizes with the target DNA to form double-stranded structure with a blunt 3'-terminus. Thus, in the presence of Exo III, only the 3'-terminus of probe is subjected to digestion. Exo III catalyzes the stepwise removal of mononucleotides from this terminus, releasing the target DNA. The released target DNA then hybridizes with another probe, whence the cycle starts anew. The signal of SYBR Green I decreases greatly. This system provides a detection limit of 160 pM, which is comparable to the existing signal amplification methods that utilized Exo III as a signal amplification nuclease. Due to the unique property of Exo III, this method shows excellent detection selectivity for single-base discrimination. More importantly, superiors to other methods based on Exo III, these probes have the advantages of easier to design, synthesize, purify and thus are much cheaper and more applicable. This new approach could be widely applied to sensitive and selective nucleic acids detection.

  18. Combination of a modified block PCR and endonuclease IV-based signal amplification system for ultra-sensitive detection of low-abundance point mutations.

    PubMed

    Xiao, Xianjin; Xu, Anqin; Zhai, Junqiu; Zhao, Meiping

    2013-12-15

    By combination of a modified block PCR and endonuclease IV-based signal amplification system, we have developed a novel approach for ultra-sensitive detection of point mutations. The method can effectively identify mutant target sequence immersed in a large background of wild-type sequences with abundance down to 0.03% (for C→A) and 0.005% (for C→G). This sensitivity is among the highest in comparison with other existing approaches and the operating procedures are simple and time saving. The method holds great potential for future application in clinical diagnosis and biomedical research.

  19. Direct RNA detection without nucleic acid purification and PCR: Combining sandwich hybridization with signal amplification based on branched hybridization chain reaction.

    PubMed

    Xu, Yao; Zheng, Zhi

    2016-05-15

    We have developed a convenient, robust and low-cost RNA detection system suitable for high-throughput applications. This system uses a highly specific sandwich hybridization to capture target RNA directly onto solid support, followed by on-site signal amplification via 2-dimensional, branched hybridizing chain polymerization through toehold-mediated strand displacement reaction. The assay uses SYBR Green to detect targets at concentrations as low as 1 pM, without involving nucleic acid purification or any enzymatic reaction, using ordinary oligonucleotides without modification or labeling. The system was demonstrated in the detection of malaria RNA in blood and GAPDH gene expression in cell lysate.

  20. Detection of biological molecules using boronate-based chemical amplification and optical sensors

    DOEpatents

    Van Antwerp, William Peter; Mastrototaro, John Joseph; Lane, Stephen M.; Satcher, Jr., Joe H.; Darrow, Christopher B.; Peyser, Thomas A.; Harder, Jennifer

    1999-01-01

    Methods are provided for the determination of the concentration of biological levels of polyhydroxylated compounds, particularly glucose. The methods utilize an amplification system that is an analyte transducer immobilized in a polymeric matrix, where the system is implantable and biocompatible. Upon interrogation by an optical system, the amplification system produces a signal capable of detection external to the skin of the patient. Quantitation of the analyte of interest is achieved by measurement of the emitted signal.

  1. Detection of biological molecules using boronate-based chemical amplification and optical sensors

    DOEpatents

    Van Antwerp, William Peter; Mastrototaro, John Joseph; Lane, Stephen M.; Satcher, Jr., Joe H.; Darrow, Christopher B.; Peyser, Thomas A.; Harder, Jennifer

    2004-06-15

    Methods are provided for the determination of the concentration of biological levels of polyhydroxylated compounds, particularly glucose. The methods utilize an amplification system that is an analyte transducer immobilized in a polymeric matrix, where the system is implantable and biocompatible. Upon interrogation by an optical system, the amplification system produces a signal capable of detection external to the skin of the patient. Quantitation of the analyte of interest is achieved by measurement of the emitted signal.

  2. Tyramine-based enzymatic conjugate repeats for ultrasensitive immunoassay accompanying tyramine signal amplification with enzymatic biocatalytic precipitation.

    PubMed

    Hou, Li; Tang, Yun; Xu, Mingdi; Gao, Zhuangqiang; Tang, Dianping

    2014-08-19

    A new impedimetric immunoassay protocol based on enzyme-triggered formation of tyramine-enzyme repeats on gold nanoparticle (AuNP) was designed for highly sensitive detection of carcinoembryonic antigen (CEA, as a model) by virtue of utilizing enzymatic biocatalytic precipitation toward 4-chloro-1-naphthol (4-CN) on anti-CEA antibody (Ab1)-modified immunosensor. Initially, AuNP was functionalized with horseradish peroxidase and detection antibody (HRP-AuNP-Ab2), and then HRP-tyramine conjugate was utilized for the formation of tyramine-HRP repeats through the triggering of the immobilized HRP on the AuNP with the aid of H2O2. In the presence of target CEA, the carried HRP-tyramine repeats accompanying the sandwiched immunocomplex catalyzed the 4-CN oxidation to produce an insoluble precipitation on the immunosensor, thus causing a local alteration of the conductivity. Three signal-transduction tags including HRP-Ab2, HRP-AuNP-Ab2, and HRP-AuNP-Ab2 with HRP-tyramine repeats were employed for target CEA evaluation, and improved analytical properties were achieved by HRP-AuNP-Ab2 with HRP-tyramine repeats. Using the unique signal-transduction tag, the analytical performance of the impedimetric immunoassay was studied in detail. Under the optimal conditions, the impedimetric immunosensor displayed a wide dynamic working range of between 0.5 pg mL(-1) and 40 ng mL(-1) with a detection limit (LOD) of 0.38 pg mL(-1) relative to target CEA. The coefficients of variation (CVs) were ≤9.3% and 13.3% for the intra-assay and interassay, respectively. The levels of CEA in eight clinical serum specimens were measured by using the developed impedimetric immunosensor. The obtained results correlated well with those from the electrochemiluminescent (ECL)-based immunoassay with a correlation coefficient of 0.998.

  3. Nicking endonuclease-assisted signal amplification of a split molecular aptamer beacon for biomolecule detection using graphene oxide as a sensing platform.

    PubMed

    Li, Xiang; Ding, Xuelian; Fan, Jing

    2015-12-07

    Sensitive and selective detection of ultralow concentrations of specific biomolecules is important in early clinical diagnoses and biomedical applications. Many types of aptasensors have been developed for the detection of various biomolecules, but usually suffer from false positive signals and high background signals. In this work, we have developed an amplified fluorescence aptasensor platform for ultrasensitive biomolecule detection based on enzyme-assisted target-recycling signal amplification and graphene oxide. By using a split molecular aptamer beacon and a nicking enzyme, the typical problem of false positive signals can be effectively resolved. Only in the presence of a target biomolecule, the sensor system is able to generate a positive signal, which significantly improves the selectivity of the aptasensor. Moreover, using graphene oxide as a super-quencher can effectively reduce the high background signal of a sensing platform. We select vascular endothelial growth factor (VEGF) and adenosine triphosphate (ATP) as model analytes in the current proof-of-concept experiments. It is shown that under optimized conditions, our strategy exhibits high sensitivity and selectivity for the quantification of VEGF and ATP with a low detection limit (1 pM and 4 nM, respectively). In addition, this biosensor has been successfully utilized in the analysis of real biological samples.

  4. Detection of biological molecules using chemical amplification and optical sensors

    DOEpatents

    Van Antwerp, William Peter; Mastrototaro, John Joseph

    2001-01-01

    Methods are provided for the determination of the concentration of biological levels of polyhydroxylated compounds, particularly glucose. The methods utilize an amplification system that is an analyte transducer immobilized in a polymeric matrix, where the system is implantable and biocompatible. Upon interrogation by an optical system, the amplification system produces a signal capable of detection external to the skin of the patient. Quantitation of the analyte of interest is achieved by measurement of the emitted signal. Specifically, the analyte transducer immobilized in a polymeric matrix can be a boronic acid moiety.

  5. Real-time colorimetric detection of target DNA using isothermal target and signaling probe amplification and gold nanoparticle cross-linking assay.

    PubMed

    Jung, Cheulhee; Chung, Ji Won; Kim, Un Ok; Kim, Min Hwan; Park, Hyun Gyu

    2011-01-15

    We describe a facile gold nanoparticle (AuNP)-mediated colorimetric method for real-time detection of target DNA in conjugation with our unique isothermal target and signaling probe amplification (iTPA) method, comprising novel ICA (isothermal chain amplification) and CPT (cycling probe technology). Under isothermal conditions, the iTPA simultaneously amplifies the target and signaling probe through two displacement events induced by a combination of four specially designed primers, the strand displacement activity of DNA polymerase, and the RNA degrading activity of RNase H. The resulting target amplicons are hybridized with gold nanoparticle cross-linking assay (GCA) probes having a DNA-RNA-DNA chimeric form followed by RNA cleavage by RNase H in the CPT step. The intact GCA probes were designed to cross-link two sets of DNA-AuNPs conjugates in the absence of target DNA, inducing aggregation (blue color) of AuNPs. On the contrary, the presence of target DNA leads to cleavage of the GCA probes in proportion to the amount of amplified target DNA and the solution remains red in color without aggregation of AuNPs. Relying on this strategy, 10(2) copies of target Chlamydia trachomatis plasmid were successfully detected in a colorimetric manner. Importantly, all the procedures employed up to the final detection of the target DNA were performed under isothermal conditions without requiring any detection instruments. Therefore, this strategy would greatly benefit convenient, real-time monitoring technology of target DNA under restricted environments.

  6. Reduced graphene oxide decorated with gold nanoparticle as signal amplification element on ultra-sensitive electrochemiluminescence determination of caspase-3 activity and apoptosis using peptide based biosensor

    PubMed Central

    Khalilzadeh, Balal; Shadjou, Nasrin; Afsharan, Hadi; Eskandani, Morteza; Nozad Charoudeh, Hojjatollah; Rashidi, Mohammad-Reza

    2016-01-01

    Introduction:Growing demands for ultrasensitive biosensing have led to the development of numerous signal amplification strategies. In this report, a novel electrochemiluminescence (ECL) method was developed for the detection and determination of caspase-3 activity based on reduced graphene oxide sheets decorated by gold nanoparticles as signal amplification element and horseradish peroxidase enzyme (HRP) as ECL intensity enhancing agent. Methods: The ECL intensity of the luminol was improved by using the streptavidin coated magnetic beads and HRP in the presence of hydrogen peroxide. The cleavage behavior of caspase-3 was characterized by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) techniques using biotinylated peptide (DEVD containing peptide) which was coated on reduced graphene oxide decorated with gold nanoparticle. The surface modification of graphene oxide was successfully confirmed by FTIR, UV-vis and x-ray spectroscopy. Results: ECL based biosensor showed that the linear dynamic range (LDR) and the lower limit of quantification (LLOQ) were 0.5-100 and 0.5 femtomolar (fM), respectively. Finally, the performance of the engineered peptide based biosensor was validated in the A549 cell line as real samples. Conclusion: The prepared peptide based biosensor could be considered as an excellent candidate for early detection of apoptosis, cell turnover, and cancer related diseases. PMID:27853677

  7. Novel electrochemical dual-aptamer-based sandwich biosensor using molybdenum disulfide/carbon aerogel composites and Au nanoparticles for signal amplification.

    PubMed

    Fang, Lin-Xia; Huang, Ke-Jing; Liu, Yang

    2015-09-15

    A new electrochemical aptamer biosensor for the platelet-derived growth factor BB (PDGF-BB) detection has been developed based on the signal amplification of MoS2/carbon aerogel composites (MoS2/CA) and sandwich assay. A facile hydrothermal route assisted by L-cysteine was applied to synthesize CA incorporated flower-like MoS2 with the large surface active sites and good conductivity. The electrochemical aptasensor was constructed by sandwiching the PDGF-BB between a glassy carbon electrode modified with thiol-terminated PDGF-BB aptamer-1 (Apt1)/gold nanoparticles (AuNPs)/MoS2/CA and the AuNPs with thiol-terminated PDGF-BB aptamer-2 (Apt2) and 6-ferrocenyl hexanethiol (Fc). Fc-AuNPs-Apt2 acted as tracer and AuNPs/MoS2/CA were utilized as the biosensor platform to immobilize a large amount of capture aptamers, owing to their layered structure and high surface-to-volume ratio. Based on the sandwich format, a dual signal amplification strategy had been successfully developed with a wide linear response in the range of 0.001-10nM and a limit of detection of 0.3 pM. The developed assay demonstrated good selectivity and high sensitivity, indicating potential applications in bioanalysis and biomedicine.

  8. Gold nanoparticles and the corresponding filter membrane as chemosensors and adsorbents for dual signal amplification detection and fast removal of mercury(ii).

    PubMed

    Chen, Gaosong; Hai, Jun; Wang, Hao; Liu, Weisheng; Chen, Fengjuan; Wang, Baodui

    2017-03-02

    Nowadays, the development of a multifunction system for the simultaneous multiple signal amplification detection and fast removal of Hg(2+) remains a major challenge. Herein, we for the first time used gold nanoparticles (Au NPs) and the corresponding filter membrane as chemosensors and adsorbents for dual signal amplification detection and fast removal of Hg(2+). Such a system was based on the formation of gold amalgam and a gold amalgam-based reaction between rhodamine B (RhB) and NaBH4 with fluorescence and colorimetric sensing functions. When the gold amalgam catalyzes the reduction of RhB, the red color and orange fluorescence of RhB gradually changed to colorless by switching the amount of Hg(2+) deposited on 13 nm Au NPs. The detection limit of the fluorescence assay and colorimetric assay is 1.16 nM and 2.54 nM for Hg(2+), respectively. Interestingly, the color and fluorescence of RhB could be recovered when the above colorless reaction solution was exposed to air for about 2 hours. Taking advantage of the above optical phenomenon, a recyclable paper-based sensor has been developed by immobilizing the Au NPs and RhB dye on filter paper and has been successfully used for detection of Hg(2+) in real water samples. In addition, the filter membrane immobilized Au NPs could allow fast removal of mercury ions in Yellow river water and tap water with the removal efficiency close to 99%.

  9. Design and construction of novel molecular conjugates for signal amplification (I): conjugation of multiple horseradish peroxidase molecules to immunoglobulin via primary amines on lysine peptide chains.

    PubMed

    Dhawan, Subhash

    2002-12-01

    Immunoconjugates are widely used for indirect detection of analytes (such as antibodies or antigens) in a variety of immunoassays. However, the availability of functional groups such as primary amines or free sulfhydryls in an immunoglobulin molecule is the limiting factor for optimal conjugation and, therefore, determines the sensitivity of an assay. In the present study, an N-terminal bromoacetylated 20 amino acid peptide containing 20 lysine residues was conjugated to N-succinimidyl-S-acetylthioacetate (SATA)-modified IgG or free sulfhydryl groups on 2-mercaptoethylamine (2-MEA)-reduced IgG molecules via a thioether (S[bond]CH(2)CONH) linkage to introduce multiple reactive primary amines per IgG. These primary amines were then covalently coupled with maleimide-activated horseradish peroxidase (HRP). The poly-HRP-antibody conjugates thus generated demonstrated greater than 15-fold signal amplification upon reaction with orthophenyldiamine substrate. The poly-HRP-antibody conjugates efficiently detected human immunodeficiency virus (HIV)-1 antibodies in plasma specimens with significantly higher sensitivity than conventionally prepared HRP-antibody conjugates in an HIV-1 solid-phase enzyme immunoassay and Western blot analysis. The signal amplification techniques reported here could have the potential for development of highly sensitive immunodiagnostic assay systems.

  10. Metal-binding sites are designed to achieve optimal mechanical and signaling properties

    PubMed Central

    Dutta, Anindita; Bahar, Ivet

    2010-01-01

    Many proteins require bound metals to achieve their function. We take advantage of increasing structural data on metal-binding proteins to elucidate three properties: the involvement of metal-binding sites in the global dynamics of the protein, predicted by elastic network models, their exposure/burial to solvent, and their signal-processing properties indicated by Markovian stochastics analysis. Systematic analysis of a dataset of 145 structures reveals that the residues that coordinate metal ions enjoy remarkably efficient and precise signal transduction properties. These properties are rationalized in terms of their physical properties: participation in hinge sites that control the softest modes collectively accessible to the protein and occupancy of central positions minimally exposed to solvent. Our observations suggest that metal-binding sites may have been evolutionary selected to achieve optimum allosteric communication. They also provide insights into basic principles for designing metal-binding sites, which are verified to be met by recently designed de novo metal-binding proteins. PMID:20826340

  11. HRP-Mimicking DNAzyme-Catalyzed in Situ Generation of Polyaniline To Assist Signal Amplification for Ultrasensitive Surface Plasmon Resonance Biosensing.

    PubMed

    Li, Haiyin; Chang, Jiafu; Hou, Ting; Li, Feng

    2017-01-03

    It is well-known that the horseradish peroxidase- (HRP-) mimicking DNAzyme, namely, hemin/G-quadruplex, can effectively catalyze the polymerization of aniline to form DNA-guided polyaniline. Meanwhile, polyaniline exhibits extraordinary electrical, electrochemical, and redox properties, as well as excellent SPR signal-enhancing ability. Herein, we report a novel ultrasensitive surface plasmon resonance (SPR) biosensor based on HRP-mimicking DNAzyme-catalyzed in situ formation of polyaniline for signal amplification, using bleomycin (BLM) as the proof-of-concept analyte. The recognition and the subsequent cleavage of DNA probe P1 by BLM switches off the hybridization between P1 and the G-rich DNA probe P2, resulting in less hemin/G-quadruplex complexes and reduced DNA-guided polyaniline deposition on the SPR Au disk surface. As compared to the case when BLM is absent, a significant shift in SPR angle is observed, which is dependent on the BLM concentration. Therefore, ultrasensitive SPR detection of the target BLM is realized, with a detection limit down to 0.35 pM, much lower than those reported in the literature. Moreover, the proposed SPR biosensor has been successfully applied for the detection of BLM spiked in human serum samples. The HRP-mimicking DNAzyme-catalyzed in situ polyaniline deposition and polyaniline-assisted signal amplification not only significantly improves the specificity and the sensitivity of the BLM assay but also allows the ultrasensitive detection of other biomolecules by simply changing the specific target recognition DNA sequences, thus providing a versatile SPR biosensing platform for the ultrasensitive detection of a variety of analytes and showing great potential for application in the fields of bioanalysis and clinical biomedicine.

  12. Isothermal amplification detection of nucleic acids by a double-nicked beacon.

    PubMed

    Shi, Chao; Zhou, Meiling; Pan, Mei; Zhong, Guilin; Ma, Cuiping

    2016-03-01

    Isothermal and rapid amplification detection of nucleic acids is an important technology in environmental monitoring, foodborne pathogen detection, and point-of-care clinical diagnostics. Here we have developed a novel method of isothermal signal amplification for single-stranded DNA (ssDNA) detection. The ssDNA target could be used as an initiator, coupled with a double-nicked molecular beacon, to originate amplification cycles, achieving cascade signal amplification. In addition, the method showed good specificity and strong anti-jamming capability. Overall, it is a one-pot and isothermal strand displacement amplification method without the requirement of a stepwise procedure, which greatly simplifies the experimental procedure and decreases the probability of contamination of samples. With its advantages, the method would be very useful to detect nucleic acids in point-of-care or field use.

  13. A label-free and high-efficient GO-based aptasensor for cancer cells based on cyclic enzymatic signal amplification.

    PubMed

    Xiao, Kunyi; Liu, Juan; Chen, Hui; Zhang, Song; Kong, Jilie

    2017-05-15

    A label-free and high-efficient graphene oxide (GO)-based aptasensor was developed for the detection of low quantity cancer cells based on cell-triggered cyclic enzymatic signal amplification (CTCESA). In the absence of target cells, hairpin aptamer probes (HAPs) and dye-labeled linker DNAs stably coexisted in solution, and the fluorescence was quenched by the GO-based FÖrster resonance energy transfer (FRET) process. In the presence of target cells, the specific binding of HAPs with the target cells triggered a conformational alternation, which resulted in linker DNA complementary pairing and cleavage by nicking endonuclease-strand scission cycles. Consequently, more cleaved fragments of linker DNAs with more the terminal labeled dyes could show the enhanced fluorescence because these cleaved DNA fragments hardly combine with GOs and prevent the FRET process. Fluorescence analysis demonstrated that this GO-based aptasensor exhibited selective and sensitive response to the presence of target CCRF-CEM cells in the concentration range from 50 to 10(5) cells. The detection limit of this method was 25 cells, which was approximately 20 times lower than the detection limit of normal fluorescence aptasensors without amplification. With high sensitivity and specificity, it provided a simple and cost-effective approach for early cancer diagnosis.

  14. Visual and highly sensitive detection of cancer cells by a colorimetric aptasensor based on cell-triggered cyclic enzymatic signal amplification.

    PubMed

    Zhang, Xianxia; Xiao, Kunyi; Cheng, Liwei; Chen, Hui; Liu, Baohong; Zhang, Song; Kong, Jilie

    2014-06-03

    Rapid and efficient detection of cancer cells at their earliest stages is one of the central challenges in cancer diagnostics. We developed a simple, cost-effective, and highly sensitive colorimetric method for visually detecting rare cancer cells based on cell-triggered cyclic enzymatic signal amplification (CTCESA). In the absence of target cells, hairpin aptamer probes (HAPs) and linker DNAs stably coexist in solution, and the linker DNA assembles DNA-AuNPs, producing a purple solution. In the presence of target cells, the specific binding of HAPs to the target cells triggers a conformational switch that results in linker DNA hybridization and cleavage by nicking endonuclease-strand scission cycles. Consequently, the cleaved fragments of linker DNA can no longer assemble into DNA-AuNPs, resulting in a red color. UV-vis spectrometry and photograph analyses demonstrated that this CTCESA-based method exhibited selective and sensitive colorimetric responses to the presence of target CCRF-CEM cells, which could be detected by the naked eye. The linear response for CCRF-CEM cells in a concentration range from 10(2) to 10(4) cells was obtained with a detection limit of 40 cells, which is approximately 20 times lower than the detection limit of normal AuNP-based methods without amplification. Given the high specificity and sensitivity of CTCESA, this colorimetric method provides a sensitive, label-free, and cost-effective approach for early cancer diagnosis and point-to-care applications.

  15. A T7exonuclease-assisted target recycling amplification with graphene oxide acting as the signal amplifier for fluorescence polarization detection of human immunodeficiency virus (HIV) DNA.

    PubMed

    Wang, Lijun; Tian, Jianniao; Yang, Wen; Zhao, Yanchun; Zhao, Shulin

    2016-03-01

    We report a fluorescence polarization (FP) platform for human immunodeficiency virus (HIV) DNA detection based on T7exonuclease-assisted target recycling amplification with graphene oxide (GO) acting as a FP signal amplifier. In the sensing method, the presence of the target DNA leads to target recycling with the assistance of T7exonuclease, furthermore, the amplification products are absorbed onto the surface of GO, so the all FP values are enhanced by GO. More importantly, this FP sensor exhibits high detection sensitivity; under optimal conditions, the change in FP is linear with the concentration of the target DNA within a concentration range of 50-2000 pmol/L, and the detection limit of this method is as low as 38.6 pmol/L. This FP sensor also exhibits high selectivity, even single-base mismatched DNA can be effectively discriminated from complementary target DNA. Above all, the proposed FP sensor may serve as a general platform for the sensitive assay of disease-related genes.

  16. DNA typing by microbead arrays and PCR-SSP: apparent false-negative or -positive hybridization or amplification signals disclose new HLA-B and -DRB1 alleles.

    PubMed

    Rahal, M; Kervaire, B; Villard, J; Tiercy, J-M

    2008-03-01

    Human leukocyte antigen (HLA) typing by polymerase chain reaction-sequence-specific oligonucleotide (PCR-SSO) hybridization on solid phase (microbead assay) or polymerase chain reaction-sequence-specific primers (PCR-SSP) requires interpretation softwares to detect all possible allele combinations. These programs propose allele calls by taking into account false-positive or false-negative signal(s). The laboratory has the option to validate typing results in the presence of strongly cross-reacting or apparent false-negative signals. Alternatively, these seemingly aberrant signals may disclose novel variants. We report here four new HLA-B (B*5620 and B*5716) and HLA-DRB1 alleles (DRB1*110107 and DRB1*1474) that were detected by apparent false-negative or -positive hybridization or amplification patterns, and ultimately resolved by sequencing. To avoid allele misassignments, a comprehensive evaluation of acquired data as documented in a quality assurance system is therefore required to confirm unambiguous typing interpretation.

  17. A functional glycoprotein competitive recognition and signal amplification strategy for carbohydrate-protein interaction profiling and cell surface carbohydrate expression evaluation

    NASA Astrophysics Data System (ADS)

    Wang, Yangzhong; Chen, Zhuhai; Liu, Yang; Li, Jinghong

    2013-07-01

    A simple and sensitive carbohydrate biosensor has been suggested as a potential tool for accurate analysis of cell surface carbohydrate expression as well as carbohydrate-based therapeutics for a variety of diseases and infections. In this work, a sensitive biosensor for carbohydrate-lectin profiling and in situ cell surface carbohydrate expression was designed by taking advantage of a functional glycoprotein of glucose oxidase acting as both a multivalent recognition unit and a signal amplification probe. Combining the gold nanoparticle catalyzed luminol electrogenerated chemiluminescence and nanocarrier for active biomolecules, the number of cell surface carbohydrate groups could be conveniently read out. The apparent dissociation constant between GOx@Au probes and Con A was detected to be 1.64 nM and was approximately 5 orders of magnitude smaller than that of mannose and Con A, which would arise from the multivalent effect between the probe and Con A. Both glycoproteins and gold nanoparticles contribute to the high affinity between carbohydrates and lectin. The as-proposed biosensor exhibits excellent analytical performance towards the cytosensing of K562 cells with a detection limit of 18 cells, and the mannose moieties on a single K562 cell were determined to be 1.8 × 1010. The biosensor can also act as a useful tool for antibacterial drug screening and mechanism investigation. This strategy integrates the excellent biocompatibility and multivalent recognition of glycoproteins as well as the significant enzymatic catalysis and gold nanoparticle signal amplification, and avoids the cell pretreatment and labelling process. This would contribute to the glycomic analysis and the understanding of complex native glycan-related biological processes.A simple and sensitive carbohydrate biosensor has been suggested as a potential tool for accurate analysis of cell surface carbohydrate expression as well as carbohydrate-based therapeutics for a variety of diseases and

  18. A upconversion luminescene biosensor based on dual-signal amplification for the detection of short DNA species of c-erbB-2 oncogene

    PubMed Central

    Lan, Jianming; Liu, Yingxin; Li, Li; Wen, Fadi; Wu, Fang; Han, Zhizhong; Sun, Weiming; Li, Chunyan; Chen, Jinghua

    2016-01-01

    High-sensitivity detection of trace amounts of c-erbB-2 oncogene was reported to be equal to or surpass the ability of CA 15-3 for early diagnosis and/or follow-up recurrent screening of breast cancer. Therefore, in the current study, by using upconversion nanoparticles (UCNPs), rare earth-doped NaYF4:Yb3+/Er3+ as the luminescent labels, a upconversion luminescent (UCL) biosensor based on dual-signal amplification of exonuclease III (ExoIII)-assisted target cycles and long-range self-assembly DNA concatamers was developed for the detection of c-erbB-2 oncogene. The proposed biosensor exhibited ultrasensitive detection with limit as low as 40 aM, which may express the potential of being used in trace analysis of c-erbB-2 oncogene and early diagnosis of breast cancer. PMID:27098295

  19. A low cost technique for synthesis of gold nanoparticles using microwave heating and its application in signal amplification for detecting Escherichia Coli O157:H7 bacteria

    NASA Astrophysics Data System (ADS)

    Thanh Ngo, Vo Ke; Giang Nguyen, Dang; Phat Huynh, Trong; Lam, Quang Vinh

    2016-09-01

    In the present work a low cost technique for preparation of gold nanoparticles (AuNPs) using microwave heating was developed. The effect of different elements (precursor reagents, irradiation time, and microwave radiation power) on the final morphology of AuNPs obtained through the microwave assisted technique has been investigated. The characterization of the samples has been carried out by transmission electron microscopy, UV-vis absorption spectroscopy, Fourier transform infrared spectroscopy, and powder x-ray diffraction. The results showed that to some extent the above-mentioned characterizations influenced the size of synthetized nanoparticles and application of microwave heating has many advantages such as low cost, rapid preparation and highly uniform particles. As an application in quartz crystal microbalance (QCM) immunosensor, AuNPs are conjugated with the Escherichia coli (E.coli) O157:H7 antibodies for signal amplification to detect E.coli O157:H7 bacteria residual in QCM system.

  20. Rapid and sensitive detection of Escherichia coli O157:H7 in milk and ground beef using magnetic bead-based immunoassay coupled with tyramide signal amplification.

    PubMed

    Aydin, Muhsin; Herzig, Gene P D; Jeong, Kwang Cheol; Dunigan, Samantha; Shah, Parth; Ahn, Soohyoun

    2014-01-01

    Escherichia coli O157:H7 is a major foodborne pathogen that has posed serious problems for food safety and public health. Recent outbreaks and recalls associated with various foods contaminated by E. coli O157:H7 clearly indicate its deleterious effect on food safety. A rapid and sensitive detection assay is needed for this harmful organism to prevent foodborne illnesses and control outbreaks in a timely manner. We developed a magnetic bead-based immunoassay for detection of E. coli O157:H7 (the most well-known Shiga toxigenic E. coli strain) with a 96-well microplate as an assay platform. Immunomagnetic separation (IMS) and tyramide signal amplification were coupled to the assay to increase its sensitivity and specificity. This immunoassay was able to detect E. coli O157:H7 in pure culture with a detection limit of 50 CFU/ml in less than 3 h without an enrichment step. The detection limit was decreased 10-fold to 5 CFU/ml with addition of a 3-h enrichment step. When this assay was tested with other nontarget foodborne pathogens and common enteric bacteria, no cross-reactivity was found. When tested with artificially contaminated ground beef and milk samples, the assay sensitivity decreased two- to fivefold, with detection limits of 250 and 100 CFU/ml, respectively, probably because of the food matrix effect. The assay results also were compared with those of a sandwich-type enzyme-linked immunosorbent assay (ELISA) and an ELISA coupled with IMS; the developed assay was 25 times and 4 times more sensitive than the standard ELISA and the IMS-ELISA, respectively. Tyramide signal amplification combined with IMS can improve sensitivity and specificity for detection of E. coli O157:H7. The developed assay could be easily adapted for other foodborne pathogens and will contribute to improved food safety and public health.

  1. Epstein-Barr Virus Interferes with the Amplification of IFNα Secretion by Activating Suppressor of Cytokine Signaling 3 in Primary Human Monocytes

    PubMed Central

    Michaud, François; Coulombe, François; Gaudreault, Eric; Paquet-Bouchard, Carine; Rola-Pleszczynski, Marek; Gosselin, Jean

    2010-01-01

    Background Epstein-Barr virus is recognized to cause lymphoproliferative disorders and is also associated with cancer. Evidence suggests that monocytes are likely to be involved in EBV pathogenesis, especially due to a number of cellular functions altered in EBV-infected monocytes, a process that may affect efficient host defense. Because type I interferons (IFNs) are crucial mediators of host defense against viruses, we investigated the effect of EBV infection on the IFNα pathway in primary human monocytes. Methodology/Principal Findings Infection of monocytes with EBV induced IFNα secretion but inhibited the positive feedback loop for the amplification of IFNα. We showed that EBV infection induced the expression of suppressor of cytokine signaling 3 (SOCS3) and, to a lesser extent, SOCS1, two proteins known to interfere with the amplification of IFNα secretion mediated by the JAK/STAT signal transduction pathway. EBV infection correlated with a blockage in the activation of JAK/STAT pathway members and affected the level of phosphorylated IFN regulatory factor 7 (IRF7). Depletion of SOCS3, but not SOCS1, by small interfering RNA (siRNA) abrogated the inhibitory effect of EBV on JAK/STAT pathway activation and significantly restored IFNα secretion. Finally, transfection of monocytes with the viral protein Zta caused the upregulation of SOCS3, an event that could not be recapitulated with mutated Zta. Conclusions/Significance We propose that EBV protein Zta activates SOCS3 protein as an immune escape mechanism that both suppresses optimal IFNα secretion by human monocytes and favors a state of type I IFN irresponsiveness in these cells. This immunomodulatory effect is important to better understand the aspects of the immune response to EBV. PMID:20689596

  2. Ultrasensitive and selective electrochemical biosensor for detection of mercury (II) ions by nicking endonuclease-assisted target recycling and hybridization chain reaction signal amplification.

    PubMed

    Hong, Minqiang; Wang, Mengyan; Wang, Jing; Xu, Xueqin; Lin, Zhenyu

    2017-02-22

    In this paper, a novel and signal-on electrochemical biosensor based on Hg(2+)- triggered nicking endonuclease-assisted target recycling and hybridization chain reaction (HCR) amplification tactics was developed for sensitive and selective detection of Hg(2+). The hairpin-shaped capture probe A (PA) contained a specific sequence which was recognized by nicking endonuclease (NEase). In the presence of Hg(2+), probe B (PB) hybridized with PA to form stand-up duplex DNA strands via the Hg(2+) mediated thymine-Hg(2+)-thymine (T-Hg(2+)-T) structure, which automatically triggered NEase to selectively digest duplex region from the recognition sites, spontaneously dissociating PB and Hg(2+) and leaving the remnant initiators. The released PB and Hg(2+) could be reused to initiate the next cycle and more initiators were generated. The long nicked double helices were formed through HCR event, which was triggered by the initiators and two hairpin-shaped signal probes labeled with methylene blue, resulting in a significant signal increase. Under optimum conditions, the resultant biosensor showed the high sensitivity and selectivity for the detection of Hg(2+) in a linear range from 10 pM to 50nM (R(2)=0.9990), and a detection limit as low as 1.6 pM (S/N=3). Moreover, the proposed biosensor was successfully applied in the detection of Hg(2+) in environment water samples with satisfactory results.

  3. Luminescence-Functionalized Metal-Organic Frameworks Based on a Ruthenium(II) Complex: A Signal Amplification Strategy for Electrogenerated Chemiluminescence Immunosensors.

    PubMed

    Xiong, Cheng-Yi; Wang, Hai-Jun; Liang, Wen-Bin; Yuan, Ya-Li; Yuan, Ruo; Chai, Ya-Qin

    2015-06-26

    Novel luminescence-functionalized metal-organic frameworks (MOFs) with superior electrogenerated chemiluminescence (ECL) properties were synthesized based on zinc ions as the central ions and tris(4,4'-dicarboxylicacid-2,2'-bipyridyl)ruthenium(II) dichloride ([Ru(dcbpy)3](2+)) as the ligands. For potential applications, the synthesized MOFs were used to fabricate a "signal-on" ECL immunosensor for the detection of N-terminal pro-B-type natriuretic peptide (NT-proBNP). As expected, enhanced ECL signals were obtained through a simple fabrication strategy because luminescence-functionalized MOFs not only effectively increased the loading of [Ru(dcbpy)3](2+), but also served as a loading platform in the ECL immunosensor. Furthermore, the proposed ECL immunosensor had a wide linear range from 5 pg mL(-1) to 25 ng mL(-1) and a relatively low detection limit of 1.67 pg mL(-1) (signal/noise=3). The results indicated that luminescence-functionalized MOFs provided a novel amplification strategy in the construction of ECL immunosensors and might have great prospects for application in bioanalysis.

  4. Early amplification options.

    PubMed

    Gabbard, Sandra Abbott; Schryer, Jennifer

    2003-01-01

    Children with permanent hearing loss have been remediated with hearing amplification devices for decades. The influx of young infants identified with hearing loss through successful newborn hearing screening programs has established a need for amplification resources for infants within the first six months of life. For the approximately two of every 1000 infants born who are identified with bilateral hearing loss [Mehl and Thomson, 1998, Pediatrics 101, p. e4], the use of amplification is commonly the first step in treating the sequella of their loss. The use of hearing aids, combined with early intervention, has been shown to significantly improve the speech and language skills of young children with hearing loss [Yoshinaga-Itano, 2000, Seminars in Hearing 21, p. 309]. Speech and language delays have contributed to compromised academic performance of school aged children with hearing loss [Johnson et al., 1997, Educational Audiology Handbook, Singular Publishing, San Diego]. Most hard-of-hearing and deaf children use hearing aids and other assistive listening devices every day throughout their lifetime and the life expectancy of a hearing aid is only five to eight years. The current challenge for pediatric audiologists is selecting and evaluating the available amplification to provide the best options for children and their families. Amplification technology has seen an explosion in growth the past few years and the options continue to expand rapidly. This article examines currently available amplification technology and reviews the selection criteria that may be used for infants and young children. Issues such as style, type, amplification features, signal processing strategies, and verification and validation tools are also discussed.

  5. Ultrasensitive sensing platform for platelet-derived growth factor BB detection based on layered molybdenum selenide-graphene composites and Exonuclease III assisted signal amplification.

    PubMed

    Huang, Ke-Jing; Shuai, Hong-Lei; Zhang, Ji-Zong

    2016-03-15

    A highly sensitive and ultrasensitive electrochemical aptasensor for platelet-derived growth factor BB (PDGF-BB) detection is fabricated based on layered molybdenum selenide-graphene (MoSe2-Gr) composites and Exonuclease III (Exo III)-aided signal amplification. MoSe2-Gr is prepared by a simple hydrothermal method and used as a promising sensing platform. Exo III has a specifical exo-deoxyribonuclease activity for duplex DNAs in the direction from 3' to 5' terminus, however its activity is limited on the duplex DNAs with more than 4 mismatched terminal bases at 3' ends. Herein, aptamer and complementary DNA (cDNA) sequences are designed with four thymine bases on 3' ends. In the presence of target protein, the aptamer associates with it and facilitates the formation of duplex DNA between cDNA and signal DNA. The duplex DNA then is digested by Exo III and releases cDNA, which hybridizes with signal DNA to perform a new cleavage process. Nevertheless, in the absence of target protein, the aptamer hybridizes with cDNA will inhibit the Exo III-assisted nucleotides cleavage. The signal DNA then hybridizes with capture DNA on the electrode. Subsequently, horse radish peroxidase is fixed on electrode by avidin-biotin reaction and then catalyzes hydrogen peroxide and hydroquinone to produce electrochemical response. Therefore, a bridge can be established between the concentration of target protein and the degree of the attenuation of the obtained signal, providing a quantitative measure of target protein with a broad detection range of 0.0001-1 nM and a detection limit of 20 fM.

  6. The Effect of Classroom Amplification on the Signal-to-Noise Ratio in Classrooms while Class Is in Session

    ERIC Educational Resources Information Center

    Larsen, Jeffery B.; Blair, James C.

    2008-01-01

    Purpose: The purpose of this study was to measure the signal-to-noise ratios in classrooms while class was in session and students were interacting with the teacher and each other. Method: Measurements of noise and reverberation were collected for 5 different classrooms in 3 different schools while class was in session. Activities taking place…

  7. Kinetic insulation as an effective mechanism for achieving pathway specificity in intracellular signaling networks

    PubMed Central

    Behar, Marcelo; Dohlman, Henrik G.; Elston, Timothy C.

    2007-01-01

    Intracellular signaling pathways that share common components often elicit distinct physiological responses. In most cases, the biochemical mechanisms responsible for this signal specificity remain poorly understood. Protein scaffolds and cross-inhibition have been proposed as strategies to prevent unwanted cross-talk. Here, we report a mechanism for signal specificity termed “kinetic insulation.” In this approach signals are selectively transmitted through the appropriate pathway based on their temporal profile. In particular, we demonstrate how pathway architectures downstream of a common component can be designed to efficiently separate transient signals from signals that increase slowly over time. Furthermore, we demonstrate that upstream signaling proteins can generate the appropriate input to the common pathway component regardless of the temporal profile of the external stimulus. Our results suggest that multilevel signaling cascades may have evolved to modulate the temporal profile of pathway activity so that stimulus information can be efficiently encoded and transmitted while ensuring signal specificity. PMID:17913886

  8. A sensitive electrochemical biosensor for detection of protein kinase A activity and inhibitors based on Phos-tag and enzymatic signal amplification.

    PubMed

    Yin, Huanshun; Wang, Mo; Li, Bingchen; Yang, Zhiqing; Zhou, Yunlei; Ai, Shiyun

    2015-01-15

    A simple, highly sensitive and selective electrochemical assay is developed for the detection of protein kinase A (PKA) activity based on the specific recognition utility of Phos-tag for kinase-induced phosphopeptides and enzymatic signal amplification. When the substrate peptide was phosphorylated by PKA reaction, they could specifically bind with Phos-tag-biotin in the presence of Zn(2+) through the formation of a specific noncovalent complex with the phosphomonoester dianion in phosphorylated peptides. Through the further specific interaction between biotin and avidin, avidin functionalized horseradish peroxidase (HRP) can be captured on the electrode surface. Under the catalytic effect of HRP, a sensitive electrochemical signal for benzoquinone was obtained, which was related to PKA activity. Under the optimal experiment conditions, the proposed electrochemical method presented dynamic range from 0.5 to 25 unit/mL with low detection limit of 0.15 unit/mL. This new detection strategy was also successfully applied to analyze the inhibition effect of inhibitors (ellagic acid and H-89) on PKA activity and monitored the PKA activity in cell lysates. Therefore, this Phos-tag-based electrochemical assay offers an alternative platform for PKA activity assay and inhibitor screening, and thus it might be a valuable tool for development of targeted therapy and clinical diagnosis.

  9. A magnetite/PMAA nanospheres-targeting SERS aptasensor for tetracycline sensing using mercapto molecules embedded core/shell nanoparticles for signal amplification.

    PubMed

    Li, Huanhuan; Chen, Quansheng; Mehedi Hassan, Md; Chen, Xiaoxing; Ouyang, Qin; Guo, Zhiming; Zhao, Jiewen

    2017-02-07

    Surface-enhanced Raman scattering (SERS) biosensors have promising potential in the field of antibiotics detection because of their ultrahigh detection sensitivity. This paper reports a rapid and sensitive SERS-based magnetic nanospheres-targeting strategy for sensing tetracycline (TTC) using aptamer-conjugated magnetite colloid nanocrystal clusters (MCNCs)-polymethacrylic acid (PMAA) magnetic nanospheres (MNs) as the recognition and the Au/PATP/SiO2 (APS) as the labels. Initially, MNs were fabricated and conjugated with the aptamers through condensation reaction. MNs possessed high saturation magnetization (Ms) value of 71.5emu/g and excellent biocompatibility, which facilitated the rapid and easy magnetic separation. Then, complementary DNA (cDNA) were loaded on the APS nanocarrier to produce a large amplification factor of Raman signals. The MNs-targeting aptasensor was thus fabricated by immobilizing the APS to the MNs' surfaces via the hybrid reaction between cDNA and aptamers. Sequel, TTC bound successfully to the aptamer upon its addition with the subsequent release of some cDNA-APS into the bulk solution. Under magnet attraction, the nanospheres were deposited together. Consequently, a display of strong SERS signals by supernatants of the resulting mixtures with increasing TTC concentrations was observed. The proposed aptasensor showed excellent performances for TTC detection along with wide linear range of 0.001-100ng/mL, low detection limit 0.001ng/mL, high sensitivity, and good selectivity to the general coexisted interferences.

  10. Electrochemical Impedance Immunosensor Based on Self-Assembled Monolayers for Rapid Detection of Escherichia coli O157:H7 with Signal Amplification Using Lectin.

    PubMed

    Li, Zhanming; Fu, Yingchun; Fang, Weihuan; Li, Yanbin

    2015-08-05

    Escherichia coli O157:H7 is a predominant foodborne pathogen with severe pathogenicity, leading to increasing attention given to rapid and sensitive detection. Herein, we propose an impedance biosensor using new kinds of screen-printed interdigitated microelectrodes (SPIMs) and wheat germ agglutinin (WGA) for signal amplification to detect E. coli O157:H7 with high sensitivity and time-efficiency. The SPIMs integrate the high sensitivity and short response time of the interdigitated electrodes and the low cost of the screen-printed electrodes. Self-assembling of bi-functional 3-dithiobis-(sulfosuccinimidyl-propionate) (DTSP) on the SPIMs was investigated and was proved to be able to improve adsorption quantity and stability of biomaterials. WGA was further adopted to enhance the signal taking advantage of the abundant lectin-binding sites on the bacteria surface. The immunosensor exhibited a detection limit of 102 cfu·mL(-1), with a linear detection range from 10(2) to 10(7) cfu·mL(-1) (r2 = 0.98). The total detection time was less than 1 h, showing its comparable sensitivity and rapid response. Furthermore, the low cost of one SPIM significantly reduced the detection cost of the biosensor. The biosensor may have great promise in food safety analysis and lead to a portable biosensing system for routine monitoring of foodborne pathogens.

  11. Facile fabrication of an ultrasensitive sandwich-type electrochemical immunosensor for the quantitative detection of alpha fetoprotein using multifunctional mesoporous silica as platform and label for signal amplification.

    PubMed

    Wang, Yulan; Li, Xiaojian; Cao, Wei; Li, Yueyun; Li, He; Du, Bin; Wei, Qin

    2014-11-01

    A novel and ultrasensitive sandwich-type electrochemical immunosensor was designed for the quantitative detection of alpha fetoprotein (AFP) using multifunctional mesoporous silica (MCM-41) as platform and label for signal amplification. MCM-41 has high specific surface area, high pore volume, large density of surface silanol groups (SiOH) and good biocompatibility. MCM-41 functionalized with 3-aminopropyltriethoxysilane (APTES), gold nanoparticles (Au NPs) and toluidine blue (TB) could enhance electrochemical signals. Moreover, primary antibodies (Ab1) and secondary antibodies (Ab2) could be effectively immobilized onto the multifunctional MCM-41 by the interaction between Au NPs and amino groups (-NH2) on antibodies. Using multifunctional MCM-41 as a platform and label could greatly simplify the fabrication process and result in a high sensitivity of the designed immunosensor. Under optimal conditions, the designed immunosensor exhibited a wide liner range from 10(-4) ng/mL to 10(3) ng/mL with a low detection limit of 0.05 pg/mL for AFP. The designed immunosensor showed acceptable selectivity, reproducibility and stability, which could provide potential applications in clinical monitoring of AFP.

  12. Amplification of a bi-phase shift-key modulated signal by a mm-wave FEL

    SciTech Connect

    Prosnitz, D.; Scharlemann, E.T.; Sheaffer, M.K.

    1991-10-01

    Bi-phase shift keying (BPSK) is a modulation scheme used in communications and radar in which the phase of a transmitted rf signal is switched in a coded pattern between discrete values differing by {pi} radians. The transmitted information rate (in communications) or resolution (in imaging radar) depends on the rate at which the transmitted signal can be modulated. Modulation rates of greater than 1 GHz are generally desired. Although the instantaneous gain bandwidth of a mm-wave FEL amplifier can be much greater than 10 GHz, slippage may limit the BPSK modulation rate that can be amplified. Qualitative slippage arguments would limit the modulation rate to relatively low values; nevertheless, simulations with a time-dependent FEL code (GINGER) indicate that rates of 2 GHz or more are amplified without much loss in modulation integrity. In this paper we describe the effects of slippage in the simulations and discuss the limits of simple arguments.

  13. Amplification of a bi-phase shift-key modulated signal by a mm-wave FEL

    NASA Astrophysics Data System (ADS)

    Prosnitz, D.; Scharlemann, E. T.; Sheaffer, M. K.

    1991-10-01

    Bi-phase shift keying (BPSK) is a modulation scheme used in communications and radar in which the phase of a transmitted RF signal is switched in a coded pattern between discrete values differing by (pi) radians. The transmitted information rate (in communications) or resolution (in imaging radar) depends on the rate at which the transmitted signal can be modulated. Modulation rates of greater than 1 GHz are generally desired. Although the instantaneous gain bandwidth of a mm-wave FEL amplifier can be much greater than 10 GHz, slippage may limit the BPSK modulation rate that can be amplified. Qualitative slippage arguments would limit the modulation rate to relatively low values; nevertheless, simulations with a time-dependent FEL code (GINGER) indicate that rates of 2 GHz or more are amplified without much loss in modulation integrity. In this paper we describe the effects of slippage in the simulations and discuss the limits of simple arguments.

  14. Amplification of a bi-phase shift-key modulated signal by a mm-wave FEL

    NASA Astrophysics Data System (ADS)

    Prosnitz, D.; Scharlemann, E. T.; Sheaffer, M. K.

    1992-07-01

    Bi-phase shift keying (BPSK) is a modulation scheme used in communications and radar in which the phase of a transmitted rf signal is switched in a coded pattern between discrete values differing by π radians. The transmitted information rate (in communications) or resolution (in imaging radar) depends on the rate at which the transmitted signal can be modulated. Modulation rates of greater than 1 GHz are generally desired. Although the instantaneous gain bandwidth of a mm-wave FEL amplifier can be much greater than 10 GHz, slippage may limit the BPSK modulation rate that can be amplified. Qualitative slippage arguments would limit the modulation rate to relatively low values; nevertheless, simulations with a time-dependent FEL code (GINGER) indicate that rates of 2 GHz or more are amplified without much loss in modulation integrity. In this paper we describe the effects of slippage in the simulations and discuss the limits of simple slippage arguments.

  15. Self-primed isothermal amplification for genomic DNA detection of human papillomavirus.

    PubMed

    Lu, Wei; Yuan, Qingpan; Yang, Zhiliu; Yao, Bo

    2017-04-15

    Rolling circle amplification (RCA) is an isothermal amplification technique with high efficiency and perfect accuracy for nucleic acids detection. However, RCA technique suffers the limitation to detect short DNA or RNA molecules. For long nucleic acid molecules, enzymatic restriction as well as heat denaturation process is usually required, which makes the amplification not effective and strictly isothermal. In this article, a simple and efficient one-pot self-primed isothermal amplification (SIA) was developed for detection of genomic DNA directly based on the combination of nicking endonuclease assisted strand displacement amplification (SDA) and exponential RCA. In virtue of numerous nicking sites on the genome, a pre-amplification of the whole genome was performed through SDA with the specific cleaving of nicking endonuclease. Meanwhile, the single strand DNA with HPV target sequence generated from SDA could hybrid with the circle probe as a primer and trigger the exponential RCA as a result of the existence of nicking endonuclease. As the reaction temperature and enzyme were the same, the amplification could be operated in one pot. The reaction solution after amplification was added on the electrode for hybridization with the sulfydryl probe to achieve the electrochemical signal. Based on the isothermal amplification, genotyping of HPV 11, 16, 18 and the detection of HPV 18 in Hela cell line were attempted with satisfied results. This approach should be a promising tool for pathogene detection in clinical diagnostics and research.

  16. Ultrasensitive electrochemical detection of protein tyrosine kinase-7 by gold nanoparticles and methylene blue assisted signal amplification.

    PubMed

    Miao, Xiangmin; Li, Zongbing; Zhu, Aihua; Feng, Zhaozhong; Tian, Jun; Peng, Xue

    2016-09-15

    We present here an ultrasensitive and simple strategy for protein tyrosine kinase-7 (PTK7) detection based on the recognition-induced structure change of sgc8 aptamer, and the signal change of methylene blue (MB) that interacted with sandwiched DNA complex. To construct such a sensor, an homogeneous nano-surface was formed firstly on the glass carbon electrode (GCE) by using negatively charged Nafion (Nf) as the inner layer and positively charged gold nanoparticles ((+)AuNPs) as the outer layer, followed by the immobilization of sgc8 aptamer based on Au-S bond. In the presence of helper probe (HP), sandwiched DNA complex was formed between the sgc8 aptamer and the DNA modified gold nanoparticle probe (DNA-AuNPs). Then, a strong current signal was produced due to the capture of abundant MB molecules by both the sandwiched DNA complex and the multiple DNAs that modified on AuNPs surface. However, the specific binding of sgc8 aptamer with PNK7 would trigger a structure transition of it, and directly prevented the following formation of sandwiched structure and the capture of MB. Thus, PTK7 detection could be realized based on monitoring the signal reduction of MB upon incubation of sgc8 aptamer with PTK7. Under optimal conditions, a low detection limit of 372 fM was obtained for PNK7 detection. Due to the employment of sgc8 aptamer, the proposed biosensor exhibited high selectivity to PNK7. Moreover, satisfactory results were obtained when the proposed method was applied for PNK7 detection in cellular debris.

  17. Electrochemical Detection of Amyloid-β Oligomers Based on the Signal Amplification of a Network of Silver Nanoparticles.

    PubMed

    Xia, Ning; Wang, Xin; Zhou, Binbin; Wu, Yangyang; Mao, Wenhui; Liu, Lin

    2016-08-03

    Amyloid-β oligomers (AβOs) are the most important toxic species in the brain of Alzheimer's disease (AD) patient. AβOs, therefore, are considered reliable molecular biomarkers for the diagnosis of AD. Herein, we reported a simple and sensitive electrochemical method for the selective detection of AβOs using silver nanoparticles (AgNPs) as the redox reporters and PrP(95-110), an AβOs-specific binding peptide, as the receptor. Specifically, adamantine (Ad)-labeled PrP(95-110), denoted as Ad-PrP(95-110), induced the aggregation and color change of AgNPs and the follow-up formation of a network of Ad-PrP(95-110)-AgNPs. Then, Ad-PrP(95-110)-AgNPs were anchored onto a β-cyclodextrin (β-CD)-covered electrode surface through the host-guest interaction between Ad and β-CD, thus producing an amplified electrochemical signal through the solid-state Ag/AgCl reaction by the AgNPs. In the presence of AβOs, Ad-PrP(95-110) interacted specifically with the AβOs, thus losing the capability to bind AgNPs and to induce the formation of an AgNPs-based network on the electrode surface. Consequently, the electrochemical signal decreased with an increase in the concentration of AβOs in the range of 20 pM to 100 nM. The biosensor had a detection limit of 8 pM and showed no response to amyloid-β monomers (AβMs) and fibrils (AβFs). On the basis of the well-defined and amplified electrochemical signal of the AgNPs-based network architecture, these results should be valuable for the design of novel electrochemical biosensors by marrying specific receptors.

  18. Amplification of the solar signal in the summer monsoon rainband in China by synergistic actions of different dynamical responses

    NASA Astrophysics Data System (ADS)

    Zhao, Liang; Wang, Jingsong; Liu, Haiwen; Xiao, Ziniu

    2017-02-01

    A rainband meridional shift index (RMSI) is defined and used to statistically prove that the East Asian summer monsoon rainband is usually significantly more northward in the early summer of solar maximum years than that of solar minimum years. By applying continuous wavelet transform, cross wavelet transform, and wavelet coherence, it is found that throughout most of the 20th century, the significant decadal oscillations of sunspot number (SSN) and the RMSI are phase-locked and since the 1960s, the SSN has led the RMSI slightly by approximately 1.4 yr. Wind and Eliassen-Palm (EP) flux analysis shows that the decadal meridional oscillation of the June rainband likely results from both a stronger or earlier onset of the tropical monsoon and poleward shift of the subtropical westerly jet in high-solar months of May and June. The dynamical responses of the lower tropical monsoon and the upper subtropical westerly jet to the 11-yr solar cycle transmit bottom-up and top-down solar signals, respectively, and the synergistic actions between the monsoon and the jet likely amplify the solar signal at the northern boundary of the monsoon to some extent.

  19. Surface plasmon resonance detection using antibody-linked magnetic nanoparticles for analyte capture, purification, concentration, and signal amplification.

    PubMed

    Soelberg, Scott D; Stevens, Richard C; Limaye, Ajit P; Furlong, Clement E

    2009-03-15

    Rapid, sensitive, and accurate detection of analytes present in low concentrations in complex matrixes is a critical challenge. One issue that affects many biosensor protocols is the number and nature of the interferences present in complex matrixes such as plasma, urine, stool, and environmental samples, resulting in loss of sensitivity and specificity. We have developed a method for rapid purification, concentration, and detection of target analytes from complex matrixes using antibody-coated superparamagnetic nanobeads (immunomagnetic beads, or IMBs). The surface plasmon resonance (SPR) detection signal from staphylococcal enterotoxin B (SEB) was dramatically increased when the IMBs were used as detection amplifiers. When SEB detection included a 10-fold concentration/purification IMB protocol, a substantial increase in detection sensitivity was observed. This procedure was used to successfully purify and concentrate SEB from serum and stool samples, then amplify the SPR detection signal. SEB at a concentration of 100 pg/mL was easily detected in both buffer and stool samples using this procedure. The IMB protocol also served to verify the analyte detection by using two different anti-SEB antibodies, mouse monoclonal antibodies attached to the magnetic nanobeads and rabbit polyclonal antibodies on the SPR sensor surface. Multiple detections of SEB in stool were performed using the same sensor surface by regenerating the sensor surfaces with a pH 2.2 buffer wash.

  20. Surface Plasmon Resonance (SPR) Detection Using Antibody-Linked Magnetic Nanoparticles for Analyte Capture, Purification, Concentration and Signal Amplification

    PubMed Central

    Soelberg, Scott D.; Stevens, Richard C.; Limaye, Ajit P.; Furlong, Clement E.

    2009-01-01

    Rapid, sensitive and accurate detection of analytes present in low concentrations in complex matrices is a critical challenge. One issue that affects many biosensor protocols is the number and nature of the interferents present in complex matrices such as plasma, urine, stool and environmental samples, resulting in loss of sensitivity and specificity. We have developed a method for rapid purification, concentration and detection of target analytes from complex matrices using antibody-coated superparamagnetic nanobeads (immunomagnetic beads, or IMBs). The SPR detection signal from Staphylococcal enterotoxin B (SEB) was dramatically increased when the IMBs were used as detection amplifiers. When SEB detection included a 10-fold concentration/purification IMB protocol, a substantial increase in detection sensitivity was observed. This procedure was used to successfully purify and concentrate SEB from serum and stool samples, then amplify the SPR detection signal. SEB at a concentration of 100 picograms/mL was easily detected in both buffer and stool samples using this procedure. The IMB protocol also served to verify the analyte detection by using two different anti-SEB antibodies, mouse monoclonal antibodies attached to the magnetic nanobeads and rabbit polyclonal antibodies on the SPR sensor surface. Multiple detections of SEB in stool were performed using the same sensor surface by regenerating the sensor surfaces with a pH 2.2 buffer wash. PMID:19215065

  1. A pH-activatable nanoparticle with signal-amplification capabilities for non-invasive imaging of tumour malignancy

    NASA Astrophysics Data System (ADS)

    Mi, Peng; Kokuryo, Daisuke; Cabral, Horacio; Wu, Hailiang; Terada, Yasuko; Saga, Tsuneo; Aoki, Ichio; Nishiyama, Nobuhiro; Kataoka, Kazunori

    2016-08-01

    Engineered nanoparticles that respond to pathophysiological parameters, such as pH or redox potential, have been developed as contrast agents for the magnetic resonance imaging (MRI) of tumours. However, beyond anatomic assessment, contrast agents that can sense these pathological parameters and rapidly amplify their magnetic resonance signals are desirable because they could potentially be used to monitor the biological processes of tumours and improve cancer diagnosis. Here, we report an MRI contrast agent that rapidly amplifies magnetic resonance signals in response to pH. We confined Mn2+ within pH-sensitive calcium phosphate (CaP) nanoparticles comprising a poly(ethylene glycol) shell. At a low pH, such as in solid tumours, the CaP disintegrates and releases Mn2+ ions. Binding to proteins increases the relaxivity of Mn2+ and enhances the contrast. We show that these nanoparticles could rapidly and selectively brighten solid tumours, identify hypoxic regions within the tumour mass and detect invisible millimetre-sized metastatic tumours in the liver.

  2. A sandwich-hybridization assay for simultaneous determination of HIV and tuberculosis DNA targets based on signal amplification by quantum dots-PowerVision™ polymer coding nanotracers.

    PubMed

    Yan, Zhongdan; Gan, Ning; Zhang, Huairong; Wang, De; Qiao, Li; Cao, Yuting; Li, Tianhua; Hu, Futao

    2015-09-15

    A novel sandwich-hybridization assay for simultaneous electrochemical detection of multiple DNA targets related to human immune deficiency virus (HIV) and tuberculosis (TB) was developed based on the different quantum dots-PowerVision(TM) polymer nanotracers. The polymer nanotracers were respectively fabricated by immobilizing SH-labeled oligonucleotides (s-HIV or s-TB), which can partially hybrid with virus DNA (HIV or TB), on gold nanoparticles (Au NPs) and then modified with PowerVision(TM) (PV) polymer-encapsulated quantum dots (CdS or PbS) as signal tags. PV is a dendrimer enzyme linked polymer, which can immobilize abundant QDs to amplify the stripping voltammetry signals from the metal ions (Pb or Cd). The capture probes were prepared through the immobilization of SH-labeled oligonucleotides, which can complementary with HIV and TB DNA, on the magnetic Fe3O4@Au (GMPs) beads. After sandwich-hybridization, the polymer nanotracers together with HIV and TB DNA targets were simultaneously introduced onto the surface of GMPs. Then the two encoding metal ions (Cd(2+) and Pb(2+)) were used to differentiate two viruses DNA due to the different subsequent anodic stripping voltammetric peaks at -0.84 V (Cd) and -0.61 V (Pb). Because of the excellent signal amplification of the polymer nanotracers and the great specificity of DNA targets, this assay could detect targets DNA as low as 0.2 femtomolar and exhibited excellent selectivity with the dynamitic range from 0.5 fM to 500 pM. Those results demonstrated that this electrochemical coding assay has great potential in applications for screening more viruses DNA while changing the probes.

  3. Sensitive ECL immunosensor for detection of retinol-binding protein based on double-assisted signal amplification strategy of multiwalled carbon nanotubes and Ru(bpy)3(2+) doped mesoporous silica nanospheres.

    PubMed

    Wu, Beina; Hu, Chenyi; Hu, Xiaoqing; Cao, Hongmei; Huang, Chusen; Shen, Hebai; Jia, Nengqin

    2013-12-15

    A novel electrochemiluminescence (ECL) strategy based on the sandwich-type immunosensor for sensitive detection of retinol-binding protein (RBP) was developed. The primary antibody anti-RBP was immobilized onto multiwalled carbon nanotubes (MWCNTs), which have large surface area and high electrical conductivity. The RBP antigen and Ru-Nafion@SiO2-labeled secondary antibody were then successively conjugated to form sandwich-type immunocomplexes through the specific interaction between antigen and antibody. The ECL signal amplification was significantly improved due to the synergistic effect of MWCNTs and mesoporous silica nanospheres (mSiO2). The developed ECL immunosensor exhibited high sensitivity and specificity for the detection of RBP and responded linearly to the clinically-relevant concentration of RBP from 78 to 5000 ng mL(-1). Moreover, the MWCNT-based ECL immunosensor displayed excellent stability and reproducibility, as well as successfully achieved the detection of RBP in patient urine samples with desirable results. The present work provided a promising technique for the clinical screening of RBP and point-of-care diagnostics.

  4. Classroom Amplification Technology: Theory and Practice.

    ERIC Educational Resources Information Center

    Crandell, Carl C.; Smaldino, Joseph J.

    2000-01-01

    This article reviews some relevant events in the development of acoustical standards for classrooms, describes classroom challenges to providing clear acoustical signals to children in classrooms, and outlines amplification solutions to some of those classroom challenges. Solutions include personal amplification devices and use of signal-to-noise…

  5. A novel sandwiched electrochemiluminescence immunosensor for the detection of carcinoembryonic antigen based on carbon quantum dots and signal amplification.

    PubMed

    Li, Nian-Lu; Jia, Li-Ping; Ma, Rong-Na; Jia, Wen-Li; Lu, Yi-Yang; Shi, Sha-Shan; Wang, Huai-Sheng

    2017-03-15

    In this study, a novel sandwiched electrochemiluminescence (ECL) immunosensor for the detection of carcinoembryonic antigen (CEA) was developed. The nanocomposite of polydopamine and Ag nanoparticles (PDA-AgNPs) was prepared by the redox reaction between Ag(+) and dopamine. This nanocomposite not only provided an effective matrix for the immobilization of primary antibody (Ab1) but also enhanced the conductivity of the electrode. Carbon quantum dots (CQDs) were immobilized on the poly(ethylenimine) functionalized graphene oxide (PEI-GO) through amido-bond. Then Au nanoparticles were decorated on the CQDs modified PEI-GO matrix, and the resulted complex AuNPs/CQDs-PEI-GO was introduced to link secondary antibody (Ab2). The CQDs can be connected to the electrode surface through the combination of CEA with Ab1 and Ab2, and then the amplified electrochemiluminescence signal of CQDs was obtained with the synergistic effect of AgNPs, polydopamine, AuNPs and PEI-GO. Under the optimal conditions, the ECL intensity was proportional to the logarithm value of CEA concentration in the linear range from 5pgmL(-1) to 500ngmL(-1) with a detection limit of 1.67pgmL(-1) for CEA detection. The immunosensor was applied for the CEA detection in real samples with satisfactory results. The proposed ECL immunosensor showed good performance with high sensitivity, specificity, reproducibility, stability and will be potential in clinical detection.

  6. Powerful Amplification Cascades of FRET-Based Two-Layer Nonenzymatic Nucleic Acid Circuits.

    PubMed

    Quan, Ke; Huang, Jin; Yang, Xiaohai; Yang, Yanjing; Ying, Le; Wang, He; Xie, Nuli; Ou, Min; Wang, Kemin

    2016-06-07

    Nucleic acid circuits have played important roles in biological engineering and have increasingly attracted researchers' attention. They are primarily based on nucleic acid hybridizations and strand displacement reactions between nucleic acid probes of different lengths. Signal amplification schemes that do not rely on protein enzyme show great potential in analytical applications. While the single amplification circuit often achieves linear amplification that may not meet the need for detection of target in a very small amount, it is very necessary to construct cascade circuits that allow for larger amplification of inputs. Herein, we have successfully engineered powerful amplification cascades of FRET-based two-layer nonenzymatic nucleic acid circuits, in which the outputs of catalyzed hairpin assembly (CHA) activate hybridization chain reactions (HCR) circuits to induce repeated hybridization, allowing real-time monitoring of self-assembly process by FRET signal. The cascades can yield 50000-fold signal amplification with the help of the well-designed and high-quality nucleic acid circuit amplifiers. Subsequently, with coupling of structure-switching aptamer, as low as 200 pM adenosine is detected in buffer, as well as in human serum. To our knowledge, we have for the first time realized real-time monitoring adaptation of HCR to CHA circuits and achieved amplified detection of nucleic acids and small molecules with relatively high sensitivity.

  7. Enzyme-free and isothermal detection of microRNA based on click-chemical ligation-assisted hybridization coupled with hybridization chain reaction signal amplification.

    PubMed

    Oishi, Motoi

    2015-05-01

    An enzyme-free and isothermal microRNA (miRNA) detection method has been developed based on click-chemical ligation-assisted hybridization coupled with hybridization chain reaction (HCR) on magnetic beads (MBs). The click-chemical ligation between an azide-modified probe DNA and a dibenzocyclooctyne-modified probe DNA occurred through the hybridization of target miRNA (miR-141). HCR on MBs was performed by the addition of DNA hairpin monomers (H1 and H2). After magnetic separation and denaturation/rehybridization of HCR products ([H1/H2] n ), the resulting HCR products were analyzed by the fluorescence emitted from an intercalative dye, allowing amplification of the fluorescent signal. The proposed assay had a limit of detection of 0.55 fmol, which was 230-fold more sensitive than that of the HCR on the MBs coupled with a conventional sandwich hybridization assay (without click-chemical ligation) (limit of detection 127 fmol). Additionally, the proposed assay could discriminate between miR-141 and other miR-200 family members. In contrast to quantitative reverse transcription polymerase chain reaction techniques using enzymes and thermal cycling, this is an enzyme-free assay that can be conducted under isothermal conditions and can specifically detect miR-141 in fetal bovine serum.

  8. Electrochemical immunosensor for N6-methyladenosine detection in human cell lines based on biotin-streptavidin system and silver-SiO2 signal amplification.

    PubMed

    Yin, Huanshun; Wang, Haiyan; Jiang, Wenjing; Zhou, Yunlei; Ai, Shiyun

    2017-04-15

    N6-methyladenosine (m6A), a kind of RNA methylation form and important epigenetic event, plays crucial roles in many biological progresses. Thus it is essential to quantitatively detect m6A in complicated biological samples. Herein, a simple and sensitive electrochemical method was developed for m6A detection using N6-methyladenosine-5'-triphosphate (m6ATP) as detection target molecule. In this detection strategy, anti-m6A antibody was selected as m6A recognition and capture reagent, silver nanoparticles and amine-PEG3-biotin functionalized SiO2 nanospheres (Ag@SiO2) was prepared and used as signal amplification label, and phos-tag-biotin played a vital role of "bridge" to link m6ATP and Ag@SiO2 through the two forms of specific interaction between phosphate group of m6ATP and phos-tag, biotin and streptavidin, respectively. Under the optimal experimental conditions, the immunosensor presented a wide linear range from 0.2 to 500nM and a low detection limit of 0.078nM (S/N=3). The reproducibility and specificity were acceptable. Moreover, the developed method was also validated for detect m6A content in human cell lines. Importantly, this detection strategy provides a promising immunodetection platform for ribonucleotides and deoxyribonucleotides with the advantages of simplicity, low-costing, specificity and sensitivity.

  9. An ultrasensitive sandwich-type electrochemical immunosensor based on signal amplification strategy of gold nanoparticles functionalized magnetic multi-walled carbon nanotubes loaded with lead ions.

    PubMed

    Li, Faying; Han, Jian; Jiang, Liping; Wang, Yulan; Li, Yueyun; Dong, Yunhui; Wei, Qin

    2015-06-15

    In this study, a novel and ultrasensitive sandwich-type electrochemical immunosensor was prepared for the quantitative detection of alpha fetoprotein (AFP), a well-known hepatocellular carcinoma biomarker. Gold nanoparticles (Au NPs) functionalized magnetic multi-walled carbon nanotubes (MWCNTs-Fe3O4) were prepared and utilized for the adsorption of lead ions (Pb(2+)) and the secondary antibodies (Ab2). The resultant nanocomposites (Pb(2+)@Au@MWCNTs-Fe3O4) were used as the label for signal amplification, showing better electrocatalytic activity towards the reduction of hydrogen peroxide (H2O2) than MWCNTs, MWCNTs-Fe3O4 or Au@MWCNTs-Fe3O4 due to the synergetic effect presented in Pb(2+)@Au@MWCNTs-Fe3O4. Moreover, Au NPs were electrodeposited on the surface of glassy carbon electrode (GCE) for the effective immobilization of primary antibodies (Ab1). Under the optimal conditions, a linear range from 10 fg/mL to 100 ng/mL and a detection limit of 3.33 fg/mL were obtained. The proposed electrochemical sandwich-type immunosensor shows high sensitivity, good selectivity and stability for the quantitative detection of AFP, holding a great potential in clinical and diagnostic applications.

  10. Au nanoparticles/hollow molybdenum disulfide microcubes based biosensor for microRNA-21 detection coupled with duplex-specific nuclease and enzyme signal amplification.

    PubMed

    Shuai, Hong-Lei; Huang, Ke-Jing; Chen, Ying-Xu; Fang, Lin-Xia; Jia, Meng-Pei

    2017-03-15

    An ultrasensitive electrochemical biosensor for detecting microRNAs is fabricated based on hollow molybdenum disulfide (MoS2) microcubes. Duplex-specific nuclease, enzyme and electrochemical-chemical-chemical redox cycling are used for signal amplification. Hollow MoS2 microcubes constructed by ultrathin nanosheets are synthesized by a facile template-assisted strategy and used as supporting substrate. For biosensor assembling, biotinylated ssDNA capture probes are first immobilized on Au nanoparticles (AuNPs)/MoS2 modified electrode in order to combine with streptavidin-conjugated alkaline phosphatase (SA-ALP). When capture probes hybridize with miRNAs, duplex-specific nuclease cleaves the formative duplexes. At the moment, the biotin group strips from the electrode surface and SA-ALP is incapacitated to attach onto electrode. Then, ascorbic acids induce the electrochemical-chemical-chemical redox cycling to produce electrochemical response in the presence of ferrocene methanol and tris (2-carboxyethyl) phosphine. Under optimum conditions, the proposed biosensor shows a good linear relationship between the current variation and logarithm of the microRNAs concentration ranging from 0.1fM to 0.1pM with a detection limit of 0.086fM (S/N=3). Furthermore, the biosensor is successfully applied to detect target miRNA-21 in human serum samples.

  11. Localization of single-copy T-DNA insertion in transgenic shallots (Allium cepa) by using ultra-sensitive FISH with tyramide signal amplification.

    PubMed

    Khrustaleva, L I; Kik, C

    2001-03-01

    The sensitivity of fluorescence in situ hybridization (FISH) for mapping plant chromosomes of single-copy DNA sequences is limited. We have adapted for plant cytogenetics a new signal-amplification method termed tyramide-FISH (Tyr-FISH). Until present this technique has only been applied to human chromosomes. The method is based on enzymatic deposition of fluorochrome-conjugated tyramide. With Tyr-FISH it was possible to detect target T-DNA sequences on plant metaphase chromosomes as small as 710 bp without using a cooled CCD camera. Short detection time and high sensitivity, in combination with a low background, make the Tyr-FISH method very suitable for routine application in plant cytogenetic research. With Tyr-FISH we analysed the position of T-DNA inserts in transgenic shallots. We found that the inserts were preferentially located in the distal region of metaphase chromosomes. Sequential fluorescence in situ hybridization with a 375 bp satellite sequence suggested that a specific T-DNA insert was located within the satellite sequence hybridization region on a metaphase chromosome. Analysis of less-condensed prophase and interphase chromosomes revealed that the T-DNA was integrated outside the satellite DNA-hybridization region in a more proximal euchromatin region.

  12. Electrochemical aptasensor for mucin 1 based on dual signal amplification of poly(o-phenylenediamine) carrier and functionalized carbon nanotubes tracing tag.

    PubMed

    Chen, Xiaojun; Zhang, Qi; Qian, Chunhua; Hao, Ning; Xu, Lin; Yao, Cheng

    2015-02-15

    Mucin 1 (MUC 1), as a most studied mucin, has become a useful marker for identifying breast cancer in the early stages. In this work, a novel method for the determination of MUC 1 in serum was developed based on a sandwich-type electrochemical aptasensor, which combined a dual signal amplification strategy of poly(o-phenylenediamine)-Au nanoparticles (PoPD-AuNPs) hybrid film as carrier and AuNPs functionalized silica/multiwalled carbon nanotubes core-shell nanocomposites (AuNPs/SiO2@MWCNTs) as tracing tag. The PoPD-AuNPs film provides a suitable microenvironment for stabilizing the primary aptamer (Apt) assembly, and the AuNPs/SiO2@MWCNTs enhances the surface area for immobilizing abundant secondary Apts as well as load large amounts of electrochemical probe thionine (Thi). In the presence of MUC 1, the sandwich-type recognition reacted on the aptasensor surface, and the Thi-AuNPs/SiO2@MWCNTs nanoprobes were captured onto the electrode surface to form biocomplex. AuNPs and MWCNTs could facilitate the electron transfer from Thi to the electrode, thus amplifying the detection response. Under the optimized experimental conditions, the proposed sensing strategy provided a wider linear dynamic range over three orders of magnitude with the detection limit down to 1 pM. Moreover, the aptasensor demonstrated good precision, acceptable stability and reproducibility.

  13. A competitive photoelectrochemical immunosensor based on a CdS-induced signal amplification strategy for the ultrasensitive detection of dexamethasone

    PubMed Central

    Wang, Xueping; Yan, Tao; Li, Yan; Liu, Yixin; Du, Bin; Ma, Hongmin; Wei, Qin

    2015-01-01

    A novel photoelectrochemical immunosensor based on the competitive strategy is proposed for the specific detection of dexamethasone (DXM). Graphitic carbon nitride coupled with bismuth sulfide are used as the sensing matrix for the immobilization of BSA-DXM on the electrode surface, while cadmium sulfide functionalized titanium dioxide (TiO2@CdS) is used as the photoelectric active labels of anti-DXM. Due to the perfect matching of energy levels between TiO2 and CdS, the in situ prepared composite labels show excellent photocurrent response under visible lights. The competitive binding of DXM in sample solutions and BSA-DXM on the electrode surface reduces the specific attachment of labels to the electrode, resulting in a decrease of the photocurrent intensity. Greatly enhanced sensitivity is achieved after the optimization of the detection conditions. Under the optimal detection condition, the well-designed immunosensor for DXM exhibits a low detection limit of 2 pg∙mL−1. Additionally, the proposed immunoassay system shows high specificity, good reproducibility and acceptable stability, which is also expected to become a promising platform for the detection of other small molecules. PMID:26648409

  14. Sensitive electrochemical aptamer cytosensor for highly specific detection of cancer cells based on the hybrid nanoelectrocatalysts and enzyme for signal amplification.

    PubMed

    Sun, Duanping; Lu, Jing; Zhong, Yuwen; Yu, Yanyan; Wang, Yu; Zhang, Beibei; Chen, Zuanguang

    2016-01-15

    Human cancer is becoming a leading cause of death in the world and the development of a straightforward strategy for early detection of cancer is urgently required. Herein, a sandwich-type electrochemical aptamer cytosensor was developed for detection of human liver hepatocellular carcinoma cells (HepG2) based on the hybrid nanoelectrocatalysts and enzyme for signal amplification. The thiolated TLS11a aptamers were used as a selective bio-recognition element, attached to the gold nanoparticles (AuNPs) modified the glassy carbon electrode (GCE) surface. Meanwhile, the electrochemical nanoprobes were fabricated through the G-quadruplex/hemin/aptamer complexes and horseradish peroxidase (HRP) immobilized on the surfaces of Au@Pd core-shell nanoparticle-modified magnetic Fe3O4/MnO2 beads (Fe3O4/MnO2/Au@Pd). After the target cells were captured, the hybrid nanoprobes were further assembled to form an aptamer-cell-nanoprobes sandwich-like system on the electrode surface. Then, hybrid Fe3O4/MnO2/Au@Pd nanoelectrocatalysts, G-quadruplex/hemin HRP-mimicking DNAzymes and the natural HRP enzyme efficiently catalyzed the oxidation of hydroquinone (HQ) with H2O2, amplifying the electrochemical signals and improving the detection sensitivity. This electrochemical cytosensor delivered a wide detection range of 1×10(2)-1×10(7)cellsmL(-1), high sensitivity with a low detection limit of 15cellsmL(-1), good selectivity and repeatability. Finally, an electrochemical reductive desorption method was performed to break gold-thiol bond and desorb the components on the AuNPs/GCE for regenerating the cytosensor. These results have demonstrated that the electrochemical cytosensor has the potential to be a feasible tool for cost-effective cancer cell detection in early cancer diagnosis.

  15. Ultrasensitive electrochemical sensing platform for microRNA based on tungsten oxide-graphene composites coupling with catalyzed hairpin assembly target recycling and enzyme signal amplification.

    PubMed

    Shuai, Hong-Lei; Huang, Ke-Jing; Xing, Ling-Li; Chen, Ying-Xu

    2016-12-15

    An ultrasensitive electrochemical biosensor for microRNA (miRNA) is developed based on tungsten oxide-graphene composites coupling with catalyzed hairpin assembly target recycling and enzyme signal amplification. WO3-Gr is prepared by a simple hydrothermal method and then coupled with gold nanoparticles to act as a sensing platform. The thiol-terminated capture probe H1 is immobilized on electrode through Au-S interaction. In the presence of target miRNA, H1 opens its hairpin structure by hybridization with target miRNA. This hybridization can be displaced from the structure by another stable biotinylated hairpin DNA (H2), and target miRNA is released back to the sample solution for next cycle. Thus, a large amount of H1-H2 duplex is produced after the cyclic process. At this point, a lot of signal indicators streptavidin-conjugated alkaline phosphatase (SA-ALP) are immobilized on the electrode by the specific binding of avidin-biotin. Then, thousands of ascorbic acid, which is the enzymatic product of ALP, induces the electrochemical-chemical-chemical redox cycling to produce a strongly electrochemical response in the presence of ferrocene methanol and tris (2-carboxyethyl) phosphine. Under the optimal experimental conditions, the established biosensor can detect target miRNA down to 0.05fM (S/N=3) with a linear range from 0.1fM to 100pM, and discriminate target miRNA from mismatched miRNA with a high selectivity.

  16. Coherent beam amplification with a photorefractive liquid crystal.

    PubMed

    Khoo, I C; Guenther, B D; Wood, M V; Chen, P; Shih, M Y

    1997-08-15

    Coherent amplification of a signal beam by a strong pump beam is observed in thin films of fullerene-doped nematic liquid crystal. Exponential gain constants as high as 2890 cm(-1) with no phase cross talk are achieved at low applied dc bias voltage and pump beam intensity. The underlying mechanism is the electro-optically induced spatially reorientation of the liquid-crystal axis and the resultant phase-shifted index grating required for two-beam coupling.

  17. Macromolecular amplification of binding response in superaptamer hydrogels.

    PubMed

    Bai, Wei; Gariano, Nicholas A; Spivak, David A

    2013-05-08

    It is becoming more important to detect ultralow concentrations of analytes for biomedical, environmental, and national security applications. Equally important is that new methods should be easy to use, inexpensive, portable, and if possible allow detection by the naked eye. By and large, detection of low concentrations of analytes cannot be achieved directly but requires signal amplification by catalysts, macromolecules, metal surfaces, or supramolecular aggregates. The rapidly progressing field of macromolecular signal amplification has been advanced using conjugated polymers, chirality in polymers, solvating polymers, and polymerization/depolymerization strategies. A new type of aptamer-based hydrogel with specific response to target proteins presented in this report demonstrates an additional category of macromolecular signal amplification. This superaptamer assembly provides the first example of using protein-specific aptamers to create volume-changing hydrogels with amplified response to the target protein. A remarkable aspect of these superaptamer hydrogels is that volume shrinking is visible to the naked eye down to femtomolar concentrations of protein. This extraordinary macromolecular amplification is attributed to a complex interplay between protein-aptamer supramolecular cross-links and the consequential reduction of excluded volume in the hydrogel. Specific recognition is even maintained in biological matrices such as urine and tears. Furthermore, the gels can be dried for long-term storage and regenerated for use without loss of activity. In practice, the ease of this biomarker detection method offers an alternative to traditional analytical techniques that require sophisticated instrumentation and highly trained personnel.

  18. DNA-based hybridization chain reaction and biotin-streptavidin signal amplification for sensitive detection of Escherichia coli O157:H7 through ELISA.

    PubMed

    Guo, Qi; Han, Jiao-Jiao; Shan, Shan; Liu, Dao-Feng; Wu, Song-Song; Xiong, Yong-Hua; Lai, Wei-Hua

    2016-12-15

    This study reported on a novel sandwich enzyme linked immunosorbent assay (ELISA) for the sensitive determination of Escherichia coli O157:H7 (E. coli O157:H7) by using DNA-based hybridization chain reaction (HCR) and biotin-streptavidin signal amplification. The anti-E. coli O157:H7 polyclonal antibody (pAb) was immobilized in the ELISA wells. The anti-E. coli O157:H7 monoclonal antibody (mAb) and initiator strand (DNA1) were labeled on gold nanoparticle (AuNP) to form a mAb-AuNP-DNA1 complex. In the presence of the target E. coli O157:H7, the sandwiched immunocomplex, which is pAb-E. coli O157:H7-mAb-AuNP-DNA1, could be formed. Two types of biotinylated hairpin were subsequently added in the ELISA well. A nicked double-stranded DNA (dsDNA) that contained abundant biotins was formed after HCR. Detection was performed after adding horseradish peroxidase-streptavidin and substrate/chromogen solution. Under optimal conditions, E. coli O157:H7 could be detected in the range of 5×10(2) CFU/mL to 1×10(7) CFU/mL; the limit of detection was 1.08×10(2) CFU/mL in pure culture. The LOD of the novel ELISA was 185 times lower than that of traditional ELISA. The proposed method is considerably specific and can be applied in the detection of whole milk samples inoculated with E. coli O157:H7. The coefficient of variation of in pure culture and in whole milk was 0.99-5.88% and 0.76-5.38%, respectively. This method offers a promising application in the detection of low concentrations of food-borne pathogens.

  19. In Situ Generation of Electron Donor to Assist Signal Amplification on Porphyrin-Sensitized Titanium Dioxide Nanostructures for Ultrasensitive Photoelectrochemical Immunoassay.

    PubMed

    Shu, Jian; Qiu, Zhenli; Zhuang, Junyang; Xu, Mingdi; Tang, Dianping

    2015-10-28

    An ultrasensitive photoelectrochemical (PEC) immunoassay protocol for quantitative detection of low-abundant proteins at a low potential was designed by utilizing porphyrin-sensitized titanium dioxide (TiO2) nanostructures. Experimental results demonstrated that the water-soluble 5,10,15,20-tetra(4-sulfophenyl)-21H,23H-porphyrin (TSPP) could be bound onto titanium dioxide via the sulfonic group. TSPP-sensitized TiO2 nanostructures exhibited better photoelectrochemical responses and stability in comparison with TiO2 nanoparticles alone under continuous illumination. Using carcinoembryonic antigen (CEA) as a model analyte, a typical PEC immunosensor by using TSPP-TiO2 as the affinity support of anti-CEA capture antibody (Ab1) to facilitate the improvement of photocurrent response was developed. Bioconjugates of secondary antibody and glucose oxidase with gold nanoparticles (Ab2/GOx-AuNPs) was introduced by an antigen-antibody immunoreaction. AuNP acted as a powerful scaffold to bind with bioactive molecules, while GOx catalyzed glucose to in situ generate hydrogen peroxide (H2O2). The generated H2O2 as a sacrificial electron donor could be oxidized by the photogenerated holes to assist the signal amplification at a low potential under light excitation, thus eliminating interference from other species coexisting in the samples. Under optimal conditions, the PEC immunosensor showed a good linear relationship ranging from 0.02 to 40 ng mL(-1) with a low detection limit of 6 pg mL(-1) CEA. The precision, reproducibility, and specificity were acceptable. In addition, the method accuracy was also evaluated for quantitatively monitoring human serum samples, giving results matching with the referenced CEA ELISA kit.

  20. Sensitive chemiluminescence immunoassay for E. coli O157:H7 detection with signal dual-amplification using glucose oxidase and laccase.

    PubMed

    Zhang, Yun; Tan, Chen; Fei, Ruihua; Liu, Xiaoxiao; Zhou, Yuan; Chen, Jing; Chen, Huanchun; Zhou, Rui; Hu, Yonggang

    2014-01-21

    A novel, sensitive chemiluminescence (CL) immunoassay for Escherichia coli O157:H7 detection with signal dual-amplification using glucose oxidase (GOx) and laccase was investigated. The method was based on the characterization of a luminol-H2O2-laccase reaction. Compared with the horseradish peroxidase-based biosensor, laccase exhibited high catalytic activity in strong alkaline medium, which was compatible with the luminol system. The capture antibody was immobilized onto the magnetic bead (MB) surfaces. The detection antibody was linked with GOx through biotin-avidin recognition. Accordingly, the bioconjugation of MB-caputure antibody- E. coli O157:H7-detection antibody-GOx catalyzed the substrate glucose, thereby generating H2O2. E. coli O157:H7 was then detected by measuring the CL intensity after H2O2 formation. Under optimal conditions, the calibration plot obtained for E. coli O157:H7 was approximately linear from 4.3 × 10(3) colony-forming unit (CFU) mL(-1) to 4.3 × 10(5) CFU mL(-1), and the total assay time was <2.0 h without any enrichment. The limit of detection for the assay was 1.2 × 10(3) CFU mL(-1) (3σ), which was considerably lower than that of enzyme-linked immunosorbent assay method (1.0 × 10(5) CFU mL(-1)) (3σ). A series of repeatability measurements of using 1.7 × 10(4) CFU mL(-1) E. coli O157:H7 exhibited reproducible results with a relative standard deviation (RSD) of 3.5% (n = 11). Moreover, the proposed method was successfully used to detect E. coli O157:H7 in synthetic samples (spring water, apple juice, and skim milk), which indicated its potential practical application. This protocol can be applied in various fields of study.

  1. Homogeneous electrochemical immunoassay of aflatoxin B1 in foodstuff using proximity-hybridization-induced omega-like DNA junctions and exonuclease III-triggered isothermal cycling signal amplification.

    PubMed

    Tang, Juan; Huang, Yapei; Liu, Huiqiong; Zhang, Cengceng; Tang, Dianping

    2016-12-01

    A new homogeneous electrochemical immunosensing platform was designed for sensitive detection of aflatoxin B1 (AFB1) in foodstuff. The system consisted of anti-AFB1 antibody labeled DNA1 (Ab-DNA1), AFB1-bovine serum albumin (BSA)-conjugated DNA2 (AFB1-DNA2), and methylene blue functionalized hairpin DNA. Owing to a specific antigen-antibody reaction between anti-AFB1 and AFB1-BSA, the immunocomplex formed assisted the proximity hybridization of DNA1 with DNA2, thus resulting in the formation of an omega-like DNA junction. Thereafter, the junction opened the hairpin DNA to construct a new double-stranded DNA, which could be readily cleaved by exonuclease III to release the omega-like DNA junction and methylene blue. The dissociated DNA junction could repeatedly hybridize with residual hairpin DNA molecules with exonuclease III-based isothermal cycling amplification, thereby releasing numerous free methylene blue molecules into the detection solution. The as-produced free methylene blue molecules could be captured by a negatively charged indium tin oxide electrode, each of which could produce an electronic signal within the applied potentials. On introduction of target AFB1, the analyte competed with AFB1-DNA2 for the conjugated anti-AFB1 on the Ab-DNA1, subsequently decreasing the amount of omega-like DNA junctions formed, hence causing methylene blue labeled hairpin DNA to move far away from the electrode surface. Under optimal conditions the detectable electrochemical signal decreased with increasing amount of target AFB1 in a dynamic working range of 0.01-30 ng mL(-1) with a detection limit of 4.8 pg mL(-1). In addition, the precision and reproducibility of this system were acceptable. Finally, the method was further evaluated for analysis of naturally contaminated or AFB1-spiked peanut samples, giving results that matched well with those obtained with a commercial AFB1 ELISA kit.

  2. A new system for the amplification of biological signals: RecA and complimentary single strand DNA probes on a leaky surface acoustic wave biosensor.

    PubMed

    Zhang, Liqun; Wang, Yunxia; Chen, Ming; Luo, Yang; Deng, Kun; Chen, Dong; Fu, Weiling

    2014-10-15

    This research describes a new amplification signals system of the leaky surface acoustic wave (LSAW) bis-peptide nucleic acid (bis-PNA) biosensor for the simple, sensitive and rapid detection of the target double-stranded DNA (dsDNA). The system consists of a RecA protein-coated complementary single-stranded DNA (cssDNA) probe complex that amplifies the biological signal to improve the sensitivity of the biosensor. The bis-PNA probe for detecting HPV was first immobilized on a gold surface membrane of the detection channel. After the probe was completely hybridized with the corresponding target DNA, different concentrations of the "RecA protein-complementary single strand DNA probe" were added to react with the bis-PNA/dsDNA complex. The phase shift of the LSAW biosensors, which was measured and found to be most significant when the RecA protein was 45 μg/mL and the ATPγS was 2.5 mmol/L. Compared with other concentrations (P<0.01) of RecA and ATPγS, the value of the phase shift was (11.74 ± 1.03) degrees and the ratio of the phase shift and hybridization time clearly outperformed that of the other concentrations. Compared to the direct hybridization of the bis-PNA probe and the target DNA sequence, the sensitivity was effectively improved and the detection time was significantly shortened. PNA binding adjacent to the area of the target sequence homologous to the probe significantly increased the yield of the hybridization reaction between the PNA/dsDNA complex and the RecA protein-coated cssDNA probe. In this condition, the phase shift was significantly obvious and the detection time was significantly shortened. In conclusion, the combination of the RecA protein-coated cssDNA probe and the LSAW bis-PNA biosensor provides sensitivity and simple and rapid detection of clinical trace pathogenic microorganisms.

  3. Characterisation of signal enhancements achieved when utilizing a photon diode in deep Raman spectroscopy of tissue

    PubMed Central

    Vardaki, Martha Z.; Matousek, Pavel; Stone, Nicholas

    2016-01-01

    We characterise the performance of a beam enhancing element (‘photon diode’) for use in deep Raman spectroscopy (DRS) of biological tissues. The optical component enhances the number of laser photons coupled into a tissue sample by returning escaping photons back into it at the illumination zone. The method is compatible with transmission Raman spectroscopy, a deep Raman spectroscopy concept, and its implementation leads to considerable enhancement of detected Raman photon rates. In the past, the enhancement concept was demonstrated with a variety of samples (pharmaceutical tablets, tissue, etc) but it was not systematically characterized with biological tissues. In this study, we investigate the enhancing properties of the photon diode in the transmission Raman geometry as a function of: a) the depth and b) the optical properties of tissue samples. Liquid tissue phantoms were employed to facilitate systematic variation of optical properties. These were chosen to mimic optical properties of human tissues, including breast and prostate. The obtained results evidence that a photon diode can enhance Raman signals of tissues by a maximum of × 2.4, although it can also decrease the signals created towards the back of samples that exhibit high scattering or absorption properties. PMID:27375932

  4. Optical pulse synthesis using brillouin selective sideband amplification

    NASA Technical Reports Server (NTRS)

    Yao, X. Steve (Inventor)

    2002-01-01

    Techniques for producing optical pulses based on Brillouin selective sideband amplification by using a common modulation control signal to modulate both a signal beam to produce multiple sideband signals and a single pump beam to produce multiple pump beams.

  5. CuO-induced signal amplification strategy for multiplexed photoelectrochemical immunosensing using CdS sensitized ZnO nanotubes arrays as photoactive material and AuPd alloy nanoparticles as electron sink.

    PubMed

    Sun, Guoqiang; Zhang, Yan; Kong, Qingkun; Zheng, Xiaoxiao; Yu, Jinghua; Song, Xianrang

    2015-04-15

    In this work, multiplexed photoelectrochemical (PEC) immunoassays are introduced into an indium tin oxide (ITO) device. Firstly, the ITO device is fabricated using a simple acid etch treatment method. Secondly, AuPd alloy nanoparticles are electro-deposited on ITO working electrodes as electron sink to construct the immunosensor platform. After that, ZnO nanotubes (ZNTs) arrays are synthesized via chemical etching of ZnO nanorods that are grown on AuPd surface by electrochemical deposition method. Subsequently, CdS is electro-deposited on ZNTs arrays and used as photoactive material. Then, CuO nanoseeds are labeled with signal antibodies and firstly used as PEC signal amplification label. The introduction of CuO brings signal amplification because of the conduction band (CB) of both CuO and ZnO are lower than that of CdS, CuO will compete the photo-induced electrons in CB of CdS with ZnO, leading to the decrease of the photocurrent intensity. Using cancer antigen 125, prostate specific antigen and α-fetoprotein as model analytes, the proposed immunoassay exhibits excellent precision and sensitivity. Meanwhile, this work provides a promising, addressable and simple strategy for the multi-detection of tumor markers.

  6. An electrochemical aptasensor for thrombin using synergetic catalysis of enzyme and porous Au@Pd core-shell nanostructures for signal amplification.

    PubMed

    Xu, Wenju; Yi, Huayu; Yuan, Yali; Jing, Pei; Chai, Yaqin; Yuan, Ruo; Wilson, George S

    2015-02-15

    In this work, a sensitive electrochemical aptasensor for thrombin (TB) based on synergetic catalysis of enzyme and porous Au@Pd core-shell nanostructure has been constructed. With the advantages of large surface area and outstanding catalytic performance, porous Au@Pd core-shell nanostructures were firstly employed as the nanocarrier for the immobilization of electroactive toluidine blue (Tb), hemin/G-quadruplex formed by intercalating hemin into the TB aptamer (TBA) and glucose oxidase (GOx). As a certain amount of glucose was added into the detection cell, GOx rapidly catalyzed the oxidation of glucose, coupling with the local generation of H2O2 in the presence of dissolved O2. Then, porous Au@Pd nanoparticles and hemin/G-quadruplex as the peroxidase mimics efficiently catalyzed the reduction of H2O2, amplifying the electrochemical signal and improving the sensitivity. Finally, a detection limit of 0.037pM for TB was achieved. The excellent performance of the aptasensor indicated its promising prospect as a valuable tool in simple and cost-effective TB detection in clinical application.

  7. Weak value amplification via second-order correlated technique

    NASA Astrophysics Data System (ADS)

    Ting, Cui; Jing-Zheng, Huang; Xiang, Liu; Gui-Hua, Zeng

    2016-02-01

    We propose a new framework combining weak measurement and second-order correlated technique. The theoretical analysis shows that weak value amplification (WVA) experiment can also be implemented by a second-order correlated system. We then build two-dimensional second-order correlated function patterns for achieving higher amplification factor and discuss the signal-to-noise ratio influence. Several advantages can be obtained by our proposal. For instance, detectors with high resolution are not necessary. Moreover, detectors with low saturation intensity are available in WVA setup. Finally, type-one technical noise can be effectively suppressed. Project supported by the Union Research Centre of Advanced Spaceflight Technology (Grant No. USCAST2013-05), the National Natural Science Foundation of China (Grant Nos. 61170228, 61332019, and 61471239), and the High-Tech Research and Development Program of China (Grant No. 2013AA122901).

  8. Orthogonal amplification of nanoparticles for improved diagnostic sensing.

    PubMed

    Peterson, Vanessa M; Castro, Cesar M; Lee, Hakho; Weissleder, Ralph

    2012-04-24

    There remains an ongoing need for fast, highly sensitive, and quantitative technologies that can detect and profile rare cells in freshly harvested samples. Recent developments in nanomaterial-based detection platforms provide advantages over traditional approaches in terms of signal sensitivity, stability, and the possibility for performing multiplexed measurements. Here, we describe a bioorthogonal, nanoparticle amplification technique capable of rapid augmentation of detection sensitivities by up to 1-2 orders of magnitude over current methods. This improvement in sensitivity was achieved by (i) significantly reducing background noise arising from nonspecific nanoparticle binding, (ii) increasing nanomaterial binding through orthogonal rounds of amplification, and (iii) implementing a cleavage step to improve assay robustness. The developed method allowed sensitive detection and molecular profiling of scant tumor cells directly in unpurified human clinical samples such as ascites. With its high sensitivity and simplified assay steps, this technique will likely have broad utility in nanomaterial-based diagnostics.

  9. Hybrid chirped pulse amplification system

    SciTech Connect

    Barty, Christopher P.; Jovanovic, Igor

    2005-03-29

    A hybrid chirped pulse amplification system wherein a short-pulse oscillator generates an oscillator pulse. The oscillator pulse is stretched to produce a stretched oscillator seed pulse. A pump laser generates a pump laser pulse. The stretched oscillator seed pulse and the pump laser pulse are directed into an optical parametric amplifier producing an optical parametric amplifier output amplified signal pulse and an optical parametric amplifier output unconverted pump pulse. The optical parametric amplifier output amplified signal pulse and the optical parametric amplifier output laser pulse are directed into a laser amplifier producing a laser amplifier output pulse. The laser amplifier output pulse is compressed to produce a recompressed hybrid chirped pulse amplification pulse.

  10. Camera-based ratiometric fluorescence transduction of nucleic acid hybridization with reagentless signal amplification on a paper-based platform using immobilized quantum dots as donors.

    PubMed

    Noor, M Omair; Krull, Ulrich J

    2014-10-21

    Paper-based diagnostic assays are gaining increasing popularity for their potential application in resource-limited settings and for point-of-care screening. Achievement of high sensitivity with precision and accuracy can be challenging when using paper substrates. Herein, we implement the red-green-blue color palette of a digital camera for quantitative ratiometric transduction of nucleic acid hybridization on a paper-based platform using immobilized quantum dots (QDs) as donors in fluorescence resonance energy transfer (FRET). A nonenzymatic and reagentless means of signal enhancement for QD-FRET assays on paper substrates is based on the use of dry paper substrates for data acquisition. This approach offered at least a 10-fold higher assay sensitivity and at least a 10-fold lower limit of detection (LOD) as compared to hydrated paper substrates. The surface of paper was modified with imidazole groups to assemble a transduction interface that consisted of immobilized QD-probe oligonucleotide conjugates. Green-emitting QDs (gQDs) served as donors with Cy3 as an acceptor. A hybridization event that brought the Cy3 acceptor dye in close proximity to the surface of immobilized gQDs was responsible for a FRET-sensitized emission from the acceptor dye, which served as an analytical signal. A hand-held UV lamp was used as an excitation source and ratiometric analysis using an iPad camera was possible by a relative intensity analysis of the red (Cy3 photoluminescence (PL)) and green (gQD PL) color channels of the digital camera. For digital imaging using an iPad camera, the LOD of the assay in a sandwich format was 450 fmol with a dynamic range spanning 2 orders of magnitude, while an epifluorescence microscope detection platform offered a LOD of 30 fmol and a dynamic range spanning 3 orders of magnitude. The selectivity of the hybridization assay was demonstrated by detection of a single nucleotide polymorphism at a contrast ratio of 60:1. This work provides an

  11. Photon number amplification/duplication through parametric conversion

    NASA Technical Reports Server (NTRS)

    Dariano, G. M.; Macchiavello, C.; Paris, M.

    1993-01-01

    The performance of parametric conversion in achieving number amplification and duplication is analyzed. It is shown that the effective maximum gains G(sub *) remain well below their integer ideal values, even for large signals. Correspondingly, one has output Fano factors F(sub *) which are increasing functions of the input photon number. On the other hand, in the inverse (deamplifier/recombiner) operating mode quasi-ideal gains G(sub *) and small factors F(sub *) approximately equal to 10 percent are obtained. Output noise and non-ideal gains are ascribed to spontaneous parametric emission.

  12. Weak Value Amplification of a Post-Selected Single Photon

    NASA Astrophysics Data System (ADS)

    Hallaji, Matin

    Weak value amplification (WVA) is a measurement technique in which the effect of a pre- and post-selected system on a weakly interacting probe is magnified. In this thesis, I present the first experimental observation of WVA of a single photon. We observed that a signal photon --- sent through a polarization interferometer and post-selected by photodetection in the almost-dark port --- can act like eight photons. The effect of this single photon is measured as a nonlinear phase shift on a separate laser beam. The interaction between the two is mediated by a sample of laser- cooled 85Rb atoms. Electromagnetically induced transparency (EIT) is used to enhance the nonlinearity and overcome resonant absorption. I believe this work to be the first demonstration of WVA where a deterministic interaction is used to entangle two distinct optical systems. In WVA, the amplification is contingent on discarding a large portion of the original data set. While amplification increases measurement sensitivity, discarding data worsens it. Questioning whether these competing effects conspire to improve or diminish measurement accuracy has resulted recently in controversy. I address this question by calculating the maximum amount of information achievable with the WVA technique. By comparing this information to that achievable by the standard technique, where no post-selection is employed, I show that the WVA technique can be advantageous under a certain class of noise models. Finally, I propose a way to optimally apply the WVA technique.

  13. Ultrasensitive photoelectrochemical immunoassay for matrix metalloproteinase-2 detection based on CdS:Mn/CdTe cosensitized TiO2 nanotubes and signal amplification of SiO2@Ab2 conjugates.

    PubMed

    Fan, Gao-Chao; Han, Li; Zhu, Hua; Zhang, Jian-Rong; Zhu, Jun-Jie

    2014-12-16

    An ultrasensitive photoelectrochemical sandwich immunoassay was developed to detect matrix metalloproteinase-2 (MMP-2, antigen, Ag) based on CdS:Mn/CdTe cosensitized TiO2 nanotubes (TiO2-NTs) and signal amplification of SiO2@Ab2 conjugates. Specifically, the TiO2-NTs electrode was first deposited with CdS:Mn by successive ionic layer adsorption and reaction technique and then further coated with CdTe quantum dots (QDs) via the layer-by-layer method, forming TiO2-NTs/CdS:Mn/CdTe cosensitized structure, which was employed as a matrix to immobilize capture MMP-2 antibodies (Ab1); whereas, SiO2 nanoparticles were coated with signal MMP-2 antibodies (Ab2) to form SiO2@Ab2 conjugates, which were used as signal amplification elements via the specific antibody-antigen immunoreaction between Ag and Ab2. The ultrahigh sensitivity of this immunoassay derived from the two major reasons as below. First, the TiO2-NTs/CdS:Mn/CdTe cosensitized structure could adequately absorb the light energy, dramatically promote electron transfer, and effectively inhibit the electron-hole recombination, resulting in significantly enhanced photocurrent intensity of the sensing electrode. However, in the presence of target Ag, the immobilized SiO2@Ab2 conjugates could evidently increase the steric hindrance of the sensing electrode and effectively depress the electron transfer, leading to obviously decreased photocurrent intensity. Accordingly, the well-designed photoelectrochemical immunoassay exhibited a low detection limit of 3.6 fg/mL and a wide linear range from 10 fg/mL to 500 pg/mL for target Ag detection. Meanwhile, it also presented good reproducibility, specificity, and stability and might open a new promising platform for the detection of other important biomarkers.

  14. Ultrasensitive DNA detection based on two-step quantitative amplification on magnetic nanoparticles

    NASA Astrophysics Data System (ADS)

    Jin, Mingliang; Liu, Xia; van den Berg, Albert; Zhou, Guofu; Shui, Lingling

    2016-08-01

    Sensitive detection of a specific deoxyribo nucleic acid (DNA) sequence is important for biomedical applications. In this report, a two-step amplification strategy is developed based on magnetic nanoparticles (MNPs) to achieve ultrasensitive DNA fluorescence detection. The first level amplification is obtained from multiple binding sites on MNPs to achieve thousands of probe DNA molecules on one nanoparticle surface. The second level amplification is gained by enzymatic reaction to achieve fluorescence signal enhancement. MNPs functionalized by probe DNA (DNAp) are bound to target DNA (t-DNA) molecules with a ratio of 1:1 on a substrate with capture DNA (DNAc). After the MNPs with DNAp are released from the substrate, alkaline phosphatase (AP) is labelled to MNPs via hybridization reaction between DNAp on MNPs and detection DNAs (DNAd) with AP. The AP on MNPs catalyses non-fluorescent 4-methylumbelliferyl phosphate (4-MUP) to fluorescent 4-methylumbelliferone (4-MU) with high intensity. Finally, fluorescence intensity of the 4-MU is detected by a conventional fluorescence spectrophotometer. With this two-step amplification strategy, the limit of detection (LOD) of 2.8 × 10-18 mol l-1 for t-DNA has been achieved.

  15. A double signal amplification platform for ultrasensitive and simultaneous detection of ascorbic acid, dopamine, uric acid and acetaminophen based on a nanocomposite of ferrocene thiolate stabilized Fe₃O₄@Au nanoparticles with graphene sheet.

    PubMed

    Liu, Meiling; Chen, Qiong; Lai, Cailang; Zhang, Youyu; Deng, Jianhui; Li, Haitao; Yao, Shouzhuo

    2013-10-15

    A double signal amplification platform for ultrasensitive and simultaneous detection of ascorbic acid (AA), dopamine (DA), uric acid (UA) and acetaminophen (AC) was fabricated by a nanocomposite of ferrocene thiolate stabilized Fe₃O₄@Au nanoparticles with graphene sheet. The platform was constructed by coating a newly synthesized phenylethynyl ferrocene thiolate (Fc-SAc) modified Fe₃O₄@Au NPs coupling with graphene sheet/chitosan (GS-chitosan) on a glassy carbon electrode (GCE) surface. The Fe₃O₄@Au-S-Fc/GS-chitosan modified GCE exhibits a synergistic catalytic and amplification effect toward AA, DA, UA and AC oxidation. The oxidation peak currents of the four compounds on the electrode were linearly dependent on AA, DA, UA and AC concentrations in the ranges of 4-400 μM, 0.5-50 μM, 1-300 μM and 0.3-250 μM in the individual detection of each component, respectively. By simultaneously changing the concentrations of AA, DA, UA and AC, their electrochemical oxidation peaks appeared at -0.03, 0.15, 0.24 and 0.35 V, and good linear current responses were obtained in the concentration ranges of 6-350, 0.5-50, 1-90 and 0.4-32 μM with the detection limits of 1, 0.1, 0.2 and 0.05 μM (S/N=3), respectively.

  16. The sandwich-type electrochemiluminescence immunosensor for α-fetoprotein based on enrichment by Fe3O4-Au magnetic nano probes and signal amplification by CdS-Au composite nanoparticles labeled anti-AFP.

    PubMed

    Zhou, Hankun; Gan, Ning; Li, Tianhua; Cao, Yuting; Zeng, Saolin; Zheng, Lei; Guo, Zhiyong

    2012-10-09

    A novel and sensitive sandwich-type electrochemiluminescence (ECL) immunosensor was fabricated on a glassy carbon electrode (GCE) for ultra trace levels of α-fetoprotein (AFP) based on sandwich immunoreaction strategy by enrichment using magnetic capture probes and quantum dots coated with Au shell (CdS-Au) as the signal tag. The capture probe was prepared by immobilizing the primary antibody of AFP (Ab1) on the core/shell Fe(3)O(4)-Au nanoparticles, which was first employed to capture AFP antigens to form Fe(3)O(4)-Au/Ab1/AFP complex from the serum after incubation. The product can be separated from the background solution through the magnetic separation. Then the CdS-Au labeled secondary antibody (Ab2) as signal tag (CdS-Au/Ab2) was conjugated successfully with Fe(3)O(4)-Au/Ab1/AFP complex to form a sandwich-type immunocomplex (Fe(3)O(4)-Au/Ab1/AFP/Ab2/CdS-Au), which can be further separated by an external magnetic field and produce ECL signals at a fixed voltage. The signal was proportional to a certain concentration range of AFP for quantification. Thus, an easy-to-use immunosensor with magnetic probes and a quantum dots signal tag was obtained. The immunosensor performed at a level of high sensitivity and a broad concentration range for AFP between 0.0005 and 5.0 ng mL(-1) with a detection limit of 0.2 pg mL(-1). The use of magnetic probes was combined with pre-concentration and separation for trace levels of tumor markers in the serum. Due to the amplification of the signal tag, the immunosensor is highly sensitive, which can offer great promise for rapid, simple, selective and cost-effective detection of effective biomonitoring for clinical application.

  17. Integrated platform with magnetic purification and rolling circular amplification for sensitive fluorescent detection of ochratoxin A.

    PubMed

    Yao, Li; Chen, Yinji; Teng, Jun; Zheng, Wanli; Wu, Jingjing; Adeloju, Samuel B; Pan, Daodong; Chen, Wei

    2015-12-15

    In this article, we report the detection of ochratoxin A (OTA) with excellent sensitivity with the two-aspect signal amplification treatments. Combining the unique property of magnetic nanoparticles and the high efficiency of the in vitro amplification of rolling circular amplification (RCA), the competitive sensing protocol for ultrasensitive detection of OTA was achieved in about 80 min. The excellent magnetic separation treatment could effectively avoid the interference of background fluorescent noise in the sensing system while the RCA could tremendously increase the hybridization sequence for the quantum dot labeled probes and further increase the sensing response signal. Afterwards, two factors affecting the final detection limit, concentration of RCA components and RCA reaction time, were all systematically optimized for the best sensing performance. The response of the optimized protocol for OTA detection is highly linear over the wider range from 10(-3) to 10 ppb, which is 3 orders improvement in sensing range, and the limit of detection is calculated to be as low as 0.13 ppt, which is 10,000 folds improvement compared with the traditional methods. More importantly, given the selected aptamer, this universal signal amplification protocol could be widely applied to other fields by just change the recognition sequence of the aptamer.

  18. Allelic drop-out and preferential amplification in single cells and human blastomeres: implications for preimplantation diagnosis of sex and cystic fibrosis.

    PubMed

    Findlay, I; Ray, P; Quirke, P; Rutherford, A; Lilford, R

    1995-06-01

    Previously the diagnosis of sex and cystic fibrosis status has been studied on single cells using the polymerase chain reaction (PCR). It has been suggested that allelic drop-out (PCR failure of one allele) and/or preferential amplification (hypo-amplification of one allele) may contribute to poor reliability and misdiagnosis, although this remains controversial as some reports suggest that allelic drop-out does not occur. We investigated an improved method of diagnosing sex and cystic fibrosis in single cells using a new technology (fluorescent PCR) to determine the base level of PCR artefacts (allelic drop-out and preferential amplification) which, in combination with improved sensitivity, should improve PCR reliability and accuracy. Fluorescent PCR gives high reliability (approximately 97%) and accuracy rates (approximately 97%) in somatic cells for both sex and cystic fibrosis diagnosis and its lower detection threshold allows allelic drop-out and preferential amplification to be easily distinguished. We also achieved high reliability and accuracy in diagnosing cystic fibrosis in human blastomeres. This study confirms earlier reports of both allelic drop-out and preferential amplification in single cell analysis. We demonstrate that both allelic drop-out and preferential amplification occur in somatic cells and suggest these are separate phenomena. Preferential amplification appeared common in single cell PCR while allelic drop-out apparently occurred at random in each allele. Preferential amplification was mainly amplification of the larger allele. We suggest that some inaccuracy/misdiagnosis may be due to both preferential amplification as well as allelic drop-out. Other findings were variability in drop-out between PCR and that amplification of signals from human blastomeres may be linked to embryo quality. We suggest that allelic drop-out is dependent on the number of cells within the sample.

  19. Experimental demonstration of joule-level non-collinear optical parametric chirped-pulse amplification in yttrium calcium oxyborate.

    PubMed

    Yu, Lianghong; Liang, Xiaoyan; Li, Jinfeng; Wu, Anhua; Zheng, Yanqing; Lu, Xiaoming; Wang, Cheng; Leng, Yuxin; Xu, Jun; Li, Ruxin; Xu, Zhizhan

    2012-05-15

    In this Letter, we report on what is, to our knowledge, the first experimental demonstration of yttrium calcium oxyborate (YCOB) for joule-level and broadband non-collinear optical parametric chirped-pulse amplification centered at 800 nm. Based on a Ti:sapphire chirped-pulse amplification front end, an amplified signal energy of 3.36 J was generated with a pump of 35 J in the crystal. Compressed pulse duration of 44.3 fs, with a bandwidth of 49 nm, was achieved. The results confirm that YCOB crystal is another potential alternative as a final amplifier besides Ti:sapphire in a petawatt laser at 800 nm.

  20. Experimental demonstration of phase-sensitive regeneration of a binary phase-shift keying channel without a phase-locked loop using Brillouin amplification.

    PubMed

    Almaiman, Ahmed; Cao, Yinwen; Ziyadi, Morteza; Mohajerin-Ariaei, Amirhossein; Liao, Peicheng; Bao, Changjing; Alishahi, Fatemeh; Fallahpour, Ahmad; Shamee, Bishara; Ahmed, Nisar; Willner, Asher J; Akasaka, Youichi; Ikeuchi, Tadashi; Takasaka, Shigehiro; Sugizaki, Ryuichi; Wilkinson, Steven; Touch, Joseph D; Tur, Moshe; Willner, Alan E

    2016-12-01

    All-optical phase regeneration of a binary phase-shift keying signal is demonstrated at 10-30 Gb/s without a phase-locked loop in a phase-sensitive amplification-based system using Brillouin amplification of the idler. The system achieves phase noise reduction of up to 56% and up to 11 dB OSNR gain at 10-5 bit error rate for the 10 Gb/s signal. The system's sensitivity to different parameters and stability is also evaluated.

  1. A waveform detector that targets template–decorrelated signals and achieves its predicted performance, Part I: Demonstration with IMS data

    SciTech Connect

    Carmichael, Joshua Daniel

    2016-01-01

    Here, waveform correlation detectors used in seismic monitoring scan multichannel data to test two competing hypotheses: that data contain (1) a noisy, amplitude-scaled version of a template waveform, or, (2) only noise. In reality, seismic wavefields include signals triggered by non-target sources (background seismicity) and targets signals that are only partially correlated with the waveform template.

  2. NAIMA: target amplification strategy allowing quantitative on-chip detection of GMOs.

    PubMed

    Morisset, Dany; Dobnik, David; Hamels, Sandrine; Zel, Jana; Gruden, Kristina

    2008-10-01

    We have developed a novel multiplex quantitative DNA-based target amplification method suitable for sensitive, specific and quantitative detection on microarray. This new method named NASBA Implemented Microarray Analysis (NAIMA) was applied to GMO detection in food and feed, but its application can be extended to all fields of biology requiring simultaneous detection of low copy number DNA targets. In a first step, the use of tailed primers allows the multiplex synthesis of template DNAs in a primer extension reaction. A second step of the procedure consists of transcription-based amplification using universal primers. The cRNA product is further on directly ligated to fluorescent dyes labelled 3DNA dendrimers allowing signal amplification and hybridized without further purification on an oligonucleotide probe-based microarray for multiplex detection. Two triplex systems have been applied to test maize samples containing several transgenic lines, and NAIMA has shown to be sensitive down to two target copies and to provide quantitative data on the transgenic contents in a range of 0.1-25%. Performances of NAIMA are comparable to singleplex quantitative real-time PCR. In addition, NAIMA amplification is faster since 20 min are sufficient to achieve full amplification.

  3. Ultrasensitive Label-Free Resonance Rayleigh Scattering Aptasensor for Hg(2+) Using Hg(2+)-Triggered Exonuclease III-Assisted Target Recycling and Growth of G-Wires for Signal Amplification.

    PubMed

    Ren, Wang; Zhang, Ying; Chen, Hong Guo; Gao, Zhong Feng; Li, Nian Bing; Luo, Hong Qun

    2016-01-19

    A novel signal-on and label-free resonance Rayleigh scattering (RRS) aptasensor was constructed for detection of Hg(2+) based on Hg(2+)-triggered Exonuclease III-assisted target recycling and growth of G-quadruplex nanowires (G-wires) for signal amplification. The hairpin DNA (H-DNA) was wisely designed with thymine-rich recognition termini and a G-quadruplex sequence in the loop and employed as a signal probe for specially recognizing trace Hg(2+) by a stable T-Hg(2+)-T structure, which automatically triggered Exonuclease III (Exo-III) digestion to recycle Hg(2+) and liberate the G-quadruplex sequence. The free G-quadruplex sequences were self-assembled into guanine nanowire (G-wire) superstructure in the presence of Mg(2+) and demonstrated by gel electrophoresis. The RRS intensity was dramatically amplified by the resultant G-wires, and the maximum RRS signal at 370 nm was linear with the logarithm of Hg(2+) concentration in the range of 50.0 pM to 500.0 nM (R = 0.9957). Selectivity experiments revealed that the as-prepared RRS sensor was specific for Hg(2+), even coexisting with high concentrations of other metal ions. This optical aptasensor was successfully applied to identify Hg(2+) in laboratory tap water and river water samples. With excellent sensitivity and selectivity, the proposed RRS aptasensor was potentially suitable for not only routine detection of Hg(2+) in environmental monitoring but also various target detection just by changing the recognition sequence of the H-DNA probe.

  4. Randomness Amplification under Minimal Fundamental Assumptions on the Devices

    NASA Astrophysics Data System (ADS)

    Ramanathan, Ravishankar; Brandão, Fernando G. S. L.; Horodecki, Karol; Horodecki, Michał; Horodecki, Paweł; Wojewódka, Hanna

    2016-12-01

    Recently, the physically realistic protocol amplifying the randomness of Santha-Vazirani sources producing cryptographically secure random bits was proposed; however, for reasons of practical relevance, the crucial question remained open regarding whether this can be accomplished under the minimal conditions necessary for the task. Namely, is it possible to achieve randomness amplification using only two no-signaling components and in a situation where the violation of a Bell inequality only guarantees that some outcomes of the device for specific inputs exhibit randomness? Here, we solve this question and present a device-independent protocol for randomness amplification of Santha-Vazirani sources using a device consisting of two nonsignaling components. We show that the protocol can amplify any such source that is not fully deterministic into a fully random source while tolerating a constant noise rate and prove the composable security of the protocol against general no-signaling adversaries. Our main innovation is the proof that even the partial randomness certified by the two-party Bell test [a single input-output pair (u* , x* ) for which the conditional probability P (x*|u*) is bounded away from 1 for all no-signaling strategies that optimally violate the Bell inequality] can be used for amplification. We introduce the methodology of a partial tomographic procedure on the empirical statistics obtained in the Bell test that ensures that the outputs constitute a linear min-entropy source of randomness. As a technical novelty that may be of independent interest, we prove that the Santha-Vazirani source satisfies an exponential concentration property given by a recently discovered generalized Chernoff bound.

  5. Amplification and modulation of fluorescent signals by using hybridization chain reactions for multiplexed sensing of biomolecules in a one-pot

    NASA Astrophysics Data System (ADS)

    Nishimura, Takahiro; Ogura, Yusuke; Yamada, Kenji; Ohno, Yuko; Tanida, Jun

    2014-02-01

    Fluorescence readout of molecular information is a promising approach for biomolecular sensing. For detection of enormous biomolecules via uorescence, biomolecular information should be converted to codes that can be readout easily and simultaneously. For the purpose, we study a biomolecule uorescence color (B/F) encoders that modulate uorescence signals by control of uorescence resonance energy transfer (FRET). The B/F encoder converts biomolecular signals into uorescent color codes represented with uorescent wavelengths and intensity levels. The combination offers a great number of codes for representing the biomolecular information. In this study, we discuss multiplexed detection of target biomolecules using B/F encoders. Use of the B/F encoders would offer a multiplexed biomolecular sensing in a one-pot without micro-fabrication like DNA microarray. In the experiments, we prepared B/F encoders based on two kinds of hybridization chain reactions (HCR) that make long double-stranded DNA polymers to control positions of uorescence and quencher molecules. In the B/F encoders, target molecules trigger to start assembling the polymer structures. The uorescent molecules in the absence of the targets are near the quenchers and the output uorescence is suppressed by FRET. The polymerization process separates the uorescent and quencher dyes and the uorescent signal increase. The experimental results show that the B/F encoders based on HCRs have linear and independent response to each target, and temporal signals during the encoding reactions are usable for multiplexed readout. This result leads to the multiplexed sensing in a one-pot by uorescent ampli cation and multiple uorescent color-coding.

  6. Multiplexed enzyme-free electrochemical immunosensor based on ZnO nanorods modified reduced graphene oxide-paper electrode and silver deposition-induced signal amplification strategy.

    PubMed

    Sun, Guoqiang; Zhang, Lina; Zhang, Yan; Yang, Hongmei; Ma, Chao; Ge, Shenguang; Yan, Mei; Yu, Jinghua; Song, Xianrang

    2015-09-15

    Herein, an origami multiplexed enzyme-free electrochemical (EC) immunodevice is developed for the first time. Typically, ZnO nanorods (ZNRs) modified reduced graphene oxide (rGO)-paper electrode is used as a sensor platform, in which rGO improves the electronic transmission rate and ZNRs provide abundant sites for capture probes binding. Furthermore, by combining the large surface area of rGO and high catalytic activity of bovine serum protein (BSA)-stabilized silver nanoparticles (Ag@BSA) toward H2O2 reduction, rGO/Ag@BSA composites can be used as an excellent signal labels. The current signal is generated from the reduction of H2O2 and further amplified by a subsequent signal labels-promoted deposition of silver. Under optimal conditions, the proposed immunoassays exhibit excellent precision, high sensitivity and a wide linear range of 0.002-120 mIU mL(-1) for human chorionic gonadotropin, 0.001-110 ng mL(-1) for prostate-specific antigen, and 0.001-100 ng mL(-1) for carcinoembryonic antigen. The results for real sample analysis demonstrate that the newly constructed immunosensor arrays provide a simple and cost-effective method for clinical applications.

  7. Reactivation of IgG-switched memory B cells by BCR-intrinsic signal amplification promotes IgG antibody production

    PubMed Central

    Lutz, Johannes; Dittmann, Kai; Bösl, Michael R; Winkler, Thomas H; Wienands, Jürgen; Engels, Niklas

    2015-01-01

    Secondary antibody responses are marked by faster kinetics, improved antibody affinity and a switch from IgM to other immunoglobulin isotypes, most notably IgG, compared with primary responses. These changes protect from reinfection and represent the principle of most vaccination strategies. Yet, the molecular mechanisms that underlie B-cell memory responses are unclear. Here we show, by inactivating the immunoglobulin tail tyrosine (ITT) signalling motif of membrane-bound IgG1 in the mouse, that the ITT facilitates maintenance and reactivation of IgG-switched memory B cells in vivo. The ITT motif equips IgG-switched cells with enhanced BCR signalling capacity, which supports their competitiveness in secondary immune reactions and drives the formation of IgG-secreting plasma cells even in the absence of T-cell help. Our results demonstrate that ITT signalling promotes the vigorous production of IgG antibodies and thus provide a molecular basis for humoral immunological memory. PMID:26815242

  8. Mass spectrometry signal amplification for ultrasensitive glycoprotein detection using gold nanoparticle as mass tag combined with boronic acid based isolation strategy.

    PubMed

    Liu, Minbo; Zhang, Lijuan; Xu, Yawei; Yang, Pengyuan; Lu, Haojie

    2013-07-25

    We describe a novel method for rapid and ultrasensitive detection of intact glycoproteins without enzymatic pretreatment which was commonly used in proteomic research. This method is based on using gold nanoparticle (AuNP) as signal tag in laser desorption/ionization mass spectrometry (LDI-MS) analysis combined with boronic acid assisted isolation strategy. Briefly speaking, target glycoproteins were firstly isolated from sample solution with boronic acid functionalized magnetic microparticles, and then the surface modified gold nanoparticles were added to covalently bind to the glycoproteins. After that, these AuNP tagged glycoproteins were eluted from magnetic microparticles and applied to LDI-MS analysis. The mass signal of AuNP rather than that of glycoprotein was detected and recorded in this strategy. Through data processing of different standard glycoproteins, we have demonstrated that the signal of AuNP could be used to quantitatively represent glycoprotein. This method allows femtomolar detection of intact glycoproteins. We believe that the successful validation of this method on three different kinds of glycoproteins suggests the potential use for tracking trace amount of target glycoproteins in real biological samples in the near future.

  9. CdTe quantum dots@luminol as signal amplification system for chrysoidine with chemiluminescence-chitosan/graphene oxide-magnetite-molecularly imprinting sensor.

    PubMed

    Duan, Huimin; Li, Leilei; Wang, Xiaojiao; Wang, Yanhui; Li, Jianbo; Luo, Chuannan

    2016-01-15

    A sensitive chemiluminescence (CL) sensor based on chemiluminescence resonance energy transfer (CRET) in CdTe quantum dots@luminol (CdTe QDs@luminol) nanomaterials combined with chitosan/graphene oxide-magnetite-molecularly imprinted polymer (Cs/GM-MIP) for sensing chrysoidine was developed. CdTe QDs@luminol was designed to not only amplify the signal of CL but also reduce luminol consumption in the detection of chrysoidine. On the basis of the abundant hydroxy and amino, Cs and graphene oxide were introduced into the GM-MIP to improve the adsorption ability. The adsorption capacities of chrysoidine by both Cs/GM-MIP and non-imprinted polymer (Cs/GM-NIP) were investigated, and the CdTe QDs@luminol and Cs/GM-MIP were characterized by UV-vis, FTIR, SEM and TEM. The proposed sensor can detect chrysoidine within a linear range of 1.0×10(-7) - 1.0×10(-5) mol/L with a detection limit of 3.2×10(-8) mol/L (3δ) due to considerable chemiluminescence signal enhancement of the CdTe quantum dots@luminol detector and the high selectivity of the Cs/GM-MIP system. Under the optimal conditions of CL, the CdTe QDs@luminol-Cs/GM-MIP-CL sensor was used for chrysoidine determination in samples with satisfactory recoveries in the range of 90-107%.

  10. Construction and 13C NMR signal-amplification efficiency of a dynamic nuclear polarizer at 6.4 T and 1.4 K

    NASA Astrophysics Data System (ADS)

    Kiswandhi, Andhika; Niedbalski, Peter; Parish, Christopher; Ferguson, Sarah; Taylor, David; McDonald, George; Lumata, Lloyd

    Dissolution dynamic nuclear polarization (DNP) is a rapidly emerging technique in biomedical and metabolic imaging since it amplifies the liquid-state nuclear magnetic resonance (NMR) and imaging (MRI) signals by >10,000-fold. Originally used in nuclear scattering experiments, DNP works by creating a non-Boltzmann nuclear spin distribution by transferring the high electron (γ = 28,000 MHz/T) thermal polarization to the nuclear spins via microwave irradiation of the sample at high magnetic field and low temperature. A dissolution device is used to rapidly dissolve the frozen sample and consequently produces an injectable ``hyperpolarized'' liquid at physiologically-tolerable temperature. Here we report the construction and performance evaluation of a dissolution DNP hyperpolarizer at 6.4 T and 1.4 K using a continuous-flow cryostat. The solid and liquid-state 13C NMR signal enhancement levels of 13C acetate samples doped with trityl OX063 and 4-oxo-TEMPO free radicals will be discussed and compared with the results from the 3.35 T commercial hyperpolarizer. This work is supported by US Dept of Defense Award No. W81XWH-14-1-0048 and Robert A. Welch Foundation Grant No. AT-1877.

  11. CdTe quantum dots@luminol as signal amplification system for chrysoidine with chemiluminescence-chitosan/graphene oxide-magnetite-molecularly imprinting sensor

    NASA Astrophysics Data System (ADS)

    Duan, Huimin; Li, Leilei; Wang, Xiaojiao; Wang, Yanhui; Li, Jianbo; Luo, Chuannan

    2016-01-01

    A sensitive chemiluminescence (CL) sensor based on chemiluminescence resonance energy transfer (CRET) in CdTe quantum dots@luminol (CdTe QDs@luminol) nanomaterials combined with chitosan/graphene oxide-magnetite-molecularly imprinted polymer (Cs/GM-MIP) for sensing chrysoidine was developed. CdTe QDs@luminol was designed to not only amplify the signal of CL but also reduce luminol consumption in the detection of chrysoidine. On the basis of the abundant hydroxy and amino, Cs and graphene oxide were introduced into the GM-MIP to improve the adsorption ability. The adsorption capacities of chrysoidine by both Cs/GM-MIP and non-imprinted polymer (Cs/GM-NIP) were investigated, and the CdTe QDs@luminol and Cs/GM-MIP were characterized by UV-vis, FTIR, SEM and TEM. The proposed sensor can detect chrysoidine within a linear range of 1.0 × 10- 7 - 1.0 × 10- 5 mol/L with a detection limit of 3.2 × 10- 8 mol/L (3δ) due to considerable chemiluminescence signal enhancement of the CdTe quantum dots@luminol detector and the high selectivity of the Cs/GM-MIP system. Under the optimal conditions of CL, the CdTe QDs@luminol-Cs/GM-MIP-CL sensor was used for chrysoidine determination in samples with satisfactory recoveries in the range of 90-107%.

  12. Physiological responses to error amplification in a robotic reaching adaptation task.

    PubMed

    Shirzad, Navid; Van der Loos, H F Machiel

    2014-01-01

    Analysis of physiological responses provides an objective measure of a person's affective state and has been proposed as a way to evaluate motivation and engagement of therapy clients during robot-assisted therapy regimens. This paper presents the analysis of three physiological responses to different levels of error amplification in a robotic reaching task to understand the feasibility of using physiological signals in order to modify therapy exercises to achieve higher participant attentiveness. In a pilot study with 22 healthy participants, we analyzed skin conductance, skin temperature, and respiration signals, with two main goals: 1) to compare physiological parameters between baseline (rest) and error-amplified reaching motion periods; and 2) to compare physiological parameters between reaching motion periods with different levels of error amplification. Results show that features extracted from skin conductance and respiration signals show significant differences between different error amplification levels. Features extracted from the skin temperature signal are not as reliable as measures of skin conductance and respiration, however they can provide supplementary information.

  13. Direct crosstalk between cancer and osteoblast lineage cells fuels metastatic growth in bone via auto-amplification of IL-6 and RANKL signaling pathways.

    PubMed

    Zheng, Yu; Chow, Shu-Oi; Boernert, Katja; Basel, Dennis; Mikuscheva, Anastasia; Kim, Sarah; Fong-Yee, Colette; Trivedi, Trupti; Buttgereit, Frank; Sutherland, Robert L; Dunstan, Colin R; Zhou, Hong; Seibel, Markus J

    2014-09-01

    The bone microenvironment and its modification by cancer and host cell interactions is a key driver of skeletal metastatic growth. Interleukin-6 (IL-6) stimulates receptor activator of NF-κB ligand (RANKL) expression in bone cells, and serum IL-6 levels are associated with poor clinical outcomes in cancer patients. We investigated the effects of RANKL on cancer cells and the role of tumor-derived IL-6 within the bone microenvironment. Using human breast cancer cell lines to induce tumors in the bone of immune-deficient mice, we first determined whether RANKL released by cells of the osteoblast lineage directly promotes IL-6 expression by cancer cells in vitro and in vivo. We then disrupted of IL-6 signaling in vivo either via knockdown of IL-6 in tumor cells or through treatment with specific anti-human or anti-mouse IL-6 receptor antibodies to investigate the tumor effect. Finally, we tested the effect of RANK knockdown in cancer cells on cancer growth. We demonstrate that osteoblast lineage-derived RANKL upregulates secretion of IL-6 by breast cancers in vivo and in vitro. IL-6, in turn, induces expression of RANK by cancer cells, which sensitizes the tumor to RANKL and significantly enhances cancer IL-6 release. Disruption in vivo of this auto-amplifying crosstalk by knockdown of IL-6 or RANK in cancer cells, or via treatment with anti-IL-6 receptor antibodies, significantly reduces tumor growth in bone but not in soft tissues. RANKL and IL-6 mediate direct paracrine-autocrine signaling between cells of the osteoblast lineage and cancer cells, significantly enhancing the growth of metastatic breast cancers within bone.

  14. Novel glucometer-based immunosensing strategy suitable for complex systems with signal amplification using surfactant-responsive cargo release from glucose-encapsulated liposome nanocarriers.

    PubMed

    Tang, Juan; Huang, Yapei; Liu, Huiqiong; Zhang, Cengceng; Tang, Dianping

    2016-05-15

    Methods based on surfactant-responsive controlled release systems of cargoes from nanocontainers have been developed for bioanalytical applications, but most were utilized for drug delivery and a few reports were focused on immunoassays. Herein we design an in situ amplified immunoassay protocol for high-efficient detection of aflatoxins (aflatoxin B1, AFB1 used in this case) based on surfactant-responsive cargo release from glucose-encapsulated liposome nanocarriers with sensitivity enhancement. Initially, biotinylated liposome nanocarrier encapsulated with glucose was synthesized using a reverse-phase evaporation method. Thereafter, the nanocarrier was utilized as the signal-generation tag on capture antibody-coating microplate through classical biotin-avidin linkage after reaction with biotinylated detection antibody. Upon addition of buffered surfactant (1X PBS-Tween 20 buffer) into the medium, the surfactant immediately hydrolyzed the conjugated liposome, and released the encapsulated glucose from the nanocarriers, which could be quantitatively determined by using a low-cost personal glucometer (PGM). The detectable signal increased with the increment of target analyte. Under the optimal conditions, the assay could allow PGM detection toward target AFB1 as low as 0.6 pg mL(-1) (0.6 ppt). Moreover, the methodology also showed good reproducibility and high specificity toward target AFB1 against other mycotoxins and proteins, and was applicable for quantitatively monitoring target AFB1 in the complex systems, e.g., naturally contaminated/spiked peanut samples and serum specimens, with the acceptable results. Taking these advantages of simplification, low cost, universality and sensitivity, our design provides a new horizon for development of advanced immunoassays in future point-of-care testing.

  15. Highly sensitive aptasensor based on synergetic catalysis activity of MoS2-Au-HE composite using cDNA-Au-GOD for signal amplification.

    PubMed

    Song, Hai-Yan; Kang, Tian-Fang; Lu, Li-Ping; Cheng, Shui-Yuan

    2017-03-01

    Single or few-layer nanosheets of MoS2 (MoS2 nanosheets) and a composite composed of MoS2 nanosheets, Au nanoparticles (AuNPs) and hemin (HE) (denoted as MoS2-Au-HE) were prepared. The composites possessed high synergetic catalysis activity towards the electroreduction of hydrogen peroxide. Furthermore, glucose oxidase (GOD) and AuNPs were used as marker of the complementary DNA (cDNA) strand of kanamycin aptamer to prepare a conjugate (reffered as cDNA-Au-GOD) that was designed as the signal probe. Both cDNA-Au-GOD and MoS2-Au-HE were applied to fabricate aptasensor for kanamycin. MoS2-Au-HE acted as solid platform for kanamycin aptamer and signal transmitters. AuNPs were employed as the supporter of cDNA and GOD which catalyze dissolved oxygen to produce hydrogen peroxide in the presence of glucose. Then cathodic peak current of H2O2 was recorded by differential pulse voltammetry (DPV). The electrochemical reduction of H2O2 was catalyzed by MoS2-Au-HE that was modified onto the surface of a glassy carbon electrode (GCE). The cathodic peak current of H2O2 was highly linearly decreased with an increase of kanamycin concentrations from 1.0ng/L to 1.0×10(5)ng/L, with a detection limit of 0.8ng/L. This aptasensor can be used to detect kanamycin in milk with high specificity, sensitivity and selectivity.

  16. BMP, Wnt and FGF signals are integrated through evolutionarily conserved enhancers to achieve robust expression of Pax3 and Zic genes at the zebrafish neural plate border.

    PubMed

    Garnett, Aaron T; Square, Tyler A; Medeiros, Daniel M

    2012-11-01

    Neural crest cells generate a range of cells and tissues in the vertebrate head and trunk, including peripheral neurons, pigment cells, and cartilage. Neural crest cells arise from the edges of the nascent central nervous system, a domain called the neural plate border (NPB). NPB induction is known to involve the BMP, Wnt and FGF signaling pathways. However, little is known about how these signals are integrated to achieve temporally and spatially specific expression of genes in NPB cells. Furthermore, the timing and relative importance of these signals in NPB formation appears to differ between vertebrate species. Here, we use heat-shock overexpression and chemical inhibitors to determine whether, and when, BMP, Wnt and FGF signaling are needed for expression of the NPB specifiers pax3a and zic3 in zebrafish. We then identify four evolutionarily conserved enhancers from the pax3a and zic3 loci and test their response to BMP, Wnt and FGF perturbations. We find that all three signaling pathways are required during gastrulation for the proper expression of pax3a and zic3 in the zebrafish NPB. We also find that, although the expression patterns driven by the pax3a and zic3 enhancers largely overlap, they respond to different combinations of BMP, Wnt and FGF signals. Finally, we show that the combination of the two pax3a enhancers is less susceptible to signaling perturbations than either enhancer alone. Taken together, our results reveal how BMPs, FGFs and Wnts act cooperatively and redundantly through partially redundant enhancers to achieve robust, specific gene expression in the zebrafish NPB.

  17. Rapid detection of avian influenza virus H5N1 in chicken tracheal samples using an impedance aptasensor with gold nanoparticles for signal amplification.

    PubMed

    Karash, Sardar; Wang, Ronghui; Kelso, Lisa; Lu, Huaguang; Huang, Tony Jun; Li, Yanbin

    2016-10-01

    Highly pathogenic avian influenza virus H5N1 is a continuous threat to public health and poultry industry. The recurrence of the H5N1 led us to develop a robust, specific, and rapid detection method for the virus. In this study, an impedance aptasensor was developed for the virus detection using specific H5N1 aptamer and a gold interdigitated microelectrode. Streptavidin was immobilized on the microelectrode surface and biotin labeled H5N1 aptamer was bound to the immobilized streptavidin. The microelectrode was blocked with the polyethylene glycol and the bound aptamer captured the virus. The impedance change caused by the captured virus was measured using an impedance analyzer. To enhance impedance signal, a nanoparticle-based amplifier was designed and implemented by forming a network-like gold nanoparticles/H5N1-aptamer/thiocyanuric acid. The detection limit of the impedance aptasensor was 0.25 HAU for the pure virus and 1 HAU for the tracheal chicken swab samples spiked with the H5N1 virus. The detection time of aptasensor without employing the amplifier was less than an hour. The amplifier increased impedance by a 57-fold for the 1 HAU samples. Only negligible impedance change was observed for non-target viruses such as H5N2, H5N3, H7N2, H1N1, and H2N2. This aptasensor provides a foundation for the development of a portable aptasensor instrument.

  18. Enhanced beam amplification in a photorefractive Bi{sub 12}TiO{sub 20} crystal by internal reflections

    SciTech Connect

    Khomenko, A.V.; Garcia-Weidner, A.; Tentori, D.

    1996-06-01

    We demonstrate experimentally that internal reflections of a signal and (or) a pump beam allow one to increase beam amplification by two-beam coupling in a long Bi{sub 12}TiO{sub 20} crystal. When fanning is negligible, we achieve an enhancement of the amplification by adjustment of the spatial period of the transformation of the beam{close_quote}s polarization states with periodic reflections of the beams on the crystal boundaries. For the case of strong fanning the fanned beam is redirected by the reflections on the crystal surface, which allows one to use it as a pump beam, thus increasing net amplification gain. {copyright} {ital 1996 Optical Society of America.}

  19. Signal Amplification Strategy of Triple-Layered Core-Shell Au@Pd@Pt Nanoparticles for Ultrasensitive Immunoassay Detection of Squamous Cell Carcinoma Antigen.

    PubMed

    Zhang, Xiaoyue; Du, Bin; Wu, Dan; Ma, Hongmin; Zhang, Yong; Li, He; Wei, Qin

    2015-02-01

    A novel and effective nonenzymatic immunosensor for the sensitive detection of squamous cell carcinoma antigen (SCC- Ag) was described based on triple-layered core-shell Au@Pd@Pt nanoparticles (Au@Pd@Pt NPs). To prepare the immunosensor, primary anti-SCC antibodies (Ab1) were immobilized onto nanoporous gold films (NPGF) of a modified glassy carbon electrode. Au@Pd@Pt NPs that possess strong catalytic activity for the reduction of H2O2 were used as catalytic labels of secondary anti-SCC antibodies (Ab2). Because of the catalytic activities of Au@Pd@Pt NPs and the large surface area of the NPGF, high sensitivity was achieved for the detection of SCC-Ag. The prepared immunosensor showed remarkable results, such as low detection limits (0.6 pg/mL), a wide linear range (0.001-10.0 ng/mL) and high stability and selectivity in the detection of SCC-Ag. Furthermore, the prepared immunosensor exhibited promising properties, which may be useful for real serum sample tests.

  20. A sensitive photoelectrochemical biosensor for AFP detection based on ZnO inverse opal electrodes with signal amplification of CdS-QDs.

    PubMed

    Xu, Ru; Jiang, Yandong; Xia, Lei; Zhang, Tianxiang; Xu, Lin; Zhang, Shuang; Liu, Dali; Song, Hongwei

    2015-12-15

    In this work, ZnO inverse opals structure (IOs) based photoelectrochemical (PEC) electrode was fabricated for alpha-fetoprotein (AFP) detection. Then, the uniform CdS quantum dots (QDs) were hydrothermally synthesized, which allowed the binding of AFP and glucose oxidase (GOD) on CdS QDs, forming the AFP-CdS-GOD composite. The competitive immunosensor of AFP and the AFP-CdS-GOD composite with anti-AFP antibodies (Ab) immobilized on FTO (fluorine-doped tin oxide) /ZnO IOs electrode was successfully applied to the detection of AFP. GOD could catalyze glucose to produce hydrogen peroxide (H2O2) acting as an electron donor to scavenge photogenerated holes in the valence band of CdS QDs, reducing the recombination of electrons and holes of CdS QDs. Also the effective energy level matching between the conduction bands of CdS QDs and ZnO widened the range of light absorption, allowing for electron injection from excited CdS QDs to ZnO upon visible light irradiation, which enhanced the photocurrent. The results show that the immunosensor of AFP possesses a large linear detection range of 0.1-500 ng/ml with a detection limit of 0.01 ng/ml. It also exhibits excellent anti-interference property and acceptable stability. This work provides a promising method for achieving excellent photoelectrochemical biosensor detection of other proteins.

  1. Sensitive electrochemical assay of alkaline phosphatase activity based on TdT-mediated hemin/G-quadruplex DNAzyme nanowires for signal amplification.

    PubMed

    Liu, Yunqing; Xiong, Erhu; Li, Xiaoyu; Li, Junjing; Zhang, Xiaohua; Chen, Jinhua

    2017-01-15

    Taking TdT-mediated hemin/G-quadruplex DNAzyme nanowires as NADH oxidase and HRP-mimicking DNAzyme, a novel DNA-based electrochemical method has been developed for sensitive and selective assay of alkaline phosphatase (AP) activity. The double-stranded DNA (dsDNA) probe consisted of thiol-functionalized DNA1 and 3'-phosphorylated DNA2, was immobilized on a gold nanoparticles (AuNPs) modified glassy carbon (GC) electrode. In the presence of AP, 3'-phosphoryl end of DNA2 was dephosphorylated. Terminal deoxynucletidyl transferase (TdT) catalyzed the sequential addition of deoxynucleotides (dTTPs) at 3'-OH end of DNA2 to extend DNA2 with a poly-T sequence. Then, G-rich DNA3 strand hybridized with the poly-T sequence of DNA2. Upon addition of hemin, the hemin/G-quadruplex DNAzyme was formed. In the presence of NADH, the hemin/G-quadruplex DNAzyme oxidased NADH to NAD(+), accompanied by the formation of H2O2 which was further catalyzed by hemin/G-quadruplex DNAzyme (served as a HRP-mimicking DNAzyme) with the thionine (Thi) as electron transfer mediator, leading to the amplified electrochemical signal. Under optimized conditions, the response peak current was linear with the concentration of AP in the range from 0.1UL(-1) to 5UL(-1) with the detection limit of 0.03UL(-1). Also, the developed biosensor possessed good selectivity, reproducibility and stability, and simple sensing structure, showing promising practical applications in AP activity assay.

  2. Focal overlap gating in velocity map imaging to achieve high signal-to-noise ratio in photo-ion pump-probe experiments

    NASA Astrophysics Data System (ADS)

    Shivaram, Niranjan; Champenois, Elio G.; Cryan, James P.; Wright, Travis; Wingard, Taylor; Belkacem, Ali

    2016-12-01

    We demonstrate a technique in velocity map imaging (VMI) that allows spatial gating of the laser focal overlap region in time resolved pump-probe experiments. This significantly enhances signal-to-noise ratio by eliminating background signal arising outside the region of spatial overlap of pump and probe beams. This enhancement is achieved by tilting the laser beams with respect to the surface of the VMI electrodes which creates a gradient in flight time for particles born at different points along the beam. By suitably pulsing our microchannel plate detector, we can select particles born only where the laser beams overlap. This spatial gating in velocity map imaging can benefit nearly all photo-ion pump-probe VMI experiments especially when extreme-ultraviolet light or X-rays are involved which produce large background signals on their own.

  3. Low-noise preamplifier for multistage photorefractive image amplification

    NASA Astrophysics Data System (ADS)

    Breugnot, S.; Rajbenbach, H.; Defour, M.; Huignard, J.-P.

    1995-07-01

    We present a two-beam coupling configuration in photorefractive BaTiO3 that provides a low-noise amplification of the signal to be detected. A two-wave mixing gain of 100 is reached, in conjunction with very low beam fanning background in the signal direction. The extensions of this configuration to photorefractive heterodyne detection and to multistage image amplification are theoretically and experimentally studied.

  4. RNA internal standard synthesis by nucleic acid sequence-based amplification for competitive quantitative amplification reactions.

    PubMed

    Lo, Wan-Yu; Baeumner, Antje J

    2007-02-15

    Nucleic acid sequence-based amplification (NASBA) reactions have been demonstrated to successfully synthesize new sequences based on deletion and insertion reactions. Two RNA internal standards were synthesized for use in competitive amplification reactions in which quantitative analysis can be achieved by coamplifying the internal standard with the wild type sample. The sequences were created in two consecutive NASBA reactions using the E. coli clpB mRNA sequence as model analyte. The primer sequences of the wild type sequence were maintained, and a 20-nt-long segment inside the amplicon region was exchanged for a new segment of similar GC content and melting temperature. The new RNA sequence was thus amplifiable using the wild type primers and detectable via a new inserted sequence. In the first reaction, the forwarding primer and an additional 20-nt-long sequence was deleted and replaced by a new 20-nt-long sequence. In the second reaction, a forwarding primer containing as 5' overhang sequence the wild type primer sequence was used. The presence of pure internal standard was verified using electrochemiluminescence and RNA lateral-flow biosensor analysis. Additional sequence deletion in order to shorten the internal standard amplicons and thus generate higher detection signals was found not to be required. Finally, a competitive NASBA reaction between one internal standard and the wild type sequence was carried out proving its functionality. This new rapid construction method via NASBA provides advantages over the traditional techniques since it requires no traditional cloning procedures, no thermocyclers, and can be completed in less than 4 h.

  5. Highly Stable and Sensitive Nucleic Acid Amplification and Cell-Phone-Based Readout.

    PubMed

    Kong, Janay E; Wei, Qingshan; Tseng, Derek; Zhang, Jingzi; Pan, Eric; Lewinski, Michael; Garner, Omai B; Ozcan, Aydogan; Di Carlo, Dino

    2017-03-02

    Key challenges with point-of-care (POC) nucleic acid tests include achieving a low-cost, portable form factor, and stable readout, while also retaining the same robust standards of benchtop lab-based tests. We addressed two crucial aspects of this problem, identifying a chemical additive, hydroxynaphthol blue, that both stabilizes and significantly enhances intercalator-based fluorescence readout of nucleic acid concentration, and developing a cost-effective fiber-optic bundle-based fluorescence microplate reader integrated onto a mobile phone. Using loop-mediated isothermal amplification on lambda DNA we achieve a 69-fold increase in signal above background, 20-fold higher than the gold standard, yielding an overall limit of detection of 25 copies/μL within an hour using our mobile-phone-based platform. Critical for a point-of-care system, we achieve a >60% increase in fluorescence stability as a function of temperature and time, obviating the need for manual baseline correction or secondary calibration dyes. This field-portable and cost-effective mobile-phone-based nucleic acid amplification and readout platform is broadly applicable to other real-time nucleic acid amplification tests by similarly modulating intercalating dye performance and is compatible with any fluorescence-based assay that can be run in a 96-well microplate format, making it especially valuable for POC and resource-limited settings.

  6. Exonuclease III-Assisted Target Recycling Amplification Coupled with Liposome-Assisted Amplification: One-Step and Dual-Amplification Strategy for Highly Sensitive Fluorescence Detection of DNA.

    PubMed

    Zhou, Fulin; Li, Baoxin

    2015-07-21

    Detection of ultralow concentration of specific DNA sequence is a central challenge in the early diagnosis of gene-related disease and biodefense application. Herein, we report a dual-amplification strategy for highly sensitive fluorescence detection of DNA. In this proposed strategy, a dumbbell-shaped DNA probe is designed to integrate target binding, magnetic separation, and signal response. In the presence of specific DNA target, the multifunctional dumbbell probe can initiate exonuclease III (Exo III)-aided target recycling amplification, and, in the meantime, generate a large number of fluorescein (FAM)-encapsulated liposomes. The developed method offers very high sensitivity due to primary amplification via numerous FAM from a liposome and secondary amplification via target recycling amplification. The detection limit of the proposed method can reach 4 aM, which is much lower than that of the Exo III-aided target recycling technique applied for DNA quantification without FAM-encapsulated liposomes amplification. Moreover, the dual-signal amplification process can be completed one-step in this system. Therefore, this method provides a simple, isothermal, and low-cost approach for sensitive detection of DNA and holds a great potential for early diagnosis in gene-related diseases.

  7. Improved PCR Amplification of Broad Spectrum GC DNA Templates.

    PubMed

    Guido, Nicholas; Starostina, Elena; Leake, Devin; Saaem, Ishtiaq

    2016-01-01

    Many applications in molecular biology can benefit from improved PCR amplification of DNA segments containing a wide range of GC content. Conventional PCR amplification of DNA sequences with regions of GC less than 30%, or higher than 70%, is complex due to secondary structures that block the DNA polymerase as well as mispriming and mis-annealing of the DNA. This complexity will often generate incomplete or nonspecific products that hamper downstream applications. In this study, we address multiplexed PCR amplification of DNA segments containing a wide range of GC content. In order to mitigate amplification complications due to high or low GC regions, we tested a combination of different PCR cycling conditions and chemical additives. To assess the fate of specific oligonucleotide (oligo) species with varying GC content in a multiplexed PCR, we developed a novel method of sequence analysis. Here we show that subcycling during the amplification process significantly improved amplification of short template pools (~200 bp), particularly when the template contained a low percent of GC. Furthermore, the combination of subcycling and 7-deaza-dGTP achieved efficient amplification of short templates ranging from 10-90% GC composition. Moreover, we found that 7-deaza-dGTP improved the amplification of longer products (~1000 bp). These methods provide an updated approach for PCR amplification of DNA segments containing a broad range of GC content.

  8. Questioning cochlear amplification

    NASA Astrophysics Data System (ADS)

    van der Heijden, Marcel; Versteegh, Corstiaen P. C.

    2015-12-01

    Thirty years ago it was hypothesized that motile processes inject mechanical energy into cochlear traveling waves. This mechanical amplification, alternatively described as negative damping, is invoked to explain both the sensitivity and the nonlinear compression of cochlear responses. There is a recent trend to present cochlear amplification as an established fact, even though the evidence is at most circumstantial and several thorny problems have remained unresolved. We analyze several of these issues, and present new basilar membrane recordings that allowed us to quantify cochlear energy flow. Specifically, we address the following questions: (1) Does auditory sensitivity require narrowband amplification? (2) Has the "RC problem" (lowpass filtering of outer hair cell receptor potential) been resolved? (3) Can OHC motility improve auditory sensitivity? (4) Is there a net power gain between neighboring locations on the basilar membrane? The analyses indicate that mechanical amplification in the cochlea is neither necessary nor useful, and that realizing it by known forms of motility would reduce sensitivity rather than enhance it. Finally, our experimental data show that the peaking of the traveling wave is realized by focusing the acoustic energy rather than amplifying it. (Abbreviations. BM: basilar membrane; CF: characteristic frequency; IHC: inner hair cell; ME: middle ear; MT; mechanotransducer; OHC: outer hair cell; SPL: sound pressure level.)

  9. Selective Inhibition of Mitochondrial JNK Signaling Achieved Using Peptide Mimicry of the Sab Kinase Interacting Motif-1 (KIM1)

    PubMed Central

    Chambers, Jeremy W.; Cherry, Lisa; Laughlin, John D.; Figuera-Losada, Mariana; LoGrasso, Philip V.

    2011-01-01

    The c-jun N-terminal kinases (JNKs) are responsive to stress stimuli leading to activation of proapoptotic proteins and transcription. Additionally, JNK mitochondrial localization has been reported. To selectively target mitochondrial JNK signaling, we exploited JNKs interaction with its mitochondrial scaffold, Sab, using small interfering RNAs (siRNAs) and a cell permeable peptide corresponding to the KIM1 domain of Sab. Gene silencing and peptide interference of this interaction disrupted JNK translocation to the mitochondria and reduced phosphorylation of Bcl-2 without significant impact on c-Jun phosphorylation or AP-1 transcription. In contrast, the JNK inhibitory peptide (TI-JIP1) prevented these three functions. Tat-SabKIM1 selectivity was also demonstrated in anisomycin-stressed HeLa cells where Tat-SabKIM1 prevented Bcl-2 phosphorylation, cell death, loss of mitochondrial membrane potential, and superoxide generation, but not c-Jun phosphorylation. Conversely, TI-JIP1 prevented all aforementioned stress-induced events. This probe introduces a means to evaluate JNK-mediated events on the mitochondria without intervening in nuclear functions of JNK. PMID:21563797

  10. Selective inhibition of mitochondrial JNK signaling achieved using peptide mimicry of the Sab kinase interacting motif-1 (KIM1).

    PubMed

    Chambers, Jeremy W; Cherry, Lisa; Laughlin, John D; Figuera-Losada, Mariana; Lograsso, Philip V

    2011-08-19

    The c-jun N-terminal kinases (JNKs) are responsive to stress stimuli leading to activation of proapoptotic proteins and transcription. Additionally, JNK mitochondrial localization has been reported. To selectively target mitochondrial JNK signaling, we exploited JNK interaction with its mitochondrial scaffold, Sab, using small interfering RNAs (siRNAs) and a cell-permeable peptide corresponding to the KIM1 domain of Sab. Gene silencing and peptide interference of this interaction disrupted JNK translocation to the mitochondria and reduced phosphorylation of Bcl-2 without significant impact on c-Jun phosphorylation or AP-1 transcription. In contrast, the JNK inhibitory peptide (TI-JIP1) prevented these three functions. Tat-Sab(KIM1) selectivity was also demonstrated in anisomycin-stressed HeLa cells where Tat-Sab(KIM1) prevented Bcl-2 phosphorylation, cell death, loss of mitochondrial membrane potential, and superoxide generation but not c-Jun phosphorylation. Conversely, TI-JIP1 prevented all aforementioned stress-induced events. This probe introduces a means to evaluate JNK-mediated events on the mitochondria without intervening in nuclear functions of JNK.

  11. Novel DNA Polymer for Amplification Pretargeting

    PubMed Central

    2015-01-01

    In this Letter, different from conventional pretargeting, an additional novel DNA polymer with multiple copies of a target was first designed to be administrated between the antitumor antibody, and the labeled effector served as an amplification pretargeting strategy. Two phosphorothioate DNA strands, a bridging and a target strand, were hybridized to form a polymer. Polymer size, as a function of molar ratios, was then monitored by size exclusion HPLC and electrophoretic mobility shift assay. Moreover, binding efficiency of polymers with the radiolabeled effector and polymer size after hybridization were measured by HPLC as well. As the polymer was expected to produce more binding sites that would be targeted by effectors, amplification pretargeting can greatly improve accumulation of effectors in tumor. This novel proof-of-concept was then well demonstrated by the in vitro test of signal amplification in antibody-binding protein L coated plate and LS174T cells. Compared to conventional pretargeting, significantly increasing radioactive signal was observed in this designed amplification pretargeting, which would serve as a useful paradigm of the potential of oligomer polymers to improve pretargeting and other related approaches. PMID:26396682

  12. Biomaterials in light amplification

    NASA Astrophysics Data System (ADS)

    Mysliwiec, Jaroslaw; Cyprych, Konrad; Sznitko, Lech; Miniewicz, Andrzej

    2017-03-01

    Biologically produced or inspired materials can serve as optical gain media, i.e. they can exhibit the phenomenon of light amplification. Some of these materials, under suitable dye-doping and optical pumping conditions, show lasing phenomena. The emerging branch of research focused on obtaining lasing action in highly disordered and highly light scattering materials, i.e. research on random lasing, is perfectly suited for biological materials. The use of biomaterials in light amplification has been extensively reported in the literature. In this review we attempt to report on progress in the development of biologically derived systems able to show the phenomena of light amplification and random lasing together with the contribution of our group to this field. The rich world of biopolymers modified with molecular aggregates and nanocrystals, and self-organized at the nanoscale, offers a multitude of possibilities for tailoring luminescent and light scattering properties that are not easily replicated in conventional organic or inorganic materials. Of particular importance and interest are light amplification and lasing, or random lasing studies in biological cells and tissues. In this review we will describe nucleic acids and their complexes employed as gain media due to their favorable optical properties and ease of manipulation. We will report on research conducted on various biomaterials showing structural analogy to nucleic acids such as fluorescent proteins, gelatins in which the first distributed feedback laser was realized, and also amyloids or silks, which, due to their dye-doped fiber-like structure, allow for light amplification. Other materials that were investigated in that respect include polysaccharides, like starch exhibiting favorable photostability in comparison to other biomaterials, and chitosan, which forms photonic crystals or cellulose. Light amplification and random lasing was not only observed in processed biomaterials but also in living

  13. Organo-erbium systems for optical amplification at telecommunications wavelengths.

    PubMed

    Ye, H Q; Li, Z; Peng, Y; Wang, C C; Li, T Y; Zheng, Y X; Sapelkin, A; Adamopoulos, G; Hernández, I; Wyatt, P B; Gillin, W P

    2014-04-01

    Modern telecommunications rely on the transmission and manipulation of optical signals. Optical amplification plays a vital part in this technology, as all components in a real telecommunications system produce some loss. The two main issues with present amplifiers, which rely on erbium ions in a glass matrix, are the difficulty in integration onto a single substrate and the need of high pump power densities to produce gain. Here we show a potential organic optical amplifier material that demonstrates population inversion when pumped from above using low-power visible light. This system is integrated into an organic light-emitting diode demonstrating that electrical pumping can be achieved. This opens the possibility of direct electrically driven optical amplifiers and optical circuits. Our results provide an alternative approach to producing low-cost integrated optics that is compatible with existing silicon photonics and a different route to an effective integrated optics technology.

  14. Cascaded multiple amplification strategy for ultrasensitive detection of HIV/HCV virus DNA.

    PubMed

    Wang, Kun; Fan, Daoqing; Liu, Yaqing; Dong, Shaojun

    2017-01-15

    Ultrasensitive detection of HIV and HCV virus DNA is of great importance for early accurate diagnostics and therapy of HIV virus-infected patients. Herein, to our best knowledge, it is the first to use DNA cascaded multiple amplification strategy for ultrasensitive detection of HIV virus DNA with G-quadruplex-specific fluorescent or colorimetric probes as signal carriers. The developed strategy also exhibited universal applicability for HCV virus DNA detection. After reaction for about 4h, high sensitivity and specificity can be achieved at both fluorescent and colorimetric strategies (limit of detection (LOD) of 10 fM and 0.5pM were reached for fluorescent and colorimetric detection, respectively). And the single-based mismatched DNA even can be distinguished by naked eyes. It is believed that the cascaded multiple amplification strategy presents a huge advance in sensing platform and potential application in future clinical diagnosis.

  15. Precocious sexual signalling and mating in Anastrepha fraterculus (Diptera: Tephritidae) sterile males achieved through juvenile hormone treatment and protein supplements.

    PubMed

    Liendo, M C; Devescovi, F; Bachmann, G E; Utgés, M E; Abraham, S; Vera, M T; Lanzavecchia, S B; Bouvet, J P; Gómez-Cendra, P; Hendrichs, J; Teal, P E A; Cladera, J L; Segura, D F

    2013-02-01

    Sexual maturation of Anastrepha fraterculus is a long process. Methoprene (a mimic of juvenile hormone) considerably reduces the time for sexual maturation in males. However, in other Anastrepha species, this effect depends on protein intake at the adult stage. Here, we evaluated the mating competitiveness of sterile laboratory males and females that were treated with methoprene (either the pupal or adult stage) and were kept under different regimes of adult food, which varied in the protein source and the sugar:protein ratio. Experiments were carried out under semi-natural conditions, where laboratory flies competed over copulations with sexually mature wild flies. Sterile, methoprene-treated males that reached sexual maturity earlier (six days old), displayed the same lekking behaviour, attractiveness to females and mating competitiveness as mature wild males. This effect depended on protein intake. Diets containing sugar and hydrolyzed yeast allowed sterile males to compete with wild males (even at a low concentration of protein), while brewer´s yeast failed to do so even at a higher concentration. Sugar only fed males were unable to achieve significant numbers of copulations. Methoprene did not increase the readiness to mate of six-day-old sterile females. Long pre-copulatory periods create an additional cost to the management of fruit fly pests through the sterile insect technique (SIT). Our findings suggest that methoprene treatment will increase SIT effectiveness against A. fraterculus when coupled with a diet fortified with protein. Additionally, methoprene acts as a physiological sexing method, allowing the release of mature males and immature females and hence increasing SIT efficiency.

  16. An investigation of signal performance enhancements achieved through innovative pixel design across several generations of indirect detection, active matrix, flat-panel arrays

    PubMed Central

    Antonuk, Larry E.; Zhao, Qihua; El-Mohri, Youcef; Du, Hong; Wang, Yi; Street, Robert A.; Ho, Jackson; Weisfield, Richard; Yao, William

    2009-01-01

    Active matrix flat-panel imager (AMFPI) technology is being employed for an increasing variety of imaging applications. An important element in the adoption of this technology has been significant ongoing improvements in optical signal collection achieved through innovations in indirect detection array pixel design. Such improvements have a particularly beneficial effect on performance in applications involving low exposures and∕or high spatial frequencies, where detective quantum efficiency is strongly reduced due to the relatively high level of additive electronic noise compared to signal levels of AMFPI devices. In this article, an examination of various signal properties, as determined through measurements and calculations related to novel array designs, is reported in the context of the evolution of AMFPI pixel design. For these studies, dark, optical, and radiation signal measurements were performed on prototype imagers incorporating a variety of increasingly sophisticated array designs, with pixel pitches ranging from 75 to 127 μm. For each design, detailed measurements of fundamental pixel-level properties conducted under radiographic and fluoroscopic operating conditions are reported and the results are compared. A series of 127 μm pitch arrays employing discrete photodiodes culminated in a novel design providing an optical fill factor of ∼80% (thereby assuring improved x-ray sensitivity), and demonstrating low dark current, very low charge trapping and charge release, and a large range of linear signal response. In two of the designs having 75 and 90 μm pitches, a novel continuous photodiode structure was found to provide fill factors that approach the theoretical maximum of 100%. Both sets of novel designs achieved large fill factors by employing architectures in which some, or all of the photodiode structure was elevated above the plane of the pixel addressing transistor. Generally, enhancement of the fill factor in either discrete or continuous

  17. Parametric nanomechanical amplification at very high frequency.

    PubMed

    Karabalin, R B; Feng, X L; Roukes, M L

    2009-09-01

    Parametric resonance and amplification are important in both fundamental physics and technological applications. Here we report very high frequency (VHF) parametric resonators and mechanical-domain amplifiers based on nanoelectromechanical systems (NEMS). Compound mechanical nanostructures patterned by multilayer, top-down nanofabrication are read out by a novel scheme that parametrically modulates longitudinal stress in doubly clamped beam NEMS resonators. Parametric pumping and signal amplification are demonstrated for VHF resonators up to approximately 130 MHz and provide useful enhancement of both resonance signal amplitude and quality factor. We find that Joule heating and reduced thermal conductance in these nanostructures ultimately impose an upper limit to device performance. We develop a theoretical model to account for both the parametric response and nonequilibrium thermal transport in these composite nanostructures. The results closely conform to our experimental observations, elucidate the frequency and threshold-voltage scaling in parametric VHF NEMS resonators and sensors, and establish the ultimate sensitivity limits of this approach.

  18. Amplification of trace amounts of nucleic acids

    DOEpatents

    Church, George M.; Zhang, Kun

    2008-06-17

    Methods of reducing background during amplification of small amounts of nucleic acids employ careful analysis of sources of low level contamination. Ultraviolet light can be used to reduce nucleic acid contaminants in reagents and equipment. "Primer-dimer" background can be reduced by judicious design of primers. We have shown clean signal-to-noise with as little as starting material as one single human cell (.about.6 picogram), E. coli cell (.about.5 femtogram) or Prochlorococcus cell (.about.3 femtogram).

  19. Gravitomagnetic amplification in cosmology

    SciTech Connect

    Tsagas, Christos G.

    2010-02-15

    Magnetic fields interact with gravitational waves in various ways. We consider the coupling between the Weyl and the Maxwell fields in cosmology and study the effects of the former on the latter. The approach is fully analytical and the results are gauge invariant. We show that the nature and the outcome of the gravitomagnetic interaction depends on the electric properties of the cosmic medium. When the conductivity is high, gravitational waves reduce the standard (adiabatic) decay rate of the B field, leading to its superadiabatic amplification. In poorly conductive environments, on the other hand, Weyl-curvature distortions can result into the resonant amplification of large-scale cosmological magnetic fields. Driven by the gravitational waves, these B fields oscillate with an amplitude that is found to diverge when the wavelengths of the two sources coincide. We present technical and physical aspects of the gravitomagnetic interaction and discuss its potential implications.

  20. Isothermal Multiple Displacement Amplification

    PubMed Central

    Luthra, Rajyalakshmi; Medeiros, L. Jeffrey

    2004-01-01

    Isothermal multiple strand displacement amplification (IMDA) of the whole human genome is a promising method for procuring abundant DNA from valuable and often limited clinical specimens. However, whether DNA generated by this method is of high quality and a faithful replication of the DNA in the original specimen, allowing for subsequent molecular diagnostic testing, requires verification. In this study, we evaluated the suitability of IMDA-generated DNA (IMDA-DNA) for detecting antigen receptor gene rearrangements, chromosomal translocations, and gene mutations using Southern blot analysis, polymerase chain reaction (PCR) methods, or sequencing methods in 28 lymphoma and leukemia clinical specimens. Molecular testing before and after whole genome amplification of these specimens using the IMDA technique showed concordance in 27 of 28 (96%) specimens. Analysis of IMDA-DNA by Southern blot analysis detected restriction fragments >12 kilobases long. No amplification bias was observed at all loci tested demonstrating that this method can be useful in generating large amounts of unbiased, high molecular weight DNA from limited clinical specimens. PMID:15269301

  1. Ultrasensitive DNA detection by cycle isothermal amplification based on nicking endonuclease and its application to logic gates.

    PubMed

    Li, Xuemei; Ding, Tianrong; Sun, Li; Mao, Changming

    2011-12-15

    In recent years, an intense interest has grown in the DNA logic gates having high potential for computation at literally the "nano-size" level. A limitation of traditional DNA logic gates is that each target strand hybridizes with only a single copy of the probe. This 1:1 hybridization radio limits the gain of the approach and thus its sensitivity. The exponential amplification of nucleic acids has become a core technology in medical diagnostics and has been widely used for the construction of DNA sensor, DNA nanomachine and DNA sequencing. It would be of great interest to develop DNA-based logic systems with exponential amplification for the output signal. In the present study, a series of three-input DNA logic gates with the cycle isothermal amplification based on nicking endonuclease (NEase) are designed. Very low concentrations of the analytes were sufficient to initiate an autocatalytic cascade, achieving a significant improvement of the detection limit, 100-fold improvement compared to the non-autocatalytic system. This was achieved by engineering a simple and flexible biological circuit designed to initiate a cascade of events to detect and amplify a specific DNA sequence. This procedure has the potential to greatly simplify the logic operation because amplification can be performed in "one-pot".

  2. Kinetic Hairpin Oligonucleotide Blockers for Selective Amplification of Rare Mutations

    PubMed Central

    Jia, Yanwei; Sanchez, J. Aquiles; Wangh, Lawrence J.

    2014-01-01

    Detection of rare mutant alleles in an excess of wild type alleles is increasingly important in cancer diagnosis. Several methods for selective amplification of a mutant allele via the polymerase chain reaction (PCR) have been reported, but each of these methods has its own limitations. A common problem is that Taq DNA polymerase errors early during amplification generate false positive mutations which also accumulate exponentially. In this paper, we described a novel method using hairpin oligonucleotide blockers that can selectively inhibit the amplification of wild type DNA during LATE-PCR amplification. LATE-PCR generates double-stranded DNA exponentially followed by linear amplification of single-stranded DNA. The efficiency of the blocker is optimized by adjusting the LATE-PCR temperature cycling profile. We also demonstrate that it is possible to minimize false positive signals caused by Taq DNA polymerase errors by using a mismatched excess primer plus a modified PCR profile to preferentially enrich for mutant target sequences prior to the start of the exponential phase of LATE-PCR amplification. In combination these procedures permit amplification of specific KRAS mutations in the presence of more than 10,000 fold excess of wild type DNA without false positive signals. PMID:25082368

  3. Non-contact photoacoustic imaging using a fiber based interferometer with optical amplification

    PubMed Central

    Hochreiner, Armin; Bauer-Marschallinger, Johannes; Burgholzer, Peter; Jakoby, Bernhard; Berer, Thomas

    2013-01-01

    In photoacoustic imaging the ultrasonic signals are usually detected by contacting transducers. For some applications contact with the tissue should be avoided. As alternatives to contacting transducers interferometric means can be used to acquire photoacoustic signals remotely. In this paper we report on non-contact three and two dimensional photoacoustic imaging using an optical fiber-based Mach-Zehnder interferometer. A detection beam is transmitted through an optical fiber network onto the surface of the specimen. Back reflected light is collected and coupled into the same optical fiber. To achieve a high signal/noise ratio the reflected light is amplified by means of optical amplification with an erbium doped fiber amplifier before demodulation. After data acquisition the initial pressure distribution is reconstructed by a Fourier domain reconstruction algorithm. We present remote photoacoustic imaging of a tissue mimicking phantom and on chicken skin. PMID:24298397

  4. Non-contact photoacoustic imaging using a fiber based interferometer with optical amplification.

    PubMed

    Hochreiner, Armin; Bauer-Marschallinger, Johannes; Burgholzer, Peter; Jakoby, Bernhard; Berer, Thomas

    2013-01-01

    In photoacoustic imaging the ultrasonic signals are usually detected by contacting transducers. For some applications contact with the tissue should be avoided. As alternatives to contacting transducers interferometric means can be used to acquire photoacoustic signals remotely. In this paper we report on non-contact three and two dimensional photoacoustic imaging using an optical fiber-based Mach-Zehnder interferometer. A detection beam is transmitted through an optical fiber network onto the surface of the specimen. Back reflected light is collected and coupled into the same optical fiber. To achieve a high signal/noise ratio the reflected light is amplified by means of optical amplification with an erbium doped fiber amplifier before demodulation. After data acquisition the initial pressure distribution is reconstructed by a Fourier domain reconstruction algorithm. We present remote photoacoustic imaging of a tissue mimicking phantom and on chicken skin.

  5. Experimental noiseless linear amplification using weak measurements

    NASA Astrophysics Data System (ADS)

    Ho, Joseph; Boston, Allen; Palsson, Matthew; Pryde, Geoff

    2016-09-01

    The viability of quantum communication schemes rely on sending quantum states of light over long distances. However, transmission loss can degrade the signal strength, adding noise. Heralded noiseless amplification of a quantum signal can provide a solution by enabling longer direct transmission distances and by enabling entanglement distillation. The central idea of heralded noiseless amplification—a conditional modification of the probability distribution over photon number of an optical quantum state—is suggestive of a parallel with weak measurement: in a weak measurement, learning partial information about an observable leads to a conditional back-action of a commensurate size. Here we experimentally investigate the application of weak, or variable-strength, measurements to the task of heralded amplification, by using a quantum logic gate to weakly couple a small single-optical-mode quantum state (the signal) to an ancilla photon (the meter). The weak measurement is carried out by choosing the measurement basis of the meter photon and, by conditioning on the meter outcomes, the signal is amplified. We characterise the gain of the amplifier as a function of the measurement strength, and use interferometric methods to show that the operation preserves the coherence of the signal.

  6. Coherent white light amplification

    DOEpatents

    Jovanovic, Igor; Barty, Christopher P.

    2004-05-25

    A system for coherent simultaneous amplification of a broad spectral range of light that includes an optical parametric amplifier and a source of a seed pulse is described. A first angular dispersive element is operatively connected to the source of a seed pulse. A first imaging telescope is operatively connected to the first angular dispersive element and operatively connected to the optical parametric amplifier. A source of a pump pulse is operatively connected to the optical parametric amplifier. A second imaging telescope is operatively connected to the optical parametric amplifier and a second angular dispersive element is operatively connected to the second imaging telescope.

  7. Weak-value amplification: state of play

    NASA Astrophysics Data System (ADS)

    Knee, George C.; Combes, Joshua; Ferrie, Christopher; Gauger, Erik M.

    2016-01-01

    Weak values arise in quantum theory when the result of a weak measurement is conditioned on a subsequent strong measurement. The majority of the trials are discarded, leaving only very few successful events. Intriguingly those can display a substantial signal amplification. This raises the question of whether weak values carry potential to improve the performance of quantum sensors, and indeed a number of impressive experimental results suggested this may be the case. By contrast, recent theoretical studies have found the opposite: using weak-values to obtain an amplification generally worsens metrological performance. This survey summarises the implications of those studies, which call for a reappraisal of weak values' utility and for further work to reconcile theory and experiment.

  8. Evidence of high-elevation amplification versus Arctic amplification

    PubMed Central

    Wang, Qixiang; Fan, Xiaohui; Wang, Mengben

    2016-01-01

    Elevation-dependent warming in high-elevation regions and Arctic amplification are of tremendous interest to many scientists who are engaged in studies in climate change. Here, using annual mean temperatures from 2781 global stations for the 1961–2010 period, we find that the warming for the world’s high-elevation stations (>500 m above sea level) is clearly stronger than their low-elevation counterparts; and the high-elevation amplification consists of not only an altitudinal amplification but also a latitudinal amplification. The warming for the high-elevation stations is linearly proportional to the temperature lapse rates along altitudinal and latitudinal gradients, as a result of the functional shape of Stefan-Boltzmann law in both vertical and latitudinal directions. In contrast, neither altitudinal amplification nor latitudinal amplification is found within the Arctic region despite its greater warming than lower latitudes. Further analysis shows that the Arctic amplification is an integrated part of the latitudinal amplification trend for the low-elevation stations (≤500 m above sea level) across the entire low- to high-latitude Northern Hemisphere, also a result of the mathematical shape of Stefan-Boltzmann law but only in latitudinal direction. PMID:26753547

  9. Isothermal Amplification of Nucleic Acids.

    PubMed

    Zhao, Yongxi; Chen, Feng; Li, Qian; Wang, Lihua; Fan, Chunhai

    2015-11-25

    Isothermal amplification of nucleic acids is a simple process that rapidly and efficiently accumulates nucleic acid sequences at constant temperature. Since the early 1990s, various isothermal amplification techniques have been developed as alternatives to polymerase chain reaction (PCR). These isothermal amplification methods have been used for biosensing targets such as DNA, RNA, cells, proteins, small molecules, and ions. The applications of these techniques for in situ or intracellular bioimaging and sequencing have been amply demonstrated. Amplicons produced by isothermal amplification methods have also been utilized to construct versatile nucleic acid nanomaterials for promising applications in biomedicine, bioimaging, and biosensing. The integration of isothermal amplification into microsystems or portable devices improves nucleic acid-based on-site assays and confers high sensitivity. Single-cell and single-molecule analyses have also been implemented based on integrated microfluidic systems. In this review, we provide a comprehensive overview of the isothermal amplification of nucleic acids encompassing work published in the past two decades. First, different isothermal amplification techniques are classified into three types based on reaction kinetics. Then, we summarize the applications of isothermal amplification in bioanalysis, diagnostics, nanotechnology, materials science, and device integration. Finally, several challenges and perspectives in the field are discussed.

  10. Self-induced parametric amplification arising from nonlinear elastic coupling in a micromechanical resonating disk gyroscope.

    PubMed

    Nitzan, Sarah H; Zega, Valentina; Li, Mo; Ahn, Chae H; Corigliano, Alberto; Kenny, Thomas W; Horsley, David A

    2015-03-12

    Parametric amplification, resulting from intentionally varying a parameter in a resonator at twice its resonant frequency, has been successfully employed to increase the sensitivity of many micro- and nano-scale sensors. Here, we introduce the concept of self-induced parametric amplification, which arises naturally from nonlinear elastic coupling between the degenerate vibration modes in a micromechanical disk-resonator, and is not externally applied. The device functions as a gyroscope wherein angular rotation is detected from Coriolis coupling of elastic vibration energy from a driven vibration mode into a second degenerate sensing mode. While nonlinear elasticity in silicon resonators is extremely weak, in this high quality-factor device, ppm-level nonlinear elastic effects result in an order-of-magnitude increase in the observed sensitivity to Coriolis force relative to linear theory. Perfect degeneracy of the primary and secondary vibration modes is achieved through electrostatic frequency tuning, which also enables the phase and frequency of the parametric coupling to be varied, and we show that the resulting phase and frequency dependence of the amplification follow the theory of parametric resonance. We expect that this phenomenon will be useful for both fundamental studies of dynamic systems with low dissipation and for increasing signal-to-noise ratio in practical applications such as gyroscopes.

  11. Measurement-based noiseless linear amplification for quantum communication

    NASA Astrophysics Data System (ADS)

    Chrzanowski, H. M.; Walk, N.; Haw, J. Y.; Thearle, O.; Assad, S. M.; Janousek, J.; Hosseini, S.; Ralph, T. C.; Symul, T.; Lam, P. K.

    2014-11-01

    Entanglement distillation is an indispensable ingredient in extended quantum communication networks. Distillation protocols are necessarily non-deterministic and require non-trivial experimental techniques such as noiseless amplification. We show that noiseless amplification could be achieved by performing a post-selective filtering of measurement outcomes. We termed this protocol measurement-based noiseless linear amplification (MBNLA). We apply this protocol to entanglement that suffers transmission loss of up to the equivalent of 100km of optical fibre and show that it is capable of distilling entanglement to a level stronger than that achievable by transmitting a maximally entangled state through the same channel. We also provide a proof-of-principle demonstration of secret key extraction from an otherwise insecure regime via MBNLA. Compared to its physical counterpart, MBNLA not only is easier in term of implementation, but also allows one to achieve near optimal probability of success.

  12. Ultrasensitive detection of low-abundance surface-marker protein using isothermal rolling circle amplification in a microfluidic nanoliter platform.

    PubMed

    Konry, Tania; Smolina, Irina; Yarmush, Joel M; Irimia, Daniel; Yarmush, Martin L

    2011-02-07

    With advances in immunology and cancer biology, there is an unmet need for increasingly sensitive systems to monitor the expression of specific cell markers for the development of new diagnostic and therapeutic tools. To address this challenge, a highly sensitive labeling method that translates antigen-antibody recognition processes into DNA detection events that can be greatly amplified via isothermal rolling circle amplification (RCA) is applied. By merging the single-molecule detection power of RCA reactions with microfluidic technology, it is demonstrated that the identification of specific protein markers can be achieved on tumor-cell surfaces in miniaturized nanoliter reaction droplets. Furthermore, this combined approach of signal amplification in a microfluidic format could extend the utility of existing methods by reducing sample and reagent consumption and enhancing the sensitivities and specificities for various applications, including early diagnosis of cancer.

  13. Laser light triggers increased Raman amplification in the regime of nonlinear Landau damping

    PubMed Central

    Depierreux, S.; Yahia, V.; Goyon, C.; Loisel, G.; Masson-Laborde, P. -E.; Borisenko, N.; Orekhov, A.; Rosmej, O.; Rienecker, T.; Labaune, C.

    2014-01-01

    Stimulated Raman backscattering (SRS) has many unwanted effects in megajoule-scale inertially confined fusion (ICF) plasmas. Moreover, attempts to harness SRS to amplify short laser pulses through backward Raman amplification have achieved limited success. In high-temperature fusion plasmas, SRS usually occurs in a kinetic regime where the nonlinear response of the Langmuir wave to the laser drive and its host of complicating factors make it difficult to predict the degree of amplification that can be achieved under given experimental conditions. Here we present experimental evidence of reduced Landau damping with increasing Langmuir wave amplitude and determine its effects on Raman amplification. The threshold for trapping effects to influence the amplification is shown to be very low. Above threshold, the complex SRS dynamics results in increased amplification factors, which partly explains previous ICF experiments. These insights could aid the development of more efficient backward Raman amplification schemes in this regime. PMID:24938756

  14. ELECTRON AMPLIFICATION IN DIAMOND.

    SciTech Connect

    SMEDLEY, J.; BEN-ZVI, I.; BURRILL, A.; CHANG, X.; GRIMES, J.; RAO, T.; SEGALOV, Z.; WU, Q.

    2006-07-10

    We report on recent progress toward development of secondary emission ''amplifiers'' for photocathodes. Secondary emission gain of over 300 has been achieved in transmission mode and emission mode for a variety of diamond samples. Techniques of sample preparation, including hydrogenation to achieve negative electron affinity (NEA), have been adapted to this application.

  15. An Intrinsically Digital Amplification Scheme for Hearing Aids

    NASA Astrophysics Data System (ADS)

    Blamey, Peter J.; Macfarlane, David S.; Steele, Brenton R.

    2005-12-01

    Results for linear and wide-dynamic range compression were compared with a new 64-channel digital amplification strategy in three separate studies. The new strategy addresses the requirements of the hearing aid user with efficient computations on an open-platform digital signal processor (DSP). The new amplification strategy is not modeled on prior analog strategies like compression and linear amplification, but uses statistical analysis of the signal to optimize the output dynamic range in each frequency band independently. Using the open-platform DSP processor also provided the opportunity for blind trial comparisons of the different processing schemes in BTE and ITE devices of a high commercial standard. The speech perception scores and questionnaire results show that it is possible to provide improved audibility for sound in many narrow frequency bands while simultaneously improving comfort, speech intelligibility in noise, and sound quality.

  16. Enhanced acoustic sensing through wave compression and pressure amplification in anisotropic metamaterials.

    PubMed

    Chen, Yongyao; Liu, Haijun; Reilly, Michael; Bae, Hyungdae; Yu, Miao

    2014-10-15

    Acoustic sensors play an important role in many areas, such as homeland security, navigation, communication, health care and industry. However, the fundamental pressure detection limit hinders the performance of current acoustic sensing technologies. Here, through analytical, numerical and experimental studies, we show that anisotropic acoustic metamaterials can be designed to have strong wave compression effect that renders direct amplification of pressure fields in metamaterials. This enables a sensing mechanism that can help overcome the detection limit of conventional acoustic sensing systems. We further demonstrate a metamaterial-enhanced acoustic sensing system that achieves more than 20 dB signal-to-noise enhancement (over an order of magnitude enhancement in detection limit). With this system, weak acoustic pulse signals overwhelmed by the noise are successfully recovered. This work opens up new vistas for the development of metamaterial-based acoustic sensors with improved performance and functionalities that are highly desirable for many applications.

  17. High-energy infrared femtosecond pulses generated by dual-chirped optical parametric amplification.

    PubMed

    Fu, Yuxi; Takahashi, Eiji J; Midorikawa, Katsumi

    2015-11-01

    We demonstrate high-energy infrared femtosecond pulse generation by a dual-chirped optical parametric amplification (DC-OPA) scheme [Opt. Express19, 7190 (2011)]. By employing a 100 mJ pump laser, a signal pulse energy exceeding 20 mJ at a wavelength of 1.4 μm was achieved before dispersion compensation. A total output energy of 33 mJ was recorded. Under a further energy scaling condition, the signal pulse was compressed to an almost transform-limited duration of 27 fs using a fused silica prism compressor. Since the DC-OPA scheme is efficient and energy scalable, design parameters for obtaining 100 mJ level infrared pulses are presented, which are suitable as driver lasers for the energy scaling of high-order harmonic generation with sub-keV photon energy.

  18. Study of Raman amplification in DPP-BOTDA sensing employing Simplex coding for sub-meter scale spatial resolution over long fiber distances

    NASA Astrophysics Data System (ADS)

    Taki, Mohammad; Soto, Marcelo A.; Bolognini, Gabriele; Di Pasquale, Fabrizio

    2013-09-01

    The impact of Raman amplification and Simplex coding is studied in combination with differential pulse-width pair Brillouin optical time-domain analysis (DPP-BOTDA) to achieve sub-meter spatial resolution over very long sensing distances. An optimization of the power levels for the Raman pumps, Brillouin pump and signal has been carried out through numerical simulations, maximizing the signal levels and avoiding at the same time nonlinear effects and pump depletion. A reduction of acoustic-wave-induced distortions in the Brillouin gain spectrum down to negligible levels has also been achieved by numerical optimization of the pulse width and duty cycle of return-to-zero Simplex coding, providing significant signal-to-noise ratio enhancement. Strain-temperature sensing over 93 km standard SMF is achieved with a strain/temperature accuracy of 34µɛ/1.7 °C, and 50 cm spatial resolution throughout the fiber length.

  19. Single primer-triggered isothermal amplification for double-stranded DNA detection.

    PubMed

    Ma, Cuiping; Han, Dianang; Deng, Meilian; Wang, Jingfei; Shi, Chao

    2015-01-11

    Here we have devised a new generation of isothermal double-stranded DNA (dsDNA) detection method, termed single primer-triggered isothermal amplification (SAMP). It is very simple only requiring one primer and a few copies of dsDNA in less than an hour are detectable with multiple signal amplification steps.

  20. Achieving high signal-to-noise in cell regulatory systems: Spatial organization of multiprotein transmembrane assemblies of FGFR and MET receptors.

    PubMed

    Blaszczyk, Michal; Harmer, Nicholas J; Chirgadze, Dimitri Y; Ascher, David B; Blundell, Tom L

    2015-09-01

    How is information communicated both within and between cells of living systems with high signal to noise? We discuss transmembrane signaling models involving two receptor tyrosine kinases: the fibroblast growth factor receptor (FGFR) and the MET receptor. We suggest that simple dimerization models might occur opportunistically giving rise to noise but cooperative clustering of the receptor tyrosine kinases observed in these systems is likely to be important for signal transduction. We propose that this may be a more general prerequisite for high signal to noise in transmembrane receptor signaling.

  1. Differential transimpedance amplifier circuit for correlated differential amplification

    DOEpatents

    Gresham, Christopher A.; Denton, M. Bonner; Sperline, Roger P.

    2008-07-22

    A differential transimpedance amplifier circuit for correlated differential amplification. The amplifier circuit increase electronic signal-to-noise ratios in charge detection circuits designed for the detection of very small quantities of electrical charge and/or very weak electromagnetic waves. A differential, integrating capacitive transimpedance amplifier integrated circuit comprising capacitor feedback loops performs time-correlated subtraction of noise.

  2. Sound-Field Amplification: Preliminary Information Regarding Special Education Referrals

    ERIC Educational Resources Information Center

    Flexer, C.; Long, S.

    2004-01-01

    In this clinical exchange, the authors discuss acoustic accessibility and sound-field amplification in general education classrooms. They bridge theory to practice by presenting preliminary information from two different school systems demonstrating how an improved signal-to-noise ratio can have a positive impact on special education referrals.

  3. Regulation of ribosomal DNA amplification by the TOR pathway

    PubMed Central

    Jack, Carmen V.; Cruz, Cristina; Hull, Ryan M.; Keller, Markus A.; Ralser, Markus; Houseley, Jonathan

    2015-01-01

    Repeated regions are widespread in eukaryotic genomes, and key functional elements such as the ribosomal DNA tend to be formed of high copy repeated sequences organized in tandem arrays. In general, high copy repeats are remarkably stable, but a number of organisms display rapid ribosomal DNA amplification at specific times or under specific conditions. Here we demonstrate that target of rapamycin (TOR) signaling stimulates ribosomal DNA amplification in budding yeast, linking external nutrient availability to ribosomal DNA copy number. We show that ribosomal DNA amplification is regulated by three histone deacetylases: Sir2, Hst3, and Hst4. These enzymes control homologous recombination-dependent and nonhomologous recombination-dependent amplification pathways that act in concert to mediate rapid, directional ribosomal DNA copy number change. Amplification is completely repressed by rapamycin, an inhibitor of the nutrient-responsive TOR pathway; this effect is separable from growth rate and is mediated directly through Sir2, Hst3, and Hst4. Caloric restriction is known to up-regulate expression of nicotinamidase Pnc1, an enzyme that enhances Sir2, Hst3, and Hst4 activity. In contrast, normal glucose concentrations stretch the ribosome synthesis capacity of cells with low ribosomal DNA copy number, and we find that these cells show a previously unrecognized transcriptional response to caloric excess by reducing PNC1 expression. PNC1 down-regulation forms a key element in the control of ribosomal DNA amplification as overexpression of PNC1 substantially reduces ribosomal DNA amplification rate. Our results reveal how a signaling pathway can orchestrate specific genome changes and demonstrate that the copy number of repetitive DNA can be altered to suit environmental conditions. PMID:26195783

  4. Active beam shaping in multi-levels amplification system

    NASA Astrophysics Data System (ADS)

    Zhao, Tianzhuo; Fan, Zhongwei; Qiu, Jisi; Tang, Xiongxin; Lin, Weiran; Zhang, Hongbo

    2016-09-01

    Using Liquid Crystal Spatial Light Modulator (LC-SLM) as a beam shaping device to improve beam quality in high-gain amplification system is reported. 1.6 nJ injected small-size signal Gaussian beam can be amplified to 5 J by 4 stages amplification, and finally output beam is a 50mm×50mm square spot with flat-top intensity distribution. In the amplification system we designed, LC-SLM is placed after the second level of amplifier, where the signal laser energy is about 20mJ, and beam size is 10mm×10mm. The structure of Fourier image transfer is also implemented in this amplifications system to be capable of maintaining high-quality image transmission in the amplification process. The LC-SLM as an object, is imaged by beam expand lenses and spatial filters lenses in the amplifications system to get good quality of imaging. By catching output spot and making a feed-back, transmission efficiency of each pixel on LC-SLM is modulated, high energy density area can be decreased to realize flat-top intensity distribution. A spot modulation function is defined as, using the maximum grey value on spot area divided by the average grey value of the image after background correction. By this, amplified laser obtains the spot modulation of 1.24 on central 90% area of the spot. Furthermore, un-uniform distribution on the full spot, soften effects of spot edge, and output beam shape can also be optimized by the LC-SLM shaping scheme in the amplification system.

  5. Spin noise amplification and giant noise in optical microcavity

    SciTech Connect

    Ryzhov, I. I.; Poltavtsev, S. V.; Kozlov, G. G.; Zapasskii, V. S.; Kavokin, A. V.; Lagoudakis, P. V.

    2015-06-14

    When studying the spin-noise-induced fluctuations of Kerr rotation in a quantum-well microcavity, we have found a dramatic increase of the noise signal (by more than two orders of magnitude) in the vicinity of anti-crossing of the polariton branches. The effect is explained by nonlinear optical instability of the microcavity giving rise to the light-power-controlled amplification of the polarization noise signal. In the framework of the developed model of built-in amplifier, we also interpret the nontrivial spectral and intensity-related properties of the observed noise signal below the region of anti-crossing of polariton branches. The discovered effect of optically controllable amplification of broadband polarization signals in microcavities in the regime of optical instability may be of interest for detecting weak oscillations of optical anisotropy in fundamental research and for other applications in optical information processing.

  6. Space Optical Communications Using Laser Beam Amplification

    NASA Technical Reports Server (NTRS)

    Agrawal, Govind

    2015-01-01

    The Space Optical Communications Using Laser Beam Amplification (SOCLBA) project will provide a capability to amplify a laser beam that is received in a modulating retro-reflector (MRR) located in a satellite in low Earth orbit. It will also improve the pointing procedure between Earth and spacecraft terminals. The technology uses laser arrays to strengthen the reflected laser beam from the spacecraft. The results of first year's work (2014) show amplification factors of 60 times the power of the signal beam. MMRs are mirrors that reflect light beams back to the source. In space optical communications, a high-powered laser interrogator beam is directed from the ground to a satellite. Within the satellite, the beam is redirected back to ground using the MMR. In the MMR, the beam passes through modulators, which encode a data signal onto the returning beam. MMRs can be used in small spacecraft for optical communications. The SOCLBA project is significant to NASA and small spacecraft due to its application to CubeSats for optical data transmission to ground stations, as well as possible application to spacecraft for optical data transmission.

  7. Isothermal strand displacement amplification (iSDA): a rapid and sensitive method of nucleic acid amplification for point-of-care diagnosis.

    PubMed

    Toley, Bhushan J; Covelli, Isabela; Belousov, Yevgeniy; Ramachandran, Sujatha; Kline, Enos; Scarr, Noah; Vermeulen, Nic; Mahoney, Walt; Lutz, Barry R; Yager, Paul

    2015-11-21

    We present a method of rapid isothermal amplification of DNA without initial heat denaturation of the template, and methods and probes for (a) real-time fluorescence detection and (b) lateral flow detection of amplicons. Isothermal strand displacement amplification (iSDA) can achieve >10(9)-fold amplification of the target sequence in <20 minutes at 49 °C, which makes it one of the fastest existing isothermal DNA amplification methods. iSDA initiates at sites where DNA base pairs spontaneously open or transiently convert into Hoogsteen pairs, i.e. "breathe", and proceeds to exponential amplification by repeated nicking, extension, and displacement of single strands. We demonstrate successful iSDA amplification and lateral flow detection of 10 copies of a Staphylococcus aureus gene, NO.-inducible l-lactate dehydrogenase (ldh1) (Richardson, Libby, and Fang, Science, 2008, 319, 1672-1676), in a clean sample and 50 copies in the presence of high concentrations of genomic DNA and mucins in <30 minutes. We also present a simple kinetic model of iSDA that incorporates competition between target and primer-dimer amplification. This is the first model that quantitates the effects of primer-dimer products in isothermal amplification reactions. Finally, we demonstrate the multiplexing capability of iSDA by the simultaneous amplification of the target gene and an engineered internal control sequence. The speed, sensitivity, and specificity of iSDA make it a powerful method for point-of-care molecular diagnosis.

  8. Sensitive detection of nucleic acids with rolling circle amplification and surface-enhanced Raman scattering spectroscopy.

    PubMed

    Hu, Juan; Zhang, Chun-yang

    2010-11-01

    Detection of specific DNA sequences is important to molecular biology research and clinical diagnostics. To improve the sensitivity of surface-enhanced Raman scattering spectroscopy (SERS), a variety of signal amplification methods has been developed, including Raman-active-dye, polymerase chain reaction (PCR) technology, molecular beacon, SERS-active substrates, and SERS-tag. However, the combination of rolling circle amplification (RCA) with SERS for nucleic acid detection has not been reported. Herein, we describe a new approach for nucleic acid detection by the combination of RCA reaction with SERS. Because of the binding of abundance repeated sequences of RCA products with gold nanoparticle (Au NP) and Rox-modified detection probes, SERS signal is significantly amplified and the detection limit of 10.0 pM might be achieved. The sensitivity of RCA-based SERS has increased by as much as 3 orders of magnitude as compared to PCR-based SERS and is also comparable with or even exceeds that of both RCA-based electrochemical and RCA-based fluorescent methods. This RCA-based SERS might discriminate perfect matched target DNA from 1-base mismatched DNA with high selectivity. The high sensitivity and selectivity of RCA-based SERS makes it a potential tool for early diagnosis of gene-related disease and also offers a great promise for multiplexed assays with DNA microarrays.

  9. A model for the nonlinear mechanism responsible for cochlear amplification.

    PubMed

    Fessel, Kimberly; Holmes, Mark H

    2014-12-01

    A nonlinear model for the mechanism responsible for the amplification of the sound wave in the ear is derived using the geometric and material properties of the system. The result is a nonlinear beam equation, with the nonlinearity appearing in a coefficient of the equation. Once derived, the beam problem is analyzed for various loading conditions. Based on this analysis it is seen that the mechanism is capable of producing a spatially localized gain, as required by any amplification mechanism, but it is also capable of increasing the spatial contrast in the signal.

  10. Influence of environmental noise on the weak value amplification

    NASA Astrophysics Data System (ADS)

    Zhu, Xuannmin; Zhang, Yu-Xiang

    2016-08-01

    Quantum systems are always disturbed by environmental noise. We have investigated the influence of the environmental noise on the amplification in weak measurements. Three typical quantum noise processes are discussed in this article. The maximum expectation values of the observables of the measuring device decrease sharply with the strength of the depolarizing and phase damping channels, while the amplification effect of weak measurement is immune to the amplitude damping noise. To obtain significantly amplified signals, we must ensure that the preselection quantum systems are kept away from the depolarizing and phase damping processes.

  11. Optimizing biased semiconductor superlattices for terahertz amplification

    SciTech Connect

    Lei, Xiaoli; Wang, Dawei; Wu, Zhaoxin; Dignam, M. M.

    2014-08-11

    Over the past 15 yr or more, researchers have been trying to achieve gain for electromagnetic fields in the terahertz frequency region using biased semiconductor superlattices, but with little success. In this work, we employ our model of the excitonic states in biased GaAs/Al{sub 0.3}Ga{sub 0.7}As semiconductor superlattices to find the optimal structures for amplification of terahertz radiation. In particular, we determine the optimum well width, barrier width, and bias field for terahertz fields with frequencies ranging from 1 to 4 terahertz. We find that gain coefficients on the order of 40 cm{sup −1} should be achievable over most of this frequency range.

  12. Chromosomal destabilization during gene amplification.

    PubMed Central

    Ruiz, J C; Wahl, G M

    1990-01-01

    Acentric extrachromosomal elements, such as submicroscopic autonomously replicating circular molecules (episomes) and double minute chromosomes, are common early, and in some cases initial, intermediates of gene amplification in many drug-resistant and tumor cell lines. In order to gain a more complete understanding of the amplification process, we investigated the molecular mechanisms by which such extrachromosomal elements are generated and we traced the fate of these amplification intermediates over time. The model system consists of a Chinese hamster cell line (L46) created by gene transfer in which the initial amplification product was shown previously to be an unstable extrachromosomal element containing an inverted duplication spanning more than 160 kilobases (J. C. Ruiz and G. M. Wahl, Mol. Cell. Biol. 8:4302-4313, 1988). In this study, we show that these molecules were formed by a process involving chromosomal deletion. Fluorescence in situ hybridization was performed at multiple time points on cells with amplified sequences. These studies reveal that the extrachromosomal molecules rapidly integrate into chromosomes, often near or at telomeres, and once integrated, the amplified sequences are themselves unstable. These data provide a molecular and cytogenetic chronology for gene amplification in this model system; an early event involves deletion to generate extrachromosomal elements, and subsequent integration of these elements precipitates a cascade of chromosome instability. Images PMID:2188107

  13. Quantum phase amplification for temporal pulse shaping and super-resolution in remote sensing

    NASA Astrophysics Data System (ADS)

    Yin, Yanchun

    QPA in the spatial domain has also been studied as a method to enhance the spatial resolution of imaging systems. A detailed model has been developed for achieving both super-resolution and detection of phase-amplified light. The imaging resolution problem considered here is treated as a binary hypotheses testing problem. Resolution enhancement is achieved by magnification of the angular separation of two targets in the sub-Rayleigh regime. The detection model includes optimization of detector segmentation, detector noise, and detection in both the spatial and the spatial frequency domain, which provide strategies for the optimization of the signal-to-noise ratio that take advantage of both the change of the field distribution and the change of energy of the signal in the QPA process. Proof-of-principle experiments have been conducted in the spatial domain. For the first time, beam angular amplification has been demonstrated, and the experimental result is in good agreement with simulations. The experimental demonstration has been achieved by observing the correlation of amplitude and angular phase in the phase-sensitive three-wave mixing process using ultrashort laser pulses and utilizing a type I three-wave mixing process. Several diagnostics have been developed and employed in the experimental measurements, including the near-field diagnostic, the far-field diagnostic, and the interferometry diagnostic. They have all been used to confirm the existence and study the properties of the QPA process on a shot-to-shot basis. Specifically, amplitude was measured in the near-field diagnostic, while the angular phase was indirectly measured in the far-field diagnostic by determining the position of the beam centroid. Interferometric measurements have been found to be of insufficient accuracy for this measurement in the way they were implemented. The demonstration of beam angular amplification by use of QPA lays the foundation for future integrated demonstration of imaging

  14. Systematic Studies of Distributed and Hybrid Raman Amplification in 10G-EPON and TWDM-PON

    NASA Astrophysics Data System (ADS)

    Kashima, N.

    2016-03-01

    We have studied distributed Raman amplification (DRA) and hybrid Raman amplification to apply both 10G-EPON and TWDM-PON systems. Two types of access networks, 50 km and 10 km, are assumed for case studies. Using the calculated optical received signal power Ps and optical signal noise ratio (OSNR), we consider the cases where the hybrid Raman amplification is necessary or not. In the case of coexistence of both systems, co-use of lumped Raman amplification (LRA) located at a central office (CO) may be possible. The co-use of LRA for both systems is discussed using the calculated Ps and OSNR.

  15. Discovery of a photoresponse amplification mechanism in compensated PN junctions

    SciTech Connect

    Zhou, Yuchun; Rahman, Samia N.; Hall, David; Lo, Yu-Hwa; Liu, Yu-Hsin; Sham, L. J.

    2015-01-19

    We report the experimental evidence of uncovering a photoresponse amplification mechanism in heavily doped, partially compensated silicon p-n junctions under very low bias voltage. We show that the observed photocurrent gain occurs at a bias that is more than an order of magnitude below the threshold voltage for conventional impact ionization. Moreover, contrary to the case of avalanche detectors and p-i-n diodes, the amplified photoresponse is enhanced rather than suppressed with increasing temperature. These distinctive characteristics lead us to hypothesize that the inelastic scattering between energetic electrons (holes) and the ionized impurities in the depletion and charge neutral regions of the p-n junction in a cyclic manner plays a significant role in the amplification process. Such an internal signal amplification mechanism, which occurs at much lower bias than impact ionization and favors room temperature over cryogenic temperature, makes it promising for practical device applications.

  16. Alternative Chemical Amplification Methods for Peroxy Radical Detection

    NASA Astrophysics Data System (ADS)

    Wood, E. C. D.

    2014-12-01

    Peroxy radicals (HO2, CH3O2, etc.) are commonly detected by the chemical amplification technique, in which ambient air is mixed with high concentrations of CO and NO, initiating a chain reaction that produces 30 - 200 NO2 molecules per sampled peroxy radical. The NO2 is then measured by one of several techniques. With the exception of CIMS-based techniques, the chemical amplification method has undergone only incremental improvements since it was first introduced in 1982. The disadvantages of the technique include the need to use high concentrations of CO and the greatly reduced sensitivity of the amplification chain length in the presence of water vapor. We present a new chemical amplification scheme in which either ethane or acetaldehyde is used in place of CO, with the NO2 product detected using Cavity Attenuated Phase Shift spectroscopy (CAPS). Under dry conditions, the amplification factor of the alternative amplifiers are approximately six times lower than the CO-based amplifier. The relative humidity "penalty" is not as severe, however, such that at typical ambient relative humidity (RH) values the amplification factor is within a factor of three of the CO-based amplifier. Combined with the NO2 sensitivity of CAPS and a dual-channel design, the detection limit of the ethane amplifier is less than 2 ppt (1 minute average, signal-to-noise ratio 2). The advantages of these alternative chemical amplification schemes are improved safety, a reduced RH correction, and increased sensitivity to organic peroxy radicals relative to HO2.

  17. Loopback rolling circle amplification for ultrasensitive detection of Kras gene.

    PubMed

    Xu, Huo; Wu, Dong; Jiang, Yifan; Zhang, Rongbo; Wu, Qingzheng; Liu, Yiyun; Li, Feng; Wu, Zai-Sheng

    2017-03-01

    Mutations in Kras gene may be used as a diagnostic marker and a target for treatment of the broad spectrum of human cancers. In this study, we developed a new class of amplification assay, double-hairpin molecular beacon (DHMB)-based cascade rolling circle amplification (RCA), for ultrasensitive and selective detection of Kras gene in a homogenous solution. Specifically, target DNA can hybridize with DHMB and activate cyclical target strand-displacement polymerization (CTDP) and nicking-mediated strand-displacement polymerization (NMDP). The resulting nicked/displaced fragments substantially outnumber target DNA and cause the cascade rolling circle amplification (C-RCA) and nicked fragment-induced strand-displacement polymerization (NFDP). Even if four amplification processes are designed, only DHMB, padlock probe and polymerization primer are involved. Under optimized conditions, this screening system exhibits a linear range of 5 orders of magnitude (from 100fM to 20nM), and the detection limit is down to 16fM. Moreover, the developed biosensing system offers a high assay specificity for perfectly matched target DNA, and the measured data from practical samples demonstrated the potential application in the cancer diagnoses. As a proof-of-concept genetic assay, the novel signaling strategy, as well as desirable analytical capability, would significantly benefit the development of versatile amplification gene profiling platforms, revealing great promise in biological studies and medical diagnostics.

  18. Sequence independent amplification of DNA

    DOEpatents

    Bohlander, Stefan K.

    1998-01-01

    The present invention is a rapid sequence-independent amplification procedure (SIA). Even minute amounts of DNA from various sources can be amplified independent of any sequence requirements of the DNA or any a priori knowledge of any sequence characteristics of the DNA to be amplified. This method allows, for example the sequence independent amplification of microdissected chromosomal material and the reliable construction of high quality fluorescent in situ hybridization (FISH) probes from YACs or from other sources. These probes can be used to localize YACs on metaphase chromosomes but also--with high efficiency--in interphase nuclei.

  19. Sequence independent amplification of DNA

    DOEpatents

    Bohlander, S.K.

    1998-03-24

    The present invention is a rapid sequence-independent amplification procedure (SIA). Even minute amounts of DNA from various sources can be amplified independent of any sequence requirements of the DNA or any a priori knowledge of any sequence characteristics of the DNA to be amplified. This method allows, for example, the sequence independent amplification of microdissected chromosomal material and the reliable construction of high quality fluorescent in situ hybridization (FISH) probes from YACs or from other sources. These probes can be used to localize YACs on metaphase chromosomes but also--with high efficiency--in interphase nuclei. 25 figs.

  20. Rapid amplification of the RM-Yplex assay.

    PubMed

    Abuidrees, Aqeela S; Alghafri, Rashed H; Hadi, Sibte

    2016-10-01

    A multiplex PCR assay consisting of 13 Rapidly Mutating Y STR loci called RM-Yplex was previously developed. Platinum(®) Taq DNA polymerase was used to amplify the 13 Y STR loci in a single reaction at an amplification time of approximately 2.5 h. In order to shorten the process with reliable results, two DNA polymerases were tested with the multiplex. Phusion(®) Flash High Fidelity, TAKARA Z-taq(TM) , and Platinum(®) Taq DNA polymerases were investigated for conducting RM-Yplex assay at various PCR cycling conditions. Rapid, robust, and efficient amplification of all the markers within the multiplex were achieved. The amplification time was reduced from 2.5 h to less than 28 min with Phusion(®) Flash High Fidelity DNA polymerase using Veriti(®) PCR thermal cycler.

  1. Danger signals in stroke.

    PubMed

    Gelderblom, Mathias; Sobey, Christopher G; Kleinschnitz, Christoph; Magnus, Tim

    2015-11-01

    Danger molecules are the first signals released from dying tissue after stroke. These danger signals bind to receptors on immune cells that will result in their activation and the release of inflammatory and neurotoxic mediators, resulting in amplification of the immune response and subsequent enlargement of the damaged brain volume. The release of danger signals is a central event that leads to a multitude of signals and cascades in the affected and neighbouring tissue, therefore providing a potential target for therapy.

  2. Non-resonant parametric amplification in biomimetic hair flow sensors: Selective gain and tunable filtering

    NASA Astrophysics Data System (ADS)

    Droogendijk, H.; Bruinink, C. M.; Sanders, R. G. P.; Krijnen, G. J. M.

    2011-11-01

    We demonstrate that the responsivity of flow sensors for harmonic flows can be improved significantly by non-resonant parametric amplification. Using electrostatic spring softening by AC-bias voltages, increased responsivity and sharp filtering are achieved in our biomimetic flow sensors. Tunable filtering is obtained for non-resonant electromechanical parametric amplification, applicable at a wide range of non-resonant frequencies while achieving highly selective gain of up to 20 dB.

  3. Ferrocenemonocarboxylic-HRP@Pt nanoparticles labeled RCA for multiple amplification of electro-immunosensing.

    PubMed

    Su, Huilan; Yuan, Ruo; Chai, Yaqin; Mao, Li; Zhuo, Ying

    2011-07-15

    A multiple amplification immunoassay was proposed to detect alpha-fetoprotein (AFP), which was based on ferrocenemonocarboxylic-HRP conjugated on Pt nanoparticles as labels for rolling circle amplification (RCA). Firstly, the capture antibody (anti-AFP) was immobilized on glass carbon electrode (GCE) deposited nano-sized gold particles. After a typical immuno-sandwich protocol, primary DNA was immobilized by labeling secondary antibody, which acted as a precursor to initiate RCA. The products of RCA provide large amount of sites to link detection DNAs, which were labeled by signal probes (ferrocenemonocarboxylic) and horseradish peroxidase (HRP). Moreover, the enzymatic amplification signals could be produced by the catalysis of HRP and Pt nanoparticles with the addition of H₂O₂. These lead to multiple amplification signals monitoring by electrochemical instrument and further resulted in high sensitivity of the immunoassay with the detection limit of 1.7 pg/mL.

  4. Multiplex detection of microRNAs by combining molecular beacon probes with T7 exonuclease-assisted cyclic amplification reaction.

    PubMed

    Liu, Yacui; Zhang, Jiangyan; Tian, Jingxiao; Fan, Xiaofei; Geng, Hao; Cheng, Yongqiang

    2017-01-01

    A simple, highly sensitive, and specific assay was developed for the homogeneous and multiplex detection of microRNAs (miRNAs) by combining molecular beacon (MB) probes and T7 exonuclease-assisted cyclic amplification. An MB probe with five base pairs in the stem region without special modification can effectively prevent the digestion by T7 exonuclease. Only in the presence of target miRNA is the MB probe hybridized with the target miRNA, and then digested by T7 exonuclease in the 5' to 3' direction. At the same time, the target miRNA is released and subsequently initiates the nuclease-assisted cyclic digestion process, generating enhanced fluorescence signal significantly. The results show that the combination of T7 exonuclease-assisted cyclic amplification reaction and MB probe possesses higher sensitivity for miRNA detection. Moreover, multiplex detection of miRNAs was successfully achieved by designing two MB probes labeled with FAM and Cy3, respectively. As a result, the method opens a new pathway for the sensitive and multiplex detection of miRNAs as well as clinical diagnosis. Graphical Abstract A simple, highly sensitive, and specific assay was developed for the detection of microRNAs by combining molecular beacon probes with T7 exonuclease-assisted cyclic amplification reaction.

  5. Ultrasensitive detection of microRNA through rolling circle amplification on a DNA tetrahedron decorated electrode.

    PubMed

    Miao, Peng; Wang, Bidou; Meng, Fanyu; Yin, Jian; Tang, Yuguo

    2015-03-18

    MicroRNAs are a class of evolutionally conserved, small noncoding RNAs involved in the regulation of gene expression and affect a variety of biological processes including cellular differentiation, immunological response, tumor development, and so on. Recently, microRNAs have been identified as promising disease biomarkers. In this work, we have fabricated a novel electrochemical method for ultrasensitive detection of microRNA. Generally, a DNA tetrahedron decorated gold electrode is employed as the recognition interface. Then, hybridizations between DNA tetrahedron, microRNA, and primer probe initiate rolling circle amplification (RCA) on the electrode surface. Silver nanoparticles attached to the RCA products provide significant electrochemical signals and a limit of detection as low as 50 aM is achieved. Moreover, homology microRNA family members with only one or two mismatches can be successfully distinguished. Therefore, this proposed method reveals great advancements toward improved disease diagnosis and prognosis.

  6. High repetition rate tunable femtosecond pulses and broadband amplification from fiber laser pumped parametric amplifier.

    PubMed

    Andersen, T V; Schmidt, O; Bruchmann, C; Limpert, J; Aguergaray, C; Cormier, E; Tünnermann, A

    2006-05-29

    We report on the generation of high energy femtosecond pulses at 1 MHz repetition rate from a fiber laser pumped optical parametric amplifier (OPA). Nonlinear bandwidth enhancement in fibers provides the intrinsically synchronized signal for the parametric amplifier. We demonstrate large tunability extending from 700 nm to 1500 nm of femtosecond pulses with pulse energies as high as 1.2 muJ when the OPA is seeded by a supercontinuum generated in a photonic crystal fiber. Broadband amplification over more than 85 nm is achieved at a fixed wavelength. Subsequent compression in a prism sequence resulted in 46 fs pulses. With an average power of 0.5 W these pulses have a peak-power above 10 MW. In particular, the average power and pulse energy scalability of both involved concepts, the fiber laser and the parametric amplifier, will enable easy up-scaling to higher powers.

  7. Experimental investigation of pulse generation with one-pump fiber optical parametric amplification.

    PubMed

    Vedadi, Armand A; Shoaie, Mohammad Amin; Brès, Camille-Sophie

    2012-11-19

    In a recent study, the theory of pulse generation with fiber optical parametric amplification using sinusoidal (clock) intensity modulated pump was revisited. This work showed that the pulses generated through such parametric interaction exhibit a shape which depends on the signal detuning with respect to the pump position (i.e. linear phase mismatch). A near Gaussian shape can only be achieved over a small region of the gain spectrum, close to the maximum gain location. Towards the extremities of the gain spectrum, the generated pulses take a near Sinc shape which can have many potential applications such as for all-optical Nyquist limited transmitters and/or receivers. In this paper we experimentally verify the theory at repetition rates up to 40 GHz. We also discuss the impact of noise, pump saturation and walk-off on the generated pulses.

  8. Efficient parametric amplification in micro-resonators with integrated piezoelectric actuation and sensing capabilities

    NASA Astrophysics Data System (ADS)

    Thomas, O.; Mathieu, F.; Mansfield, W.; Huang, C.; Trolier-McKinstry, S.; Nicu, L.

    2013-04-01

    We report, in this work, on unprecedented levels of parametric amplification in microelectromechanical resonators, operated in air, with integrated piezoelectric actuation and sensing capabilities. The method relies on an analytical/numerical understanding of the influence of geometrical nonlinearities inherent to the bridge-like configuration of the resonators. We provide analytical formulae to predict the performances of the parametric amplifier below the nonlinearity threshold, in terms of gain and quality factor (Q) enhancement. The analysis explains how to overcome this nonlinearity threshold by controlling the drive signals. It predicts that in theory, any Q-factor enhancement can be achieved. Experimental validation demonstrates a Q-factor enhancement by up to a factor 14 in air.

  9. Repressor logic modules assembled by rolling circle amplification platform to construct a set of logic gates

    PubMed Central

    Wei, Hua; Hu, Bo; Tang, Suming; Zhao, Guojie; Guan, Yifu

    2016-01-01

    Small molecule metabolites and their allosterically regulated repressors play an important role in many gene expression and metabolic disorder processes. These natural sensors, though valuable as good logic switches, have rarely been employed without transcription machinery in cells. Here, two pairs of repressors, which function in opposite ways, were cloned, purified and used to control DNA replication in rolling circle amplification (RCA) in vitro. By using metabolites and repressors as inputs, RCA signals as outputs, four basic logic modules were constructed successfully. To achieve various logic computations based on these basic modules, we designed series and parallel strategies of circular templates, which can further assemble these repressor modules in an RCA platform to realize twelve two-input Boolean logic gates and a three-input logic gate. The RCA-output and RCA-assembled platform was proved to be easy and flexible for complex logic processes and might have application potential in molecular computing and synthetic biology. PMID:27869177

  10. Optical chirped beam amplification and propagation

    DOEpatents

    Barty, Christopher P.

    2004-10-12

    A short pulse laser system uses dispersive optics in a chirped-beam amplification architecture to produce high peak power pulses and high peak intensities without the potential for intensity dependent damage to downstream optical components after amplification.

  11. Parametric amplification by coupled flux qubits

    SciTech Connect

    Rehák, M.; Neilinger, P.; Grajcar, M.; Oelsner, G.; Hübner, U.; Meyer, H.-G.; Il'ichev, E.

    2014-04-21

    We report parametric amplification of a microwave signal in a Kerr medium formed from superconducting qubits. Two mutually coupled flux qubits, embedded in the current antinode of a superconducting coplanar waveguide resonator, are used as a nonlinear element. Shared Josephson junctions provide the qubit-resonator coupling, resulting in a device with a tunable Kerr constant (up to 3 × 10{sup −3}) and a measured gain of about 20 dB. This arrangement represents a unit cell which can be straightforwardly extended to a quasi one-dimensional quantum metamaterial with large tunable Kerr nonlinearity, providing a basis for implementation of wide-band travelling wave parametric amplifiers.

  12. Amplification sans bruit d'images optiques

    NASA Astrophysics Data System (ADS)

    Gigan, S.; Delaubert, V.; Lopez, L.; Treps, N.; Maitre, A.; Fabre, C.

    2004-11-01

    Nous utilisons un Oscillateur Paramétrique Optique (OPO) pompé sous le seuil dans le but d'amplifier une image multimode transverse sans dégradation du rapport signal à bruit. Le dispositif expérimental met en œuvre un OPO de type II triplement résonant et semi-confocal pour le faisceau amplifié. L'existence d'effets quantiques lors de l'amplification multimode dans un tel dispositif a été montrée expérimentalement. Plus généralement, ceci nous a amené à étudier les propriétés quantiques transverses des faisceaux lumineux amplifiés. Une telle étude peut trouver des applications non seulement en imagerie, mais également dans le traitement quantique de l'information.

  13. Metformin inhibits age-related centrosome amplification in Drosophila midgut stem cells through AKT/TOR pathway.

    PubMed

    Na, Hyun-Jin; Park, Joung-Sun; Pyo, Jung-Hoon; Jeon, Ho-Jun; Kim, Young-Shin; Arking, Robert; Yoo, Mi-Ae

    2015-07-01

    We delineated the mechanism regulating the inhibition of centrosome amplification by metformin in Drosophila intestinal stem cells (ISCs). Age-related changes in tissue-resident stem cells may be closely associated with tissue aging and age-related diseases, such as cancer. Centrosome amplification is a hallmark of cancers. Our recent work showed that Drosophila ISCs are an excellent model for stem cell studies evaluating age-related increase in centrosome amplification. Here, we showed that metformin, a recognized anti-cancer drug, inhibits age- and oxidative stress-induced centrosome amplification in ISCs. Furthermore, we revealed that this effect is mediated via down-regulation of AKT/target of rapamycin (TOR) activity, suggesting that metformin prevents centrosome amplification by inhibiting the TOR signaling pathway. Additionally, AKT/TOR signaling hyperactivation and metformin treatment indicated a strong correlation between DNA damage accumulation and centrosome amplification in ISCs, suggesting that DNA damage might mediate centrosome amplification. Our study reveals the beneficial and protective effects of metformin on centrosome amplification via AKT/TOR signaling modulation. We identified a new target for the inhibition of age- and oxidative stress-induced centrosome amplification. We propose that the Drosophila ISCs may be an excellent model system for in vivo studies evaluating the effects of anti-cancer drugs on tissue-resident stem cell aging.

  14. Classroom Amplification To Enhance Student Performance.

    ERIC Educational Resources Information Center

    DiSarno, Neil J.; Schowalter, Melissa; Grassa, Patricia

    2002-01-01

    Discussion of classroom amplification systems to improve the performance of students with hearing loss or learning disabilities addresses the auditory challenges of inclusive classrooms, changing the classroom environment to reduce noise, types of amplification systems, and what teachers observe about amplification. (Contains references.) (DB)

  15. Field-Applicable Recombinase Polymerase Amplification Assay for Rapid Detection of Mycoplasma capricolum subsp. capripneumoniae.

    PubMed

    Liljander, Anne; Yu, Mingyan; O'Brien, Elizabeth; Heller, Martin; Nepper, Julia F; Weibel, Douglas B; Gluecks, Ilona; Younan, Mario; Frey, Joachim; Falquet, Laurent; Jores, Joerg

    2015-09-01

    Contagious caprine pleuropneumonia (CCPP) is a highly contagious disease caused by Mycoplasma capricolum subsp. capripneumoniae that affects goats in Africa and Asia. Current available methods for the diagnosis of Mycoplasma infection, including cultivation, serological assays, and PCR, are time-consuming and require fully equipped stationary laboratories, which make them incompatible with testing in the resource-poor settings that are most relevant to this disease. We report a rapid, specific, and sensitive assay employing isothermal DNA amplification using recombinase polymerase amplification (RPA) for the detection of M. capricolum subsp. capripneumoniae. We developed the assay using a specific target sequence in M. capricolum subsp. capripneumoniae, as found in the genome sequence of the field strain ILRI181 and the type strain F38 and that was further evidenced in 10 field strains from different geographical regions. Detection limits corresponding to 5 × 10(3) and 5 × 10(4) cells/ml were obtained using genomic DNA and bacterial culture from M. capricolum subsp. capripneumoniae strain ILRI181, while no amplification was obtained from 71 related Mycoplasma isolates or from the Acholeplasma or the Pasteurella isolates, demonstrating a high degree of specificity. The assay produces a fluorescent signal within 15 to 20 min and worked well using pleural fluid obtained directly from CCPP-positive animals without prior DNA extraction. We demonstrate that the diagnosis of CCPP can be achieved, with a short sample preparation time and a simple read-out device that can be powered by a car battery, in <45 min in a simulated field setting.

  16. Parametric amplification of a superconducting plasma wave

    NASA Astrophysics Data System (ADS)

    Rajasekaran, S.; Casandruc, E.; Laplace, Y.; Nicoletti, D.; Gu, G. D.; Clark, S. R.; Jaksch, D.; Cavalleri, A.

    2016-11-01

    Many applications in photonics require all-optical manipulation of plasma waves, which can concentrate electromagnetic energy on sub-wavelength length scales. This is difficult in metallic plasmas because of their small optical nonlinearities. Some layered superconductors support Josephson plasma waves, involving oscillatory tunnelling of the superfluid between capacitively coupled planes. Josephson plasma waves are also highly nonlinear, and exhibit striking phenomena such as cooperative emission of coherent terahertz radiation, superconductor-metal oscillations and soliton formation. Here, we show that terahertz Josephson plasma waves can be parametrically amplified through the cubic tunnelling nonlinearity in a cuprate superconductor. Parametric amplification is sensitive to the relative phase between pump and seed waves, and may be optimized to achieve squeezing of the order-parameter phase fluctuations or terahertz single-photon devices.

  17. Parametric amplification of a superconducting plasma wave

    SciTech Connect

    Rajasekaran, S.; Casandruc, E.; Laplace, Y.; Nicoletti, D.; Gu, G. D.; Clark, S. R.; Jaksch, D.; Cavalleri, A.

    2016-07-11

    Many applications in photonics require all-optical manipulation of plasma waves, which can concentrate electromagnetic energy on sub-wavelength length scales. This is difficult in metallic plasmas because of their small optical nonlinearities. Some layered superconductors support Josephson plasma waves, involving oscillatory tunnelling of the superfluid between capacitively coupled planes. Josephson plasma waves are also highly nonlinear, and exhibit striking phenomena such as cooperative emission of coherent terahertz radiation, superconductor–metal oscillations and soliton formation. In this paper, we show that terahertz Josephson plasma waves can be parametrically amplified through the cubic tunnelling nonlinearity in a cuprate superconductor. Finally, parametric amplification is sensitive to the relative phase between pump and seed waves, and may be optimized to achieve squeezing of the order-parameter phase fluctuations or terahertz single-photon devices.

  18. Parametric amplification of a superconducting plasma wave

    DOE PAGES

    Rajasekaran, S.; Casandruc, E.; Laplace, Y.; ...

    2016-07-11

    Many applications in photonics require all-optical manipulation of plasma waves, which can concentrate electromagnetic energy on sub-wavelength length scales. This is difficult in metallic plasmas because of their small optical nonlinearities. Some layered superconductors support Josephson plasma waves, involving oscillatory tunnelling of the superfluid between capacitively coupled planes. Josephson plasma waves are also highly nonlinear, and exhibit striking phenomena such as cooperative emission of coherent terahertz radiation, superconductor–metal oscillations and soliton formation. In this paper, we show that terahertz Josephson plasma waves can be parametrically amplified through the cubic tunnelling nonlinearity in a cuprate superconductor. Finally, parametric amplification is sensitivemore » to the relative phase between pump and seed waves, and may be optimized to achieve squeezing of the order-parameter phase fluctuations or terahertz single-photon devices.« less

  19. Separate TRP channels mediate amplification and transduction in drosophila

    NASA Astrophysics Data System (ADS)

    Lehnert, Brendan P.; Baker, Allison E.; Wilson, Rachel I.

    2015-12-01

    Auditory receptor cells rely on mechanically-gated channels to transform sound stimuli into neural activity. Several TRP channels have been implicated in Drosophila auditory transduction, but mechanistic studies have been hampered by the inability to record subthreshold signals from receptor neurons. We developed a non-invasive method for measuring these signals by recording from a central neuron that is electrically coupled to a genetically-defined population of auditory receptors. We find that the TRPN family member NompC, which is necessary for the active amplification of motion by the auditory organ, is not required for transduction. Instead, NompC sensitizes the transduction complex to movement and precisely regulates the static forces on the complex. In contrast, the TRPV channels Nanchung and Inactive are required for responses to sound, suggesting they are components of the transduction complex. Thus, transduction and active amplification are genetically separable processes in Drosophila hearing.

  20. PCR-free and label-free fluorescent detection of telomerase activity at single-cell level based on triple amplification.

    PubMed

    Gao, Yanfang; Xu, Jing; Li, Baoxin; Jin, Yan

    2016-07-15

    As a universal biomarker for cancer diagnostics and cancer therapeutics, telomerase has attracted extensive attention concerning its detection and discovery of its inhibitors. Herein, we developed a PCR-free and label-free fluorescent strategy for facile, reliable and highly sensitive assay of human telomerase activity from crude cancer cell extracts. A G-quadruplex-selective fluorescent dye, N-methyl mesoporphyrin IX (NMM), was utilized as signal probe. Two hairpin probes with hidden G-quadruplex strand in their stem were designed as assembly components of strand displacement reaction (SDR). In this strategy, one telomerase elongation product contains several hexamer repeats which can hybridize with numerous assistant DNA to release a lot of trigger DNA (T-DNA) of SDR for achieving first step amplification. Then, strand displacement reaction led to the formation of G-quadruplex at the both end of two hairpin DNA probes for realizing second step amplification. Finally, the re-released T-DNA initiated another cycle of SDR, resulting in a significant increase in the fluorescence intensity of NMM. By taking advantage of triple signal amplification, the telomerase activity in the HeLa extracts equivalent to 1-3000 cells was detected in homogeneous solution. Telomerase activities of different cell lines, including cancer cells and normal cell, were also successfully evaluated. Meanwhile, the inhibition effect of 3'-azido-3'-deoxythymidine (AZT) was also investigated. Therefore, it offers a simple and reliable method for detecting telomerase activity at single-cell level without complex pre-modification of probe and enzyme auxiliary signal amplification, which has the merits of simplicity, rapid response, low cost and high reliability.

  1. A geostatistical approach to mapping site response spectral amplifications

    USGS Publications Warehouse

    Thompson, E.M.; Baise, L.G.; Kayen, R.E.; Tanaka, Y.; Tanaka, H.

    2010-01-01

    If quantitative estimates of the seismic properties do not exist at a location of interest then the site response spectral amplifications must be estimated from data collected at other locations. Currently, the most common approach employs correlations of site class with maps of surficial geology. Analogously, correlations of site class with topographic slope can be employed where the surficial geology is unknown. Our goal is to identify and validate a method to estimate site response with greater spatial resolution and accuracy for regions where additional effort is warranted. This method consists of three components: region-specific data collection, a spatial model for interpolating seismic properties, and a theoretical method for computing spectral amplifications from the interpolated seismic properties. We consider three spatial interpolation schemes: correlations with surficial geology, termed the geologic trend (GT), ordinary kriging (OK), and kriging with a trend (KT). We estimate the spectral amplifications from seismic properties using the square root of impedance method, thereby linking the frequency-dependent spectral amplifications to the depth-dependent seismic properties. Thus, the range of periods for which this method is applicable is limited by the depth of exploration. A dense survey of near-surface S-wave slowness (Ss) throughout Kobe, Japan shows that the geostatistical methods give more accurate estimates of Ss than the topographic slope and GT methods, and the OK and KT methods perform equally well. We prefer the KT model because it can be seamlessly integrated with geologic maps that cover larger regions. Empirical spectral amplifications show that the region-specific data achieve more accurate estimates of observed median short-period amplifications than the topographic slope method. ?? 2010 Elsevier B.V.

  2. Frequency domain optical parametric amplification

    PubMed Central

    Schmidt, Bruno E.; Thiré, Nicolas; Boivin, Maxime; Laramée, Antoine; Poitras, François; Lebrun, Guy; Ozaki, Tsuneyuki; Ibrahim, Heide; Légaré, François

    2014-01-01

    Today’s ultrafast lasers operate at the physical limits of optical materials to reach extreme performances. Amplification of single-cycle laser pulses with their corresponding octave-spanning spectra still remains a formidable challenge since the universal dilemma of gain narrowing sets limits for both real level pumped amplifiers as well as parametric amplifiers. We demonstrate that employing parametric amplification in the frequency domain rather than in time domain opens up new design opportunities for ultrafast laser science, with the potential to generate single-cycle multi-terawatt pulses. Fundamental restrictions arising from phase mismatch and damage threshold of nonlinear laser crystals are not only circumvented but also exploited to produce a synergy between increased seed spectrum and increased pump energy. This concept was successfully demonstrated by generating carrier envelope phase stable, 1.43 mJ two-cycle pulses at 1.8 μm wavelength. PMID:24805968

  3. Deterministic noiseless amplification of coherent states

    NASA Astrophysics Data System (ADS)

    Hu, Meng-Jun; Zhang, Yong-Sheng

    2015-08-01

    A universal deterministic noiseless quantum amplifier has been shown to be impossible. However, probabilistic noiseless amplification of a certain set of states is physically permissible. Regarding quantum state amplification as quantum state transformation, we show that deterministic noiseless amplification of coherent states chosen from a proper set is attainable. The relation between input coherent states and gain of amplification for deterministic noiseless amplification is thus derived. Furthermore, we extend our result to more general situation and show that deterministic noiseless amplification of Gaussian states is also possible. As an example of application, we find that our amplification model can obtain better performance in homodyne detection to measure the phase of state selected from a certain set. Besides, other possible applications are also discussed.

  4. An ultrasensitive sandwich-type electrochemical immunosensor based on the signal amplification system of double-deck gold film and thionine unite with platinum nanowire inlaid globular SBA-15 microsphere.

    PubMed

    Wang, Ping; Li, Mingdang; Pei, Fubin; Li, Yueyun; Liu, Qing; Dong, Yunhui; Chu, Qingyan; Zhu, Hongjun

    2017-05-15

    A novel thionine unites with platinum nanowire inlaid globular SBA-15 (Pt NWs@g-SBA-15/Thi) not only utilizes as an efficient electrical signal probe but also constitutes an amplifying system with double-deck gold film (D-Au film) have been applied to the fabrication of sandwich-type immunosensor for detecting hepatitis B surface antigen (HBs Ag). The D-Au film can accelerate the electron transfer on the electrode interface due to the tunneling effect between the two Au films and can improve the load capacity of primary antibodies (Ab1) because of the good biocompatibility. The Pt NWs@g-SBA-15/Thi with uniform globular morphology not only can effectively reduce the spatial limitation for loading the secondary antibodies (Ab2) but also can provide outstanding pore accessibility of guest species from outside and offer catalytically active sites in a large scale. Besides, the presence of Thi can well enhance the electrical conductivity of Pt NWs@g-SBA-15/Thi. With the good cooperation between D-Au film and Pt NWs@g-SBA-15/Thi, a linear relationship between current signals and the concentrations of HBs Ag was obtained in the wide range from 10 fg/mL to 100ng/mL and the detection limit of HBs Ag was 3.3 fg/mL (signal-to-noise ratio of 3). Furthermore, the designed immunosensor with excellent selectivity, reproducibility and stability shows excellent performance in detection of human serum samples and provides a promising capacity for detecting a wide range of other tumor markers in clinical application.

  5. An RNA transcription-based amplification technique (NASBA) for the detection of viable Salmonella enterica.

    PubMed

    Simpkins, S A; Chan, A B; Hays, J; Pöpping, B; Cook, N

    2000-01-01

    Possession of mRNA is indicative of cell viability. RTPCR is not appropriate for mRNA detection as it cannot unambiguously detect mRNA in a DNA background. The alternative amplification technique, NASBA, avoids the disadvantages of RTPCR. We have devised a method for detection of viable Salmonella enterica. This involves NASBA amplification of mRNA transcribed from the dnaK gene. Amplification of mRNA extracted from viable and heat-killed cells from the same population produced consistent and highly significant (P > 0.01) differences between the respective signals. The signal obtained from viable cells was completely eradicated by RNase treatment, while PCR amplification of treated and untreated samples was unaffected, indicating that NASBA was unaffected by background DNA.

  6. Parametric amplification in AgGaSe2

    NASA Technical Reports Server (NTRS)

    Barnes, Norman P.; Gettemy, Donald J.; Hietanen, Jack R.; Iannini, Rebecca A.

    1989-01-01

    AgGaSe2 has been grown, annealed, and characterized for the mid-IR. Characterization includes measurement of the average power-limiting factors including absorption and the variation of the refractive indices with temperature. Using specially annealed crystals 20 mm long and a Ho:YAG pump, parametric amplification at 3.39 microns has achieved a gain of 2.9 with a peak power input of only 8 MW/sq cm.

  7. Enhanced solid-phase recombinase polymerase amplification and electrochemical detection.

    PubMed

    Del Río, Jonathan Sabaté; Lobato, Ivan Magriñà; Mayboroda, Olena; Katakis, Ioanis; O'Sullivan, Ciara K

    2017-03-02

    Recombinase polymerase amplification (RPA) is an elegant method for the rapid, isothermal amplification of nucleic acids. Here, we elucidate the optimal surface chemistry for rapid and efficient solid-phase RPA, which was fine-tuned in order to obtain a maximum signal-to-noise ratio, defining the optimal DNA probe density, probe-to-lateral spacer ratio (1:0, 1:1, 1:10 and 1:100) and length of a vertical spacer of the probe as well as investigating the effect of different types of lateral spacers. The use of different labelling strategies was also examined in order to reduce the number of steps required for the analysis, using biotin or horseradish peroxidase-labelled reverse primers. Optimisation of the amplification temperature used and the use of surface blocking agents were also pursued. The combination of these changes facilitated a significantly more rapid amplification and detection protocol, with a lowered limit of detection (LOD) of 1 · 10(-15) M. The optimised protocol was applied to the detection of Francisella tularensis in real samples from hares and a clear correlation with PCR and qPCR results observed and the solid-phase RPA demonstrated to be capable of detecting 500 fM target DNA in real samples. Graphical abstract Relative size of thiolated lateral spacers tested versus the primer and the uvsx recombinase protein.

  8. Methods and devices based on brillouin selective sideband amplification

    NASA Technical Reports Server (NTRS)

    Yao, X. Steve (Inventor)

    2003-01-01

    Opto-electronic devices and techniques using Brillouin scattering to select a sideband in a modulated optical carrier signal for amplification. Two lasers respectively provide a carrier signal beam and a Brillouin pump beam which are fed into an Brillouin optical medium in opposite directions. The relative frequency separation between the lasers is adjusted to align the frequency of the backscattered Brillouin signal with a desired sideband in the carrier signal to effect a Brillouin gain on the sideband. This effect can be used to implement photonic RF signal mixing and conversion with gain, conversion from phase modulation to amplitude modulation, photonic RF frequency multiplication, optical and RF pulse generation and manipulation, and frequency-locking of lasers.

  9. High peak-power kilohertz laser system employing single-stage multi-pass amplification

    DOEpatents

    Shan, Bing; Wang, Chun; Chang, Zenghu

    2006-05-23

    The present invention describes a technique for achieving high peak power output in a laser employing single-stage, multi-pass amplification. High gain is achieved by employing a very small "seed" beam diameter in gain medium, and maintaining the small beam diameter for multiple high-gain pre-amplification passes through a pumped gain medium, then leading the beam out of the amplifier cavity, changing the beam diameter and sending it back to the amplifier cavity for additional, high-power amplification passes through the gain medium. In these power amplification passes, the beam diameter in gain medium is increased and carefully matched to the pump laser's beam diameter for high efficiency extraction of energy from the pumped gain medium. A method of "grooming" the beam by means of a far-field spatial filter in the process of changing the beam size within the single-stage amplifier is also described.

  10. Brillouin Amplification--A Powerful New Scheme for Microwave Photonic Communications

    NASA Technical Reports Server (NTRS)

    Yao, S.; Maleki, L.

    1997-01-01

    We introduce the Brillouin selective sideband amplification technique and demonstrate many important applications of this technique in photonic microwave systems, including efficient phase modulation to amplitude modulation conversion, photonic frequency multiplication, photonic signal mixing with gain, and frequency multiplied signal up conversion.

  11. Low noise, low heat dissipation, high gain AC-DC front end amplification for scanning probe microscopy.

    PubMed

    Messina, Paolo; Fradin, F Y; Pittana, Paolo

    2009-02-04

    We report here on the design, construction and testing of a vacuum compatible AC-DC amplification system for low signal measurements with scanning probes. The most important feature of this new amplification system is incorporated within the head of a scanning tunneling microscope (STM). This is achieved with a very low thermal dissipation radio frequency amplifier at the STM head. The amplifier gain is higher than 40 dB and has a 50 dB maximum. Further, the AC noise figure is 0.7 dB between 100 and 1000 MHz. The noise induced in the DC amplifier is less than 2 pA RMS (root mean square), which enables the microscope to scan over soft insulating molecular layers. Thermal drift at the STM tip-sample interface is below 0.1 nm min(-1) both in air and in vacuum operation. Atomic resolution on highly oriented pyrolytic graphite surfaces is reliably achieved. Spin noise measurements are provided as an example of an application.

  12. Low noise, low heat dissipation, high gain AC-DC front end amplification for scanning probe microscopy.

    SciTech Connect

    Messina, P.; Fradin, F. Y.; Pittana, P.

    2009-01-01

    We report here on the design, construction and testing of a vacuum compatible AC-DC amplification system for low signal measurements with scanning probes. The most important feature of this new amplification system is incorporated within the head of a scanning tunneling microscope (STM). This is achieved with a very low thermal dissipation radio frequency amplifier at the STM head. The amplifier gain is higher than 40 dB and has a 50 dB maximum. Further, the AC noise figure is 0.7 dB between 100 and 1000 MHz. The noise induced in the DC amplifier is less than 2 pA RMS (root mean square), which enables the microscope to scan over soft insulating molecular layers. Thermal drift at the STM tip-sample interface is below 0.1 nm min{sup -1} both in air and in vacuum operation. Atomic resolution on highly oriented pyrolytic graphite surfaces is reliably achieved. Spin noise measurements are provided as an example of an application.

  13. Chemical Amplification with Encapsulated Reagents

    NASA Technical Reports Server (NTRS)

    Chen, Jian; Koemer, Steffi; Craig, Stephen; Lin, Shirley; Rudkevich, Dmitry M.; Rebek, Julius, Jr.

    2002-01-01

    Autocatalysis and chemical amplification are characteristic properties of living systems, and they give rise to behaviors such as increased sensitivity, responsiveness, and self-replication. Here we report a synthetic system in which a unique form of compartmentalization leads to nonlinear, autocatalytic behavior. The compartment is a reversibly formed capsule in which a reagent is sequestered. Reaction products displace the reagent from the capsule into solution and the reaction rate is accelerated. The resulting self-regulation is sensitive to the highly selective molecular recognition properties of the capsule.

  14. Bacteriophage Amplification-Coupled Detection and Identification of Bacterial Pathogens

    NASA Astrophysics Data System (ADS)

    Cox, Christopher R.; Voorhees, Kent J.

    Current methods of species-specific bacterial detection and identification are complex, time-consuming, and often require expensive specialized equipment and highly trained personnel. Numerous biochemical and genotypic identification methods have been applied to bacterial characterization, but all rely on tedious microbiological culturing practices and/or costly sequencing protocols which render them impractical for deployment as rapid, cost-effective point-of-care or field detection and identification methods. With a view towards addressing these shortcomings, we have exploited the evolutionarily conserved interactions between a bacteriophage (phage) and its bacterial host to develop species-specific detection methods. Phage amplification-coupled matrix assisted laser desorption time-of-flight mass spectrometry (MALDI-TOF-MS) was utilized to rapidly detect phage propagation resulting from species-specific in vitro bacterial infection. This novel signal amplification method allowed for bacterial detection and identification in as little as 2 h, and when combined with disulfide bond reduction methods developed in our laboratory to enhance MALDI-TOF-MS resolution, was observed to lower the limit of detection by several orders of magnitude over conventional spectroscopy and phage typing methods. Phage amplification has been combined with lateral flow immunochromatography (LFI) to develop rapid, easy-to-operate, portable, species-specific point-of-care (POC) detection devices. Prototype LFI detectors have been developed and characterized for Yersinia pestis and Bacillus anthracis, the etiologic agents of plague and anthrax, respectively. Comparable sensitivity and rapidity was observed when phage amplification was adapted to a species-specific handheld LFI detector, thus allowing for rapid, simple, POC bacterial detection and identification while eliminating the need for bacterial culturing or DNA isolation and amplification techniques.

  15. Amplification of ultra-short laser pulses via resonant backward Raman amplification in plasma

    NASA Astrophysics Data System (ADS)

    Mishra, S. K.; Andreev, A.

    2016-08-01

    In this paper, we have examined the possibility of using resonant backward Raman amplification (BRA) as an efficient mechanism in amplifying the low intensity ultra-short ( ≤ fs ) pulses using plasma as intermediate amplifying medium; such pulses are anticipated to get produced in the form of the secondary sources at ALPS (Attosecond Light Pulse Source) center of ELI (Extreme Light Infrastructure). In preliminary assessment of the scheme, the analytical expressions for the pump/seed laser pulses and plasma characteristic features are obtained which concisely describe the parameter regime of resonant BRA applicability in achieving significant amplification. The consistency of the scheme in the context of ELI-ALPS sources has been validated through particle in cell (PIC) simulations. The peak intensity of the amplified seed pulse predicted via simulation results is found in reasonable agreement with the analytical estimates. Utilizing these analytical expressions as a basis in perspective of ELI-ALPS parameter access, a specific example displaying the key plasma and laser parameters for amplifying weak seed pulse has been configured; the limitations and conceivable remedies in resonant BRA implementation have also been highlighted.

  16. Trophic amplification of climate warming.

    PubMed

    Kirby, Richard R; Beaugrand, Gregory

    2009-12-07

    Ecosystems can alternate suddenly between contrasting persistent states due to internal processes or external drivers. It is important to understand the mechanisms by which these shifts occur, especially in exploited ecosystems. There have been several abrupt marine ecosystem shifts attributed either to fishing, recent climate change or a combination of these two drivers. We show that temperature has been an important driver of the trophodynamics of the North Sea, a heavily fished marine ecosystem, for nearly 50 years and that a recent pronounced change in temperature established a new ecosystem dynamic regime through a series of internal mechanisms. Using an end-to-end ecosystem approach that included primary producers, primary, secondary and tertiary consumers, and detritivores, we found that temperature modified the relationships among species through nonlinearities in the ecosystem involving ecological thresholds and trophic amplifications. Trophic amplification provides an alternative mechanism to positive feedback to drive an ecosystem towards a new dynamic regime, which in this case favours jellyfish in the plankton and decapods and detritivores in the benthos. Although overfishing is often held responsible for marine ecosystem degeneration, temperature can clearly bring about similar effects. Our results are relevant to ecosystem-based fisheries management (EBFM), seen as the way forward to manage exploited marine ecosystems.

  17. Trophic amplification of climate warming

    PubMed Central

    Kirby, Richard R.; Beaugrand, Gregory

    2009-01-01

    Ecosystems can alternate suddenly between contrasting persistent states due to internal processes or external drivers. It is important to understand the mechanisms by which these shifts occur, especially in exploited ecosystems. There have been several abrupt marine ecosystem shifts attributed either to fishing, recent climate change or a combination of these two drivers. We show that temperature has been an important driver of the trophodynamics of the North Sea, a heavily fished marine ecosystem, for nearly 50 years and that a recent pronounced change in temperature established a new ecosystem dynamic regime through a series of internal mechanisms. Using an end-to-end ecosystem approach that included primary producers, primary, secondary and tertiary consumers, and detritivores, we found that temperature modified the relationships among species through nonlinearities in the ecosystem involving ecological thresholds and trophic amplifications. Trophic amplification provides an alternative mechanism to positive feedback to drive an ecosystem towards a new dynamic regime, which in this case favours jellyfish in the plankton and decapods and detritivores in the benthos. Although overfishing is often held responsible for marine ecosystem degeneration, temperature can clearly bring about similar effects. Our results are relevant to ecosystem-based fisheries management (EBFM), seen as the way forward to manage exploited marine ecosystems. PMID:19740882

  18. Dynamics and control of DNA sequence amplification

    NASA Astrophysics Data System (ADS)

    Marimuthu, Karthikeyan; Chakrabarti, Raj

    2014-10-01

    DNA amplification is the process of replication of a specified DNA sequence in vitro through time-dependent manipulation of its external environment. A theoretical framework for determination of the optimal dynamic operating conditions of DNA amplification reactions, for any specified amplification objective, is presented based on first-principles biophysical modeling and control theory. Amplification of DNA is formulated as a problem in control theory with optimal solutions that can differ considerably from strategies typically used in practice. Using the Polymerase Chain Reaction as an example, sequence-dependent biophysical models for DNA amplification are cast as control systems, wherein the dynamics of the reaction are controlled by a manipulated input variable. Using these control systems, we demonstrate that there exists an optimal temperature cycling strategy for geometric amplification of any DNA sequence and formulate optimal control problems that can be used to derive the optimal temperature profile. Strategies for the optimal synthesis of the DNA amplification control trajectory are proposed. Analogous methods can be used to formulate control problems for more advanced amplification objectives corresponding to the design of new types of DNA amplification reactions.

  19. Dynamics and control of DNA sequence amplification

    SciTech Connect

    Marimuthu, Karthikeyan; Chakrabarti, Raj E-mail: rajc@andrew.cmu.edu

    2014-10-28

    DNA amplification is the process of replication of a specified DNA sequence in vitro through time-dependent manipulation of its external environment. A theoretical framework for determination of the optimal dynamic operating conditions of DNA amplification reactions, for any specified amplification objective, is presented based on first-principles biophysical modeling and control theory. Amplification of DNA is formulated as a problem in control theory with optimal solutions that can differ considerably from strategies typically used in practice. Using the Polymerase Chain Reaction as an example, sequence-dependent biophysical models for DNA amplification are cast as control systems, wherein the dynamics of the reaction are controlled by a manipulated input variable. Using these control systems, we demonstrate that there exists an optimal temperature cycling strategy for geometric amplification of any DNA sequence and formulate optimal control problems that can be used to derive the optimal temperature profile. Strategies for the optimal synthesis of the DNA amplification control trajectory are proposed. Analogous methods can be used to formulate control problems for more advanced amplification objectives corresponding to the design of new types of DNA amplification reactions.

  20. The application of distributed Raman amplification in an all optical network

    NASA Astrophysics Data System (ADS)

    Zheng, Xiaoping; Feng, Feifei; Zhang, Hanyi; Li, Yanhe

    2003-04-01

    The effect of distributed Raman amplification (DRA) on the optical signal to noise ratio (OSNR) of an all optical network (AON) is examined by analyzing two types of node isolated principal (NIP). Based on the parameters used in calculation, it is found that in the first case of NIP, the OSNR of a signal passing through such AON can be improved by about 8 dB compared with no DRA. Whereas in the second case of NIP, the OSNR of the signal can be reduced by 11 dB. This kind of phenomena is analyzed and attributed to the dependence of noise figure of amplification system on the way of the active amplification utilization.

  1. A Label-Free and Sensitive Fluorescent Qualitative Assay for Bisphenol A Based on Rolling Circle Amplification/Exonuclease III-Combined Cascade Amplification

    PubMed Central

    Li, Xia; Song, Juan; Xue, Qing-Wang; You, Fu-Heng; Lu, Xia; Kong, Yan-Cong; Ma, Shu-Yi; Jiang, Wei; Li, Chen-Zhong

    2016-01-01

    Bisphenol A (BPA) detection in drinking water and food packaging materials has attracted much attention since the discovery that BPA can interfere with normal physiological processes and cause adverse health effects. Here, we constructed a label-free aptamer fluorescent assay for selective and sensitive detection of BPA based on the rolling circle amplification (RCA)/Exonuclease III (Exo III)-combined cascade amplification strategy. First, the duplex DNA probe (RP) with anti-BPA aptamer and trigger sequence was designed for BPA recognition and signal amplification. Next, under the action of BPA, the trigger probe was liberated from RP to initiate RCA reaction as primary amplification. Subsequently, the RCA products were used to trigger Exo III assisted secondary amplification with the help of hairpin probes, producing plenty of “G-quadruplex” in lantern-like structures. Finally, the continuously enriched “G-quadruplex lanterns” were lightened by zinc(II)-protoporphyrin IX (ZnPPIX) generating enhanced fluorescence signals. By integrating the primary RCA and secondary Exo III mediated cascade amplification strategy, this method displayed an excellent sensitivity with the detection limits of 5.4 × 10−17 M. In addition, the anti-BPA aptamer exhibits high recognition ability with BPA, guaranteeing the specificity of detection. The reporter signal probe (G-quadruplex with ZnPPIX) provides a label-free fluorescence signals readout without complicated labeling procedures, making the method simple in design and cost-effective in operation. Moreover, environmental samples analysis was also performed, suggesting that our strategy was reliable and had a great potential application in environmental monitoring. PMID:28335318

  2. Divided-pulse nonlinear amplification and simultaneous compression

    SciTech Connect

    Hao, Qiang; Zhang, Qingshan; Sun, Tingting; Chen, Jie; Wang, Yuqing; Guo, Zhengru; Yang, Kangwen; Guo, Zhanhua; Zeng, Heping

    2015-03-09

    We report on a fiber laser system delivering 122 fs pulse duration and 600 mW average power at 1560 nm by the interplay between divided pulse amplification and nonlinear pulse compression. A small-core double-clad erbium-doped fiber with anomalous dispersion carries out the pulse amplification and simultaneously compresses the laser pulses such that a separate compressor is no longer necessary. A numeric simulation reveals the existence of an optimum fiber length for producing transform-limited pulses. Furthermore, frequency doubling to 780 nm with 240 mW average power and 98 fs pulse duration is achieved by using a periodically poled lithium niobate crystal at room temperature.

  3. Measurement-based noiseless linear amplification for quantum communication

    NASA Astrophysics Data System (ADS)

    Chrzanowski, Helen M.; Walk, Nathan; Assad, Syed M.; Janousek, Jiri; Hosseini, Sara; Ralph, Timothy C.; Symul, Thomas; Lam, Ping Koy

    2014-04-01

    Entanglement distillation is an indispensable ingredient in extended quantum communication networks. Distillation protocols are necessarily non-deterministic and require advanced experimental techniques such as noiseless amplification. Recently, it was shown that the benefits of noiseless amplification could be extracted by performing a post-selective filtering of the measurement record to improve the performance of quantum key distribution. We apply this protocol to entanglement degraded by transmission loss of up to the equivalent of 100 km of optical fibre. We measure an effective entangled resource stronger than that achievable by even a maximally entangled resource passively transmitted through the same channel. We also provide a proof-of-principle demonstration of secret key extraction from an otherwise insecure regime. The measurement-based noiseless linear amplifier offers two advantages over its physical counterpart: ease of implementation and near-optimal probability of success. It should provide an effective and versatile tool for a broad class of entanglement-based quantum communication protocols.

  4. Digital Droplet Multiple Displacement Amplification (ddMDA) for Whole Genome Sequencing of Limited DNA Samples

    PubMed Central

    Rhee, Minsoung; Light, Yooli K.; Meagher, Robert J.; Singh, Anup K.

    2016-01-01

    Multiple displacement amplification (MDA) is a widely used technique for amplification of DNA from samples containing limited amounts of DNA (e.g., uncultivable microbes or clinical samples) before whole genome sequencing. Despite its advantages of high yield and fidelity, it suffers from high amplification bias and non-specific amplification when amplifying sub-nanogram of template DNA. Here, we present a microfluidic digital droplet MDA (ddMDA) technique where partitioning of the template DNA into thousands of sub-nanoliter droplets, each containing a small number of DNA fragments, greatly reduces the competition among DNA fragments for primers and polymerase thereby greatly reducing amplification bias. Consequently, the ddMDA approach enabled a more uniform coverage of amplification over the entire length of the genome, with significantly lower bias and non-specific amplification than conventional MDA. For a sample containing 0.1 pg/μL of E. coli DNA (equivalent of ~3/1000 of an E. coli genome per droplet), ddMDA achieves a 65-fold increase in coverage in de novo assembly, and more than 20-fold increase in specificity (percentage of reads mapping to E. coli) compared to the conventional tube MDA. ddMDA offers a powerful method useful for many applications including medical diagnostics, forensics, and environmental microbiology. PMID:27144304

  5. Digital droplet multiple displacement amplification (ddMDA) for whole genome sequencing of limited DNA samples

    SciTech Connect

    Rhee, Minsoung; Light, Yooli K.; Meagher, Robert J.; Singh, Anup K.; Kumar-Sinha, Chandan

    2016-05-04

    Here, multiple displacement amplification (MDA) is a widely used technique for amplification of DNA from samples containing limited amounts of DNA (e.g., uncultivable microbes or clinical samples) before whole genome sequencing. Despite its advantages of high yield and fidelity, it suffers from high amplification bias and non-specific amplification when amplifying sub-nanogram of template DNA. Here, we present a microfluidic digital droplet MDA (ddMDA) technique where partitioning of the template DNA into thousands of sub-nanoliter droplets, each containing a small number of DNA fragments, greatly reduces the competition among DNA fragments for primers and polymerase thereby greatly reducing amplification bias. Consequently, the ddMDA approach enabled a more uniform coverage of amplification over the entire length of the genome, with significantly lower bias and non-specific amplification than conventional MDA. For a sample containing 0.1 pg/μL of E. coli DNA (equivalent of ~3/1000 of an E. coli genome per droplet), ddMDA achieves a 65-fold increase in coverage in de novo assembly, and more than 20-fold increase in specificity (percentage of reads mapping to E. coli) compared to the conventional tube MDA. ddMDA offers a powerful method useful for many applications including medical diagnostics, forensics, and environmental microbiology.

  6. Digital droplet multiple displacement amplification (ddMDA) for whole genome sequencing of limited DNA samples

    DOE PAGES

    Rhee, Minsoung; Light, Yooli K.; Meagher, Robert J.; ...

    2016-05-04

    Here, multiple displacement amplification (MDA) is a widely used technique for amplification of DNA from samples containing limited amounts of DNA (e.g., uncultivable microbes or clinical samples) before whole genome sequencing. Despite its advantages of high yield and fidelity, it suffers from high amplification bias and non-specific amplification when amplifying sub-nanogram of template DNA. Here, we present a microfluidic digital droplet MDA (ddMDA) technique where partitioning of the template DNA into thousands of sub-nanoliter droplets, each containing a small number of DNA fragments, greatly reduces the competition among DNA fragments for primers and polymerase thereby greatly reducing amplification bias. Consequently,more » the ddMDA approach enabled a more uniform coverage of amplification over the entire length of the genome, with significantly lower bias and non-specific amplification than conventional MDA. For a sample containing 0.1 pg/μL of E. coli DNA (equivalent of ~3/1000 of an E. coli genome per droplet), ddMDA achieves a 65-fold increase in coverage in de novo assembly, and more than 20-fold increase in specificity (percentage of reads mapping to E. coli) compared to the conventional tube MDA. ddMDA offers a powerful method useful for many applications including medical diagnostics, forensics, and environmental microbiology.« less

  7. A fluorescence biosensor for VEGF detection based on DNA assembly structure switching and isothermal amplification.

    PubMed

    Li, Junlong; Sun, Kexin; Chen, Zhongping; Shi, Jifei; Zhou, Dandan; Xie, Guoming

    2017-03-15

    Vascular endothelial growth factor (VEGF) is an important biomarker in cancer angiogenesis. Here, we develop a aptasensor method for VEGF detection based on DNA assembly structure switching and isothermal amplification. The design employs a DNA assembly made of a isothermal amplification template, a aptamer, a primer and a protector chain. The DNA assembly is unable to undergo isothermal amplification in the absence of target. The presence of the target, however, triggers a structure switching event that causes hybridization of primer with template to facilitate isothermal amplification. Whereafter, toehold-mediated DNA strand displacement reaction between the generated (single-stranded DNA) ssDNA and fluorescent/quencher labeled probe are performed. Then, the increase in fluorescence provides an analytical signal. This strategy opens up a sensitive, selective and simple sensing platform for detection of VEGF. The system was also implemented to analyze the VEGF in human serum samples with satisfactory results.

  8. Amplification without instability: applying fluid dynamical insights in chemistry and biology

    NASA Astrophysics Data System (ADS)

    McCoy, Jonathan H.

    2013-11-01

    While amplification of small perturbations often arises from instability, transient amplification is possible locally even in asymptotically stable systems. That is, knowledge of a system's stability properties can mislead one's intuition for its transient behaviors. This insight, which has an interesting history in fluid dynamics, has more recently been rediscovered in ecology. Surprisingly, many nonlinear fluid dynamical and ecological systems share linear features associated with transient amplification of noise. This paper aims to establish that these features are widespread in many other disciplines concerned with noisy systems, especially chemistry, cell biology and molecular biology. Here, using classic nonlinear systems and the graphical language of network science, we explore how the noise amplification problem can be reframed in terms of activatory and inhibitory interactions between dynamical variables. The interaction patterns considered here are found in a great variety of systems, ranging from autocatalytic reactions and activator-inhibitor systems to influential models of nerve conduction, glycolysis, cell signaling and circadian rhythms.

  9. Multiscale image contrast amplification (MUSICA)

    NASA Astrophysics Data System (ADS)

    Vuylsteke, Pieter; Schoeters, Emile P.

    1994-05-01

    This article presents a novel approach to the problem of detail contrast enhancement, based on multiresolution representation of the original image. The image is decomposed into a weighted sum of smooth, localized, 2D basis functions at multiple scales. Each transform coefficient represents the amount of local detail at some specific scale and at a specific position in the image. Detail contrast is enhanced by non-linear amplification of the transform coefficients. An inverse transform is then applied to the modified coefficients. This yields a uniformly contrast- enhanced image without artefacts. The MUSICA-algorithm is being applied routinely to computed radiography images of chest, skull, spine, shoulder, pelvis, extremities, and abdomen examinations, with excellent acceptance. It is useful for a wide range of applications in the medical, graphical, and industrial area.

  10. Third Sound Amplification and Detailed Balance

    SciTech Connect

    Eddinger, J. D.; Ellis, F. M.

    2006-09-07

    Condensation of atoms from the vapor into a third sound resonance is expected to be capable of acoustic amplification. This results from normal to superfluid conversion that coherently accommodates atoms into the third sound velocity field. Consideration of third sound in light of the equilibrium detailed balance between vapor particles and the superfluid film provides further evidence that acoustic amplification is attainable.

  11. Influence of interfacial Dzyaloshinskii-Moriya interaction on the parametric amplification of spin waves

    SciTech Connect

    Verba, Roman; Tiberkevich, Vasil; Slavin, Andrei

    2015-09-14

    The influence of the interfacial Dzyaloshinskii-Moriya interaction (IDMI) on the parametric amplification of spin waves propagating in ultrathin ferromagnetic film is considered theoretically. It is shown that the IDMI changes the relation between the group velocities of the signal and idler spin waves in a parametric amplifier, which may result in the complete vanishing of the reversed idler wave. In the optimized case, the idler spin wave does not propagate from the pumping region at all, which increases the efficiency of the amplification of the signal wave and suppresses the spurious impact of the idler waves on neighboring spin-wave processing devices.

  12. Nondegenerate Parametric Amplification in Superlattices and the Limits of Strong and Weak Dissipation

    NASA Astrophysics Data System (ADS)

    Hyart, Timo; Alekseev, Kirill N.

    We develop a semiclassical theory of the nondegenerate parametric amplification in a single miniband of superlattice. We present the formulas describing absorption and gain of signal and idler fields in superlattice and analyze the limiting cases of strong and weak dissipation. We show how the well-known Manley-Rowe relations arise in the tight-binding lattice in the weak dissipation limit. Our results can be applied to an amplification of THz signals in semiconductor superlattices and a control of nonlinear transport of cold atoms in optical lattices.

  13. Nondegenerate Parametric Amplification in Superlattices and the Limits of Strong and Weak Dissipation

    NASA Astrophysics Data System (ADS)

    Hyart, Timo; Alekseev, Kirill N.

    2010-12-01

    We develop a semiclassical theory of the nondegenerate parametric amplification in a single miniband of superlattice. We present the formulas describing absorption and gain of signal and idler fields in superlattice and analyze the limiting cases of strong and weak dissipation. We show how the well-known Manley-Rowe relations arise in the tight-binding lattice in the weak dissipation limit. Our results can be applied to an amplification of THz signals in semiconductor superlattices and a control of nonlinear transport of cold atoms in optical lattices.

  14. Isothermal solid-phase amplification system for detection of Yersinia pestis.

    PubMed

    Mayboroda, Olena; Gonzalez Benito, Angel; Sabaté del Rio, Jonathan; Svobodova, Marketa; Julich, Sandra; Tomaso, Herbert; O'Sullivan, Ciara K; Katakis, Ioanis

    2016-01-01

    DNA amplification is required for most molecular diagnostic applications, but conventional polymerase chain reaction (PCR) has disadvantages for field testing. Isothermal amplification techniques are being developed to respond to this problem. One of them is the recombinase polymerase amplification (RPA) that operates at isothermal conditions without sacrificing specificity and sensitivity in easy-to-use formats. In this work, RPA was used for the optical detection of solid-phase amplification of the potential biowarfare agent Yersinia pestis. Thiolated forward primers were immobilized on the surface of maleimide-activated microtitre plates for the quantitative detection of synthetic and genomic DNA, with elongation occurring only in the presence of the specific template DNA and solution phase reverse primers. Quantitative detection was achieved via the use of biotinylated reverse primers and post-amplification addition of streptavidin-HRP conjugate. The overall time of amplification and detection was less than 1 h at a constant temperature of 37 °C. Single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA) sequences were detected, achieving detection limits of 4.04*10(-13) and 3.14*10(-16) M, respectively. The system demonstrated high specificity with negligible responses to non-specific targets.

  15. Focal amplification and oncogene dependency of GAB2 in breast cancer.

    PubMed

    Bocanegra, M; Bergamaschi, A; Kim, Y H; Miller, M A; Rajput, A B; Kao, J; Langerød, A; Han, W; Noh, D-Y; Jeffrey, S S; Huntsman, D G; Børresen-Dale, A-L; Pollack, J R

    2010-02-04

    DNA amplifications in breast cancer are frequent on chromosome 11q, in which multiple driver oncogenes likely reside in addition to cyclin D1 (CCND1). One such candidate, the scaffolding adapter protein, GRB2-associated binding protein 2 (GAB2), functions in ErbB signaling and was recently shown to enhance mammary epithelial cell proliferation, and metastasis of ERBB2 (HER2/neu)-driven murine breast cancer. However, the amplification status and function of GAB2 in the context of amplification remain undefined. In this study, by genomic profiling of 172 breast tumors, and fluorescence in situ hybridization validation in an independent set of 210 scorable cases, we observed focal amplification spanning GAB2 (11q14.1) independent of CCND1 (11q13.2) amplification, consistent with a driver role. Further, small interfering RNA (siRNA)-mediated knockdown of GAB2 in breast cancer lines (SUM52, SUM44PE and MDA468) with GAB2 amplification revealed a dependency on GAB2 for cell proliferation, cell-cycle progression, survival and invasion, likely mediated through altered phosphatidylinositol 3-kinase (PI3K) and mitogen-activated protein kinase (MAPK) signaling. GAB2 knockdown also reduced proliferation and survival in a cell line (BT474) with ERBB2 amplification, consistent with the possibility that GAB2 can function downstream of ERBB2. Our studies implicate focal amplification of GAB2 in breast carcinogenesis, and underscore an oncogenic role of scaffolding adapter proteins, and a potential new point of therapeutic intervention.

  16. Technique for extending the range of a signal measuring circuit

    DOEpatents

    Chaprnka, Anthony G.; Sun, Shan C.; Vercellotti, Leonard C.

    1978-01-01

    An input signal supplied to a signal measuring circuit is either amplified or attenuated as necessary to establish the magnitude of the input signal within the defined dynamic range of the measuring circuit and the output signal developed by the measuring circuit is subsequently readjusted through amplification or attenuation to develop an output signal which corresponds to the magnitude of the initial input signal.

  17. Enhanced Amplification and Fan-Out Operation in an All-Magnetic Transistor.

    PubMed

    Barman, Saswati; Saha, Susmita; Mondal, Sucheta; Kumar, Dheeraj; Barman, Anjan

    2016-09-14

    Development of all-magnetic transistor with favorable properties is an important step towards a new paradigm of all-magnetic computation. Recently, we showed such possibility in a Magnetic Vortex Transistor (MVT). Here, we demonstrate enhanced amplification in MVT achieved by introducing geometrical asymmetry in a three vortex sequence. The resulting asymmetry in core to core distance in the three vortex sequence led to enhanced amplification of the MVT output. A cascade of antivortices travelling in different trajectories including a nearly elliptical trajectory through the dynamic stray field is found to be responsible for this amplification. This asymmetric vortex transistor is further used for a successful fan-out operation, which gives large and nearly equal gains in two output branches. This large amplification in magnetic vortex gyration in magnetic vortex transistor is proposed to be maintained for a network of vortex transistor. The above observations promote the magnetic vortex transistors to be used in complex circuits and logic operations.

  18. Enhanced Amplification and Fan-Out Operation in an All-Magnetic Transistor

    PubMed Central

    Barman, Saswati; Saha, Susmita; Mondal, Sucheta; Kumar, Dheeraj; Barman, Anjan

    2016-01-01

    Development of all-magnetic transistor with favorable properties is an important step towards a new paradigm of all-magnetic computation. Recently, we showed such possibility in a Magnetic Vortex Transistor (MVT). Here, we demonstrate enhanced amplification in MVT achieved by introducing geometrical asymmetry in a three vortex sequence. The resulting asymmetry in core to core distance in the three vortex sequence led to enhanced amplification of the MVT output. A cascade of antivortices travelling in different trajectories including a nearly elliptical trajectory through the dynamic stray field is found to be responsible for this amplification. This asymmetric vortex transistor is further used for a successful fan-out operation, which gives large and nearly equal gains in two output branches. This large amplification in magnetic vortex gyration in magnetic vortex transistor is proposed to be maintained for a network of vortex transistor. The above observations promote the magnetic vortex transistors to be used in complex circuits and logic operations. PMID:27624662

  19. KASER: Knowledge Amplification by Structured Expert Randomization.

    PubMed

    Rubin, Stuart H; Murthy, S N Jayaram; Smith, Michael H; Trajković, Ljiljana

    2004-12-01

    In this paper and attached video, we present a third-generation expert system named Knowledge Amplification by Structured Expert Randomization (KASER) for which a patent has been filed by the U.S. Navy's SPAWAR Systems Center, San Diego, CA (SSC SD). KASER is a creative expert system. It is capable of deductive, inductive, and mixed derivations. Its qualitative creativity is realized by using a tree-search mechanism. The system achieves creative reasoning by using a declarative representation of knowledge consisting of object trees and inheritance. KASER computes with words and phrases. It possesses a capability for metaphor-based explanations. This capability is useful in explaining its creative suggestions and serves to augment the capabilities provided by the explanation subsystems of conventional expert systems. KASER also exhibits an accelerated capability to learn. However, this capability depends on the particulars of the selected application domain. For example, application domains such as the game of chess exhibit a high degree of geometric symmetry. Conversely, application domains such as the game of craps played with two dice exhibit no predictable pattern, unless the dice are loaded. More generally, we say that domains whose informative content can be compressed to a significant degree without loss (or with relatively little loss) are symmetric. Incompressible domains are said to be asymmetric or random. The measure of symmetry plus the measure of randomness must always sum to unity.

  20. Directional Amplification with a Josephson Circuit

    NASA Astrophysics Data System (ADS)

    Abdo, Baleegh; Sliwa, Katrina; Frunzio, Luigi; Devoret, Michel

    2013-07-01

    Nonreciprocal devices perform crucial functions in many low-noise quantum measurements, usually by exploiting magnetic effects. In the proof-of-principle device presented here, on the other hand, two on-chip coupled Josephson parametric converters (JPCs) achieve directionality by exploiting the nonreciprocal phase response of the JPC in the transmission-gain mode. The nonreciprocity of the device is controlled in situ by varying the amplitude and phase difference of two independent microwave pump tones feeding the system. At the desired working point and for a signal frequency of 8.453 GHz, the device achieves a forward power gain of 15 dB within a dynamical bandwidth of 9 MHz, a reverse gain of -6dB, and suppression of the reflected signal by 8 dB. We also find that the amplifier adds a noise equivalent to less than 1.5 photons at the signal frequency (referred back to the input). It can process up to 3 photons at the signal frequency per inverse dynamical bandwidth. With a directional amplifier operating along the principles of this device, qubit and readout preamplifier could be integrated on the same chip.

  1. High-Level Clonal FGFR Amplification and Response to FGFR Inhibition in a Translational Clinical Trial

    PubMed Central

    Babina, Irina S.; Herrera-Abreu, Maria Teresa; Tarazona, Noelia; Peckitt, Clare; Kilgour, Elaine; Smith, Neil R.; Geh, Catherine; Rooney, Claire; Cutts, Ros; Campbell, James; Ning, Jian; Fenwick, Kerry; Swain, Amanda; Brown, Gina; Chua, Sue; Thomas, Anne; Johnston, Stephen R.D.; Ajaz, Mazhar; Sumpter, Katherine; Gillbanks, Angela; Watkins, David; Chau, Ian; Popat, Sanjay; Cunningham, David; Turner, Nicholas C.

    2017-01-01

    FGFR1 and FGFR2 are amplified in many tumor types, yet what determines response to FGFR inhibition in amplified cancers is unknown. In a translational clinical trial, we show that gastric cancers with high-level clonal FGFR2 amplification have a high response rate to the selective FGFR inhibitor AZD4547, whereas cancers with subclonal or low-level amplification did not respond. Using cell lines and patient-derived xenograft models, we show that high-level FGFR2 amplification initiates a distinct oncogene addiction phenotype, characterized by FGFR2-mediated transactivation of alternative receptor kinases, bringing PI3K/mTOR signaling under FGFR control. Signaling in low-level FGFR1-amplified cancers is more restricted to MAPK signaling, limiting sensitivity to FGFR inhibition. Finally, we show that circulating tumor DNA screening can identify high-level clonally amplified cancers. Our data provide a mechanistic understanding of the distinct pattern of oncogene addiction seen in highly amplified cancers and demonstrate the importance of clonality in predicting response to targeted therapy. Significance Robust single-agent response to FGFR inhibition is seen only in high-level FGFR-amplified cancers, with copy-number level dictating response to FGFR inhibition in vitro, in vivo, and in the clinic. High-level amplification of FGFR2 is relatively rare in gastric and breast cancers, and we show that screening for amplification in circulating tumor DNA may present a viable strategy to screen patients. PMID:27179038

  2. Digital selection and analogue amplification coexist in a cortex-inspired silicon circuit

    NASA Astrophysics Data System (ADS)

    Hahnloser, Richard H. R.; Sarpeshkar, Rahul; Mahowald, Misha A.; Douglas, Rodney J.; Seung, H. Sebastian

    2000-06-01

    Digital circuits such as the flip-flop use feedback to achieve multi-stability and nonlinearity to restore signals to logical levels, for example 0 and 1. Analogue feedback circuits are generally designed to operate linearly, so that signals are over a range, and the response is unique. By contrast, the response of cortical circuits to sensory stimulation can be both multistable and graded. We propose that the neocortex combines digital selection of an active set of neurons with analogue response by dynamically varying the positive feedback inherent in its recurrent connections. Strong positive feedback causes differential instabilities that drive the selection of a set of active neurons under the constraints embedded in the synaptic weights. Once selected, the active neurons generate weaker, stable feedback that provides analogue amplification of the input. Here we present our model of cortical processing as an electronic circuit that emulates this hybrid operation, and so is able to perform computations that are similar to stimulus selection, gain modulation and spatiotemporal pattern generation in the neocortex.

  3. Numerical investigation of output beam quality in efficient broadband optical parametric chirped pulse amplification

    NASA Astrophysics Data System (ADS)

    Liu, Xiao-Di; Xu, Lu; Liang, Xiao-Yan

    2017-01-01

    We theoretically analyzed output beam quality of broad bandwidth non-collinear optical parametric chirped pulse amplification (NOPCPA) in LiB3O5 (LBO) centered at 800 nm. With a three-dimensional numerical model, the influence of the pump intensity, pump and signal spatial modulations, and the walk-off effect on the OPCPA output beam quality are presented, together with conversion efficiency and the gain spectrum. The pump modulation is a dominant factor that affects the output beam quality. Comparatively, the influence of signal modulation is insignificant. For a low-energy system with small beam sizes, walk-off effect has to be considered. Pump modulation and walk-off effect lead to asymmetric output beam profile with increased modulation. A special pump modulation type is found to optimize output beam quality and efficiency. For a high-energy system with large beam sizes, the walk-off effect can be neglected, certain back conversion is beneficial to reduce the output modulation. A trade-off must be made between the output beam quality and the conversion efficiency, especially when the pump modulation is large since. A relatively high conversion efficiency and a low output modulation are both achievable by controlling the pump modulation and intensity.

  4. Ligation with nucleic acid sequence-based amplification.

    PubMed

    Ong, Carmichael; Tai, Warren; Sarma, Aartik; Opal, Steven M; Artenstein, Andrew W; Tripathi, Anubhav

    2012-01-01

    This work presents a novel method for detecting nucleic acid targets using a ligation step along with an isothermal, exponential amplification step. We use an engineered ssDNA with two variable regions on the ends, allowing us to design the probe for optimal reaction kinetics and primer binding. This two-part probe is ligated by T4 DNA Ligase only when both parts bind adjacently to the target. The assay demonstrates that the expected 72-nt RNA product appears only when the synthetic target, T4 ligase, and both probe fragments are present during the ligation step. An extraneous 38-nt RNA product also appears due to linear amplification of unligated probe (P3), but its presence does not cause a false-positive result. In addition, 40 mmol/L KCl in the final amplification mix was found to be optimal. It was also found that increasing P5 in excess of P3 helped with ligation and reduced the extraneous 38-nt RNA product. The assay was also tested with a single nucleotide polymorphism target, changing one base at the ligation site. The assay was able to yield a negative signal despite only a single-base change. Finally, using P3 and P5 with longer binding sites results in increased overall sensitivity of the reaction, showing that increasing ligation efficiency can improve the assay overall. We believe that this method can be used effectively for a number of diagnostic assays.

  5. Highly efficient cascaded amplification using Pr(3+)-doped mid-infrared chalcogenide fiber amplifiers.

    PubMed

    Hu, Jonathan; Menyuk, Curtis R; Wei, Chengli; Brandon Shaw, L; Sanghera, Jasbinder S; Aggarwal, Ishwar D

    2015-08-15

    We computationally investigate cascaded amplification in a three-level mid-infrared (IR) Pr(3+)-doped chalcogenide fiber amplifier. The overlap of the cross-sections in the transitions (3)H(6)→(3)H(5) and (3)H(5)→(3)H(4) enable both transitions to simultaneously amplify a single wavelength in the range between 4.25 μm and 4.55 μm. High gain and low noise are achieved simultaneously if the signal is at 4.5 μm. We show that 45% of pump power that is injected at 2 μm can be shifted to 4.5 μm. The efficiency of using a mid-IR fiber amplifier is higher than what can be achieved by using mid-IR supercontinuum generation, which has been estimated at 25%. This mid-IR fiber amplifier can be used in conjunction with quantum cascade lasers to obtain a tunable, high-power mid-IR source.

  6. Offset-Free Gigahertz Midinfrared Frequency Comb Based on Optical Parametric Amplification in a Periodically Poled Lithium Niobate Waveguide

    NASA Astrophysics Data System (ADS)

    Mayer, A. S.; Phillips, C. R.; Langrock, C.; Klenner, A.; Johnson, A. R.; Luke, K.; Okawachi, Y.; Lipson, M.; Gaeta, A. L.; Fejer, M. M.; Keller, U.

    2016-11-01

    We report the generation of an optical-frequency comb in the midinfrared region with 1-GHz comb-line spacing and no offset with respect to absolute-zero frequency. This comb is tunable from 2.5 to 4.2 μ m and covers a critical spectral region for important environmental and industrial applications, such as molecular spectroscopy of trace gases. We obtain such a comb using a highly efficient frequency conversion of a near-infrared frequency comb. The latter is based on a compact diode-pumped semiconductor saturable absorber mirror-mode-locked ytterbium-doped calcium-aluminum gadolynate (Yb:CALGO) laser operating at 1 μ m . The frequency-conversion process is based on optical parametric amplification (OPA) in a periodically poled lithium niobate (PPLN) chip containing buried waveguides fabricated by reverse proton exchange. The laser with a repetition rate of 1 GHz is the only active element of the system. It provides the pump pulses for the OPA process as well as seed photons in the range of 1.4 - 1.8 μ m via supercontinuum generation in a silicon-nitride (Si3 N4 ) waveguide. Both the PPLN and Si3 N4 waveguides represent particularly suitable platforms for low-energy nonlinear interactions; they allow for mid-IR comb powers per comb line at the microwatt level and signal amplification levels up to 35 dB, with 2 orders of magnitude less pulse energy than reported in OPA systems using bulk devices. Based on numerical simulations, we explain how high amplification can be achieved at low energy using the interplay between mode confinement and a favorable group-velocity mismatch configuration where the mid-IR pulse moves at the same velocity as the pump.

  7. A novel, sensitive and label-free loop-mediated isothermal amplification detection method for nucleic acids using luminophore dyes.

    PubMed

    Roy, Sharmili; Wei, Sim Xiao; Ying, Jean Liew Zhi; Safavieh, Mohammadali; Ahmed, Minhaz Uddin

    2016-12-15

    Electrochemiluminescence (ECL) has been widely rendered for nucleic acid testing. Here, we integrate loop-mediated isothermal amplification (LAMP) with ECL technique for DNA detection and quantification. The target LAMP DNA bound electrostatically with [Ru(bpy)3](+2) on the carbon electrode surface, and an ECL reaction was triggered by tripropylamine (TPrA) to yield luminescence. We illustrated this method as a new and highly sensitive strategy for the detection of sequence-specific DNA from different meat species at picogram levels. The proposed strategy renders the signal amplification capacities of TPrA and combines LAMP with inherently high sensitivity of the ECL technique, to facilitate the detection of low quantities of DNA. By leveraging this technique, target DNA of Sus scrofa (pork) meat was detected as low as 1pg/µL (3.43×10(-1)copies/µL). In addition, the proposed technique was applied for detection of Bacillus subtilis DNA samples and detection limit of 10pg/µL (2.2×10(3)copies/µL) was achieved. The advantages of being isothermal, sensitive and robust with ability for multiplex detection of bio-analytes makes this method a facile and appealing sensing modality in hand-held devices to be used at the point-of-care (POC).

  8. Precision phase estimation based on weak-value amplification

    NASA Astrophysics Data System (ADS)

    Qiu, Xiaodong; Xie, Linguo; Liu, Xiong; Luo, Lan; Li, Zhaoxue; Zhang, Zhiyou; Du, Jinglei

    2017-02-01

    In this letter, we propose a precision method for phase estimation based on the weak-value amplification (WVA) technique using a monochromatic light source. The anomalous WVA significantly suppresses the technical noise with respect to the intensity difference signal induced by the phase delay when the post-selection procedure comes into play. The phase measured precision of this method is proportional to the weak-value of a polarization operator in the experimental range. Our results compete well with the wide spectrum light phase weak measurements and outperform the standard homodyne phase detection technique.

  9. Demonstrating a Multi-drug Resistant Mycobacterium tuberculosis Amplification Microarray

    PubMed Central

    Linger, Yvonne; Kukhtin, Alexander; Golova, Julia; Perov, Alexander; Qu, Peter; Knickerbocker, Christopher; Cooney, Christopher G.; Chandler, Darrell P.

    2014-01-01

    Simplifying microarray workflow is a necessary first step for creating MDR-TB microarray-based diagnostics that can be routinely used in lower-resource environments. An amplification microarray combines asymmetric PCR amplification, target size selection, target labeling, and microarray hybridization within a single solution and into a single microfluidic chamber. A batch processing method is demonstrated with a 9-plex asymmetric master mix and low-density gel element microarray for genotyping multi-drug resistant Mycobacterium tuberculosis (MDR-TB). The protocol described here can be completed in 6 hr and provide correct genotyping with at least 1,000 cell equivalents of genomic DNA. Incorporating on-chip wash steps is feasible, which will result in an entirely closed amplicon method and system. The extent of multiplexing with an amplification microarray is ultimately constrained by the number of primer pairs that can be combined into a single master mix and still achieve desired sensitivity and specificity performance metrics, rather than the number of probes that are immobilized on the array. Likewise, the total analysis time can be shortened or lengthened depending on the specific intended use, research question, and desired limits of detection. Nevertheless, the general approach significantly streamlines microarray workflow for the end user by reducing the number of manually intensive and time-consuming processing steps, and provides a simplified biochemical and microfluidic path for translating microarray-based diagnostics into routine clinical practice. PMID:24796567

  10. Demonstrating a multi-drug resistant Mycobacterium tuberculosis amplification microarray.

    PubMed

    Linger, Yvonne; Kukhtin, Alexander; Golova, Julia; Perov, Alexander; Qu, Peter; Knickerbocker, Christopher; Cooney, Christopher G; Chandler, Darrell P

    2014-04-25

    Simplifying microarray workflow is a necessary first step for creating MDR-TB microarray-based diagnostics that can be routinely used in lower-resource environments. An amplification microarray combines asymmetric PCR amplification, target size selection, target labeling, and microarray hybridization within a single solution and into a single microfluidic chamber. A batch processing method is demonstrated with a 9-plex asymmetric master mix and low-density gel element microarray for genotyping multi-drug resistant Mycobacterium tuberculosis (MDR-TB). The protocol described here can be completed in 6 hr and provide correct genotyping with at least 1,000 cell equivalents of genomic DNA. Incorporating on-chip wash steps is feasible, which will result in an entirely closed amplicon method and system. The extent of multiplexing with an amplification microarray is ultimately constrained by the number of primer pairs that can be combined into a single master mix and still achieve desired sensitivity and specificity performance metrics, rather than the number of probes that are immobilized on the array. Likewise, the total analysis time can be shortened or lengthened depending on the specific intended use, research question, and desired limits of detection. Nevertheless, the general approach significantly streamlines microarray workflow for the end user by reducing the number of manually intensive and time-consuming processing steps, and provides a simplified biochemical and microfluidic path for translating microarray-based diagnostics into routine clinical practice.

  11. High field side lower hybrid launch leads to wave amplification on alpha particles

    NASA Astrophysics Data System (ADS)

    Ochs, Ian; Bertelli, Nicola; Fisch, Nathaniel

    2015-11-01

    Although lower hybrid waves have been shown to be effective in driving plasma current in present-day tokamaks, they are predicted to strongly interact with the energetic α particles born from fusion reactions in eventual tokamak reactors. However, in the presence of the expected steep α particle birth gradient, this interaction can produce wave amplification rather than wave damping. Here, we identify the flexibilities in achieving this amplification effect through a consideration of symmetries in the channeling interaction, in the wave propagation, and in the tokamak field configuration. Interestingly, for current drive that supports the poloidal magnetic field, we find that wave amplification through α channeling is fundamentally coupled to the elusive | kl | upshift. In so doing, we show that wave launch from the tokamak high-field side is favorable both for α-channeling and for achieving the | kl | upshift. We then present a simple linear model to calculate the required radial gradients to achieve amplification. Combining this model with ray tracing simulations, we demonstrate the potential for substantial wave amplification in a regime consistent with a hot-ion-mode fusion reactor.

  12. Nanoparticle-bridge assay for amplification-free electrical detection of oligonucleotides

    NASA Astrophysics Data System (ADS)

    Teimouri, Manouchehr

    The aim of this research is to investigate a highly sensitive, fast, inexpensive, and field-applicable amplification-free nanoparticle-based oligonucleotide detection method which does not rely on any enzymatic or signal amplification process. In this approach, target oligonucleotide strands are detected through the formation of nanoparticle satellites which make an electrical path between two electrodes. This method enables an extremely sensitive oligonucleotide detection because even a few oligonucleotide strands can form a single nanoparticle satellite which can solely generates an electrical output signal. Results showed that this oligonucleotide detection method can detect oligonucleotide single strands at concentrations as low as 50 femtomolar without any amplification process. This detection method can be implemented in many fields such as biodefense, food safety, clinical research, and forensics.

  13. A cascade amplification strategy based on rolling circle amplification and hydroxylamine amplified gold nanoparticles enables chemiluminescence detection of adenosine triphosphate.

    PubMed

    Wang, Ping; Zhang, Tonghuan; Yang, Taoyi; Jin, Nan; Zhao, Yanjun; Fan, Aiping

    2014-08-07

    A highly sensitive and selective chemiluminescent (CL) biosensor for adenosine triphosphate (ATP) was developed by taking advantage of the ATP-dependent enzymatic reaction (ATP-DER), the powerful signal amplification capability of rolling circle amplification (RCA), and hydroxylamine-amplified gold nanoparticles (Au NPs). The strategy relies on the ability of ATP, a cofactor of T4 DNA ligase, to trigger the ligation-RCA reaction. In the presence of ATP, the T4 DNA ligase catalyzes the ligation reaction between the two ends of the padlock probe, producing a closed circular DNA template that initiates the RCA reaction with phi29 DNA polymerase and dNTP. Therein, many complementary copies of the circular template can be generated. The ATP-DER is eventually converted into a detectable CL signal after a series of processes, including gold probe hybridization, hydroxylamine amplification, and oxidative gold metal dissolution coupled with a simple and sensitive luminol CL reaction. The CL signal is directly proportional to the ATP level. The results showed that the detection limit of the assay is 100 pM of ATP, which compares favorably with those of other ATP detection techniques. In addition, by taking advantage of ATP-DER, the proposed CL sensing system exhibits extraordinary specificity towards ATP and could distinguish the target molecule ATP from its analogues. The proposed method provides a new and versatile platform for the design of novel DNA ligation reaction-based CL sensing systems for other cofactors. This novel ATP-DER based CL sensing system may find wide applications in clinical diagnosis as well as in environmental and biomedical fields.

  14. High-power Femtosecond Optical Parametric Amplification at 1 kHz in BiB(3)O(6) pumped at 800 nm.

    PubMed

    Petrov, Valentin; Noack, Frank; Tzankov, Pancho; Ghotbi, Masood; Ebrahim-Zadeh, Majid; Nikolov, Ivailo; Buchvarov, Ivan

    2007-01-22

    Substantial power scaling of a travelling-wave femtosecond optical parametric amplifier, pumped near 800 nm by a 1 kHz Ti:sapphire laser amplifier, is demonstrated using monoclinic BiB(3)O(6) in a two stage scheme with continuum seeding. Total energy output (signal plus idler) exceeding 1 mJ is achieved, corresponding to an intrinsic conversion efficiency of approximately 32% for the second stage. The tunability extends from 1.1 to 2.9 microm. The high parametric gain and broad amplification bandwidth of this crystal allowed the maintenance of the pump pulse duration, leading to pulse lengths less than 140 fs, both for the signal and idler pulses, even at such high output levels.

  15. Preferential Amplification of Pathogenic Sequences.

    PubMed

    Ge, Fang; Parker, Jayme; Chul Choi, Sang; Layer, Mark; Ross, Katherine; Jilly, Bernard; Chen, Jack

    2015-06-11

    The application of next generation sequencing (NGS) technology in the diagnosis of human pathogens is hindered by the fact that pathogenic sequences, especially viral, are often scarce in human clinical specimens. This known disproportion leads to the requirement of subsequent deep sequencing and extensive bioinformatics analysis. Here we report a method we called "Preferential Amplification of Pathogenic Sequences (PATHseq)" that can be used to greatly enrich pathogenic sequences. Using a computer program, we developed 8-, 9-, and 10-mer oligonucleotides called "non-human primers" that do not match the most abundant human transcripts, but instead selectively match transcripts of human pathogens. Instead of using random primers in the construction of cDNA libraries, the PATHseq method recruits these short non-human primers, which in turn, preferentially amplifies non-human, presumably pathogenic sequences. Using this method, we were able to enrich pathogenic sequences up to 200-fold in the final sequencing library. This method does not require prior knowledge of the pathogen or assumption of the infection; therefore, it provides a fast and sequence-independent approach for detection and identification of human viruses and other pathogens. The PATHseq method, coupled with NGS technology, can be broadly used in identification of known human pathogens and discovery of new pathogens.

  16. Toughness amplification in natural composites

    NASA Astrophysics Data System (ADS)

    Barthelat, Francois; Rabiei, Reza

    2011-04-01

    Natural structural materials such as bone and seashells are made of relatively weak building blocks, yet they exhibit remarkable combinations of stiffness, strength and toughness. This performance can be largely explained by their "staggered microstructure": stiff inclusions of high aspect ratio are laid parallel to each other with some overlap, and bonded by a softer matrix. While stiffness and strength are now well understood for staggered composites, the mechanisms involved in fracture are still largely unknown. This is a significant lack since the amplification of toughness with respect to their components is by far the most impressive feature in natural staggered composites such as nacre or bone. Here a model capturing the salient mechanisms involved in the cracking of a staggered structure is presented. We show that the pullout of inclusions and large process zones lead to tremendous toughness by far exceeding that of individual components. The model also suggests that a material like nacre cannot reach steady state cracking, with the implication that the toughness increases indefinitely with crack advance. These findings agree well with existing fracture data, and for the first time relate microstructural parameters with overall toughness. These insights will prove useful in the design of biomimetic materials, and provide clues on how bone fractures at the nano and microscales.

  17. Solid-phase PCR for rapid multiplex detection of Salmonella spp. at the subspecies level, with amplification efficiency comparable to conventional PCR.

    PubMed

    Chin, Wai Hoe; Sun, Yi; Høgberg, Jonas; Hung, Tran Quang; Wolff, Anders; Bang, Dang Duong

    2017-04-01

    Solid-phase PCR (SP-PCR) has attracted considerable interest in different research fields since it allows parallel DNA amplification on the surface of a solid substrate. However, the applications of SP-PCR have been hampered by the low efficiency of the solid-phase amplification. In order to increase the yield of the solid-phase amplification, we studied various parameters including the length, the density, as well as the annealing position of the solid support primer. A dramatic increase in the signal-to-noise (S/N) ratio was observed when increasing the length of solid support primers from 45 to 80 bp. The density of the primer on the surface was found to be important for the S/N ratio of the SP-PCR, and the optimal S/N was obtained with a density of 1.49 × 10(11) molecules/mm(2). In addition, the use of solid support primers with a short overhang at the 5' end would help improve the S/N ratio of the SP-PCR. With optimized conditions, SP-PCR can achieve amplification efficiency comparable to conventional PCR, with a limit of detection of 1.5 copies/μl (37.5 copies/reaction). These improvements will pave the way for wider applications of SP-PCR in various fields such as clinical diagnosis, high-throughput DNA sequencing, and single-nucleotide polymorphism analysis. Graphical abstract Schematic representation of solid-phase PCR.

  18. Limits of Femtosecond Fiber Amplification by Parabolic Pre-Shaping

    PubMed Central

    Fu, Walter; Tang, Yuxing; McComb, Timothy S.; Lowder, Tyson L.; Wise, Frank W.

    2017-01-01

    We explore parabolic pre-shaping as a means of generating and amplifying ultrashort pulses. We develop a theoretical framework for modeling the technique and use its conclusions to design a femtosecond fiber amplifier. Starting from 9 ps pulses, we obtain 4.3 μJ, nearly transform-limited pulses 275 fs in duration, simultaneously achieving over 40 dB gain and 33-fold compression. Finally, we show that this amplification scheme is limited by Raman scattering, and outline a method by which the pulse duration and energy may be further improved and tailored for a given application. PMID:28331242

  19. Electromagnetically induced transparency with amplification in superconducting circuits.

    PubMed

    Joo, Jaewoo; Bourassa, Jérôme; Blais, Alexandre; Sanders, Barry C

    2010-08-13

    We show that controlling relative phases of electromagnetic fields driving an atom with a Δ-configuration energy-level structure enables optical susceptibility to be engineered in novel ways. In particular, relative-phase control can yield electromagnetically induced transparency but with the benefit that the transparency window is sandwiched between an absorption and an amplification band rather than between two absorption bands in typical electromagnetically induced transparency. We show that this new phenomenon is achievable for a microwave field interacting with a fluxonium superconducting circuit.

  20. Earthquake ground motion amplification for surface waves

    NASA Astrophysics Data System (ADS)

    Bowden, Daniel C.; Tsai, Victor C.

    2017-01-01

    Surface waves from earthquakes are known to cause strong damage, especially for larger structures such as skyscrapers and bridges. However, common practice in characterizing seismic hazard at a specific site considers the effect of near-surface geology on only vertically propagating body waves. Here we show that surface waves have a unique and different frequency-dependent response to known geologic structure and that this amplification can be analytically calculated in a manner similar to current hazard practices. Applying this framework to amplification in the Los Angeles Basin, we find that peak ground accelerations for certain large regional earthquakes are underpredicted if surface waves are not properly accounted for and that the frequency of strongest ground motion amplification can be significantly different. Including surface-wave amplification in hazards calculations is therefore essential for accurate predictions of strong ground motion for future San Andreas Fault ruptures.

  1. AMPLIFICATION OF RIBOSOMAL RNA SEQUENCES - Book Chapter

    EPA Science Inventory

    This book chapter contains the following headings and subheadings: Introduction; Experimental Approach - Precautions, Template, Primers, Reaction Conditions, Enhancers, Post Amplification; Procedures - Template DNA, Basic PCR, Thermal Cycle Parameters, Enzyme Addition, Agarose Ge...

  2. Coupled isothermal polynucleotide amplification and translation system

    NASA Technical Reports Server (NTRS)

    Joyce, Gerald F. (Inventor)

    1998-01-01

    A cell-free system for polynucleotide amplification and translation is disclosed. Also disclosed are methods for using the system and a composition which allows the various components of the system to function under a common set of reaction conditions.

  3. Mode group specific amplification length in an asymmetric LPG assisted few-mode EDFA

    NASA Astrophysics Data System (ADS)

    Rastogi, Vipul; Gaur, Ankita; Aschieri, Pierre; Dussardier, Bernard

    2017-01-01

    This article presents a scheme for few-mode EDFA, which allows to choose independent amplification lengths for different mode groups. The EDF is a dual concentric core fiber, where the central core is connected to the line FMF and the ring core is doped with erbium to provide amplification. The modes of FMF are launched into the central core of the EDF, are converted into ring modes using LPG for amplification and then converted back into central core modes using another LPG. The distance between the LPGs determines the amplification length. The amplification length, can thus, be chosen for a given mode group. We demonstrate the working of this concept by choosing LP11 and LP21 mode groups of the FMF and show that a suitable choice of amplification lengths for the two mode groups can tailor the differential modal gain (DMG) to any desired value. We demonstrate achieving zero DMG among all the mode of LP11 and LP21 mode groups using this concept while having gain in excess of 20 dB. The study should be useful for optical fiber communication system employing space-division multiplexing (SDM).

  4. Amplification of a seed pumped by a chirped laser in the strong coupling Brillouin regime

    SciTech Connect

    Schluck, F.; Lehmann, G.; Spatschek, K. H.

    2015-09-15

    Seed amplification via Brillouin backscattering of a long pump laser-pulse is considered. The interaction takes place in the so called strong coupling regime. Pump chirping is applied to mitigate spontaneous Raman backscattering of the pump before interacting with the seed. The strong coupling regime facilitates stronger exponential growth and narrower seeds compared to the so called weak coupling regime, although in the latter the scaling with pump amplitude is stronger. Strong coupling is achieved when the pump laser amplitude exceeds a certain threshold. It is shown how the chirp influences both the linear as well as the nonlinear amplification process. First, linear amplification as well as the seed profiles are determined in dependence of the chirping rate. In contrast to the weak coupling situation, the evolution is not symmetric with respect to the sign of the chirping rate. In the nonlinear stage of the amplification, we find an intrinsic chirp of the seed pulse even for an un-chirped pump. We show that chirping the pump may have a strong influence on the shape of the seed in the nonlinear amplification phase. Also, the influence of pump chirp on the efficiency of Brillouin seed amplification is discussed.

  5. Improving the classroom listening skills of children with Down syndrome by using sound-field amplification.

    PubMed

    Bennetts, Lee K; Flynn, Mark C

    2002-03-01

    Many children with Down syndrome have fluctuating conductive hearing losses further reducing their speech, language and academic development. It is within the school environment where access to auditory information is crucial that many children with Down syndrome are especially disadvantaged. Conductive hearing impairment which is often fluctuating and undetected reduces the child's ability to extract the important information from the auditory signal. Unfortunately, the design and acoustics of the classroom leads to problems in extracting the speech signal through reduced speech intensity due to the increased distance of the student from the teacher in addition to masking from excessive background noise. One potential solution is the use of sound-field amplification which provides a uniform amplification to the teacher's voice through the use of a microphone and loudspeakers. This investigation examined the efficacy of sound-field amplification for 4 children with Down syndrome. Measures of speech perception were taken with and without the sound-field system and found that the children perceived significantly more speech in all conditions where the sound-field system was used (p < .0001). Importantly, listening performance with the sound-field system was not affected by reducing the signal-to-noise ratio through increasing the level of background noise. In summary, sound-field amplification provides improved access to the speech signal for children with Down syndrome and as a consequence leads to improved classroom success.

  6. In-field Raman amplification on coherent optical fiber links for frequency metrology.

    PubMed

    Clivati, C; Bolognini, G; Calonico, D; Faralli, S; Mura, A; Levi, F

    2015-04-20

    Distributed Raman amplification (DRA) is widely exploited for the transmission of broadband, modulated signals used in data links, but not yet in coherent optical links for frequency metrology, where the requirements are rather different. After preliminary tests on fiber spools, in this paper we deeper investigate Raman amplification on deployed in-field optical metrological links. We actually test a Doppler-stabilized optical link both on a 94 km-long metro-network implementation with multiplexed ITU data channels and on a 180 km-long dedicated fiber haul connecting two cities, where DRA is employed in combination with Erbium-doped fiber amplification (EDFA). The performance of DRA is detailed in both experiments, indicating that it does not introduce noticeable penalties for the metrological signal or for the ITU data channels. We hence show that Raman amplification of metrological signals can be compatible with a wavelength division multiplexing architecture and that it can be used as an alternative or in combination with dedicated bidirectional EDFAs. No deterioration is noticed in the coherence properties of the delivered signal, which attains frequency instability at the 10(-19) level in both cases. This study can be of interest also in view of the undergoing deployment of continental fiber networks for frequency metrology.

  7. Rolling circle amplification of metazoan mitochondrialgenomes

    SciTech Connect

    Simison, W. Brian; Lindberg, D.R.; Boore, J.L.

    2005-07-31

    Here we report the successful use of rolling circle amplification (RCA) for the amplification of complete metazoan mt genomes to make a product that is amenable to high-throughput genome sequencing techniques. The benefits of RCA over PCR are many and with further development and refinement of RCA, the sequencing of organellar genomics will require far less time and effort than current long PCR approaches.

  8. Amplification of chromosomal DNA in situ

    DOEpatents

    Christian, Allen T.; Coleman, Matthew A.; Tucker, James D.

    2002-01-01

    Amplification of chromosomal DNA in situ to increase the amount of DNA associated with a chromosome or chromosome region is described. The amplification of chromosomal DNA in situ provides for the synthesis of Fluorescence in situ Hybridization (FISH) painting probes from single dissected chromosome fragments, the production of cDNA libraries from low copy mRNAs and improved in Comparative Genomic Hybridization (CGH) procedures.

  9. Chemical amplification based on fluid partitioning

    DOEpatents

    Anderson, Brian L.; Colston, Jr., Billy W.; Elkin, Chris

    2006-05-09

    A system for nucleic acid amplification of a sample comprises partitioning the sample into partitioned sections and performing PCR on the partitioned sections of the sample. Another embodiment of the invention provides a system for nucleic acid amplification and detection of a sample comprising partitioning the sample into partitioned sections, performing PCR on the partitioned sections of the sample, and detecting and analyzing the partitioned sections of the sample.

  10. Amplification uncertainty relation for probabilistic amplifiers

    NASA Astrophysics Data System (ADS)

    Namiki, Ryo

    2015-09-01

    Traditionally, quantum amplification limit refers to the property of inevitable noise addition on canonical variables when the field amplitude of an unknown state is linearly transformed through a quantum channel. Recent theoretical studies have determined amplification limits for cases of probabilistic quantum channels or general quantum operations by specifying a set of input states or a state ensemble. However, it remains open how much excess noise on canonical variables is unavoidable and whether there exists a fundamental trade-off relation between the canonical pair in a general amplification process. In this paper we present an uncertainty-product form of amplification limits for general quantum operations by assuming an input ensemble of Gaussian-distributed coherent states. It can be derived as a straightforward consequence of canonical uncertainty relations and retrieves basic properties of the traditional amplification limit. In addition, our amplification limit turns out to give a physical limitation on probabilistic reduction of an Einstein-Podolsky-Rosen uncertainty. In this regard, we find a condition that probabilistic amplifiers can be regarded as local filtering operations to distill entanglement. This condition establishes a clear benchmark to verify an advantage of non-Gaussian operations beyond Gaussian operations with a feasible input set of coherent states and standard homodyne measurements.

  11. Onshore seismic amplifications due to bathymetric features

    NASA Astrophysics Data System (ADS)

    Rodríguez-Castellanos, A.; Carbajal-Romero, M.; Flores-Guzmán, N.; Olivera-Villaseñor, E.; Kryvko, A.

    2016-08-01

    We perform numerical calculations for onshore seismic amplifications, taking into consideration the effect of bathymetric features on the propagation of seismic movements. To this end, the boundary element method is applied. Boundary elements are employed to irradiate waves and, consequently, force densities can be obtained for each boundary element. From this assumption, Huygens’ principle is applied, and since the diffracted waves are built at the boundary from which they are radiated, this idea is equivalent to Somigliana’s representation theorem. The application of boundary conditions leads to a linear system being obtained (Fredholm integral equations). Several numerical models are analyzed, with the first one being used to verify the proposed formulation, and the others being used to estimate onshore seismic amplifications due to the presence of bathymetric features. The results obtained show that compressional waves (P-waves) generate onshore seismic amplifications that can vary from 1.2 to 5.2 times the amplitude of the incident wave. On the other hand, the shear waves (S-waves) can cause seismic amplifications of up to 4.0 times the incident wave. Furthermore, an important result is that in most cases the highest seismic amplifications from an offshore earthquake are located on the shoreline and not offshore, despite the seafloor configuration. Moreover, the influence of the incident angle of seismic waves on the seismic amplifications is highlighted.

  12. Molecular architectures for electrocatalytic amplification of oligonucleotide hybridization.

    PubMed

    Mir, Mònica; Alvarez, Marta; Azzaroni, Omar; Tiefenauer, Louis; Knoll, Wolfgang

    2008-09-01

    In this work, we describe a new platform suitable for electrocatalytic amplification of oligonucleotide hybridization based on the use of supramolecular bioconjugates incorporating ferrocene-labeled streptavidin. Our goals were aimed at designing a biosensing platform which could support highly reproducible and stable electrocatalytic amplification with maximum efficiency. The use of nonlabeled streptavidin as an underlying layer promotes a major improvement on the characteristics of the amplified electrochemical signal. In addition, the electrocatalytic current can be easily amplified by tuning the concentration of electron donor species in solution. Because of the fact that the redox labels are bioconjugated to the DNA strands, increasing the ionic strength does not lead to the loss of redox labels. More importantly, increasing the concentration of donors only involves the magnification of the signal without implying the permeation of donors (thus reducing the efficient electrocatalysis). This approach represents a major improvement on the use of electrocatalytically amplified DNA-sensing platforms, thus overcoming any possible limitation in connection with the reproducibility and reliability of this well-established method.

  13. Heat induces gene amplification in cancer cells

    SciTech Connect

    Yan, Bin; Ouyang, Ruoyun; Huang, Chenghui; Liu, Franklin; Neill, Daniel; Li, Chuanyuan; Dewhirst, Mark

    2012-10-26

    Highlights: Black-Right-Pointing-Pointer This study discovered that heat exposure (hyperthermia) results in gene amplification in cancer cells. Black-Right-Pointing-Pointer Hyperthermia induces DNA double strand breaks. Black-Right-Pointing-Pointer DNA double strand breaks are considered to be required for the initiation of gene amplification. Black-Right-Pointing-Pointer The underlying mechanism of heat-induced gene amplification is generation of DNA double strand breaks. -- Abstract: Background: Hyperthermia plays an important role in cancer therapy. However, as with radiation, it can cause DNA damage and therefore genetic instability. We studied whether hyperthermia can induce gene amplification in cancer cells and explored potential underlying molecular mechanisms. Materials and methods: (1) Hyperthermia: HCT116 colon cancer cells received water-submerged heating treatment at 42 or 44 Degree-Sign C for 30 min; (2) gene amplification assay using N-(phosphoacetyl)-L-aspartate (PALA) selection of cabamyl-P-synthetase, aspartate transcarbarmylase, dihydro-orotase (cad) gene amplified cells; (3) southern blotting for confirmation of increased cad gene copies in PALA-resistant cells; (4) {gamma}H2AX immunostaining to detect {gamma}H2AX foci as an indication for DNA double strand breaks. Results: (1) Heat exposure at 42 or 44 Degree-Sign C for 30 min induces gene amplification. The frequency of cad gene amplification increased by 2.8 and 6.5 folds respectively; (2) heat exposure at both 42 and 44 Degree-Sign C for 30 min induces DNA double strand breaks in HCT116 cells as shown by {gamma}H2AX immunostaining. Conclusion: This study shows that heat exposure can induce gene amplification in cancer cells, likely through the generation of DNA double strand breaks, which are believed to be required for the initiation of gene amplification. This process may be promoted by heat when cellular proteins that are responsible for checkpoints, DNA replication, DNA repair and

  14. Prospects of obtaining terawatt class infrared pulses using standard optical parametric amplification

    NASA Astrophysics Data System (ADS)

    Guo, Xiaoyang; Tokita, Shigeki; Tu, Xiaoniu; Zheng, Yanqing; Kawanaka, Junji

    2017-02-01

    We conceptually propose a standard optical parametric amplification system based on YCOB crystal to achieve terawatt (TW) class infrared (IR) pulses with 100 mJ level energy, which would be one order of magnitude more energetic and powerful than currently available IR pulses and suitable to generate high photon flux water window x-rays.

  15. Advanced unrepeatered systems using novel Raman amplification schemes

    NASA Astrophysics Data System (ADS)

    Chang, Do-il; Pelouch, Wayne; Burtsev, Sergey; Perrier, Philippe; Fevrier, Herve

    2015-01-01

    Unrepeatered transmission systems provide a cost-effective solution to transmit high capacity channels in submarine networks to communicate between coastal population centers or in terrestrial networks to connect remote areas where service access is difficult. The main goal of unrepeatered systems has traditionally been to achieve the longest reach, however, increasing traffic demands now require unrepeatered systems to support both longer reach and higher transport capacity. As a result, transmission rate of unrepeatered systems has quickly moved from 10 Gb/s to 40 Gb/s or 100 Gb/s. This paper reviews the key basic technologies, with a specific focus on Raman amplification, required for long-reach, high-capacity unrepeatered optical transmission systems. We will discuss novel Raman amplification schemes, enhanced remote optically pumped amplifiers (ROPA), ultra-low loss / large effective area fibers, and coherent transmission with advanced modulation format and high FEC coding gain. We will also report recent experimental demonstrations that show how these technologies have been combined to achieve industry's leading capacity and reach transmission.

  16. Holographic Storage in Electrooptic Crystals.: II. Beam COUPLING—LIGHT Amplification

    NASA Astrophysics Data System (ADS)

    Kukhtarev, N. V.; Markov, V. B.; Odulov, S. G.; Soskin, M. S.; Vinetskii, V. L.

    The theory of energy transfer between two coupled beams writing holograms in electrooptic crystals is developed. The gain is calculated for different processes causing holographic recording in the crystal. The amplifications of the light beam in the course of recording holograms in nominally pure reduced LiNbO3 crystals is investigated. The amplification of about 10 cm-1 is obtained for small signals. The gain is found to be independent of the total light intensity and intensity ratio of the writing beams and decreases linearly with the grating spacing.

  17. Ultrahigh contrast from a frequency-doubled chirped-pulse-amplification beamline.

    PubMed

    Hillier, David; Danson, Colin; Duffield, Stuart; Egan, David; Elsmere, Stephen; Girling, Mark; Harvey, Ewan; Hopps, Nicholas; Norman, Michael; Parker, Stefan; Treadwell, Paul; Winter, David; Bett, Thomas

    2013-06-20

    This paper describes frequency-doubled operation of a high-energy chirped-pulse-amplification beamline. Efficient type-I second-harmonic generation was achieved using a 3 mm thick 320 mm aperture KDP crystal. Shots were fired at a range of energies achieving more than 100 J in a subpicosecond, 527 nm laser pulse with a power contrast of 10(14).

  18. Label-Free Isothermal Amplification Assay for Specific and Highly Sensitive Colorimetric miRNA Detection

    PubMed Central

    2016-01-01

    We describe a new method for the detection of miRNA in biological samples. This technology is based on the isothermal nicking enzyme amplification reaction and subsequent hybridization of the amplification product with gold nanoparticles and magnetic microparticles (barcode system) to achieve naked-eye colorimetric detection. This platform was used to detect a specific miRNA (miRNA-10b) associated with breast cancer, and attomolar sensitivity was demonstrated. The assay was validated in cell culture lysates from breast cancer cells and in serum from a mouse model of breast cancer. PMID:27713932

  19. Temperature- and wavelength-insensitive parametric amplification enabled by noncollinear achromatic phase-matching

    PubMed Central

    Tang, Daolong; Ma, Jingui; Wang, Jing; Zhou, Bingjie; Xie, Guoqiang; Yuan, Peng; Zhu, Heyuan; Qian, Liejia

    2016-01-01

    Optical parametric chirped-pulse amplification (OPCPA) has been demonstrated to be a promising approach for pushing femtosecond pulses towards ultra-high peak powers. However, the future success of OPCPA strongly relies on the ability to manipulate its phase-matching (PM) configuration. When a high average power pump laser is involved, the thermal effects in nonlinear crystals induce phase-mismatch distortions that pose an inherent limitation on the conversion efficiency. Here, we demonstrate that the noncollinear configuration previously adopted for wavelength-insensitive PM can be employed for temperature-insensitive PM when the noncollinear angle is properly reset. Simultaneous temperature- and wavelength-insensitive PM is realized for the first time by imposing such a temperature-insensitive noncollinear configuration with an angularly dispersed seed signal. Based on the lithium triborate crystal, the proposed noncollinear achromatic PM has a thermal acceptance 6 times larger than that of the conventional wavelength-insensitive noncollinear PM and has a sufficient spectral acceptance to support pulse durations of ~20 fs at 800 nm. These achievements open new possibilities for generating ultra-high peak power lasers with high average power. PMID:27786299

  20. Demonstration of a chip-based optical isolator with parametric amplification

    NASA Astrophysics Data System (ADS)

    Hua, Shiyue; Wen, Jianming; Jiang, Xiaoshun; Hua, Qian; Jiang, Liang; Xiao, Min

    2016-11-01

    Despite being fundamentally challenging in integrated (nano)photonics, achieving chip-based light non-reciprocity becomes increasingly urgent in signal processing and optical communications. Because of material incompatibilities in conventional approaches based on the Faraday effect, alternative solutions have resorted to nonlinear processes to obtain one-way transmission. However, dynamic reciprocity in a recent theoretical analysis has pinned down the functionalities of these nonlinear isolators. To bypass such dynamic reciprocity, we here demonstrate an optical isolator on a silicon chip enforced by phase-matched parametric amplification in four-wave mixing. Using a high-Q microtoroid resonator, we realize highly non-reciprocal transport at the 1,550 nm wavelength when waves are injected from both directions in two different operating configurations. Our design, compatible with current complementary metal-oxide-semiconductor (CMOS) techniques, yields convincing isolation performance with sufficiently low insertion loss for a wide range of input power levels. Moreover, our work demonstrates the possibility of designing chip-based magnetic-free optical isolators for information processing and laser protection.

  1. Demonstration of a chip-based optical isolator with parametric amplification.

    PubMed

    Hua, Shiyue; Wen, Jianming; Jiang, Xiaoshun; Hua, Qian; Jiang, Liang; Xiao, Min

    2016-11-25

    Despite being fundamentally challenging in integrated (nano)photonics, achieving chip-based light non-reciprocity becomes increasingly urgent in signal processing and optical communications. Because of material incompatibilities in conventional approaches based on the Faraday effect, alternative solutions have resorted to nonlinear processes to obtain one-way transmission. However, dynamic reciprocity in a recent theoretical analysis has pinned down the functionalities of these nonlinear isolators. To bypass such dynamic reciprocity, we here demonstrate an optical isolator on a silicon chip enforced by phase-matched parametric amplification in four-wave mixing. Using a high-Q microtoroid resonator, we realize highly non-reciprocal transport at the 1,550 nm wavelength when waves are injected from both directions in two different operating configurations. Our design, compatible with current complementary metal-oxide-semiconductor (CMOS) techniques, yields convincing isolation performance with sufficiently low insertion loss for a wide range of input power levels. Moreover, our work demonstrates the possibility of designing chip-based magnetic-free optical isolators for information processing and laser protection.

  2. Demonstration of a chip-based optical isolator with parametric amplification

    PubMed Central

    Hua, Shiyue; Wen, Jianming; Jiang, Xiaoshun; Hua, Qian; Jiang, Liang; Xiao, Min

    2016-01-01

    Despite being fundamentally challenging in integrated (nano)photonics, achieving chip-based light non-reciprocity becomes increasingly urgent in signal processing and optical communications. Because of material incompatibilities in conventional approaches based on the Faraday effect, alternative solutions have resorted to nonlinear processes to obtain one-way transmission. However, dynamic reciprocity in a recent theoretical analysis has pinned down the functionalities of these nonlinear isolators. To bypass such dynamic reciprocity, we here demonstrate an optical isolator on a silicon chip enforced by phase-matched parametric amplification in four-wave mixing. Using a high-Q microtoroid resonator, we realize highly non-reciprocal transport at the 1,550 nm wavelength when waves are injected from both directions in two different operating configurations. Our design, compatible with current complementary metal-oxide-semiconductor (CMOS) techniques, yields convincing isolation performance with sufficiently low insertion loss for a wide range of input power levels. Moreover, our work demonstrates the possibility of designing chip-based magnetic-free optical isolators for information processing and laser protection. PMID:27886189

  3. Temperature- and wavelength-insensitive parametric amplification enabled by noncollinear achromatic phase-matching

    NASA Astrophysics Data System (ADS)

    Tang, Daolong; Ma, Jingui; Wang, Jing; Zhou, Bingjie; Xie, Guoqiang; Yuan, Peng; Zhu, Heyuan; Qian, Liejia

    2016-10-01

    Optical parametric chirped-pulse amplification (OPCPA) has been demonstrated to be a promising approach for pushing femtosecond pulses towards ultra-high peak powers. However, the future success of OPCPA strongly relies on the ability to manipulate its phase-matching (PM) configuration. When a high average power pump laser is involved, the thermal effects in nonlinear crystals induce phase-mismatch distortions that pose an inherent limitation on the conversion efficiency. Here, we demonstrate that the noncollinear configuration previously adopted for wavelength-insensitive PM can be employed for temperature-insensitive PM when the noncollinear angle is properly reset. Simultaneous temperature- and wavelength-insensitive PM is realized for the first time by imposing such a temperature-insensitive noncollinear configuration with an angularly dispersed seed signal. Based on the lithium triborate crystal, the proposed noncollinear achromatic PM has a thermal acceptance 6 times larger than that of the conventional wavelength-insensitive noncollinear PM and has a sufficient spectral acceptance to support pulse durations of ~20 fs at 800 nm. These achievements open new possibilities for generating ultra-high peak power lasers with high average power.

  4. A label-free fluorescent direct detection of live Salmonella typhimurium using cascade triple trigger sequences-regenerated strand displacement amplification and hairpin template-generated-scaffolded silver nanoclusters.

    PubMed

    Zhang, Peng; Liu, Hui; Li, Xiaocheng; Ma, Suzhen; Men, Shuai; Wei, Heng; Cui, Jingjing; Wang, Hongning

    2017-01-15

    The harm of Salmonella typhimurium (S. typhimurium) to public health mainly by the consumption of contaminated agricultural products or water stresses an urgent need for rapid detection methods to help control the spread of S. typhimurium. In this work, an intelligently designed sensor system took creative advantage of triple trigger sequences-regenerated strand displacement amplification and self-protective hairpin template-generated-scaffolded silver nanoclusters (AgNCs) for the first time. In the presence of live S. typhimurium, single-stranded trigger sequences were released from aptamer-trigger sequences complex, initiating a branch migration to open the hairpin template I containing complementary scaffolds of AgNCs. Then the first strand displacement amplification was induced to produce numerous scaffolds of AgNCs and reporter strands which initiated a branch migration to open the hairpin template II containing complementary scaffolds of AgNCs. Then the second strand displacement amplification was induced to generate numerous scaffolds of AgNCs and trigger sequences which initiated the third branch migration and strand displacement amplification to produce numerous scaffolds of AgNCs and reporter strands in succession. Cyclically, the reproduction of the trigger sequences and cascade successive production of scaffolds were achieved successfully, forming highly fluorescent AgNCs, thus providing significantly enhanced fluorescent signals to achieve ultrasensitive detection of live S. typhimurium down to 50 CFU/mL with a linear range from 10(2) to 10(7)CFU/mL. It is the first report on a fluorescent biosensor for detecting viable S. typhimurium directly, which can distinguish from heat denatured S. typhimurium. And it develops a new strategy to generate the DNA-scaffolds for forming AgNCs.

  5. Amplification properties of vacuum ultraviolet Ar2* produced by infrared high-intensity laser

    NASA Astrophysics Data System (ADS)

    Kaku, Masanori; Harano, Shinya; Matsumoto, Ryota; Katto, Masahito; Kubodera, Shoichi

    2011-07-01

    We report optical amplification of Ar2* at 126nm, pumped by optical-field-induced ionization (OFI) created by an infrared high-intensity laser. A gain--length product of 0.84 was obtained by using multipass amplification with a vacuum ultraviolet (VUV) cavity. The gain--length product was increased up to 4.3 through the use of single-pass amplification with a VUV reflector and a hollow 5.0cm-long fiber. Similar small signal gain coefficients of 0.84 and 0.86cm-1 were obtained in two different experiments, in which OFI Ar plasma gain media were produced in free space filled with Ar and inside an Ar-filled hollow fiber.

  6. 2D dynamic studies combined with the surface curvature analysis to predict Arias Intensity amplification

    NASA Astrophysics Data System (ADS)

    Torgoev, Almaz; Havenith, Hans-Balder

    2016-07-01

    A 2D elasto-dynamic modelling of the pure topographic seismic response is performed for six models with a total length of around 23.0 km. These models are reconstructed from the real topographic settings of the landslide-prone slopes situated in the Mailuu-Suu River Valley, Southern Kyrgyzstan. The main studied parameter is the Arias Intensity (Ia, m/sec), which is applied in the GIS-based Newmark method to regionally map the seismically-induced landslide susceptibility. This method maps the Ia values via empirical attenuation laws and our studies investigate a potential to include topographic input into them. Numerical studies analyse several signals with varying shape and changing central frequency values. All tests demonstrate that the spectral amplification patterns directly affect the amplification of the Ia values. These results let to link the 2D distribution of the topographically amplified Ia values with the parameter called as smoothed curvature. The amplification values for the low-frequency signals are better correlated with the curvature smoothed over larger spatial extent, while those values for the high-frequency signals are more linked to the curvature with smaller smoothing extent. The best predictions are provided by the curvature smoothed over the extent calculated according to Geli's law. The sample equations predicting the Ia amplification based on the smoothed curvature are presented for the sinusoid-shape input signals. These laws cannot be directly implemented in the regional Newmark method, as 3D amplification of the Ia values addresses more problem complexities which are not studied here. Nevertheless, our 2D results prepare the theoretical framework which can potentially be applied to the 3D domain and, therefore, represent a robust basis for these future research targets.

  7. Achieving metrological precision limits through postselection

    NASA Astrophysics Data System (ADS)

    Alves, G. Bié; Pimentel, A.; Hor-Meyll, M.; Walborn, S. P.; Davidovich, L.; Filho, R. L. de Matos

    2017-01-01

    Postselection strategies have been proposed with the aim of amplifying weak signals, which may help to overcome detection thresholds associated with technical noise in high-precision measurements. Here we use an optical setup to experimentally explore two different postselection protocols for the estimation of a small parameter: a weak-value amplification procedure and an alternative method that does not provide amplification but nonetheless is shown to be more robust for the sake of parameter estimation. Each technique leads approximately to the saturation of quantum limits for the estimation precision, expressed by the Cramér-Rao bound. For both situations, we show that parameter estimation is improved when the postselection statistics are considered together with the measurement device.

  8. Efficient Heterostructures for Combined Interference and Plasmon Resonance Raman Amplification.

    PubMed

    Alvarez-Fraga, Leo; Climent-Pascual, Esteban; Aguilar-Pujol, Montserrat; Ramírez-Jiménez, Rafael; Jiménez-Villacorta, Félix; Prieto, Carlos; de Andrés, Alicia

    2017-02-01

    The detection, identification, and quantification of different types of molecules and the optical imaging of, for example, cellular processes are important challenges. Here, we present how interference-enhanced Raman scattering (IERS) in adequately designed heterostructures can provide amplification factors relevant for both detection and imaging. Calculations demonstrate that the key factor is maximizing the absolute value of the refractive indices' difference between dielectric and metal layers. Accordingly, Si/Al/Al2O3/graphene heterostructures have been fabricated by optimizing the thickness and roughness and reaching enhancement values up to 700 for 488 nm excitation. The deviation from the calculated enhancement, 1200, is mainly due to reflectivity losses and roughness of the Al layer. The IERS platforms are also demonstrated to improve significantly the quality of white light images of graphene and are foreseen to be adequate to reveal the morphology of 2D and biological materials. A graphene top layer is adequate for most organic molecule deposition and often quenches possible fluorescence, permitting Raman signal detection, which, for a rhodamine 6G (R6G) monolayer, presents a gain of 400. Without graphene, the nonquenched R6G fluorescence is similarly amplified. The wavelength dependence of the involved refractive indices predicts much higher amplification (around 10(4)) for NIR excitation. These interference platforms can therefore be used to gain contrast and intensity in white light, Raman, and fluorescence imaging. We also demonstrate that surface-enhanced Raman scattering and IERS amplifications can be efficiently combined, leading to a gain of >10(5) (at 488 nm) by depositing a Ag nanostructured transparent film on the IERS platform. When the plasmonic structures deposited on the IERS platforms are optimized, single-molecule detection can be actively envisaged.

  9. Time varying arctic climate change amplification

    SciTech Connect

    Chylek, Petr; Dubey, Manvendra K; Lesins, Glen; Wang, Muyin

    2009-01-01

    During the past 130 years the global mean surface air temperature has risen by about 0.75 K. Due to feedbacks -- including the snow/ice albedo feedback -- the warming in the Arctic is expected to proceed at a faster rate than the global average. Climate model simulations suggest that this Arctic amplification produces warming that is two to three times larger than the global mean. Understanding the Arctic amplification is essential for projections of future Arctic climate including sea ice extent and melting of the Greenland ice sheet. We use the temperature records from the Arctic stations to show that (a) the Arctic amplification is larger at latitudes above 700 N compared to those within 64-70oN belt, and that, surprisingly; (b) the ratio of the Arctic to global rate of temperature change is not constant but varies on the decadal timescale. This time dependence will affect future projections of climate changes in the Arctic.

  10. Amplification, Redundancy, and Quantum Chernoff Information

    NASA Astrophysics Data System (ADS)

    Zwolak, Michael; Riedel, C. Jess; Zurek, Wojciech H.

    2014-04-01

    Amplification was regarded, since the early days of quantum theory, as a mysterious ingredient that endows quantum microstates with macroscopic consequences, key to the "collapse of the wave packet," and a way to avoid embarrassing problems exemplified by Schrödinger's cat. Such a bridge between the quantum microworld and the classical world of our experience was postulated ad hoc in the Copenhagen interpretation. Quantum Darwinism views amplification as replication, in many copies, of the information about quantum states. We show that such amplification is a natural consequence of a broad class of models of decoherence, including the photon environment we use to obtain most of our information. This leads to objective reality via the presence of robust and widely accessible records of selected quantum states. The resulting redundancy (the number of copies deposited in the environment) follows from the quantum Chernoff information that quantifies the information transmitted by a typical elementary subsystem of the environment.

  11. Amplification, redundancy, and quantum Chernoff information.

    PubMed

    Zwolak, Michael; Riedel, C Jess; Zurek, Wojciech H

    2014-04-11

    Amplification was regarded, since the early days of quantum theory, as a mysterious ingredient that endows quantum microstates with macroscopic consequences, key to the "collapse of the wave packet," and a way to avoid embarrassing problems exemplified by Schrödinger's cat. Such a bridge between the quantum microworld and the classical world of our experience was postulated ad hoc in the Copenhagen interpretation. Quantum Darwinism views amplification as replication, in many copies, of the information about quantum states. We show that such amplification is a natural consequence of a broad class of models of decoherence, including the photon environment we use to obtain most of our information. This leads to objective reality via the presence of robust and widely accessible records of selected quantum states. The resulting redundancy (the number of copies deposited in the environment) follows from the quantum Chernoff information that quantifies the information transmitted by a typical elementary subsystem of the environment.

  12. Wavelength preserved phase erasure and PSK to conventional OOK data format conversion based on phase sensitive amplification

    NASA Astrophysics Data System (ADS)

    Yu, Kan; Yang, Weili; Yu, Yu

    2016-10-01

    In this paper, a phase erasure and format conversion of phase-shift keying (PSK) to conventional on-off keying (OOK) is proposed and demonstrated theoretically and experimentally. Using a single-pump nondegenerate phase sensitive amplification process in a highly nonlinear fiber, the 0 and 1-bits of the PSK signal obtain different gains through amplification and de-amplification. As a result, the modulation information is transferred onto the amplitude. With an optimized input power difference between the signal and idler, the signal phase information is erased with wavelength preservation after the PSA. The output constellation and eye diagrams show an effective phase erasure and format conversion of PSK to conventional OOK. The error vector magnitude is utilized to evaluate the scheme performance. The proposed scheme provides the flexibility and resiliency for future photonic networks.

  13. Parametric amplification of orbital angular momentum beams based on light-acoustic interaction

    SciTech Connect

    Gao, Wei E-mail: zhuzhihandd@sina.com; Mu, Chunyuan; Yang, Yuqiang; Li, Hongwei; Zhu, Zhihan E-mail: zhuzhihandd@sina.com

    2015-07-27

    A high fidelity amplification of beams carrying orbital angular momentum (OAM) is very crucial for OAM multiplexing and other OAM-based applications. Here, we report a demonstration of stimulated Brillouin amplification for OAM beams, and the energy conversion efficiency of photon-phonon coupling and the phase structure of amplified signals are investigated in collinear and noncollinear frame systems, respectively. Our results demonstrate that the OAM signals can be efficiently amplified without obvious noise introduced, and the modes of output signal are independent of the pump modes or the geometrical frames. Meanwhile, an OAM state depending on the optical modes and the geometrical frames is loaded into phonons by coherent light-acoustic interaction, which reveals more fundamental significance and a great application potential in OAM-multiplexing.

  14. Theory and practice of enzyme bioaffinity electrodes. Chemical, enzymatic, and electrochemical amplification of in situ product detection.

    PubMed

    Limoges, Benoît; Marchal, Damien; Mavré, François; Savéant, Jean-Michel

    2008-06-11

    The two articles in this series are dedicated to bioaffinity electrodes with in situ detection of the product of the enzyme label after recognition by its conjugate immobilized on the electrode. Part 1 was devoted to direct electrochemical detection, whereas the present contribution deals with homogeneous chemical and enzymatic amplification of the primary electrochemical signal. The theoretical relationships that are established for these modes of amplification are applied to the avidin-biotin recognition in a system that involves alkaline phosphatase as enzyme label and 4-amino-2,6-dichloro-phenyl phosphate as substrate, generating 2,6-dichloro-4-aminophenol as electrochemically active product. Chemical amplification then results from the addition of NADH, which reduces the 2,6-dichloro-quinonimine resulting from the electrochemical oxidation of 2,6-dichloro-4-aminophenol. An increased amplification is obtained when the reduction of 2,6-dichloro-quinonimine involves diaphorase in solution with NADH as substrate. The excellent agreement between theoretical predictions and experimental data required a detailed theoretical analysis and the independent determination of the key kinetic parameters of the system. The theoretical analysis was extended to monolayer and multilayered films of auxiliary enzyme as well as to electrochemical amplification by means of closely spaced dual electrodes so as to offer a rational comparative panorama of the amplification capabilities of the various possible strategies. Confinement of the profile of the product, and/or its oxidized form, in the vicinity the electrode surface appears as a key parameter of amplification.

  15. Photoacoustic imaging using lock-in amplification and pulsed fiber lasers

    NASA Astrophysics Data System (ADS)

    Shi, Wei; Hajireza, Parsin; Zemp, Roger

    2016-03-01

    Photoacoustic (PA) imaging is a non-invasive, non-ionizing imaging technology with high optical contrast between blood and tissue, and with high sensitivity of hemoglobin concentration and oxygen saturation due to different optical absorption spectra resulting from different oxygenation of hemoglobin. Most PA imaging systems implement a nanosecond pulsed laser source as excitation source to induce PA signal, and rely on broadband amplifiers to record time-domain PA signals [1-6]. Some groups, however, have reported using modulated continuous-wave lasers as an excitation source for frequency-domain imaging [7-9]. Frequency-domain imaging offers the potential of lock-in amplification which has sensitivities as low as nV even in noise orders of magnitude higher than the signal. However, although modulated CW sources works for low cost and compact PA imaging, it does not satisfy thermal and stress confinement conditions required for optimal PA signal strength. Here, we investigate a PA methodology using pulsed fiber lasers as excitation laser source combined with lock-in amplification technology. For comparison, we also studied time-domain PA methodology. Phantom studies show that signal-to-noise ratio (SNR) obtained with frequency domain PA imaging is significantly more sensitive than that obtained using time-domain PA imaging when the laser pulse repetition rate (PRR) matches the bandwidth of ultrasound transducer. Therefore, high sensitive PA imaging technology using pulsed fiber laser sources with lock-in amplification may potentially greatly extend the depth of PA imaging.

  16. Optical amplification enhancement in photonic crystals

    SciTech Connect

    Sapienza, R.; Leonetti, M.; Froufe-Perez, L. S.; Galisteo-Lopez, J. F.; Lopez, C.; Conti, C.

    2011-02-15

    Improving and controlling the efficiency of a gain medium is one of the most challenging problems of laser research. By measuring the gain length in an opal-based photonic crystal doped with laser dye, we demonstrate that optical amplification is more than twenty-fold enhanced along the {Gamma}-K symmetry directions of the face-centered-cubic photonic crystal. These results are theoretically explained by directional variations of the density of states, providing a quantitative connection between density of the states and light amplification.

  17. A Simple, Low-Cost Platform for Real-Time Isothermal Nucleic Acid Amplification

    PubMed Central

    Craw, Pascal; Mackay, Ruth E.; Naveenathayalan, Angel; Hudson, Chris; Branavan, Manoharanehru; Sadiq, S. Tariq; Balachandran, Wamadeva

    2015-01-01

    Advances in microfluidics and the introduction of isothermal nucleic acid amplification assays have resulted in a range of solutions for nucleic acid amplification tests suited for point of care and field use. However, miniaturisation of instrumentation for such assays has not seen such rapid advances and fluorescence based assays still depend on complex, bulky and expensive optics such as fluorescence microscopes, photomultiplier tubes and sensitive lens assemblies. In this work we demonstrate a robust, low cost platform for isothermal nucleic acid amplification on a microfluidic device. Using easily obtainable materials and commercial off-the-shelf components, we show real time fluorescence detection using a low cost photodiode and operational amplifier without need for lenses. Temperature regulation on the device is achieved using a heater fabricated with standard printed circuit board fabrication methods. These facile construction methods allow fabrications at a cost compatible with widespread deployment to resource poor settings. PMID:26389913

  18. Optical parametric amplification of X-shaped localised wave-packets

    SciTech Connect

    Dubietis, A; Smilgevicius, V; Stabinis, A; Valiulis, G; Piskarskas, A

    2009-07-31

    The general concepts for generation and amplification of the X-pulses in optical parametric amplifiers under the plane-wave and localised (Bessel beam, or more generally, X-pulse) pump are reviewed. It is shown numerically and experimentally that X-pulse phase-matching gives rise to spontaneous emergence of the localised light structures in the regime of the parametric frequency down-conversion. The parametric amplification technique of localised waves is extended to the chirped X-pulse optical parametric amplification concept, which allows one to achieve few optical cycle, high-peak power localised wave packets for laser-matter interactions. (special issue devoted to the 80th birthday of s.a. akhmanov)

  19. A dual amplification fluorescent strategy for sensitive detection of DNA methyltransferase activity based on strand displacement amplification and DNAzyme amplification.

    PubMed

    Cui, Wanling; Wang, Lei; Jiang, Wei

    2016-03-15

    DNA methyltransferase (MTase) plays a critical role in many biological processes and has been regarded as a predictive cancer biomarker and a therapeutic target in cancer treatment. Sensitive detection of DNA MTase activity is essential for early cancer diagnosis and therapeutics. Here, we developed a dual amplification fluorescent strategy for sensitive detection of DNA MTase activity based on strand displacement amplification (SDA) and DNAzyme amplification. A trifunctional double-stranded DNA (dsDNA) probe was designed including a methylation site for DNA MTase recognition, a complementary sequence of 8-17 DNAzyme for synthesizing DNAzyme, and a nicking site for nicking enzyme cleavage. Firstly, the trifunctional dsDNA probe was methylated by DNA MTase to form the methylated dsDNA. Subsequently, HpaII restriction endonuclease specifically cleaved the residue of unmethylated dsDNA. Next, under the action of polymerase and nicking enzyme, the methylared dsDNA initiated SDA, releasing numbers of 8-17 DNAzymes. Finally, the released 8-17 DNAzymes triggered DNAzyme amplification reaction to induce a significant fluorescence enhancement. This strategy could detect DNA MTase activity as low as 0.0082U/mL. Additionally, the strategy was successfully applied for evaluating the inhibitions of DNA MTase using two anticancer drugs, 5-azacytidine and 5-aza-2'-deoxycytidine. The results indicate the proposed strategy has a potential application in early cancer diagnosis and therapeutics.

  20. Parametric dispersion and amplification of acoustohelicon waves in piezoelectric semiconductors

    NASA Astrophysics Data System (ADS)

    Neogi, A.; Ghosh, S.

    1991-01-01

    Assuming that the origin of the nonlinear interaction lies in the second-order optical susceptibility arising from the nonlinear induced current density and using the coupled-mode theory, the parametric dispersion and amplification of acoustohelicon waves is analytically investigated in a longitudinally magnetized piezoelectric semiconductor of noncentrosymmetric nature. The relevant experiments have not been reported. The threshold value of the pump electric field E0th and its corresponding excitation intensity is obtained. The longitudinal magnetic field decreases the required magnitude of E0th for the excitation of parametric amplification. The phenomenon of self-defocusing of the signal in the prevailing case is found to be a consequence of the negative dispersive characteristics exhibited by the acoustohelicon waves. Numerical analyses are performed for an InSb crystal at 77 K, duly irradiated by frequency-doubled pulsed 10.6-μm CO2 lasers. The parametric gain constant is observed to be maximum when the cyclotron frequency ωc attains the magnitude equal to that of ω0, the incident laser frequency (=1.78×1014 s-1 ).

  1. Short-Pulse Amplification by Strongly-Coupled Stimulated Brillouin Scattering

    NASA Astrophysics Data System (ADS)

    Edwards, Matthew; Jia, Qing; Mikhailova, Julia; Fisch, Nathaniel

    2016-10-01

    We examine the feasibility of strongly-coupled stimulated Brillouin scattering as a mechanism for the plasma-based amplification of sub-picosecond pulses. Fluid theory and particle-in-cell calculations are used to compare the relative advantages of Raman and Brillouin amplification over a broad range of parameters, with a focus on determining the maximum amplified pulse intensities and minimum durations that can be achieved. Amplification of short-wavelength pulses is considered in detail, with particular emphasis on the practical development of plasma-based x-ray amplifiers. Our results suggest that Brillouin scattering may allow amplification of shorter wavelength light than Raman scattering, but that at optical frequencies better performance is generally realized with Raman amplification, as strongly-coupled Brillouin scattering has limited capacity for amplifying sub-picosecond pulses. This work was supported by NNSA Grant No. DENA0002948 and AFOSR Grant No. FA9550-15-1-0391. M.R.E. gratefully acknowledges the support of the NSF through a Graduate Research Fellowship.

  2. Desert Amplification in a Warming Climate

    PubMed Central

    Zhou, Liming

    2016-01-01

    Here I analyze the observed and projected surface temperature anomalies over land between 50°S-50°N for the period 1950–2099 by large-scale ecoregion and find strongest warming consistently and persistently seen over driest ecoregions such as the Sahara desert and the Arabian Peninsula during various 30-year periods, pointing to desert amplification in a warming climate. This amplification enhances linearly with the global mean greenhouse gases(GHGs) radiative forcing and is attributable primarily to a stronger GHGs-enhanced downward longwave radiation forcing reaching the surface over drier ecoregions as a consequence of a warmer and thus moister atmosphere in response to increasing GHGs. These results indicate that desert amplification may represent a fundamental pattern of global warming associated with water vapor feedbacks over land in low- and mid- latitudes where surface warming rates depend inversely on ecosystem dryness. It is likely that desert amplification might involve two types of water vapor feedbacks that maximize respectively in the tropical upper troposphere and near the surface over deserts, with both being very dry and thus extremely sensitive to changes of water vapor. PMID:27538725

  3. Nondeterministic Noiseless Linear Amplification of Quantum Systems

    NASA Astrophysics Data System (ADS)

    Ralph, T. C.; Lund, A. P.

    2009-04-01

    We introduce the concept of non-deterministic noiseless linear amplification. We propose a linear optical realization of this transformation that could be built with current technology. We discuss the application of the device to distillation of continuous variable entanglement. We demonstrate that highly pure entanglement can be distilled from transmission over a lossy channel.

  4. Detection of cochlear amplification and its activation.

    PubMed

    Dong, Wei; Olson, Elizabeth S

    2013-08-20

    The operation of the mammalian cochlea relies on a mechanical traveling wave that is actively boosted by electromechanical forces in sensory outer hair cells (OHCs). This active cochlear amplifier produces the impressive sensitivity and frequency resolution of mammalian hearing. The cochlear amplifier has inspired scientists since its discovery in the 1970s, and is still not well understood. To explore cochlear electromechanics at the sensory cell/tissue interface, sound-evoked intracochlear pressure and extracellular voltage were measured using a recently developed dual-sensor with a microelectrode attached to a micro-pressure sensor. The resulting coincident in vivo observations of OHC electrical activity, pressure at the basilar membrane and basilar membrane displacement gave direct evidence for power amplification in the cochlea. Moreover, the results showed a phase shift of voltage relative to mechanical responses at frequencies slightly below the peak, near the onset of amplification. Based on the voltage-force relationship of isolated OHCs, the shift would give rise to effective OHC pumping forces within the traveling wave peak. Thus, the shift activates the cochlear amplifier, serving to localize and thus sharpen the frequency region of amplification. These results are the most concrete evidence for cochlear power amplification to date and support OHC somatic forces as its source.

  5. Topographic amplification across a taiwanese ridge

    NASA Astrophysics Data System (ADS)

    Rault, Claire; Meunier, Patrick; Burtin, Arnaud; Marc, Odin; Weian Chao, Vvn; Wu, Yih-Min; Hovius, Niels

    2016-04-01

    A line of 6 broadband seismometers have been deployed across a ridge in the Hualien County (Eastern Taiwan) in order to study topographic amplification. Since March 2015, the network has been continuously recording waves incoming from the Taiwanese regional seismicity. The hill is well approximated by a triangular topography of 3600m in length by 900m in height. We present a preliminary analysis performed over a dozen of earthquakes selected from the Seismic Taiwanese catalog (CWBSN). We show that most of the Uphill records exhibit a systematic amplification of seismic waves (peak to peak of particle velocity) in the relevant frequency band [0.5-2Hz]. By contrast, energy within the larger frequency band [6-20Hz] reflects local site effects induced by the soil layer. We report amplification ratios ranging from ranging from 1.2 to 3 and from 1.8 to 4 for P and S waves respectively. We show that amplification processes at the top strongly depend on the parameter α defined as the angle between the azimuth of incoming wave and the azimuth of the ridge divide.

  6. Desert Amplification in a Warming Climate

    NASA Astrophysics Data System (ADS)

    Zhou, Liming

    2016-08-01

    Here I analyze the observed and projected surface temperature anomalies over land between 50°S-50°N for the period 1950–2099 by large-scale ecoregion and find strongest warming consistently and persistently seen over driest ecoregions such as the Sahara desert and the Arabian Peninsula during various 30-year periods, pointing to desert amplification in a warming climate. This amplification enhances linearly with the global mean greenhouse gases(GHGs) radiative forcing and is attributable primarily to a stronger GHGs-enhanced downward longwave radiation forcing reaching the surface over drier ecoregions as a consequence of a warmer and thus moister atmosphere in response to increasing GHGs. These results indicate that desert amplification may represent a fundamental pattern of global warming associated with water vapor feedbacks over land in low- and mid- latitudes where surface warming rates depend inversely on ecosystem dryness. It is likely that desert amplification might involve two types of water vapor feedbacks that maximize respectively in the tropical upper troposphere and near the surface over deserts, with both being very dry and thus extremely sensitive to changes of water vapor.

  7. Detection of Cochlear Amplification and Its Activation

    PubMed Central

    Dong, Wei; Olson, Elizabeth S.

    2013-01-01

    The operation of the mammalian cochlea relies on a mechanical traveling wave that is actively boosted by electromechanical forces in sensory outer hair cells (OHCs). This active cochlear amplifier produces the impressive sensitivity and frequency resolution of mammalian hearing. The cochlear amplifier has inspired scientists since its discovery in the 1970s, and is still not well understood. To explore cochlear electromechanics at the sensory cell/tissue interface, sound-evoked intracochlear pressure and extracellular voltage were measured using a recently developed dual-sensor with a microelectrode attached to a micro-pressure sensor. The resulting coincident in vivo observations of OHC electrical activity, pressure at the basilar membrane and basilar membrane displacement gave direct evidence for power amplification in the cochlea. Moreover, the results showed a phase shift of voltage relative to mechanical responses at frequencies slightly below the peak, near the onset of amplification. Based on the voltage-force relationship of isolated OHCs, the shift would give rise to effective OHC pumping forces within the traveling wave peak. Thus, the shift activates the cochlear amplifier, serving to localize and thus sharpen the frequency region of amplification. These results are the most concrete evidence for cochlear power amplification to date and support OHC somatic forces as its source. PMID:23972858

  8. Desert Amplification in a Warming Climate.

    PubMed

    Zhou, Liming

    2016-08-19

    Here I analyze the observed and projected surface temperature anomalies over land between 50°S-50°N for the period 1950-2099 by large-scale ecoregion and find strongest warming consistently and persistently seen over driest ecoregions such as the Sahara desert and the Arabian Peninsula during various 30-year periods, pointing to desert amplification in a warming climate. This amplification enhances linearly with the global mean greenhouse gases(GHGs) radiative forcing and is attributable primarily to a stronger GHGs-enhanced downward longwave radiation forcing reaching the surface over drier ecoregions as a consequence of a warmer and thus moister atmosphere in response to increasing GHGs. These results indicate that desert amplification may represent a fundamental pattern of global warming associated with water vapor feedbacks over land in low- and mid- latitudes where surface warming rates depend inversely on ecosystem dryness. It is likely that desert amplification might involve two types of water vapor feedbacks that maximize respectively in the tropical upper troposphere and near the surface over deserts, with both being very dry and thus extremely sensitive to changes of water vapor.

  9. A novel piezoelectrically actuated 2-DoF compliant micro/nano-positioning stage with multi-level amplification

    NASA Astrophysics Data System (ADS)

    Zhu, Wu-Le; Zhu, Zhiwei; Shi, Yi; Chen, Xiangfan; He, Yu; Ehmann, Kornel F.; Ju, Bing-Feng

    2016-10-01

    This article presents a novel two-degrees-of-freedom (2-DoF) piezo-actuated parallel-kinematic micro/nano-positioning stage with multi-level amplification. The mirror symmetric stage consists of four leverage mechanisms, two Scott-Russell mechanisms, and a Z-shaped flexure hinge (ZFH) mechanism. Taking advantage of the ZFH mechanism, 2-DoF motions with final-level flexural amplification and decoupled motion guidance are achieved. Analytical models of the stage are developed and validated through finite element analysis to characterize its working performance. Practical testing of a prototype stage is conducted to demonstrate the design process and to quantify its response characteristics. Due to the utilized multi-level amplification, a practical amplification ratio of 13.0 is realized by the prototype. The decoupled output motion guidance feature of the stage makes it amenable for implementation in raster scanning type of measurements.

  10. A novel piezoelectrically actuated 2-DoF compliant micro/nano-positioning stage with multi-level amplification.

    PubMed

    Zhu, Wu-Le; Zhu, Zhiwei; Shi, Yi; Chen, Xiangfan; He, Yu; Ehmann, Kornel F; Ju, Bing-Feng

    2016-10-01

    This article presents a novel two-degrees-of-freedom (2-DoF) piezo-actuated parallel-kinematic micro/nano-positioning stage with multi-level amplification. The mirror symmetric stage consists of four leverage mechanisms, two Scott-Russell mechanisms, and a Z-shaped flexure hinge (ZFH) mechanism. Taking advantage of the ZFH mechanism, 2-DoF motions with final-level flexural amplification and decoupled motion guidance are achieved. Analytical models of the stage are developed and validated through finite element analysis to characterize its working performance. Practical testing of a prototype stage is conducted to demonstrate the design process and to quantify its response characteristics. Due to the utilized multi-level amplification, a practical amplification ratio of 13.0 is realized by the prototype. The decoupled output motion guidance feature of the stage makes it amenable for implementation in raster scanning type of measurements.

  11. Rapid detection of Mycobacterium tuberculosis by recombinase polymerase amplification.

    PubMed

    Boyle, David S; McNerney, Ruth; Teng Low, Hwee; Leader, Brandon Troy; Pérez-Osorio, Ailyn C; Meyer, Jessica C; O'Sullivan, Denise M; Brooks, David G; Piepenburg, Olaf; Forrest, Matthew S

    2014-01-01

    Improved access to effective tests for diagnosing tuberculosis (TB) has been designated a public health priority by the World Health Organisation. In high burden TB countries nucleic acid based TB tests have been restricted to centralised laboratories and specialised research settings. Requirements such as a constant electrical supply, air conditioning and skilled, computer literate operators prevent implementation of such tests in many settings. Isothermal DNA amplification technologies permit the use of simpler, less energy intensive detection platforms more suited to low resource settings that allow the accurate diagnosis of a disease within a short timeframe. Recombinase Polymerase Amplification (RPA) is a rapid, low temperature isothermal DNA amplification reaction. We report here RPA-based detection of Mycobacterium tuberculosis complex (MTC) DNA in <20 minutes at 39 °C. Assays for two MTC specific targets were investigated, IS6110 and IS1081. When testing purified MTC genomic DNA, limits of detection of 6.25 fg (IS6110) and 20 fg (IS1081)were consistently achieved. When testing a convenience sample of pulmonary specimens from suspected TB patients, RPA demonstrated superior accuracy to indirect fluorescence microscopy. Compared to culture, sensitivities for the IS1081 RPA and microscopy were 91.4% (95%CI: 85, 97.9) and 86.1% (95%CI: 78.1, 94.1) respectively (n = 71). Specificities were 100% and 88.6% (95% CI: 80.8, 96.1) respectively. For the IS6110 RPA and microscopy sensitivities of 87.5% (95%CI: 81.7, 93.2) and 70.8% (95%CI: 62.9, 78.7) were obtained (n = 90). Specificities were 95.4 (95% CI: 92.3,98.1) and 88% (95% CI: 83.6, 92.4) respectively. The superior specificity of RPA for detecting tuberculosis was due to the reduced ability of fluorescence microscopy to distinguish Mtb complex from other acid fast bacteria. The rapid nature of the RPA assay and its low energy requirement compared to other amplification technologies suggest RPA-based TB assays

  12. Site amplifications for generic rock sites

    USGS Publications Warehouse

    Boore, D.M.; Joyner, W.B.

    1997-01-01

    Seismic shear-wave velocity as a function of depth for generic rock sites has been estimated from borehole data and studies of crustal velocities, and these velocities have been used to compute frequency-dependent amplifications for zero attenuation for use in simulations of strong ground motion. We define a generic rock site as one whose velocity at shallow depths equals the average of those from the rock sites sampled by the borehole data. Most of the boreholes are in populated areas; for that reason, the rock sites sampled are of particular engineering significance. We consider two generic rock sites: rock, corresponding to the bulk of the borehole data, and very hard rock, such as is found in glaciated regions in large areas of eastern North America or in portions of western North America. The amplifications on rock sites can be in excess of 3.5 at high frequencies, in contrast to the amplifications of less than 1.2 on very hard rock sites. The consideration of unattenuated amplification alone is computationally convenient, but what matters for ground-motion estimation is the combined effect of amplification and attenuation. For reasonable values of the attenuation parameter K0, the combined effect of attenuation and amplification for rock sites peaks between about 2 and 5 Hz with a maximum level of less than 1.8. The combined effect is about a factor of 1.5 at 1 Hz and is less than unity for frequencies in the range of 10 to 20 Hz (depending on K0). Using these amplifications, we find provisional values of about ???? = 70 bars and K0 = 0.035 sec for rock sites in western North America by fitting our empirically determined response spectra for an M 6.5 event to simulated values. The borehole data yield shear velocities (V??30) of 618 and 306 m/sec for "rock" and "soil" sites, respectively, when averaged over the upper 30 m. From this, we recommend that V??30 equals 620 and 310 m/sec for applications requiring the average velocity for rock and soil sites in

  13. Identification and functional characterization of a type I signal peptidase gene of Bacillus megaterium DSM319.

    PubMed

    Nahrstedt, H; Wittchen, K- D; Rachman, M A; Meinhardt, F

    2004-04-01

    The sipM gene of Bacillus megaterium encoding a type I signal peptidase (SPase) was isolated and structurally characterized. RNA analysis revealed a transcript size in accordance with a bicistronic operon comprising sipM and an adjacent open reading frame. Inactivation of sipM by targeted gene disruption could not be achieved, indicating its essential role for cell viability since there might be no other type I SPase of major importance present in B. megaterium. Plasmid-assisted amplification of the gene resulted in an increase in activity of the heterologous glucanase used as an extracellular reporter, suggesting a potential bottleneck for protein secretion within this species.

  14. Unity-Efficiency Parametric Down-Conversion via Amplitude Amplification

    NASA Astrophysics Data System (ADS)

    Niu, Murphy Yuezhen; Sanders, Barry C.; Wong, Franco N. C.; Shapiro, Jeffrey H.

    2017-03-01

    We propose an optical scheme, employing optical parametric down-converters interlaced with nonlinear sign gates (NSGs), that completely converts an n -photon Fock-state pump to n signal-idler photon pairs when the down-converters' crystal lengths are chosen appropriately. The proof of this assertion relies on amplitude amplification, analogous to that employed in Grover search, applied to the full quantum dynamics of single-mode parametric down-conversion. When we require that all Grover iterations use the same crystal, and account for potential experimental limitations on crystal-length precision, our optimized conversion efficiencies reach unity for 1 ≤n ≤5 , after which they decrease monotonically for n values up to 50, which is the upper limit of our numerical dynamics evaluations. Nevertheless, our conversion efficiencies remain higher than those for a conventional (no NSGs) down-converter.

  15. Improved optical amplification using metamaterial based split ring structures in optical fibres

    NASA Astrophysics Data System (ADS)

    Prakash, Geetha; Nigam, Raaghvam; Das, Sovan; Chellappa, Sharath

    2016-04-01

    Optical fibres provide the best solutions for transmitting high speed, large amounts of data with good power efficiency. However such transmission would also need amplification for transmission over large distances. Erbium Doped Fibre Amplifiers(EDFAs) are currently being used for optical amplification. But good amplification is achievable with multiple stages and considerable length of EDFA fibres. In this paper we compare the use of Silver Split Ring Resonators(SRRs) , Gold Nano Rods and Silver Fishnet structures which give metamaterial properties to be used in optical fibres to give better amplification than EDFA based fibres. Metamaterials belong to a new class of materials with negative values for permittivity and permeability. Such materials would exhibit negative refractive index leading to these materials being called as left handed media.If such left handed media have an internal structure made of dimensions much smaller than the wavelength but sufficiently thick to exhibit bulk properties, using other optical domains such as plasmonics, it is possible to control light interactions and propagation. Artificial structures smaller than the wavelength of light can be used to enhance electric and magnetic fields. Surface plasmons can be excited on a metal and this can enhance the electric field at the surface. Our paper proposes the use of this phenomenon of achieving gain at optical frequencies by using SRRs, Fishnet structures , Nano Rods. We compare the performance of these structures and observe that they provide gain which is much more than that provided by EDFAs.

  16. Graded Achievement, Tested Achievement, and Validity

    ERIC Educational Resources Information Center

    Brookhart, Susan M.

    2015-01-01

    Twenty-eight studies of grades, over a century, were reviewed using the argument-based approach to validity suggested by Kane as a theoretical framework. The review draws conclusions about the meaning of graded achievement, its relation to tested achievement, and changes in the construct of graded achievement over time. "Graded…

  17. Bioanalytical applications of isothermal nucleic acid amplification techniques.

    PubMed

    Deng, Huimin; Gao, Zhiqiang

    2015-01-01

    The most popular in vitro nucleic acid amplification techniques like polymerase chain reaction (PCR) including real-time PCR are costly and require thermocycling, rendering them unsuitable for uses at point-of-care. Highly efficient in vitro nucleic acid amplification techniques using simple, portable and low-cost instruments are crucial in disease diagnosis, mutation detection and biodefense. Toward this goal, isothermal amplification techniques that represent a group of attractive in vitro nucleic acid amplification techniques for bioanalysis have been developed. Unlike PCR where polymerases are easily deactivated by thermally labile constituents in a sample, some of the isothermal nucleic acid amplification techniques, such as helicase-dependent amplification and nucleic acid sequence-based amplification, enable the detection of bioanalytes with much simplified protocols and with minimal sample preparations since the entire amplification processes are performed isothermally. This review focuses on the isothermal nucleic acid amplification techniques and their applications in bioanalytical chemistry. Starting off from their amplification mechanisms and significant properties, the adoption of isothermal amplification techniques in bioanalytical chemistry and their future perspectives are discussed. Representative examples illustrating the performance and advantages of each isothermal amplification technique are discussed along with some discussion on the advantages and disadvantages of each technique.

  18. Real-time monitoring of mycobacterium genomic DNA with target-primed rolling circle amplification by a Au nanoparticle-embedded SPR biosensor.

    PubMed

    Xiang, Yang; Zhu, Xiaoyan; Huang, Qing; Zheng, Junsong; Fu, Weiling

    2015-04-15

    In this study, we developed a surface plasmon resonance (SPR) DNA biosensor array based on target-primed rolling circle amplification (RCA) for isothermal and rapid detection of two pathogenic mycobacteria, Mycobacterium tuberculosis complex (MTBC) and Mycobacterium avium complex (MAC).The species-specific padlock probe (PLP) was designed to target the sequence in 16S-23S rRNA gene internal transcribed spacer (ITS). After ligation, the circularized PLP could be primed by the target sequence to initial RCA. The RCA performed simultaneously with the cleavage reaction to produce small fragments of single strand DNA which immediately hybridized with the probe immobilized on the sensor chip without denaturation. This process caused SPR angle changes on the chip surface, which made the detection for analysis from the solution achievable, and dynamic real-time RCA monitoring of mycobacterium possible. Besides, Au nanoparticles (AuNPs) were directly assembled onto the surface of the sensor chip via hexanedithiol (HDT) for the enhancement of sensitivity as a label-free detection system. Experimental results show that the signal enhancement by the target-primed RCA together with AuNPs-embedded surface caused at least10-fold increased sensitivity as compared with conventional RCA on bare SPR chip method. Within 40min amplification duration as low as 20amol of synthetic targets and 10(4)CFUmL(-1) of genomic DNA from clinical samples can be detected. The proposed method not only provides a simple design idea for liquid-phase amplification monitoring, but also apply it in clinical pathogen detection, which holds great promise in ultrasensitive bioassay in the future.

  19. Highly sensitive photoelectrochemical immunoassay with enhanced amplification using horseradish peroxidase induced biocatalytic precipitation on a CdS quantum dots multilayer electrode.

    PubMed

    Zhao, Wei-Wei; Ma, Zheng-Yuan; Yu, Pei-Pei; Dong, Xiao-Ya; Xu, Jing-Juan; Chen, Hong-Yuan

    2012-01-17

    Herein we demonstrate the protocol of a biocatalytic precipitation (BCP)-based sandwich photoelectrochemical (PEC) horseradish peroxidase (HRP)-linked immunoassay on the basis of their synergy effect for the ultrasensitive detection of mouse IgG (antigen, Ag) as a model protein. The hybrid film consisting of oppositely charged polyelectrolytes and CdS quantum dots (QDs) is developed by the classic layer by layer (LbL) method and then employed as the photoactive antibody (Ab) immobilization matrix for the subsequent sandwich-type Ab-Ag affinity interactions. Improved sensitivity is achieved through using the bioconjugates of HRP-secondary antibodies (Ab(2)). In addition to the much enhanced steric hindrance compared with the original one, the presence of HRP would further stimulate the BCP onto the electrode surface for signal amplification, concomitant to a competitive nonproductive absorption that lowers the photocurrent intensity. As a result of the multisignal amplification in this HRP catalyzed BCP-based PEC immunoassay, it possesses excellent analytical performance. The antigen could be detected from 0.5 pg/mL to 5.0 ng/mL with a detection limit of 0.5 pg/mL.

  20. Amplification and reversal of Knudsen force by thermoelectric heating

    SciTech Connect

    O'Neill, William J.; Wada, Mizuki; Strongrich, Andrew D.; Cofer, Anthony; Alexeenko, Alina A.

    2014-12-09

    We show that the Knudsen thermal force generated by a thermally-induced flow over a heated beam near a colder wall could be amplified significantly by thermoelectric heating. Bidirectional actuation is achieved by switching the polarity of the thermoelectric device bias voltage. The measurements of the resulting thermal forces at different rarefaction regimes, realized by changing geometry and gas pressure, are done using torsional microbalance. The repulsive or attractive forces between a thermoelectrically heated or cooled plate and a substrate are shown to be up to an order of magnitude larger than for previously studied configurations and heating methods due to favorable coupling of two thermal gradients. The amplification and reversal of the Knudsen force is confirmed by numerical solution of the Boltzmann-ESBGK kinetic model equation. Because of the favorable scaling with decreasing system size, the Knudsen force with thermoelectric heating offers a novel actuation and sensing mechanism for nano/microsystems.

  1. Amplification and reversal of Knudsen force by thermoelectric heating

    NASA Astrophysics Data System (ADS)

    O'Neill, William J.; Wada, Mizuki; Strongrich, Andrew D.; Cofer, Anthony; Alexeenko, Alina A.

    2014-12-01

    We show that the Knudsen thermal force generated by a thermally-induced flow over a heated beam near a colder wall could be amplified significantly by thermoelectric heating. Bidirectional actuation is achieved by switching the polarity of the thermoelectric device bias voltage. The measurements of the resulting thermal forces at different rarefaction regimes, realized by changing geometry and gas pressure, are done using torsional microbalance. The repulsive or attractive forces between a thermoelectrically heated or cooled plate and a substrate are shown to be up to an order of magnitude larger than for previously studied configurations and heating methods due to favorable coupling of two thermal gradients. The amplification and reversal of the Knudsen force is confirmed by numerical solution of the Boltzmann-ESBGK kinetic model equation. Because of the favorable scaling with decreasing system size, the Knudsen force with thermoelectric heating offers a novel actuation and sensing mechanism for nano/microsystems.

  2. Quantum-limited amplification and entanglement in coupled nonlinear resonators.

    PubMed

    Eichler, C; Salathe, Y; Mlynek, J; Schmidt, S; Wallraff, A

    2014-09-12

    We demonstrate a coupled cavity realization of a Bose-Hubbard dimer to achieve quantum-limited amplification and to generate frequency entangled microwave fields with squeezing parameters well below -12  dB. In contrast to previous implementations of parametric amplifiers, our dimer can be operated both as a degenerate and as a nondegenerate amplifier. The large measured gain-bandwidth product of more than 250 MHz for the nondegenerate operation and the saturation at input photon numbers as high as 2000 per μs are both expected to be improvable even further, while maintaining wide frequency tunability of about 2 GHz. Featuring flexible control over all relevant system parameters, the presented Bose-Hubbard dimer based on lumped element circuits has significant potential as an elementary cell in nonlinear cavity arrays for quantum simulations.

  3. Detection of MYCN Amplification in Serum DNA Using Conventional Polymerase Chain Reaction

    PubMed Central

    2016-01-01

    Neuroblastoma (NB) is the most common extra-cranial solid tumor of childhood and is characterized by a wide range of clinical behaviors. Amplification of MYCN is a well-known poor prognostic factor in NB patients. As the MYCN amplification status is usually tested using tumor specimens, lengthy and invasive procedures are unavoidable. To evaluate the possibility of detecting MYCN amplification without invasive procedure, we performed conventional polymerase chain reaction (PCR) analysis to identify MYCN amplification using the preserved serum DNA. PCR of serum DNA was done in 105 NB patients whose MYCN status had been confirmed by fluorescence in situ hybridization. MYCN amplification was evaluated as the ratio of signal intensities between MYCN and NAGK (M/N ratio). When regarding the tissue FISH results as a reference, 10 patients had MYCN-amplified (MNA) NB, and 95 had non-MNA NB. The M/N ratio of the MNA group (median 2.56, range 1.01-3.58) was significantly higher than that of the non-MNA group (median 0.97, range 0.67-5.18) (P < 0.001). In the receiver operating characteristic curve analysis, the area under the curve was 0.957 (95% confidence interval 0.898–1.000; P < 0.001), and it showed 90.9% sensitivity and 97.9% specificity with the selected cut-off value set as 1.6. The detection of MYCN amplification using conventional PCR analysis of serum samples seems to be a simple and promising method to evaluate the MYCN status of NB patients. Further study with a larger set of patients is needed to confirm the accuracy of this result. PMID:27510381

  4. Biomass changes and trophic amplification of plankton in a warmer ocean.

    PubMed

    Chust, Guillem; Allen, J Icarus; Bopp, Laurent; Schrum, Corinna; Holt, Jason; Tsiaras, Kostas; Zavatarelli, Marco; Chifflet, Marina; Cannaby, Heather; Dadou, Isabelle; Daewel, Ute; Wakelin, Sarah L; Machu, Eric; Pushpadas, Dhanya; Butenschon, Momme; Artioli, Yuri; Petihakis, George; Smith, Chris; Garçon, Veronique; Goubanova, Katerina; Le Vu, Briac; Fach, Bettina A; Salihoglu, Baris; Clementi, Emanuela; Irigoien, Xabier

    2014-07-01

    Ocean warming can modify the ecophysiology and distribution of marine organisms, and relationships between species, with nonlinear interactions between ecosystem components potentially resulting in trophic amplification. Trophic amplification (or attenuation) describe the propagation of a hydroclimatic signal up the food web, causing magnification (or depression) of biomass values along one or more trophic pathways. We have employed 3-D coupled physical-biogeochemical models to explore ecosystem responses to climate change with a focus on trophic amplification. The response of phytoplankton and zooplankton to global climate-change projections, carried out with the IPSL Earth System Model by the end of the century, is analysed at global and regional basis, including European seas (NE Atlantic, Barents Sea, Baltic Sea, Black Sea, Bay of Biscay, Adriatic Sea, Aegean Sea) and the Eastern Boundary Upwelling System (Benguela). Results indicate that globally and in Atlantic Margin and North Sea, increased ocean stratification causes primary production and zooplankton biomass to decrease in response to a warming climate, whilst in the Barents, Baltic and Black Seas, primary production and zooplankton biomass increase. Projected warming characterized by an increase in sea surface temperature of 2.29 ± 0.05 °C leads to a reduction in zooplankton and phytoplankton biomasses of 11% and 6%, respectively. This suggests negative amplification of climate driven modifications of trophic level biomass through bottom-up control, leading to a reduced capacity of oceans to regulate climate through the biological carbon pump. Simulations suggest negative amplification is the dominant response across 47% of the ocean surface and prevails in the tropical oceans; whilst positive trophic amplification prevails in the Arctic and Antarctic oceans. Trophic attenuation is projected in temperate seas. Uncertainties in ocean plankton projections, associated to the use of single global and

  5. An isothermal electrochemical biosensor for the sensitive detection of microRNA based on a catalytic hairpin assembly and supersandwich amplification.

    PubMed

    Zhang, Hua; Wang, Qing; Yang, Xiaohai; Wang, Kemin; Li, Qing; Li, Zhiping; Gao, Lei; Nie, Wenyan; Zheng, Yan

    2017-01-16

    A novel isothermal electrochemical biosensor was proposed for the sensitive detection of microRNA (miRNA) based on the ingenious combination of the target-catalyzed hairpin assembly (CHA) and supersandwich amplification strategies. Since miRNA-221 has been reported to be overexpressed in cancers and has been a potentially useful biomarker for the diagnosis of the related diseases, miRNA-221 was chosen as a model target miRNA. The target miRNA-221 triggered a toehold strand displacement assembly of the two hairpin substrates, which led to the cyclicality of the target miRNA and the CHA products. Subsequently, the CHA products hybridized with a capture probe on the electrode and the exposed stem of the CHA products was further used to propagate the supersandwich. After this, the signal probe was modified with horseradish peroxidase (HRP) to form a supersandwich multiplex HRP-DNA label, which could achieve an amplified electrochemical signal. Using the isothermal dual signal amplification strategies, miRNA-221 as low as 0.6 pM (3σ) could be detected. In addition, this biosensor showed high selectivity and could discriminate miRNA-221 from the homologous miRNAs. Note that human miRNA from cancer cells could also be detected and the results were in excellent agreement with those obtained using qRT-PCR. Given that the biosensor avoided the introduction of nanoparticles, the limitation of using the nanoparticles was overcome. The proposed biosensor has great potential for broad applications in the field of clinical analysis.

  6. Most probable number - loop mediated isothermal amplification (MPN-LAMP) for quantifying waterborne pathogens in <25min.

    PubMed

    Ahmad, Farhan; Stedtfeld, Robert D; Waseem, Hassan; Williams, Maggie R; Cupples, Alison M; Tiedje, James M; Hashsham, Syed A

    2017-01-01

    We are reporting a most probable number approach integrated to loop mediated isothermal technique (MPN-LAMP) focusing on Gram-negative Escherichia coli and Gram-positive Enterococcus faecalis bacterial cells without nucleic acids extraction. LAMP assays for uidA from E. coli and gelE from E. faecalis were successfully performed directly on cells up to single digit concentration using a commercial real time PCR instrument. Threshold time values of LAMP assays of bacterial cells, heat treated bacterial cells (95°C for 5min), and their purified genomic DNA templates were similar, implying that amplification could be achieved directly from bacterial cells at 63°C. Viability of bacterial cells was confirmed by using propidium monoazide in a LAMP assay with E. faecalis. To check its functionality on a microfluidic platform, MPN-LAMP assays targeting <10CFU of bacteria were also translated onto polymeric microchips and monitored by a low-cost fluorescence imaging system. The overall system provided signal-to-noise (SNR) ratios up to 800, analytical sensitivity of <10CFU, and time to positivity of about 20min. MPN-LAMP assays were performed for cell concentrations in the range of 10(5)CFU to <10CFU. MPN values from LAMP assays confirmed that the amplifications were from <10CFU. The method described here, applicable directly on cells at 63°C, eliminates the requirement of complex nucleic acids extraction steps, facilitating the development of sensitive, rapid, low-cost, and field-deployable systems. This rapid MPN-LAMP approach has the potential to replace conventional MPN method for waterborne pathogens.

  7. Investigation of the amplification properties of Ho:YAG single crystal fiber

    NASA Astrophysics Data System (ADS)

    Li, Yuan; Zhang, Zeyu; Buckley, Ian; Miller, Jerome K.; Johnson, Eric G.; Nie, Craig D.; Harrington, James A.; Shori, Ramesh K.

    2015-02-01

    0.5% Holmium (Ho) doped YAG single crystal fiber (SCF) was fabricated using the laser heated pedestal growth (LHPG) method and amplification properties of the fabricated Ho:YAG SCF were studied. The relatively large lengthto- diameter ratio provides guiding for both the pump and signal beams propagating in the SCF. The propagation and gain of signals with different modes were studied. A numerical method based on finite difference (FD) beam propagation method (BPM) combined with the rate equations was developed for theoretical simulation. The results are encouraging to demonstrate the advantages of SCF for its fiber-like beam guiding property and solid state material gain property. The simulation tool provides details about how the fiber shape and launched mode affect the gain and output beam shape as well as predicts the amplification behavior of such unique specialty fibers.

  8. Distinct roles of TRP channels in auditory transduction and amplification in Drosophila.

    PubMed

    Lehnert, Brendan P; Baker, Allison E; Gaudry, Quentin; Chiang, Ann-Shyn; Wilson, Rachel I

    2013-01-09

    Auditory receptor cells rely on mechanically gated channels to transform sound stimuli into neural activity. Several TRP channels have been implicated in Drosophila auditory transduction, but mechanistic studies have been hampered by the inability to record subthreshold signals from receptor neurons. Here, we develop a non-invasive method for measuring these signals by recording from a central neuron that is electrically coupled to a genetically defined population of auditory receptor cells. We find that the TRPN family member NompC, which is necessary for the active amplification of sound-evoked motion by the auditory organ, is not required for transduction in auditory receptor cells. Instead, NompC sensitizes the transduction complex to movement and precisely regulates the static forces on the complex. In contrast, the TRPV channels Nanchung and Inactive are required for responses to sound, suggesting they are components of the transduction complex. Thus, transduction and active amplification are genetically separable processes in Drosophila hearing.

  9. Resonant amplification of vortex-core oscillations by coherent magnetic-field pulses

    PubMed Central

    Yu, Young-Sang; Han, Dong-Soo; Yoo, Myoung-Woo; Lee, Ki-Suk; Choi, Youn-Seok; Jung, Hyunsung; Lee, Jehyun; Im, Mi-Young; Fischer, Peter; Kim, Sang-Koog

    2013-01-01

    Vortex structures in soft magnetic nanodisks are highly attractive due to their scientific beauty and potential technological applications. Here, we experimentally demonstrated the resonant amplification of vortex oscillations by application of simple coherent field pulses tuned to optimal width and time intervals. In order to investigate vortex excitations on the sub-ns time scale, we employed state-of-the-art time-resolved full-field soft X-ray microscopy of 70 ps temporal and 25 nm lateral resolution. We found that, due to the resonant enhancement of the vortex gyration motion, the signal input power can be significantly reduced to ~ 1 Oe in field strength, while increasing signal gains, by increasing the number of the optimal field pulses. We identified the origin of this behavior as the forced resonant amplification of vortex gyration. This work represents an important milestone towards the potential implementation of vortex oscillations in future magnetic vortex devices. PMID:23416729

  10. Resonant amplification of vortex-core oscillations by coherent magnetic-field pulses.

    PubMed

    Yu, Young-Sang; Han, Dong-Soo; Yoo, Myoung-Woo; Lee, Ki-Suk; Choi, Youn-Seok; Jung, Hyunsung; Lee, Jehyun; Im, Mi-Young; Fischer, Peter; Kim, Sang-Koog

    2013-01-01

    Vortex structures in soft magnetic nanodisks are highly attractive due to their scientific beauty and potential technological applications. Here, we experimentally demonstrated the resonant amplification of vortex oscillations by application of simple coherent field pulses tuned to optimal width and time intervals. In order to investigate vortex excitations on the sub-ns time scale, we employed state-of-the-art time-resolved full-field soft X-ray microscopy of 70 ps temporal and 25 nm lateral resolution. We found that, due to the resonant enhancement of the vortex gyration motion, the signal input power can be significantly reduced to ~ 1 Oe in field strength, while increasing signal gains, by increasing the number of the optimal field pulses. We identified the origin of this behavior as the forced resonant amplification of vortex gyration. This work represents an important milestone towards the potential implementation of vortex oscillations in future magnetic vortex devices.

  11. Experimental demonstration of spatially coherent beam combining using optical parametric amplification.

    PubMed

    Kurita, Takashi; Sueda, Keiichi; Tsubakimoto, Koji; Miyanaga, Noriaki

    2010-07-05

    We experimentally demonstrated coherent beam combining using optical parametric amplification with a nonlinear crystal pumped by random-phased multiple-beam array of the second harmonic of a Nd:YAG laser at 10-Hz repetition rate. In the proof-of-principle experiment, the phase jump between two pump beams was precisely controlled by a motorized actuator. For the demonstration of multiple-beam combining a random phase plate was used to create random-phased beamlets as a pump pulse. Far-field patterns of the pump, the signal, and the idler indicated that the spatially coherent signal beams were obtained on both cases. This approach allows scaling of the intensity of optical parametric chirped pulse amplification up to the exa-watt level while maintaining diffraction-limited beam quality.

  12. Next-generation repeat-free FISH probes for DNA amplification in glioblastoma in vivo: Improving patient selection to MDM2-targeted inhibitors.

    PubMed

    Brunelli, Matteo; Eccher, Albino; Cima, Luca; Trippini, Tobia; Pedron, Serena; Chilosi, Marco; Barbareschi, Mattia; Scarpa, Aldo; Pinna, Giampietro; Cabrini, Giulio; Pilotto, Sara; Carbognin, Luisa; Bria, Emilio; Tortora, Giampaolo; Fioravanzo, Adele; Schiavo, Nicola; Meglio, Mario; Sava, Teodoro; Belli, Laura; Martignoni, Guido; Ghimenton, Claudio

    2017-01-01

    A next-generation FISH probe mapping to the MDM2 locus-specific region has recently been designed. The level of MDM2 gene amplification (high versus low) may allow selection of patients for cancer treatment with MDM2 inhibitors and may predict their responsiveness. We investigated the spectrum of MDM2 gene alterations using the new probes in vivo after visualizing single neoplastic cells in situ from a series of glioblastomas. Signals from next-generation repeat-free FISH interphase probes were identified in tissue microarrays that included 3 spots for each of the 48 cases. The murine double minutes (MDM2)-specific DNA probe and the satellite enumeration probe for chromosome 12 were used. Three cases (6%) showed more than 25 signals (high gene amplification), and 7 (15%) showed 3-10 signals (gains); among these, 4 cases (8%) had an equal number of MDM2 and centromeric signals on chromosome 12 (polyploidy). Genomic heterogeneity was observed only in 3 cases with low gene amplification. In our series, 6% of glioblastomas exhibited high MDM2 amplification (in vivo) with a pattern related to the known double minutes/chromothripsis phenomenon (in situ), and only cases with low amplification showed genomic heterogeneity. We concluded that the rate of MDM2 gene amplification can be a useful predictive biomarker to improve patient selection.

  13. Parallel parametric amplification of coherently excited propagating spin waves in a microscopic Ni{sub 81}Fe{sub 19} waveguide

    SciTech Connect

    Brächer, T.; Pirro, P.; Meyer, T.; Heussner, F.; Lägel, B.; Serga, A. A.; Hillebrands, B.

    2014-05-19

    We present parallel parametric amplification of coherently excited, propagating spin waves in a microstructured magnonic Ni{sub 81}Fe{sub 19} waveguide. Amplification is achieved by the pumping field generated by a microwave current flowing through a Cu micro-stripline underneath the waveguide. By employing microfocussed Brillouin light scattering spectroscopy, we investigate the spatial decay of the propagating spin waves and their amplification by means of parallel pumping. We analyze the dependence of the intensity of the amplified spin waves on the spin-wave excitation power, pumping power, and pumping duration, revealing the most efficient working point for a noise-free amplification. This paves the way for a frequency selective amplification of spin waves in microstructured magnonic circuits.

  14. Performance scaling via passive pulse shaping in cavity-enhanced optical parametric chirped-pulse amplification.

    PubMed

    Siddiqui, Aleem M; Moses, Jeffrey; Hong, Kyung-Han; Lai, Chien-Jen; Kärtner, Franz X

    2010-06-15

    We show that an enhancement cavity seeded at the full repetition rate of the pump laser can automatically reshape small-signal gain across the interacting pulses in an optical parametric chirped-pulse amplifier for close-to-optimal operation, significantly increasing both the gain bandwidth and the conversion efficiency, in addition to boosting gain for high-repetition-rate amplification. Applied to a degenerate amplifier, the technique can provide an octave-spanning gain bandwidth.

  15. Amplification of postwildfire peak flow by debris

    USGS Publications Warehouse

    Kean, Jason W.; Mcguire, Luke; Rengers, Francis; Smith, Joel B.; Staley, Dennis M.

    2016-01-01

    In burned steeplands, the peak depth and discharge of postwildfire runoff can substantially increase from the addition of debris. Yet methods to estimate the increase over water flow are lacking. We quantified the potential amplification of peak stage and discharge using video observations of postwildfire runoff, compiled data on postwildfire peak flow (Qp), and a physically based model. Comparison of flood and debris flow data with similar distributions in drainage area (A) and rainfall intensity (I) showed that the median runoff coefficient (C = Qp/AI) of debris flows is 50 times greater than that of floods. The striking increase in Qp can be explained using a fully predictive model that describes the additional flow resistance caused by the emergence of coarse-grained surge fronts. The model provides estimates of the amplification of peak depth, discharge, and shear stress needed for assessing postwildfire hazards and constraining models of bedrock incision.

  16. Amplification of postwildfire peak flow by debris

    NASA Astrophysics Data System (ADS)

    Kean, J. W.; McGuire, L. A.; Rengers, F. K.; Smith, J. B.; Staley, D. M.

    2016-08-01

    In burned steeplands, the peak depth and discharge of postwildfire runoff can substantially increase from the addition of debris. Yet methods to estimate the increase over water flow are lacking. We quantified the potential amplification of peak stage and discharge using video observations of postwildfire runoff, compiled data on postwildfire peak flow (Qp), and a physically based model. Comparison of flood and debris flow data with similar distributions in drainage area (A) and rainfall intensity (I) showed that the median runoff coefficient (C = Qp/AI) of debris flows is 50 times greater than that of floods. The striking increase in Qp can be explained using a fully predictive model that describes the additional flow resistance caused by the emergence of coarse-grained surge fronts. The model provides estimates of the amplification of peak depth, discharge, and shear stress needed for assessing postwildfire hazards and constraining models of bedrock incision.

  17. Gene amplification in thyroid cancer: A new mechanism defined by comparative genomic hybridization

    SciTech Connect

    Chen, X.N.; Lai, E.; Fagin, J.A.

    1994-09-01

    More than 12,000 new cases of thyroid cancer develop each year in the USA for which scant information is available on cytogenetic abnormalities. Comparative genomic hybridization (CGH) now permits the detection and mapping of amplified regions in DNAs without the need for metaphase tumor preparations. Using CGH, we have analyzed the DNA copy number changes in 10 human thyroid tumors. Further, the procedure of CGH was modified by using a chromomycin and sitamycin reverse-banding technique to provide a high resolution assignment of amplified regions. The results revealed a series of copy number changes including amplifications of chromosome bands 1p36, 1q42, 2p13, 2p21, 19q13.1 and losses of chromosome bands 16q12-13 and 16q22-23. The most striking regions of amplifications, band 2p21 and 2p13 were further analyzed by constructing a 90-member chromosome 2 BAC map and using 9 members of this to analyze WRO, a thyroid follicular carcinoma cell line containing double minutes (dm). Eight BACs revealed three copies of chromosome 2; one BAC mapping at the 2p21-22 border revealed significant and variable amplification in all of the metaphase dm`s suggesting a local amplification. Interphase analyses revealed about 40-70 signals per nucleus. The variable signals seen in the WRO cell population suggest the existence of a minor population with higher copy number and smaller amplicon size. In summary, high resolution CGH has been combined with BAC contig construction for the analysis of thyroid tumor to reveal a very specific region amplification on chromosome 2p21 likely to contain a new gene involved in thyroid cancer tumorigenesis.

  18. A model for amplification of hair-bundle motion by cyclical binding of Ca2+ to mechanoelectrical-transduction channels

    PubMed Central

    Choe, Yong; Magnasco, Marcelo O.; Hudspeth, A. J.

    1998-01-01

    Amplification of auditory stimuli by hair cells augments the sensitivity of the vertebrate inner ear. Cell-body contractions of outer hair cells are thought to mediate amplification in the mammalian cochlea. In vertebrates that lack these cells, and perhaps in mammals as well, active movements of hair bundles may underlie amplification. We have evaluated a mathematical model in which amplification stems from the activity of mechanoelectrical-transduction channels. The intracellular binding of Ca2+ to channels is posited to promote their closure, which increases the tension in gating springs and exerts a negative force on the hair bundle. By enhancing bundle motion, this force partially compensates for viscous damping by cochlear fluids. Linear stability analysis of a six-state kinetic model reveals Hopf bifurcations for parameter values in the physiological range. These bifurcations signal conditions under which the system’s behavior changes from a damped oscillatory response to spontaneous limit-cycle oscillation. By varying the number of stereocilia in a bundle and the rate constant for Ca2+ binding, we calculate bifurcation frequencies spanning the observed range of auditory sensitivity for a representative receptor organ, the chicken’s cochlea. Simulations using prebifurcation parameter values demonstrate frequency-selective amplification with a striking compressive nonlinearity. Because transduction channels occur universally in hair cells, this active-channel model describes a mechanism of auditory amplification potentially applicable across species and hair-cell types. PMID:9860967

  19. Quantum amplification effect in a horizon fluctuation

    SciTech Connect

    Ansari, Mohammad H.

    2010-05-15

    The appearance of a few unevenly spaced bright flashes of light on top of Hawking radiation is the sign of the amplification effect in black hole horizon fluctuations. Previous studies on this problem suffer from the lack of considering all emitted photons in the theoretical spectroscopy of these fluctuations. In this paper, we include all of the physical transition weights and present a consistent intensity formula. This modifies a black hole radiation pattern.

  20. Hormonal Involvement in Breast Cancer Gene Amplification

    DTIC Science & Technology

    2008-10-01

    re-replication creates extra copies of the gene. This in turn will also increase production of the protein encoded by the amplified gene. Hormonal... increases in MCM proteins and Cdt1 have been shown to induce DNA amplification in yeast (Gopalakrishnan et al., 2001; Nguyen et al., 2001; Green et al...2006) and increased Cdt1 results in re-replication in human cells (Dorn et al., 2008). The N- terminus of Cdt1 is important for re-replication