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Sample records for achieve signal amplification

  1. Thermoacoustic amplification of photoacoustic signal

    NASA Astrophysics Data System (ADS)

    Bijnen, F. G. C.; Dongen, J. v.; Reuss, J.; Harren, F. J. M.

    1996-06-01

    The thermoacoustic effect is used to amplify the photoacoustic signal induced by trace gas absorption of CO2 laser radiation. The acoustic wave pattern in a thermoacoustic amplifier coupled to a photoacoustic cell is represented in terms of electric transmission lines. Predictions of this model have resulted in a prototype thermoacoustic-photoacoustic (TAPA) detector to get a better understanding of this combination. The photoacoustic signal strength of the TAPA cell was linear with the trace gas density in the cell. Within this study we observed for the TAPA cell a higher PA signal than generated by a normal PA cell. Design criteria for better thermoacoustic amplification of photoacoustic signal are discussed.

  2. 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

  3. Parametric Amplification For Detecting Weak Optical Signals

    NASA Technical Reports Server (NTRS)

    Hemmati, Hamid; Chen, Chien; Chakravarthi, Prakash

    1996-01-01

    Optical-communication receivers of proposed type implement high-sensitivity scheme of optical parametric amplification followed by direct detection for reception of extremely weak signals. Incorporates both optical parametric amplification and direct detection into optimized design enhancing effective signal-to-noise ratios during reception in photon-starved (photon-counting) regime. Eliminates need for complexity of heterodyne detection scheme and partly overcomes limitations imposed on older direct-detection schemes by noise generated in receivers and by limits on quantum efficiencies of photodetectors.

  4. Optically Controlled Signal Amplification for DNA Computation.

    PubMed

    Prokup, Alexander; Hemphill, James; Liu, Qingyang; Deiters, Alexander

    2015-10-16

    The hybridization chain reaction (HCR) and fuel-catalyst cycles have been applied to address the problem of signal amplification in DNA-based computation circuits. While they function efficiently, these signal amplifiers cannot be switched ON or OFF quickly and noninvasively. To overcome these limitations, a light-activated initiator strand for the HCR, which enabled fast optical OFF → ON switching, was developed. Similarly, when a light-activated version of the catalyst strand or the inhibitor strand of a fuel-catalyst cycle was applied, the cycle could be optically switched from OFF → ON or ON → OFF, respectively. To move the capabilities of these devices beyond solution-based operations, the components were embedded in agarose gels. Irradiation with customizable light patterns and at different time points demonstrated both spatial and temporal control. The addition of a translator gate enabled a spatially activated signal to travel along a predefined path, akin to a chemical wire. Overall, the addition of small light-cleavable photocaging groups to DNA signal amplification circuits enabled conditional control as well as fast photocontrol of signal amplification. PMID:25621535

  5. 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

  6. Enzymatic signal amplification of molecular beacons for sensitive DNA detection.

    PubMed

    Li, Jianwei Jeffery; Chu, Yizhuo; Lee, Benjamin Yi-Hung; Xie, Xiaoliang Sunney

    2008-04-01

    Molecular beacons represent a new family of fluorescent probes for nucleic acids, and have found broad applications in recent years due to their unique advantages over traditional probes. Detection of nucleic acids using molecular beacons has been based on hybridization between target molecules and molecular beacons in a 1:1 stoichiometric ratio. The stoichiometric hybridization, however, puts an intrinsic limitation on detection sensitivity, because one target molecule converts only one beacon molecule to its fluorescent form. To increase the detection sensitivity, a conventional strategy has been target amplification through polymerase chain reaction. Instead of target amplification, here we introduce a scheme of signal amplification, nicking enzyme signal amplification, to increase the detection sensitivity of molecular beacons. The mechanism of the signal amplification lies in target-dependent cleavage of molecular beacons by a DNA nicking enzyme, through which one target DNA can open many beacon molecules, giving rise to amplification of fluorescent signal. Our results indicate that one target DNA leads to cleavage of hundreds of beacon molecules, increasing detection sensitivity by nearly three orders of magnitude. We designed two versions of signal amplification. The basic version, though simple, requires that nicking enzyme recognition sequence be present in the target DNA. The extended version allows detection of target of any sequence by incorporating rolling circle amplification. Moreover, the extended version provides one additional level of signal amplification, bringing the detection limit down to tens of femtomolar, nearly five orders of magnitude lower than that of conventional hybridization assay.

  7. 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.

  8. Signal amplification on microarrays: techniques and advances in tyramide signal amplification (TSA)

    NASA Astrophysics Data System (ADS)

    Adler, Karl E.; Tyler, Mary C.; Mikulskis, Alvydas; O'Malley, Mike; Broadbent, Jeff J.; Golenko, Eva E.; Johnson, Andy L.; Lott, Steve; Khimani, Anis H.; Bobrow, Mark N.

    2001-06-01

    Increased sensitivity for differential mRNA expression analysis on microarrays is rapidly becoming a serious need as the technology matures. Current techniques using direct cyanine labeled targets are effective for expression analysis of abundant mRNA sources but have limited utility for analysis where mRNA quantities are limited. Tyramide signal amplification (TSATM) applied to microarray detection provides dramatic improvements in sensitivity, allowing the reduction of sample sizes by as much as 200-fold. The technique includes hapten labeling of two separate RNA populations, microarray hybridization and detection of each hapten with sequential signal amplification steps. The system uses fluorescein and biotin nucleotide analogs as the hapten pair. Hybridized fluorescein and biotin labeled targets are sequentially reacted with horseradish peroxidase and cyanine 3 and cyanine 5 tyramides, resulting in the numerous depositions of these fluorophors on the array. Differential gene expression analysis of LNCaP and PC3 prostate cancer cell lines using one microgram of total RNA and TSA detection, indicates good correlation with results obtained starting with 100 micrograms ((mu) g) of total RNA in a conventional cyanine 3 and cyanine 5 nucleotide analog labeling and detection system (i.e., the direct method).

  9. Reversible Gating of Plasmonic Coupling for Optical Signal Amplification.

    PubMed

    Khoury, Christopher G; Fales, Andrew M; Vo-Dinh, Tuan

    2016-07-20

    Amplification of optical signals is useful for a wide variety of applications, ranging from data signal transmission to chemical sensing and biomedical diagnostics. One such application in chemical sensing is surface-enhanced Raman scattering (SERS), an important technique for increasing the Raman signal using the plasmonic effect of enhanced electromagnetic fields associated with metallic nanostructures. One of the most important limitations of SERS-based amplification is the difficulty to reproducibly control the SERS signal. Here, we describe the design and implementation of a unique hybrid system capable of producing reversible gating of plasmonic coupling for Raman signal amplification. The hybrid system is composed of two subsystems: (1) colloidal magneto-plasmonic nanoparticles for SERS enhancement and (2) a micromagnet substrate with an externally applied magnetic field to modulate the colloidal nanoparticles. For this proof of concept demonstration, the nanoparticles were labeled with a Raman-active dye, and it was shown that the detected SERS signal could be reproducibly modulated by controlling the externally applied magnetic field. The developed system provides a simple, robust, inexpensive, and reusable device for SERS signal modulation. These properties will open up new possibilities for optical signal amplification and gating as well for high-throughput, reproducible SERS detection.

  10. Analytical model of signal amplification in silicon waveguides

    NASA Astrophysics Data System (ADS)

    Meng, Fan; Yu, Chong-Xiu; Yuan, Jin-Hui

    2012-07-01

    In this paper, an analytical model to investigate the parametric amplification (PA) and the PA + stimulated Raman scattering (SRS) in silicon waveguides is put forward. When two pump signals are employed, the PA bandwidth of the probe signal is so large that the Raman contribution has to be considered. When Raman contribution fraction f is set to be 0, only the PA occurs to amplify the probe signal, and when f is set to be 0.043, the PA and the SRS amplify the probe signal at the same time. The signal amplifications of both single and dual pump schemes are investigated by using this model. With this model, three main affecting factors, i.e., zero dispersion wavelength (ZDWL), third-order dispersion (TOD), and fourth-order dispersion (FOD), are discussed in detail.

  11. Introduction to parametric amplification of quantum signals with Josephson circuits

    NASA Astrophysics Data System (ADS)

    Roy, Ananda; Devoret, Michel

    2016-08-01

    This short and opinionated review starts with the concept of quantum signals at microwave frequencies and focuses on the principle of linear parametric amplification. This process emerges from the dispersive nonlinearity of Josephson junctions driven with appropriate tones. We discuss two defining characteristics of the corresponding amplifying devices: i) the number of modes excited by the signal, idler and pump waves and ii) the number of independent ports through which these waves enter into the circuit.

  12. DNAzyme amplification of molecular beacon signal.

    PubMed

    Tian, Ye; Mao, Chengde

    2005-09-15

    This paper reports an improved catalytic molecular beacon. Addition of the target oligonucleotide activates a DNA enzyme (DNAzyme), which, in turn, activates multiple copies of molecular beacons (MB) and gives rise to a strong fluorescence signal. In a previous design, the activated DNAzyme could oligomerize, especially dimerize, and result in inactivation of the DNAzyme. The current design avoids this problem, upon activated by the target DNA, the DNAzyme will stay constantly active. With the improved method, a detection of 10pM DNA has been demonstrated, which is 1000 times more sensitive than the method previously reported.

  13. 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.

  14. 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.

  15. 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.

  16. 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. PMID:27209276

  17. Cascade Signal Amplification Based on Copper Nanoparticle-Reported Rolling Circle Amplification for Ultrasensitive Electrochemical Detection of the Prostate Cancer Biomarker.

    PubMed

    Zhu, Ye; Wang, Huijuan; Wang, Lin; Zhu, Jing; Jiang, Wei

    2016-02-01

    An ultrasensitive and highly selective electrochemical assay was first attempted by combining the rolling circle amplification (RCA) reaction with poly(thymine)-templated copper nanoparticles (CuNPs) for cascade signal amplification. As proof of concept, prostate specific antigen (PSA) was selected as a model target. Using a gold nanoparticle (AuNP) as a carrier, we synthesized the primer-AuNP-aptamer bioconjugate for signal amplification by increasing the primer/aptamer ratio. The specific construction of primer-AuNP-aptamer/PSA/anti-PSA sandwich structure triggered the effective RCA reaction, in which thousands of tandem poly(thymine) repeats were generated and directly served as the specific templates for the subsequent CuNP formation. The signal readout was easily achieved by dissolving the RCA product-templated CuNPs and detecting the released copper ions with differential pulse stripping voltammetry. Because of the designed cascade signal amplification strategy, the newly developed method achieved a linear range of 0.05-500 fg/mL, with a remarkable detection limit of 0.020 ± 0.001 fg/mL PSA. Finally, the feasibility of the developed method for practical application was investigated by analyzing PSA in the real clinical human serum samples. The ultrasensitivity, specificity, convenience, and capability for analyzing the clinical samples demonstrate that this method has great potential for practical disease diagnosis applications. PMID:26765624

  18. 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. PMID:26592629

  19. Optomechanical correlations and signal self-amplification in interferometric measurements

    NASA Astrophysics Data System (ADS)

    Cohadon, P.-F.; Verlot, P.; Tavernarakis, A.; Briant, T.; Heidmann, A.

    2010-05-01

    Radiation pressure exerted by light in interferometric measurements is responsible for displacements of mirrors which appear as an additional back-action noise and limit the sensitivity of the measurement. We experimentally study these effects by monitoring in a very high-finesse optical cavity the displacements of a mirror with a sensitivity at the 10-20 m/ level. This very high sensitivity is a step towards the observation of fundamental quantum effects of radiation pressure such as the standard quantum limit in interferometric measurements. We report the observation of optomechanical correlations between two optical beams sent into the same moving mirror cavity. We also observed a self-amplification of a signal, which is a consequence of dynamical back-action of radiation pressure in a detuned cavity, and may improve the interferometric measurement sensitivity beyond the standard quantum limit.

  20. Liposomes with High Refractive Index Encapsulants as Tunable Signal Amplification Tools in Surface Plasmon Resonance Spectroscopy.

    PubMed

    Fenzl, Christoph; Hirsch, Thomas; Baeumner, Antje J

    2015-11-01

    One major goal in the surface plasmon resonance (SPR) technique is the reliable detection of small molecules as well as low analyte concentrations. This can be achieved by a viable signal amplification strategy. We therefore investigated optimal liposome characteristics for use as a signal enhancement system for SPR sensors, as liposomes excel not only at versatility but also at colloidal stability and ease of functionalization. These characteristics include the encapsulation of high refractive index markers, lipid composition, liposome size, and surface modifications to best match the requirements of the SPR system. Our studies of the binding of biotinylated liposomes to surface-immobilized streptavidin show that the refractive index of the encapsulant has a major influence on the SPR signal and outweighs the influence of the thin lipid bilayer. Thus, the signal amplification properties of liposomes can be adjusted to the respective needs of any analytical task by simply exchanging the encapsulant solution. In this work, a maximum enhancement factor of 23 was achieved by encapsulating a 500 mM sucrose solution. Dose-response studies with and without liposome enhancement revealed an improvement of the limit of detection from 10 nmol L(-1) to 320 pmol L(-1) streptavidin concentration with a much higher sensitivity of 3 mRIU per logarithmic unit of the concentration between 500 pmol L(-1) and 10 nmol L(-1). PMID:26455696

  1. Sensitive SERS detection of DNA methyltransferase by target triggering primer generation-based multiple signal amplification strategy.

    PubMed

    Li, Ying; Yu, Chuanfeng; Han, Huixia; Zhao, Caisheng; Zhang, Xiaoru

    2016-07-15

    A novel and sensitive surface-enhanced Raman scattering (SERS) method is proposed for the assay of DNA methyltransferase (MTase) activity and evaluation of inhibitors by developing a target triggering primer generation-based multiple signal amplification strategy. By using of a duplex substrate for Dam MTase, two hairpin templates and a Raman probe, multiple signal amplification mode is achieved. Once recognized by Dam MTase, the duplex substrate can be cleaved by Dpn I endonuclease and two primers are released for triggering the multiple signal amplification reaction. Consequently, a wide dynamic range and remarkably high sensitivity are obtained under isothermal conditions. The detection limit is 2.57×10(-4)UmL(-1). This assay exhibits an excellent selectivity and is successfully applied in the screening of inhibitors for Dam MTase. In addition, this novel sensing system is potentially universal as the recognition element can be conveniently designed for other target analytes by changing the substrate of DNA MTase.

  2. Amplification and Damping of Environmental Signals in Intensively Managed Landscapes

    NASA Astrophysics Data System (ADS)

    Belmont, P.; Kumarasamy, K.; Kelly, S. A.; Vaughan, A. A.; Call, B.

    2015-12-01

    Landscapes transmit pulses of water, sediment and solutes through the terrestrial environment and network of stream channels. The timing, frequency and magnitude of these pulses depend on inputs of water, energy and rock/soil as well as a multitude of critical zone processes that may modulate the signal. Therefore, the potential for a landscape to generate and transmit these pulses changes over long timescales, primarily as a function of climate and local base level rise or fall. Humans have profoundly altered many critical zone processes that govern these environmental signals, often targeting rate-limiting processes for the purpose of enhancing economic productivity and/or reducing financial risk. These alterations are especially evident in the intensively managed landscapes of the Midwestern US, where vegetation change, soil tillage, and pervasive modifications of the surface and subsurface drainage system have substantially changed water, sediment and nutrient fluxes. Effects, in terms of amplification or damping of environmental signals, are strongly dependent on landscape setting, and often non-linear. Hysteresis and sensitivity may hinder the return to the initial state when perturbations have reached a certain threshold. We draw from multiple examples in the upper Midwestern US to illustrate, at a basic level, the mechanisms by which landscape evolution establishes the template for generation and transmission of environmental signals and furthermore how humans have altered critical zone processes to optimize specific landscape outputs, often at the expense of modifying the rate and/or magnitude of many other landscape outputs. We present a conceptual model and discuss implications for mathematical modeling of water, sediment and nutrient fluxes to guide watershed management and restoration.

  3. Invasive reaction assisted strand-displacement signal amplification for sensitive DNA detection.

    PubMed

    Zou, Bingjie; Song, Qinxin; Wang, Jianping; Liu, Yunlong; Zhou, Guohua

    2014-11-18

    A novel DNA detection assay was proposed by invasive reaction coupled with molecular beacon assisted strand-displacement signal amplification (IRASA). Target DNAs are firstly hybridized to two probes to initiate invasive reaction to produce amplified flaps. Then these flaps are further amplified by strand-displacement signal amplification. The detection limit was around 0.2 pM.

  4. Nanoparticle-based signal generation and amplification in microfluidic devices for bioanalysis.

    PubMed

    Hu, Chong; Yue, Wanqing; Yang, Mengsu

    2013-11-21

    Signal generation and amplification based on nanomaterials and microfluidic techniques have both attracted considerable attention separately due to the demands for ultrasensitive and high-throughput detection of biomolecules. This article reviews the latest development of signal amplification strategies based on nanoparticles for bioanalysis and their integration and applications in microfluidic systems. The applications of nanoparticles in bioanalysis were categorized based on the different approaches of signal amplification, and the microfluidic techniques were summarized based on cell analysis and biomolecule detection with a focus on the integration of nanoparticle-based amplification in microfluidic devices for ultrasensitive bioanalysis. The advantages and limitations of the combination of nanoparticles-based amplification with microfluidic techniques were evaluated, and the possible developments for future research were discussed.

  5. 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.

  6. Ultrasensitive detection of uranyl by graphene oxide-based background reduction and RCDzyme-based enzyme strand recycling signal amplification.

    PubMed

    Li, Ming-Hui; Wang, Yong-Sheng; Cao, Jin-Xiu; Chen, Si-Han; Tang, Xian; Wang, Xiao-Feng; Zhu, Yu-Feng; Huang, Yan-Qin

    2015-10-15

    We proposed a novel strategy which combines graphene oxide-based background reduction with RCDzyme-based enzyme strand recycling amplification for ultrahigh sensitive detection of uranyl. The RCDzyme is designed to contain a guanine (G)-rich sequence that replaces the partial sequence in an uranyl-specific DNAzyme. This multifunctional probe can act as the target recognition element, DNAzyme and the primer of signal amplification. The presence of UO2(2+) can induce the cleavage of the substrate strands in RCDzyme. Then, each released enzyme strand can hybridize with another substrate strands to trigger many cycles of the cleavage by binding uranyl, leading to the formation of more G-quadruplexes by split guanine-rich oligonucleotide fragments. The resulting G-quadruplexes could bind to N-methyl-mesoporphyrin IX (NMM), causing an amplified detection signal for the target uranyl. Next, graphene oxide-based background reduction strategy was further employed for adsorbing free ssDNA and NMM, thereby providing a proximalis zero-background signal. The combination of RCDzyme signal amplification and proximalis zero-background signal remarkably improves the sensitivity of this method, achieving a dynamic range of two orders of magnitude and giving a detection limit down to 86 pM, which is much lower than those of related literature reports. These achievements might be helpful in the design of highly sensitive analytical platform for wide applications in environmental and biomedical fields. PMID:26000462

  7. Enzyme-free signal amplification for electrochemical detection of Mycobacterium lipoarabinomannan antibody on a disposable chip.

    PubMed

    Wang, Lisong; Leng, Chuan; Tang, Sheng; Lei, Jianping; Ju, Huangxian

    2012-01-01

    A simple, rapid, and disposable immunosensor at screen printed carbon electrode (SPCE) was developed by using gold nanoparticles (AuNPs) labeled Staphylococcal protein A (Au-SPA) as the electrochemical tag for detection of lipoarabinomannan antibody (anti-LAM). The immunosensor as the disposable chip was prepared by immobilizing capture antigen on screen printed carbon working electrode by passive adsorption, and characterized with scanning electron microscopy. After binding with the anti-LAM for further capture of Au-SPA, AuNPs were introduced as an electrochemical tag by the eletrooxidation of AuNPs in 0.1M HCl to produce strong electroactive substance for signal amplification. Compared with the enzyme-based immunosensor, AuNPs as enzyme-free tag for signal amplification exhibited many advantages such as no requirement of deoxygenation, and high stability. Under optimal detection conditions and at a preoxidation potential of +1.3 V for 30s, this method achieved the linear concentration of anti-LAM from 15.6 to 1000 ng mL(-1) with a detection limit of 5.3 ng mL(-1). The immunosensor showed a good performance with high selectivity, acceptable stability, and simple operation, providing a promising application as an adjunctive tool in early tuberculosis diagnosis. PMID:22709935

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

    PubMed Central

    Ang, Yan Shan; Yung, Lin-Yue Lanry

    2014-01-01

    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. PMID:25056307

  9. E2F Activators Signal and Maintain Centrosome Amplification in Breast Cancer Cells

    PubMed Central

    Lee, Mi-Young; Moreno, Carlos S.

    2014-01-01

    Centrosomes ensure accurate chromosome segregation by directing spindle bipolarity. Loss of centrosome regulation results in centrosome amplification, multipolar mitosis and aneuploidy. Since centrosome amplification is common in premalignant lesions and breast tumors, it is proposed to play a central role in breast tumorigenesis, a hypothesis that remains to be tested. The coordination between the cell and centrosome cycles is of paramount importance to maintain normal centrosome numbers, and the E2Fs may be responsible for regulating these cycles. However, the role of E2F activators in centrosome amplification is unclear. Because E2Fs are deregulated in Her2+ cells displaying centrosome amplification, we addressed whether they signal this abnormal process. Knockdown of E2F1 or E2F3 in Her2+ cells decreased centrosome amplification without significantly affecting cell cycle progression, whereas the overexpression of E2F1, E2F2, or E2F3 increased centrosome amplification in MCF10A mammary epithelial cells. Our results revealed that E2Fs affect the expression of proteins, including Nek2 and Plk4, known to influence the cell/centrosome cycles and mitosis. Downregulation of E2F3 resulted in cell death and delays/blocks in cytokinesis, which was reversed by Nek2 overexpression. Nek2 overexpression enhanced centrosome amplification in Her2+ breast cancer cells silenced for E2F3, revealing a role for the E2F activators in maintaining centrosome amplification in part through Nek2. PMID:24797070

  10. 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.

  11. 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. PMID:26274239

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

    NASA Astrophysics Data System (ADS)

    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.

  13. Combined tyramide signal amplification and quantum dots for sensitive and photostable immunofluorescence detection.

    PubMed

    Ness, Jayne M; Akhtar, Rizwan S; Latham, Cecelia B; Roth, Kevin A

    2003-08-01

    Conventional immunofluorescence detection of biologically relevant proteins and antigens in tissue sections is often limited by relatively weak signals that fade rapidly on illumination. We have developed an immunohistochemical protocol that combines the sensitivity of tyramide signal amplification with the photostability of quantum dots to overcome these limitations. This simple method provides a sensitive and stable fluorescence immunohistochemical alternative to standard chromogen detection.

  14. Sensitive SERS detection of DNA methyltransferase by target triggering primer generation-based multiple signal amplification strategy.

    PubMed

    Li, Ying; Yu, Chuanfeng; Han, Huixia; Zhao, Caisheng; Zhang, Xiaoru

    2016-07-15

    A novel and sensitive surface-enhanced Raman scattering (SERS) method is proposed for the assay of DNA methyltransferase (MTase) activity and evaluation of inhibitors by developing a target triggering primer generation-based multiple signal amplification strategy. By using of a duplex substrate for Dam MTase, two hairpin templates and a Raman probe, multiple signal amplification mode is achieved. Once recognized by Dam MTase, the duplex substrate can be cleaved by Dpn I endonuclease and two primers are released for triggering the multiple signal amplification reaction. Consequently, a wide dynamic range and remarkably high sensitivity are obtained under isothermal conditions. The detection limit is 2.57×10(-4)UmL(-1). This assay exhibits an excellent selectivity and is successfully applied in the screening of inhibitors for Dam MTase. In addition, this novel sensing system is potentially universal as the recognition element can be conveniently designed for other target analytes by changing the substrate of DNA MTase. PMID:26926592

  15. Low noise amplification of an optically carried microwave signal: application to atom interferometry

    NASA Astrophysics Data System (ADS)

    Lévèque, T.; Gauguet, A.; Chaibi, W.; Landragin, A.

    2010-12-01

    In this paper, we report a new scheme to amplify a microwave signal carried on a laser light at λ=852 nm. The amplification is done via a semiconductor tapered amplifier and this scheme is used to drive stimulated Raman transitions in an atom interferometer. Sideband generation in the amplifier, due to self-phase and amplitude modulation, is investigated and characterized. We also demonstrate that the amplifier does not induce any significant phase-noise on the beating signal. Finally, the degradation of the performances of the interferometer due to the amplification process is shown to be negligible.

  16. 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

  17. An immunochromatographic biosensor combined with a water-swellable polymer for automatic signal generation or amplification.

    PubMed

    Kim, Kahee; Joung, Hyou-Arm; Han, Gyeo-Re; Kim, Min-Gon

    2016-11-15

    An immunochromatographic assay (ICA) strip is one of the most widely used platforms in the field of point-of-care biosensors for the detection of various analytes in a simple, fast, and inexpensive manner. Currently, several approaches for sequential reactions in ICA platforms have improved their usability, sensitivity, and versatility. In this study, a new, simple, and low-cost approach using automatic sequential-reaction ICA strip is described. The automatic switching of a reagent pad from separation to attachment to the test membrane was achieved using a water-swellable polymer. The reagent pad was dried with an enzyme substrate for signal generation or with signal-enhancing materials. The strip design and system operation were confirmed by the characterization of the raw materials and flow analysis. We demonstrated the operation of the proposed sensor by using various chemical reaction-based assays, including metal-ion amplification, enzyme-colorimetric reaction, and enzyme-catalyzed chemiluminescence. Furthermore, by employing C-reactive protein as a model, we successfully demonstrated that the new water-swellable polymer-based ICA sensor can be utilized to detect biologically relevant analytes in human serum. PMID:27203463

  18. An immunochromatographic biosensor combined with a water-swellable polymer for automatic signal generation or amplification.

    PubMed

    Kim, Kahee; Joung, Hyou-Arm; Han, Gyeo-Re; Kim, Min-Gon

    2016-11-15

    An immunochromatographic assay (ICA) strip is one of the most widely used platforms in the field of point-of-care biosensors for the detection of various analytes in a simple, fast, and inexpensive manner. Currently, several approaches for sequential reactions in ICA platforms have improved their usability, sensitivity, and versatility. In this study, a new, simple, and low-cost approach using automatic sequential-reaction ICA strip is described. The automatic switching of a reagent pad from separation to attachment to the test membrane was achieved using a water-swellable polymer. The reagent pad was dried with an enzyme substrate for signal generation or with signal-enhancing materials. The strip design and system operation were confirmed by the characterization of the raw materials and flow analysis. We demonstrated the operation of the proposed sensor by using various chemical reaction-based assays, including metal-ion amplification, enzyme-colorimetric reaction, and enzyme-catalyzed chemiluminescence. Furthermore, by employing C-reactive protein as a model, we successfully demonstrated that the new water-swellable polymer-based ICA sensor can be utilized to detect biologically relevant analytes in human serum.

  19. 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.

  20. 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.

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

    NASA Astrophysics Data System (ADS)

    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.

  2. 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. PMID:27176316

  3. 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.

  4. Quinone Methide Signal Amplification: Covalent Reporter Labeling of Cancer Epitopes using Alkaline Phosphatase Substrates.

    PubMed

    Polaske, Nathan W; Kelly, Brian D; Ashworth-Sharpe, Julia; Bieniarz, Christopher

    2016-03-16

    Diagnostic assays with the sensitivity required to improve cancer therapeutics depend on the development of new signal amplification technologies. Herein, we report the development and application of a novel amplification system which utilizes latent quinone methides (QMs) activated by alkaline phosphatase (AP) for signal amplification in solid-phase immunohistochemical (IHC) assays. Phosphate-protected QM precursor substrates were prepared and conjugated to either biotin or a fluorophore through an amine-functionalized linker group. Upon reaction with AP, the phosphate group is cleaved, followed by elimination of the leaving group and formation of the highly reactive and short-lived QM. The QMs either react with tissue nucleophiles in close proximity to their site of generation, or are quenched by nucleophiles in the reaction media. The reporter molecules that covalently bind to the tissue were then detected visually by fluorescence microscopy in the case of fluorophore reporters, or brightfield microscopy using diaminobenzidine (DAB) in the case of biotin reporters. With multiple reporters deposited per enzyme, significant signal amplification was observed utilizing QM precursor substrates containing either benzyl difluoro or benzyl monofluoro leaving group functionalities. However, the benzyl monofluoro leaving group gave superior results with respect to both signal intensity and discretion, the latter of which was found to be imperative for use in diagnostic IHC assays. PMID:26731201

  5. 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...

  6. Signal amplification of streptavidin-horseradish peroxidase functionalized carbon nanotubes for amperometric detection of attomolar DNA.

    PubMed

    Gao, Wenchao; Dong, Haifeng; Lei, Jianping; Ji, Hanxu; Ju, Huangxian

    2011-05-14

    A novel biosensing strategy for selective electrochemical detection of DNA down to the attomolar level with a linear range of 5 orders of magnitude was developed by the specific recognitions of target DNA and streptavidin to biotin labelled molecular beacon and signal amplification of streptavidin-horseradish peroxidase functionalized carbon nanotubes.

  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. PMID:23112677

  8. A Fast and Sensitive Quantitative Lateral Flow Immunoassay for Cry1Ab Based on a Novel Signal Amplification Conjugate

    PubMed Central

    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. PMID:23112677

  9. Development of a near-infrared fluorescence ELISA method using tyramide signal amplification.

    PubMed

    Gong, Haibiao; Cradduck, Mark; Cheung, Lael; Olive, D Michael

    2012-07-01

    In this study, we applied tyramide signal amplification (TSA) to fluorescence enzyme-linked immunosorbent assay (ELISA) employing horseradish peroxidase (HRP) as the detection enzyme. When used with a human epidermal growth factor ELISA kit, the TSA method led to a >100-fold increase in fluorescence signal intensity in comparison to an unamplified method. It also showed wider dynamic range and better sensitivity compared to a conventional method using tetramethylbenzidine as the HRP substrate. PMID:22490466

  10. 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.

  11. 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.

  12. Increased sensitivity of HIV-1 p24 ELISA using a photochemical signal amplification system

    PubMed Central

    Bystryak, Simon; Santockyte, Rasa

    2016-01-01

    In this study we describe a photochemical signal amplification method (PSAM) for increasing of the sensitivity of enzyme-linked immunosorbent assay (ELISA) for determination of HIV-1 p24 antigen. This method can be used for both commercially available and in-house ELISA tests, and has the advantage of being considerably simpler and less costly than alternative signal amplification methods. The photochemical signal amplification method is based on an autocatalytic photochemical reaction of a horseradish peroxidase (HRP) substrate, orthophenylenediamine (OPD). To compare the performance of PSAM-boosted ELISA with a conventional colorimetric ELISA for determination of HIV-1 p24 antigen we employed a PerkinElmer HIV-1 p24 ELISA kit, using conventional ELISA alongside ELISA + PSAM. In the present study, we show that PSAM technology allows one to increase the analytical sensitivity and dynamic range of a commercial HIV-1 p24 ELISA kit, with and without immune-complex disruption (ICD and Non-ICD ELISA), by a factor of approximately 40-fold. ELISA + PSAM is compatible with commercially available microtiter plate readers, requires only an inexpensive illumination device, and the PSAM amplification step takes no longer than 15 min. PMID:26090753

  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. Sensitive detection of multiple mycotoxins by SPRi with gold nanoparticles as signal amplification tags.

    PubMed

    Hu, Weihua; Chen, Hongming; Zhang, Huanhuan; He, Guangli; Li, Xin; Zhang, Xiaoxing; Liu, Yang; Li, Chang Ming

    2014-10-01

    Detection of multiple toxic mycotoxins is of importance in food quality control. Surface plasmon resonance imaging (SPRi) is an advanced tool for simultaneously multiple detections with accuracy; however, it suffers from limited sensitivity due to the instrumental constraint and small sizes of mycotoxins with only one epitope for an insensitive competitive immunoassay. In this work a gold nanoparticle (AuNP)-enhanced SPRi chip is designed to sensitively detect multiple mycotoxins using a competitive immunoassay format. The sensing surface is constructed by uniformly attaching dense mycotoxin antigens on poly[oligo(ethylene glycol) methacrylate-co-glycidyl methacrylate] (POEGMA-co-GMA) brush modified SPRi gold chip. After competitive binding in a sample solution containing respective monoclonal antibodies, secondary antibody-conjugated AuNPs are employed to bind with the captured monoclonal antibodies for further amplification of the SPRi signal. Highly specific and sensitive simultaneous detection is achieved for three typical mycotoxins including Aflatoxin B1 (AFB1), Ochratoxin A (OTA) and Zearalenone (ZEN) with low detection limits of 8, 30 and 15 pg mL(-1) and dynamic ranges covering three orders of magnitude. PMID:24992296

  15. Sensitive detection of multiple mycotoxins by SPRi with gold nanoparticles as signal amplification tags.

    PubMed

    Hu, Weihua; Chen, Hongming; Zhang, Huanhuan; He, Guangli; Li, Xin; Zhang, Xiaoxing; Liu, Yang; Li, Chang Ming

    2014-10-01

    Detection of multiple toxic mycotoxins is of importance in food quality control. Surface plasmon resonance imaging (SPRi) is an advanced tool for simultaneously multiple detections with accuracy; however, it suffers from limited sensitivity due to the instrumental constraint and small sizes of mycotoxins with only one epitope for an insensitive competitive immunoassay. In this work a gold nanoparticle (AuNP)-enhanced SPRi chip is designed to sensitively detect multiple mycotoxins using a competitive immunoassay format. The sensing surface is constructed by uniformly attaching dense mycotoxin antigens on poly[oligo(ethylene glycol) methacrylate-co-glycidyl methacrylate] (POEGMA-co-GMA) brush modified SPRi gold chip. After competitive binding in a sample solution containing respective monoclonal antibodies, secondary antibody-conjugated AuNPs are employed to bind with the captured monoclonal antibodies for further amplification of the SPRi signal. Highly specific and sensitive simultaneous detection is achieved for three typical mycotoxins including Aflatoxin B1 (AFB1), Ochratoxin A (OTA) and Zearalenone (ZEN) with low detection limits of 8, 30 and 15 pg mL(-1) and dynamic ranges covering three orders of magnitude.

  16. 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

  17. SERS assay of telomerase activity at single-cell level and colon cancer tissues via quadratic signal amplification.

    PubMed

    Shi, Muling; Zheng, Jing; Liu, Changhui; Tan, Guixiang; Qing, Zhihe; Yang, Sheng; Yang, Jinfeng; Tan, Yongjun; Yang, Ronghua

    2016-03-15

    As an important biomarker and therapeutic target, telomerase has attracted extensive attention concerning its detection and monitoring. Recently, enzyme-assisted amplification approaches have provided useful platforms for the telomerase activity detection, however, further improvement in sensitivity is still hindered by the single-step signal amplification. Herein, we develop a quadratic signal amplification strategy for ultrasensitive surface-enhanced Raman scattering (SERS) detection of telomerase activity. The central idea of our design is using telomerase-induced silver nanoparticles (AgNPs) assembly and silver ions (Ag(+))-mediated cascade amplification. In our approach, each telomerase-aided DNA sequence extension could trigger the formation of a long double-stranded DNA (dsDNA), making numerous AgNPs assembling along with this long strand through specific Ag-S bond, to form a primary amplification element. For secondary amplification, each conjugated AgNP was dissolved into Ag(+), which can effectively induce the 4-aminobenzenethiol (4-ABT) modified gold nanoparticles (AuNPs@4-ABT) to undergo aggregation to form numerous "hot-spots". Through quadratic amplifications, a limit of detection down to single HeLa cell was achieved. More importantly, this method demonstrated good performance when applied to tissues from colon cancer patients, which exhibits great potential in the practical application of telomerase-based cancer diagnosis in early stages. To demonstrate the potential in screening the telomerase inhibitors and telomerase-targeted drugs, the proposed design is successfully employed to measure the inhibition of telomerase activity by 3'-azido-3'-deoxythymidine.

  18. Asymmetric signal amplification for simultaneous SERS detection of multiple cancer markers with significantly different levels.

    PubMed

    Ye, Sujuan; Wu, Yanying; Zhai, Xiaomo; Tang, Bo

    2015-08-18

    Simultaneous detection of cancer biomarkers holds great promise for the early diagnosis of different cancers. However, in the presence of high-concentration biomarkers, the signals of lower-expression biomarkers are overlapped. Existing techniques are not suitable for simultaneously detecting multiple biomarkers at concentrations with significantly different orders of magnitude. Here, we propose an asymmetric signal amplification method for simultaneously detecting multiple biomarkers with significantly different levels. Using the bifunctional probe, a linear amplification mode responds to high-concentration markers, and quadratic amplification mode responds to low-concentration markers. With the combined biobarcode probe and hybridization chain reaction (HCR) amplification method, the detection limits of microRNA (miRNA) and ATP via surface-enhanced Raman scattering (SERS) detection are 0.15 fM and 20 nM, respectively, with a breakthrough of detection concentration difference over 11 orders of magnitude. Furthermore, successful determination of miRNA and ATP in cancer cells supports the practicability of the assay. This methodology promises to open an exciting new avenue for the detection of various types of biomolecules. PMID:26218034

  19. Signal-to-noise ratio enhancement of silicon nanowires biosensor with rolling circle amplification.

    PubMed

    Gao, Anran; Zou, Nengli; Dai, Pengfei; Lu, Na; Li, Tie; Wang, Yuelin; Zhao, Jianlong; Mao, Hongju

    2013-09-11

    Herein, we describe a novel approach for rapid, label-free and specific DNA detection by applying rolling circle amplification (RCA) based on silicon nanowire field-effect transistor (SiNW-FET) for the first time. Highly responsive SiNWs were fabricated with a complementary metal oxide semiconductor (CMOS) compatible anisotropic self-stop etching technique which eliminated the need for hybrid method. The probe DNA was immobilized on the surface of SiNW, followed by sandwich hybridization with the perfectly matched target DNA and RCA primer that acted as a primer to hybridize the RCA template. The RCA reaction created a long single-stranded DNA (ssDNA) product and thus enhanced the electronic responses of SiNW significantly. The signal-to-noise ratio (SNR) as a figure-of-merit was analyzed to estimate the signal enhancement and possible detection limit. The nanosensor showed highly sensitive concentration-dependent conductance change in response to specific target DNA sequences. Because of the binding of an abundance of repeated sequences of RCA products, the SNR of >20 for 1 fM DNA detection was achieved, implying a detection floor of 50 aM. This RCA-based SiNW biosensor also discriminated perfectly matched target DNA from one-base mismatched DNA with high selectivity due to the substantially reduced nonspecific binding onto the SiNW surface through RCA. The combination of SiNW FET sensor with RCA will increase diagnostic capacity and the ability of laboratories to detect unexpected viruses, making it a potential tool for early diagnosis of gene-related diseases. PMID:23937430

  20. Signal-Switchable Electrochemiluminescence System Coupled with Target Recycling Amplification Strategy for Sensitive Mercury Ion and Mucin 1 Assay.

    PubMed

    Jiang, Xinya; Wang, Huijun; Wang, Haijun; Yuan, Ruo; Chai, Yaqin

    2016-09-20

    In the present work, we first found that mercury ion (Hg(2+)) has an efficient quenching effect on the electrochemiluminescence (ECL) of N-(aminobutyl)-N-(ethylisoluminol) (ABEI). Since we were inspired by this discovery, an aptamer-based ECL sensor was fabricated based on a Hg(2+) triggered signal switch coupled with an exonuclease I (Exo I)-stimulated target recycling amplification strategy for ultrasensitive determination of Hg(2+) and mucin 1 (MUC1). Concretely, the ECL intensity of ABEI-functionalized silver nanoparticles decorated graphene oxide nanocomposite (GO-AgNPs-ABEI) was initially enhanced by ferrocene labeled ssDNA (Fc-S1) (first signal switch "on" state) in the existence of H2O2. With the aid of aptamer, assistant ssDNA (S2) and full thymine (T) bases ssDNA (S3) modified Au nanoparticles (AuNPs-S2-S3) were immobilized on the sensing surface through the hybridization reaction. Then, via the strong and stable T-Hg(2+)-T interaction, an abundance of Hg(2+) was successfully captured on the AuNPs-S2-S3 and effectively inhibited the ECL reaction of ABEI (signal switch "off" state). Finally, the signal switch "on" state was executed by utilizing MUC1 as an aptamer-specific target to bind aptamer, leading to the large decrease of the captured Hg(2+). To further improve the sensitivity of the aptasensor, Exo I was implemented to digest the binded aptamer, which resulted in the release of MUC1 for achieving target recycling with strong detectable ECL signal even in a low level of MUC1. By integrating the quenching effect of Hg(2+) to reduce the background signal and target recycling for signal amplification, this proposed ECL aptasensor was successfully used to detect Hg(2+) and MUC1 sensitively with a wide linear response. PMID:27529728

  1. 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.

  2. 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. PMID:27394946

  3. Hollow PDA-Au nanoparticles-enabled signal amplification for sensitive nonenzymatic colorimetric immunodetection of carbohydrate antigen 125.

    PubMed

    Zhao, Yan; Zheng, Yiqun; Zhao, Caiyun; You, Jinmao; Qu, Fengli

    2015-09-15

    A novel colorimetric immunoassay was designed for the sensitive detection of carbohydrate antigen 125 (CA125). The success of this immunoassay relies on the use of hollow polydopamine-gold nanoparticles (PDA-Au) for signal amplification to achieve sensitive nonenzymatic colorimetric detection. In particular, PDA-Au was used as a stable and sensitive label and aminated-Fe3O4 magnetic nanoparticles (Fe3O4 NPs) were employed to immobilize capture antibody (Ab1) and acted as a separable immunosensing probe. PDA-Au exhibited high catalytic performance towards p-nitrophenol reduction and thus resulted in significant color change and UV/vis signal variations. The immunoassay was performed based on sandwich protocol. As compared to pure Au nanoparticles, the signal amplification and sensitivity of PDA-Au-based assay was significantly improved. For instance, the dynamic range of the developed colorimetric immunoassay for CA125 was 0.1-100 U/mL with a detection limit of 0.1 U/mL at S/N=3. In addition, this immunoassay was also tested for the analysis of clinical serum samples, which demonstrated its potential for practical diagnostic applications.

  4. 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

    2014-03-01

    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.

  5. Tyramide Signal Amplification: Fluorescence In Situ Hybridization for Identifying Homoeologous Chromosomes.

    PubMed

    Fominaya, Araceli; Loarce, Yolanda; González, Juan M; Ferrer, Esther

    2016-01-01

    Tyramide signal amplification (TSA) fluorescence in situ hybridization (FISH) has been shown as a valuable molecular tool for visualizing specific amplified DNA sequences in chromosome preparations. This chapter describes how to perform TSA-FISH, paying special interest to its two critical steps: probe generation and metaphase plate generation. The potential of physically mapping 12S-globulin sequences by TSA-FISH as a means of identifying homeology among chromosome regions of Avena species was tested and is discussed. PMID:27511165

  6. 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.

  7. 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.

  8. Aided Electrophysiology Using Direct Audio Input: Effects of Amplification and Absolute Signal Level

    PubMed Central

    Billings, Curtis J.; Miller, Christi W.; Tremblay, Kelly L.

    2016-01-01

    Purpose This study investigated (a) the effect of amplification on cortical auditory evoked potentials (CAEPs) at different signal levels when signal-to-noise ratios (SNRs) were equated between unaided and aided conditions, and (b) the effect of absolute signal level on aided CAEPs when SNR was held constant. Method CAEPs were recorded from 13 young adults with normal hearing. A 1000-Hz pure tone was presented in unaided and aided conditions with a linear analog hearing aid. Direct audio input was used, allowing recorded hearing aid noise floor to be added to unaided conditions to equate SNRs between conditions. An additional stimulus was created through scaling the noise floor to study the effect of signal level. Results Amplification resulted in delayed N1 and P2 peak latencies relative to the unaided condition. An effect of absolute signal level (when SNR was constant) was present for aided CAEP area measures, such that larger area measures were found at higher levels. Conclusion Results of this study further demonstrate that factors in addition to SNR must also be considered before CAEPs can be used to clinically to measure aided thresholds. PMID:26953543

  9. An electrochemical peptide cleavage-based biosensor for matrix metalloproteinase-2 detection with exonuclease III-assisted cycling signal amplification.

    PubMed

    Wang, Ding; Yuan, Yali; Zheng, Yingning; Chai, Yaqin; Yuan, Ruo

    2016-05-01

    In this work, an electrochemical peptide biosensor was developed for matrix metalloproteinase-2 (MMP-2) detection by conversion of a peptide cleavage event into DNA detection with exonuclease III (Exo III)-assisted cycling signal amplification.

  10. Electrochemiluminescence detection of near single DNA molecules by using quantum dots-dendrimer nanocomposites for signal amplification.

    PubMed

    Divsar, Faten; Ju, Huangxian

    2011-09-21

    An ultrasensitive electrochemiluminescent biosensor was developed for detection of near single DNA molecules with a linear range of 7 orders of magnitude by combining the specific recognition of a molecular beacon with signal amplification of quantum dots-dendrimer nanocomposites.

  11. 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

  12. Hydroquinone-quinone oxidation by molecular oxygen: a simple tool for signal amplification through auto-generation of hydrogen peroxide.

    PubMed

    Sella, Eran; Shabat, Doron

    2013-08-21

    Signal amplification methods are of obvious importance for various diagnostic assays. We have developed a new small-molecule-based probe that, upon activation with sub-stoichiometric amounts of hydrogen peroxide, produces an auto-inductive amplification reaction. The signal is produced through the oxidation reaction of hydroquinone to the corresponding quinone derivative by molecular oxygen. This oxidation is accompanied by the formation of hydrogen peroxide, which can enter the amplification sequence and initiate a new diagnostic cycle. The generated quinone is composed of a donor-acceptor conjugated pair and fluoresces at a distinct wavelength, allowing the formation to be monitored by a convenient fluorescence assay.

  13. Lable-free quadruple signal amplification strategy for sensitive electrochemical p53 gene biosensing.

    PubMed

    Wang, Zonghua; Xia, Jianfei; Song, Daimin; Zhang, Feifei; Yang, Min; Gui, Rijun; Xia, Lin; Bi, Sai; Xia, Yanzhi

    2016-03-15

    A versatile label-free quadruple signal amplification biosensing platform for p53 gene (target DNA) detection was proposed. The chitosan-graphene (CS-GR) modified electrode with excellent electron transfer ability could provide a large specific surface for high levels of AuNPs-DNA attachment. The large amount of AuNPs could immobilize more capture probes and enhance the electrochemical signal with the excellent electrocatalytic activity. Furthermore, with the assist of N.BstNB I (the nicking endonuclease), target DNA could be reused and more G-quadruplex-hemin DNAzyme could be formed, allowing significant signal amplification in the presence of H2O2. Such strategy can enhance the oxidation-reduction reaction of adsorbed methylene blue (MB) and efficiently improve the sensitivity of the proposed biosensor. The morphologies of materials and the stepwise biosensor were characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM) and cyclic voltammetry (CV). Differential pulse voltammetry (DPV) signals of MB provided quantitative measures of the concentrations of target DNA, with a linear calibration range of 1.0 × 10(-15)-1.0 × 10(-9)M and a detection limit of 3.0 × 10(-16)M. Moreover, the resulting biosensor also exhibited good specificity, acceptable reproducibility and stability, indicating that the present strategy was promising for broad potential application in clinic assay.

  14. Signal amplification based on the local nonlinear Mach-Zehnder interferometer

    NASA Astrophysics Data System (ADS)

    Srivastava, Arpita; Gupta, Man Mohan; Medhekar, S.

    2012-03-01

    Using the phase modulation of spatial solitons, a new scheme for all-optical signal amplification has been proposed in this paper. The considered structure is composed of the nonlinear Mach-Zehnder interferometer (NMZI) with the straight control waveguide (CWG), the uniform nonlinear medium (NLM) and the linear output waveguide. The local NMZI functions like a phase shifter. The light-induced index changes in the local nonlinear MZI are responsible for the input beam routing in the uniform nonlinear medium. The coupling of the input beam to the output waveguide depends on its propagation direction in the NLM. It is shown that the signal launched at CWG can deflect the beam launched at the NMZI (input beam) and a modulated (amplified) output could be obtained at the output waveguide. Further, signal pulse may be reshaped by appropriately increasing the NLM length. In addition, amplification factor may be enhanced by increasing the NLM length and injecting an appropriate continuous wave beam along with the signal beam at CWG.

  15. 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

  16. 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

  17. 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.}

  18. Harmonic signal generation and frequency upconversion using selective sideband Brillouin amplification in single-mode fiber.

    PubMed

    Lee, Kwang-Hyun; Choi, Woo-Young

    2007-06-15

    Harmonic signal generation and frequency upconversion at millimeter-wave bands are experimentally demonstrated by using selective sideband Brillouin amplification induced by stimulated Brillouin scattering in a single-mode fiber. The harmonic signals and frequency upconverted signals are simultaneously generated by the beating of optical sidebands, one of which is Brillouin amplified. By using this method, we successfully demonstrate generation of third-harmonic millimeter waves at 32.55 GHz with f(LO) of 10.85 GHz and upconversion of 10 Mbps quadrature-shift keyed data at f(IF) of 1.55 GHz into a 30 GHz band with more than 17 dB RF power gain.

  19. Arsenic-induced instrumental genes of apoptotic signal amplification in death-survival interplay

    PubMed Central

    Roy, Sonali; Narzary, Bardwi; Ray, Atish; Bordoloi, Manobjyoti

    2016-01-01

    Arsenic is a global health concern at present and it is well reported for causing systemic toxicity. It is also well known for generation of free radical and inducing apoptosis in different cell types. Paradoxically arsenic is reported to be a susceptible carcinogen as well. There are several reports demonstrating diverse mechanism of apoptosis in different cell types. However, the universal scenario of instrumental genes and their interaction leading to amplification of apoptotic signal are yet to be completely uncovered, which is predicted here. Conventional studies on signaling pathway aided by time and concentration kinetics data are inadequate for prediction of anchored genes for apoptotic signal amplification. Therefore, expression profile-based approach is adopted. Core apoptosis related and glutathione metabolism genes in 1 and 10 μM of arsenic-treated HepG2 cells were analyzed after 12 h of incubation. An arsenic treatment of 1 μM exhibits no cell death at 12 h, whereas 10 μM arsenic treatment reveals around 50% cell death at 12 h. Results depict 28 and 44 affected genes in 1 and 10 μM arsenic-treated cells, respectively. Early initiation of apoptotic signaling is detected in no cell death regimens (at 1 μM), whereas amplified apoptotic signal is demonstrated at 50% cell death regimens (at 10 μM). Instrumental genes involved in progression of apoptosis in the concourse of cell death and survival is designated from the responsive genes common to both the condition. We predict the initiation process is fairly aided by the activation of intrinsic pathway, which is amplified via TNF signaling and extrinsic pathway. Furthermore, regulatory genes involved in interplay between apoptosis/anti-apoptosis and their interactions are demonstrated here. PMID:27785370

  20. Photorefractive amplification of dynamic light signals using photoconductive ferroelectric liquid crystals

    NASA Astrophysics Data System (ADS)

    Sasaki, Takeo

    2015-05-01

    The photorefractive effect in photoconductive ferroelectric liquid crystal blends containing photoconductive chiral compounds was investigated. Terthiophene compounds possessing chiral structures were mixed with an achiral smectic C liquid crystal mixture. The blends exhibit the ferroelectric chiral smectic C phase. The photorefractivity of the liquid crystal blends was investigated by two-beam coupling experiments. The photoconductive ferroelectric liquid crystal blends prepared in this study exhibited a large gain coefficient of over 1200 cm-1 and a fast response time shorter than 1 ms. Amplification of an moving optical image signal of over 30 fps using the photorefractive ferroelectric liquid crystal was demonstrated.

  1. Photorefractive amplification of moving light signals by photoconductive ferroelectric liquid crystal blends

    NASA Astrophysics Data System (ADS)

    Sasaki, Takeo; Yoshino, Masanori

    2015-09-01

    Ferroelectric liquid crystal blends composed of a smectic liquid crystalline mixture, a photoconductive chiral dopant, and an electron trap reagent exhibit significant photorefractivity together with rapid responses. As such, they allow the dynamic amplification of moving optical signals. In the present work, the durability of a photorefractive ferroelectric liquid crystal blend was investigated. A series of photoconductive chiral dopants was prepared and the durability of blends incorporating these dopants during laser irradiation was examined. In addition, the effect of the conduction of photogenerated ionic species on the photorefractivity decay was clarified.

  2. Dynamic amplification of light signals in photorefractive ferroelectric liquid crystalline mixtures.

    PubMed

    Sasaki, Takeo; Kajikawa, Satoshi; Naka, Yumiko

    2014-01-01

    The photorefractive effect in photoconductive ferroelectric liquid crystals that contain photoconductive chiral compounds was investigated. Terthiophene compounds with chiral structures were chosen as the photoconductive chiral compounds, and they were mixed with an achiral smectic C liquid crystal. The mixtures exhibit the ferroelectric chiral smectic C phase. The photorefractivity of the mixtures was investigated by two-beam coupling experiments. It was found that the ferroelectric liquid crystals containing the photoconductive chiral compound exhibit a large gain coefficient of over 1200 cm(-1) and a fast response time of 1 ms. Real-time dynamic amplification of an optical image signal of over 30 fps using the photorefractive ferroelectric liquid crystal was demonstrated.

  3. Parametric Amplification Protocol for Frequency-Modulated Magnetic Resonance Force Microscopy Signals

    NASA Astrophysics Data System (ADS)

    Harrell, Lee; Moore, Eric; Lee, Sanggap; Hickman, Steven; Marohn, John

    2011-03-01

    We present data and theoretical signal and noise calculations for a protocol using parametric amplification to evade the inherent tradeoff between signal and detector frequency noise in force-gradient magnetic resonance force microscopy signals, which are manifested as a modulated frequency shift of a high- Q microcantilever. Substrate-induced frequency noise has a 1 / f frequency dependence, while detector noise exhibits an f2 dependence on modulation frequency f . Modulation of sample spins at a frequency that minimizes these two contributions typically results in a surface frequency noise power an order of magnitude or more above the thermal limit and may prove incompatible with sample spin relaxation times as well. We show that the frequency modulated force-gradient signal can be used to excite the fundamental resonant mode of the cantilever, resulting in an audio frequency amplitude signal that is readily detected with a low-noise fiber optic interferometer. This technique allows us to modulate the force-gradient signal at a sufficiently high frequency so that substrate-induced frequency noise is evaded without subjecting the signal to the normal f2 detector noise of conventional demodulation.

  4. An electrochemical biosensor for sensitive detection of microRNA-155: combining target recycling with cascade catalysis for signal amplification.

    PubMed

    Wu, Xiaoyan; Chai, Yaqin; Zhang, Pu; Yuan, Ruo

    2015-01-14

    In this work, a new electrochemical biosensor based on catalyzed hairpin assembly target recycling and cascade electrocatalysis (cytochrome c (Cyt c) and alcohol oxidase (AOx)) for signal amplification was constructed for highly sensitive detection of microRNA (miRNA). It is worth pointing out that target recycling was achieved only based on strand displacement process without the help of nuclease. Moreover, porous TiO2 nanosphere was synthesized, which could offer more surface area for Pt nanoparticles (PtNPs) enwrapping and enhance the amount of immobilized DNA strand 1 (S1) and Cyt c accordingly. With the mimicking sandwich-type reaction, the cascade catalysis amplification strategy was carried out by AOx catalyzing ethanol to acetaldehyde with the concomitant formation of high concentration of H2O2, which was further electrocatalyzed by PtNPs and Cyt c. This newly designed biosensor provided a sensitive detection of miRNA-155 from 0.8 fM to 1 nM with a relatively low detection limit of 0.35 fM.

  5. Weak signal amplification and detection by higher-order sensory neurons.

    PubMed

    Jung, Sarah N; Longtin, Andre; Maler, Leonard

    2016-04-01

    Sensory systems must extract behaviorally relevant information and therefore often exhibit a very high sensitivity. How the nervous system reaches such high sensitivity levels is an outstanding question in neuroscience. Weakly electric fish (Apteronotus leptorhynchus/albifrons) are an excellent model system to address this question because detailed background knowledge is available regarding their behavioral performance and its underlying neuronal substrate. Apteronotus use their electrosense to detect prey objects. Therefore, they must be able to detect electrical signals as low as 1 μV while using a sensory integration time of <200 ms. How these very weak signals are extracted and amplified by the nervous system is not yet understood. We studied the responses of cells in the early sensory processing areas, namely, the electroreceptor afferents (EAs) and pyramidal cells (PCs) of the electrosensory lobe (ELL), the first-order electrosensory processing area. In agreement with previous work we found that EAs cannot encode very weak signals with a spike count code. However, PCs can encode prey mimic signals by their firing rate, revealing a huge signal amplification between EAs and PCs and also suggesting differences in their stimulus encoding properties. Using a simple leaky integrate-and-fire (LIF) model we predict that the target neurons of PCs in the midbrain torus semicircularis (TS) are able to detect very weak signals. In particular, TS neurons could do so by assuming biologically plausible convergence rates as well as very simple decoding strategies such as temporal integration, threshold crossing, and combining the inputs of PCs.

  6. Signal amplification using colloidal gold in a biolayer interferometry-based immunosensor for the mycotoxin deoxynivalenol.

    PubMed

    Maragos, C M

    2012-01-01

    Deoxynivalenol (DON) is a toxin produced by certain species of Fusarium fungi that can infest wheat, barley and corn. The fungi cause diseases in crops worldwide and some of the secondary metabolites, such as DON, can adversely affect animal health and food safety. To monitor DON in wheat rapidly, a biosensor using the principle of biolayer interferometry (BLI) was developed. The signal from the sensor was substantially amplified through the use of a primary antibody-colloidal gold conjugate. The amplification was much greater in the presence of wheat matrix than in buffered solution, suggesting matrix components may have contributed to the enhancement. The improved signal provided by the amplification allowed for the development of rapid qualitative and quantitative assays. The limit of detection of the method was 0.09 mg kg(-1); the limit of quantitation was 0.35 mg kg(-1). Recovery from wheat spiked over the range from 0.2 to 5 mg kg(-1) averaged 103% (RSD = 12%). The quantitative assay compared favourably (r(2) = 0.9698) with a reference chromatographic method for 40 naturally contaminated wheats. The qualitative assay was able to classify accurately the same group of 40 samples as either above or below a 0.5 mg kg(-1) threshold. These results suggest that the BLI technique can be used to measure DON in wheat rapidly.

  7. Enzymatic amplification of DNA/RNA hybrid molecular beacon signaling in nucleic acid detection.

    PubMed

    Jacroux, Thomas; Rieck, Daniel C; Cui, Rong; Ouyang, Yexin; Dong, Wen-Ji

    2013-01-15

    A rapid assay operable under isothermal or nonisothermal conditions is described, where the sensitivity of a typical molecular beacon (MB) system is improved by using thermostable RNase H to enzymatically cleave an MB composed of a DNA stem and an RNA loop (R/D-MB). On hybridization of the R/D-MB to target DNA, there was a modest increase in fluorescence intensity (~5.7× above background) due to an opening of the probe and a concomitant reduction in the Förster resonance energy transfer efficiency. The addition of thermostable RNase H resulted in the cleavage of the RNA loop, which eliminated energy transfer. The cleavage step also released bound target DNA, enabling it to bind to another R/D-MB probe and rendering the approach a cyclic amplification scheme. Full processing of R/D-MBs maximized the fluorescence signal to the fullest extent possible (12.9× above background), resulting in an approximately 2- to 2.8-fold increase in the signal-to-noise ratio observed isothermally at 50 °C following the addition of RNase H. The probe was also used to monitor real-time polymerase chain reactions by measuring enhancement of donor fluorescence on R/D-MB binding to amplified pUC19 template dilutions. Hence, the R/D-MB-RNase H scheme can be applied to a broad range of nucleic acid amplification methods.

  8. A graphene oxide-based enzyme-free signal amplification platform for homogeneous DNA detection.

    PubMed

    Zhang, Zhen; Liu, Yufei; Ji, Xinghu; Xiang, Xia; He, Zhike

    2014-10-01

    A graphene oxide (GO) based enzyme-free signal amplification platform for homogeneous DNA sensing is developed with simplicity and high sensitivity. In the absence of the target DNA, labeled hairpin probe 1 (H1) and probe 2 (H2) were adsorbed on the surface of GO, resulting in the fluorescence quenching of the dyes and minimizing the background fluorescence. The addition of the target DNA facilitated the formation of double-stranded DNA (dsDNA) between H1 and H2, causing the probes to separate from GO and release the target DNA through a strand displacement reaction. Meanwhile, the whole reaction started anew. This is an excellent isothermal signal amplification technique without the involvement of enzymes. By monitoring the change of the fluorescence intensity, the target DNA not only can be determined in buffer solution, but also can be detected in 1% serum solution spiked with a series of concentrations of the target DNA. In addition, the consumption amount of the probes in this method is lower than that in traditional molecular beacon methods.

  9. A cascade signal amplification strategy for surface enhanced Raman spectroscopy detection of thrombin based on DNAzyme assistant DNA recycling and rolling circle amplification.

    PubMed

    Gao, Fenglei; Du, Lili; Tang, Daoquan; Lu, Yao; Zhang, Yanzhuo; Zhang, Lixian

    2015-04-15

    A sensitive protocol for surface enhanced Raman spectroscopy (SERS) detection of thrombin is designed with R6G-Ag NPs as a signal tag by combining DNAzyme assistant DNA recycling and rolling circle amplification (RCA). Molecular beacon (MB) as recognition probe immobilizes on the glass slides and performs the amplification procedure. After thrombin-induced structure-switching DNA hairpins of probe 1, the DNAzyme is liberated from the caged structure, which hybridizes with the MB for cleavage of the MB in the presence of cofactor Zn(2+) and initiates the DNA recycling process, leading to the cleavage of a large number of MB and the generation of numerous primers for triggering RCA reaction. The long amplified RCA product which contained hundreds of tandem-repeat sequences, which can bind with oligonucleotide functionalized Ag NPs reporters. The attached signal tags can be easily read out by SERS. Because of the cascade signal amplification, these newly designed protocols provides a sensitive SERS detection of thrombin down to the femolar level (2.3fM) with a linear range of 5 orders of magnitude (from 10(-14) to 10(-9)M) and have high selectivity toward its target protein. The proposed method is expected to be a good clinical tool for the diagnosis of a thrombotic disease.

  10. Signal amplification by adsorption-induced catalytic reduction of dissolved oxygen on nitrogen-doped carbon nanotubes for electrochemiluminescent immunoassay.

    PubMed

    Deng, Shengyuan; Hou, Zhentao; Lei, Jianping; Lin, Dajie; Hu, Zheng; Yan, Feng; Ju, Huangxian

    2011-11-28

    A signal amplification strategy was proposed for quantum dot-based electrochemiluminescence by an adsorption-induced catalytic reduction of dissolved oxygen at the sidewall of nitrogen-doped carbon nanotubes, which led to a 'signal-on' sandwich immunoassay with a linear range of 6 orders of magnitude.

  11. [Methods for the detection of ventricular late potentials. High amplification ECG, signal averaging technic, frequency analysis and intracardiac mapping].

    PubMed

    Hombach, V; Eggeling, T; Höher, M; Höpp, H W; Kochs, M; Giel, I; Emsermann, P; Hirche, H; Hilger, H H

    1988-06-01

    Circumscribed areas of injured myocardium which lead to late ventricular depolarization represent the pathologic-anatomic substrate for reentry mechanisms potentially capable of propagating ventricular tachycardia at the ventricular level. If the myocardial area from which delayed ventricular depolarization and, consequently, late potentials eminate, exceeds a critical minimal size, documentation of such signals can not only be achieved with direct endocardial mapping or catheter mapping but also by means of special high-resolution ECG techniques from the body surface. Since high amplification of the conventional ECG results in registration of noise signals in amplitude of up to 50 microV, late potentials with their amplitudes at the body surface ranging from 5 to a maximum of 20 microV, can only be discriminated after substantial enhancement of the signal-to-noise ratio. The noise arises from no less than three sources: physiologic noise, for example, from muscle activity; electronic noise from amplifiers and background noise of 50 or 60 Hz, respectively. To improve the signal-to-noise ratio, currently three methods are employed: sequential or temporal signal averaging, spatial signal averaging and fast Fourier transformation analysis of the frequency spectrum of the highly-amplified ECG. Temporal signal averaging has the purpose of smoothing randomly-occurring background noise and, at a specified point in time of the ECG cycle, to sum the signal incurred. The effectivity of this technique, however, is subject to certain conditions: the signal to be registered and the background noise must be independent from each other, the noise must be stationary and show normal random distribution, the signal of interest must be periodic and/or coupled with a fixed interval to a point in the ECG cycle which can be used as a trigger. The quality of the averaged signal is dependent on trigger stability. There are three approaches to trigger processing: voltage threshold

  12. Electrochemical sensor for glutathione detection based on mercury ion triggered hybridization chain reaction signal amplification.

    PubMed

    Wang, Yonghong; Jiang, Lun; Leng, Qinggang; Wu, Yaohui; He, Xiaoxiao; Wang, Kemin

    2016-03-15

    In this work, we design a new simple and highly sensitive strategy for electrochemical detection of glutathione (GSH) via mercury ion (Hg(2+)) triggered hybridization chain reaction (HCR) signal amplification. It is observed that in the absence of GSH, a specific thymine-Hg(2+)-thymine (T-Hg(2+)-T) coordination can fold into hairpin structures. While in the presence of GSH, it thus can be chelated with Hg(2+), resulting in Hg(2+) released from the T-Hg(2+)-T hairpin complex which then forms into ssDNA structure to further hybridize with the surface-immobilized capture DNA probe on the gold electrode with a sticky tail left. The presence of two hairpin helper probes through HCR leads to the formation of extended dsDNA superstructure on the electrode surface, which therefore causes the intercalation of numerous electroactive species ([Ru(NH3)6](3+)) into the dsDNA grooves, followed by a significantly amplified signal output whose intensity is related to the concentration of the GSH. Taking advantage of merits of enzyme-free amplification power of the HCR, the inherent high sensitivity of the electrochemical technique, and label-free detection which utilizes an electroactive species as a signaling molecule that binds to the anionic phosphate backbone of DNA strands via electrostatic force, not only does the proposed strategy enable sensitive detection of GSH, but show high selectivity against other amino acid, making our method a simple and sensitive addition to the amplified GSH detection. PMID:26528805

  13. Probing signal amplification by reversible exchange using an NMR flow system

    PubMed Central

    Mewis, Ryan E; Atkinson, Kevin D; Cowley, Michael J; Duckett, Simon B; Green, Gary G R; Green, Richard A; Highton, Louise A R; Kilgour, David; Lloyd, Lyrelle S; Lohman, Joost A B; Williamson, David C

    2014-01-01

    Hyperpolarization methods are used in NMR to overcome its inherent sensitivity problem. Herein, the biologically relevant target nicotinamide is polarized by the hyperpolarization technique signal amplification by reversible exchange. We illustrate how the polarization transfer field, and the concentrations of parahydrogen, the polarization-transfer-catalyst and substrate can be used to maximize signal amplification by reversible exchange effectiveness by reference to the first-order spin system of this target. The catalyst is shown to be crucial in this process, first by facilitating the transfer of hyperpolarization from parahydrogen to nicotinamide and then by depleting the resulting polarized states through further interaction. The 15 longitudinal one, two, three and four spin order terms produced are rigorously identified and quantified using an automated flow apparatus in conjunction with NMR pulse sequences based on the only parahydrogen spectroscopy protocol. The rates of build-up of these terms were shown to follow the order four∼three > two > single spin; this order parallels their rates of relaxation. The result of these competing effects is that the less-efficiently formed single-spin order terms dominate at the point of measurement with the two-spin terms having amplitudes that are an order of magnitude lower. We also complete further measurements to demonstrate that 13C NMR spectra can be readily collected where the long-lived quaternary 13C signals appear with significant intensity. These are improved upon by using INEPT. In summary, we dissect the complexity of this method, highlighting its benefits to the NMR community and its applicability for high-sensitivity magnetic resonance imaging detection in the future. © 2014 The Authors. Magnetic Resonance in Chemistry by John Wiley & Sons, Ltd. PMID:24801201

  14. 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.

  15. 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).

  16. Sub-femtomolar electrochemical detection of DNA using surface circular strand-replacement polymerization and gold nanoparticle catalyzed silver deposition for signal amplification.

    PubMed

    Gao, Fenglei; Zhu, Zhu; Lei, Jianping; Geng, Yao; Ju, Huangxian

    2013-01-15

    A highly sensitive method was developed for detection of target DNA. This method combined circular strand-displacement polymerization (CSRP) with silver enhancement to achieve dual signal amplification. After molecular beacon (MB) hybridized with target DNA, the reporter gold nanoparticle (Au NPs) was attached to an electrode surface by hybridization between Au NP labeled primer and stem part of the MB to initiate a polymerization of DNA strand, which led to the release of target and another polymerization cycle. Thus the CSRP produced the multiplication of target-related reporter Au NPs on the surface. The Au NPs then catalyzed silver deposition for subsequent stripping analysis of silver. The dual signal amplification offered a dramatic enhancement of the stripping response. This signal could discriminate perfect matched target DNA from 1-base mismatch DNA. The dynamic range of the sequence-specific DNA detection was from 10(-16) to 10(-12)mol L(-1) with a detection limit down to sub-femtomolar level. This proposed method exhibited an efficient amplification performance, and would open new opportunities for sensitive detection of other biorecognition events.

  17. "Signal-on" photoelectrochemical biosensor for sensitive detection of human T-Cell lymphotropic virus type II DNA: dual signal amplification strategy integrating enzymatic amplification with terminal deoxynucleotidyl transferase-mediated extension.

    PubMed

    Shen, Qingming; Han, Li; Fan, Gaochao; Zhang, Jian-Rong; Jiang, Liping; Zhu, Jun-Jie

    2015-01-01

    A novel "signal-on" photoelectrochemical (PEC) biosensor for sensitive detection of human T-cell lymphotropic virus type II (HTLV-II) DNA was developed on the basis of enzymatic amplification coupled with terminal deoxynucleotidyl transferase (TdT)-mediated extension strategy. The intensity of the photocurrent signal was proportional to the concentration of the HTLV-II DNA-target DNA (tDNA) by dual signal amplification. In this protocol, GR-CdS:Mn/ZnS nanocomposites were used as photoelectric conversion material, while pDNA was used as the tDNA recognizing unit. Moreover, the TdT-mediated extension and the enzymatic signal amplification technique were used to enhance the sensitivity of detection. Using this novel dual signal amplification strategy, the prototype of PEC DNA sensor can detect as low as ∼0.033 fM of HTLV-II DNA with a linear range of 0.1-5000 fM, with excellent differentiation ability even for single-base mismatches. This PEC DNA assay opens a promising platform to detect various DNA targets at ultralow levels for early diagnoses of different diseases. PMID:25871300

  18. 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

  19. NMR Signal Amplification by Reversible Exchange of Sulfur-Heterocyclic Compounds Found In Petroleum

    PubMed Central

    Coffey, Aaron M.; Goodson, Boyd M.; Chekmenev, Eduard Y.

    2016-01-01

    NMR hyperpolarization via Signal Amplification by Reversible Exchange (SABRE) was employed to investigate the feasibility of enhancing the NMR detection sensitivity of sulfur-heterocycles (specifically 2-methylthiophene and dibenzothiophenes), a family of compounds typically found in petroleum and refined petroleum products. SABRE hyperpolarization of sulfur-heterocycles (conducted in seconds) offers potential advantages of providing structural information about sulfur-containing contaminants in petroleum, thereby informing petroleum purification and refining to minimize sulfur content in refined products such as gasoline. Moreover, NMR spectroscopy sensitivity gains endowed by hyperpolarization potentially allows for performing structural assays using inexpensive, low-magnetic-field (ca. 1 T) high-resolution NMR spectrometers ideally suited for industrial applications in the field. PMID:27500206

  20. Dual-primer self-generation SERS signal amplification assay for PDGF-BB using label-free aptamer.

    PubMed

    Ye, SuJuan; Zhai, XiaoMo; Wu, YanYing; Kuang, ShaoPing

    2016-05-15

    Highly sensitive detection of proteins, especially those associated with cancers, is essential to biomedical research as well as clinical diagnosis. In this work, a simple and novel one-two-three signal amplification surface-enhanced Raman scattering (SERS) method for the detection of protein is fabricated by using label-free aptamer and dual-primer self-generation. Platelet-derived growth factor B-chain (PDGF-BB) is selected as the model protein. The one-two-three cascade DNA amplification means one target-aptamer binding event, two hairpin DNA switches and three DNA amplification reactions. This strategy possesses some remarkable features compared to conventional signal amplification methods: (i) A smart probe including a label-free aptamer is fabricated, for suitable hybridization without hindering the affinity of the aptamer toward its target. (ii) Using the unique structure switch of the aptamer and cooperator, a one-two-three working mode is developed to amplify the SERS signal. The amplification efficiency is enhanced. Given the unique and attractive characteristics, a simple and universal strategy is designed to accomplish ultrasensitive detection of proteins. The detection limit of PDGF-BB via SERS detection is 0.42 pM, with the linear range from 1.0×10(-12)M to 10(-8)M. It is potentially universal because the aptamer can be easily designed for biomolecules whose aptamers undergo similar conformational changes.

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

    PubMed

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

    2015-07-01

    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. PMID:26012841

  2. 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.

  3. Porous platinum nanotubes modified with dendrimers as nanocarriers and electrocatalysts for sensitive electrochemical aptasensors based on enzymatic signal amplification.

    PubMed

    Xu, Wenju; Wu, Yongmei; Yi, Huayu; Bai, Lijuan; Chai, Yaqin; Yuan, Ruo

    2014-02-11

    A highly sensitive electrochemical aptasensor for thrombin detection is developed and demonstrated by using porous platinum nanotubes modified with polyamidoamine dendrimers as nanocarriers and electrocatalysts. The proposed strategy affords a low detection limit of 0.03 pM based on enzyme-based signal amplification.

  4. Droplet-Free Digital Enzyme-Linked Immunosorbent Assay Based on a Tyramide Signal Amplification System.

    PubMed

    Akama, Kenji; Shirai, Kentaro; Suzuki, Seigo

    2016-07-19

    Digital enzyme-linked immunosorbent assay (ELISA) is a single molecule counting technology and is one of the most sensitive immunoassay methods. The key aspect of this technology is to concentrate enzyme reaction products from a single target molecule in femtoliter droplets. This study presents a novel Digital ELISA that does not require droplets; instead, enzyme reaction products are concentrated using a tyramide signal amplification system. In our method, tyramide substrate reacts with horseradish peroxidase (HRP) labeled with an immunocomplex on beads, and the substrate is converted into short-lived radical intermediates. By adjusting the bead concentration in the HRP-tyramide reaction and conducting the reaction using freely moving beads, tyramide radicals are deposited only on beads labeled with HRP and there is no diffusion to other beads. Consequently, the fluorescence signal is localized on a portion of the beads, making it possible to count the number of labeled beads digitally. The performance of our method was demonstrated by detecting hepatitis B surface antigen with a limit of detection of 0.09 mIU/mL (139 aM) and a dynamic range of over 4 orders of magnitude. The obtained limit of detection represents a >20-fold higher sensitivity than conventional ELISA. Our method has potential applications in simple in vitro diagnostic systems for detecting ultralow concentrations of protein biomarkers.

  5. A theophylline quartz crystal microbalance biosensor based on recognition of RNA aptamer and amplification of signal.

    PubMed

    Dong, Zong-Mu; Zhao, Guang-Chao

    2013-04-21

    A quartz crystal microbalance (QCM) biosensor for theophylline was developed by recognition of RNA aptamer and gold nanoparticle amplification technique. Firstly, a designed small single-stranded RNA, RNA1, was immobilized onto the QCM electrode through a thiol linker. Then, the complementary stranded RNA2, which can combine with RNA1 to form a double-stranded RNA with a recognition unit of theophylline, could be self-assembled on the QCM electrode surface through a hybrid reaction in the presence of theophylline. The recognition process could cause a frequency change of QCM to give the signal related to theophylline. When RNA2 was tethered to gold nanoparticles, the signal could be amplified to further enhance the sensitivity of the designed sensor. Under the optimal conditions, the QCM-based biosensor showed excellent sensitivity (limit of detection, 8.2 nM) and specificity with a dissociation constant of Kd = 5.26 × 10(-7) M. The sensor can be used to quantitatively detect theophylline in serum, suggesting that it can be applied in complex biological samples.

  6. 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.

  7. Alternative configuration scheme for signal amplification with scanning ion conductance microscopy.

    PubMed

    Kim, Joonhui; Kim, Seong-Oh; Cho, Nam-Joon

    2015-02-01

    Scanning Ion Conductance Microscopy (SICM) is an emerging nanotechnology tool to investigate the morphology and charge transport properties of nanomaterials, including soft matter. SICM uses an electrolyte filled nanopipette as a scanning probe and detects current changes based on the distance between the nanopipette apex and the target sample in an electrolyte solution. In conventional SICM, the pipette sensor is excited by applying voltage as it raster scans near the surface. There have been attempts to improve upon raster scanning because it can induce collisions between the pipette sidewalls and target sample, especially for soft, dynamic materials (e.g., biological cells). Recently, Novak et al. demonstrated that hopping probe ion conductance microscopy (HPICM) with an adaptive scan method can improve the image quality obtained by SICM for such materials. However, HPICM is inherently slower than conventional raster scanning. In order to optimize both image quality and scanning speed, we report the development of an alternative configuration scheme for SICM signal amplification that is based on applying current to the nanopipette. This scheme overcomes traditional challenges associated with low bandwidth requirements of conventional SICM. Using our alternative scheme, we demonstrate successful imaging of L929 fibroblast cells and discuss the capabilities of this instrument configuration for future applications. PMID:25725851

  8. 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). PMID:26920780

  9. Alternative configuration scheme for signal amplification with scanning ion conductance microscopy

    NASA Astrophysics Data System (ADS)

    Kim, Joonhui; Kim, Seong-Oh; Cho, Nam-Joon

    2015-02-01

    Scanning Ion Conductance Microscopy (SICM) is an emerging nanotechnology tool to investigate the morphology and charge transport properties of nanomaterials, including soft matter. SICM uses an electrolyte filled nanopipette as a scanning probe and detects current changes based on the distance between the nanopipette apex and the target sample in an electrolyte solution. In conventional SICM, the pipette sensor is excited by applying voltage as it raster scans near the surface. There have been attempts to improve upon raster scanning because it can induce collisions between the pipette sidewalls and target sample, especially for soft, dynamic materials (e.g., biological cells). Recently, Novak et al. demonstrated that hopping probe ion conductance microscopy (HPICM) with an adaptive scan method can improve the image quality obtained by SICM for such materials. However, HPICM is inherently slower than conventional raster scanning. In order to optimize both image quality and scanning speed, we report the development of an alternative configuration scheme for SICM signal amplification that is based on applying current to the nanopipette. This scheme overcomes traditional challenges associated with low bandwidth requirements of conventional SICM. Using our alternative scheme, we demonstrate successful imaging of L929 fibroblast cells and discuss the capabilities of this instrument configuration for future applications.

  10. 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

  11. Signal processing for high granularity calorimeter: amplification, filtering, memorization and digitalization

    NASA Astrophysics Data System (ADS)

    Royer, L.; Manen, S.; Gay, P.

    2010-12-01

    A very-front-end electronics dedicated to high granularity calorimeters has been designed and its performance measured. This electronics performs the amplification of the charge delivered by the detector thanks to a low-noise Charge Sensitive Amplifier. The dynamic range is improved using a bandpass filter based on a Gated Integrator. Studying its weighting function, we show that this filter is more efficient than standard CRRC shaper, thanks to the integration time which can be expand near the bunch interval time, whereas the peaking time of the CRRC shaper is limited to pile-up consideration. Moreover, the Gated Integrator performs intrinsically the analog memorization of the signal before its delayed digital conversion. The analog-to-digital conversion is performed through a 12-bit cyclic ADC specifically developed for this application. The very-front-end channel has been fabricated using a 0.35 μm CMOS technology. Measurements show a global non-linearity better than 0.1%. The Equivalent Noise Charge at the input of the channel is evaluated to 1.8 fC, compare to the maximum input charge of 10 pC. The power consumption of the complete channel is limited to 6.5 mW.

  12. Amplification of small signals by voltage-gated sodium channels in drone photoreceptors.

    PubMed

    Coles, J A; Schneider-Picard, G

    1989-04-01

    Photoreceptor cells of the drone, Apis mellifera male, have a voltage-gated Na+ membrane conductance that can be blocked by tetrodotoxin (TTX) and generates an action potential on abrupt depolarization: an action potential is triggered by the rising phase of a receptor potential evoked by an intense light flash (Autrum and von Zwehl 1964; Baumann 1968). We measured the intracellular voltage response to a small (9%), brief (30 ms) decrease in light intensity from a background, and found that its amplitude was decreased by 1 microM TTX. The response amplitude was maximal when the background intensity depolarized the cell to -38 mV. With intensities depolarizing the cell membrane to -45 to -33 mV the average response amplitude was decreased by TTX from 1.2 mV to 0.5 mV. TTX is also known to decrease the voltage noise during steady illumination (Ferraro et al. 1983) but, despite this, the ratio of peak-to-peak signal to noise was, on average, decreased by TTX. The results suggest that drone photoreceptors use voltage-gated Na+ channels for graded amplification of responses to small, rapid changes in light intensity.

  13. Signal-amplification and real-time fluorescence anisotropy detection of apyrase by carbon nanoparticle.

    PubMed

    Liu, Jinhua; Yu, Jing; Chen, Jianrong; Yang, Ronghua; Shih, Kaimin

    2014-05-01

    Carbon nanomaterial combined with aptamer has been developed as an efficient bioanalytical method in sensor design. Herein, depending on carbon nanoparticle (cCNP)-enhanced fluorescence anisotropy (FA), a novel aptamer-based sensor (aptasensor) enabling signal-amplification and real-time detection of apyrase is reported. The foundation of our sensor design based on ATP-aptamer(P) can be adsorbed on the surface of cCNPs, resulting in the increase of FA due to the mass of cCNPs, and P-ATP complex has weak binding ability to cCNPs with minimal change of FA. Apyrase, being an integral membrane protein, can hydrolyze ATP and make P-ATP complex disassemble, and thus lead to the increasing of FA. Therefore, this approach is demonstrated to be a novel candidate for the detection of apyrase, with high sensitivity and selectivity. The linear dynamic range for the concentrations of apyrase is between 0.1 and 0.5 U/μL along with a detection limit of 0.05 U/μL. Furthermore, these results indicated that our design is a flexible and sensitive method for biomolecule analysis, which makes it promising for practical biomolecule analyses.

  14. Hybrid Chirped Pulse Amplification

    SciTech Connect

    Jovanovic, I; Barty, C P J

    2002-05-07

    We present a novel chirped pulse amplification method which combines optical parametric amplification and laser amplification. We have demonstrated this hybrid CPA concept with a combination of beta-barium borate and Ti:sapphire. High-efficiency, multi-terawatt compatible amplification is achieved without gain narrowing and without electro-optic modulators using a simple commercial pump laser.

  15. 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.

  16. Direct visualization of the reaction transformation and signal amplification in a DNA molecular machine with total internal reflection fluorescence microscopy

    PubMed Central

    Ren, Rui; Wang, Haiyan; Liu, Rui; Zhang, Shusheng

    2013-01-01

    In this study, as a proof of concept, the signal amplification in an artificial DNA molecular machine was directly visualized via total internal reflection fluorescence microscopy (TIRFM). The molecular machine brought about obvious morphology change in DNA nanostructures as well as signal amplifications. On one hand, through a triggered and autonomically repeated RCA, a DNA nano-complex featuring a “locked” circular DNA template (serving as raw feed) was converted into a long periodically repeated strand, i.e., the RCA products. On the other hand, this RCA was repeated in three controllable reaction phases, bring about progressive signal amplification. It was testified that the RCA products (presented as long thread-like fluorescent objects) can be easily distinguished from the inputted DNA probes (presented as fluorescent dots), thus the transformation in reaction can be visualized. Also, by quantitive counting of the aforementioned fluorescence objects, the progress of the reaction through the phases, along with time, and over the lysozyme concentration can be demonstrated through TIRFM visualization. Overall, it was demonstrated that TIRFM is an efficient approach to quantitatively visualize the biochemical processes at single-molecule level. PMID:24790951

  17. 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

  18. One-step, multiplexed fluorescence detection of microRNAs based on duplex-specific nuclease signal amplification.

    PubMed

    Yin, Bin-Cheng; Liu, Yu-Qiang; Ye, Bang-Ce

    2012-03-21

    Traditional molecular beacons, widely applied for detection of nucleic acids, have an intrinsic limitation on sensitivity, as one target molecule converts only one beacon molecule to its fluorescent form. Herein, we take advantage of the duplex-specific nuclease (DSN) to create a new signal-amplifying mechanism, duplex-specific nuclease signal amplification (DSNSA), to increase the detection sensitivity of molecular beacons (Taqman probes). DSN nuclease is employed to recycle the process of target-assisted digestion of Taqman probes, thus, resulting in a significant fluorescence signal amplification through which one target molecule cleaves thousands of probe molecules. We further demonstrate the efficiency of this DSNSA strategy for rapid direct quantification of multiple miRNAs in biological samples. Our experimental results showed a quantitative measurement of sequence-specific miRNAs with the detection limit in the femtomolar range, nearly 5 orders of magnitude lower than that of conventional molecular beacons. This amplification strategy also demonstrated a high selectivity for discriminating differences between miRNA family members. Considering the superior sensitivity and specificity, as well as the multiplex and simple-to-implement features, this method promises a great potential of becoming a routine tool for simultaneously quantitative analysis of multiple miRNAs in tissues or cells, and supplies valuable information for biomedical research and clinical early diagnosis.

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

    PubMed

    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

  20. 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.

  1. Approaches towards molecular amplification for sensing.

    PubMed

    Goggins, Sean; Frost, Christopher G

    2016-06-01

    Diagnostic assays that rely on molecular interactions have come a long way; from initial reversible detection systems towards irreversible reaction indicator-based methods. More recently, the emergence of innovative molecular amplification methodologies has revolutionised sensing, allowing diagnostic assays to achieve ultra-low limits of detection. There have been a significant number of molecular amplification approaches developed over recent years to accommodate the wide variety of analytes that require sensitive detection. To celebrate this achievement, this comprehensive critical review has been compiled to give a broad overview of the many different approaches used to attain amplification in sensing with an aim to inspire the next generation of diagnostic assays looking to achieve the ultimate detection limit. This review has been created with the focus on how each conceptually unique molecular amplification methodology achieves amplification, not just its sensitivity, while highlighting any key processes. Excluded are any references that were not found to contain an obvious molecular amplifier or amplification component, or that did not use an appropriate signal readout that could be incorporated into a sensing application. Additionally, methodologies where amplification is achieved through advances in instrumentation are also excluded. Depending upon the type of approach employed, amplification strategies are divided into four categories: target, label, signal or receptor amplification. More recent, more complex protocols combine a number of approaches and are therefore categorised by which amplification component described within was considered as the biggest advancement. The advantages and disadvantages of each methodology are discussed along with any limits of detection, if stated in the original article. Any subsequent use of the methodology within sensing or any other application is also mentioned to draw attention to its practicality. The importance of

  2. 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

  3. 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.

  4. Sensitive detection of T4 polynucleotide kinase activity based on coupled exonuclease reaction and nicking enzyme-assisted fluorescence signal amplification.

    PubMed

    Hou, Ting; Wang, Xiuzhong; Lu, Tingting; Liu, Xiaojuan; Li, Feng

    2014-05-01

    As a prominent member of the 5'-kinase family, T4 polynucleotide kinase (PNK) plays an important role in gene function regulations, and the study of PNK activity and its potential inhibitors is significant for research related to the DNA phosphorylation process. Here, we proposed a novel strategy for the detection of PNK activity and its inhibition, which combines exonuclease enzyme reaction and nicking enzyme-assisted fluorescence signal amplification. Through recycling cleavage of DNA fluorescence probe for signal amplification, a highly sensitive PNK sensing platform is developed, and a very low detection limit of 0.05 mU/mL is achieved, which is better than or comparable to that of the previously reported PNK assays. The present approach adopts a simple separation-free procedure in which the enzyme assay is conducted in homogeneous solutions. Additionally, the inhibitory effects of several known kinase inhibitors on PNK have been successfully detected. Since the proposed assay exhibits the advantages of high sensitivity and simplicity, it holds great potential in providing a promising platform for convenient and highly sensitive detection of PNK activity and its inhibitors.

  5. Sensitive electrochemical immunoassay of carcinoembryonic antigen with signal dual-amplification using glucose oxidase and an artificial catalase.

    PubMed

    Tang, Juan; Tang, Dianping; Li, Qunfang; Su, Biling; Qiu, Bin; Chen, Guonan

    2011-07-01

    A new dual-amplification strategy of electrochemical signal based on the catalytic recycling of the product was developed for the antigen-antibody interaction by glucose oxidase (GOD)- conjugated gold-silver hollow microspheres (AuAgHSs) coupled with an artificial catalase, Prussian blue nanoparticles (PB), on a graphene-based immunosensing platform. The first signal amplification introduced in this study was based on the labeled GOD on the AuAgHSs toward the catalytic oxidation of glucose. The generated H(2)O(2) was catalytically reduced by the immobilized PB on the graphene nanosheets with the second amplification. With a sandwich-type immunoassay format, carcinoembryonic antigen (CEA) was monitored as a model analyte by using the synthesized AuAgHSs as labels in pH 6.0 phosphate buffer containing 10mM glucose. Under optimal conditions, the electrochemical immunosensor exhibited a wide dynamic range of 0.005-50 ng mL(-1) with a low detection limit (LOD) of 1.0 pg mL(-1) CEA (at 3σ). Both the intra- and inter-assay coefficients of variation (CVs) were lower than 10%. The specificity and stability of the immunosensor were acceptable. In addition, the assay was evaluated for clinical serum specimens, and received a good correlation with those obtained by the referenced electrochemiluminescent (ECL). PMID:21641413

  6. Signal amplification architecture for electrochemical aptasensor based on network-like thiocyanuric acid/gold nanoparticle/ssDNA.

    PubMed

    Chen, Zhengbo; Li, Lidong; Tian, Yu; Mu, Xiaojiao; Guo, Lin

    2012-01-01

    In this work, we described signal amplification architecture for electronic aptamer-based sensor (E-AB), which is applicable to a wide range of aptamers. Herein, we only take lysozyme as the representative sensing target. The amplification method was based on the network of thiocyanuric acid (TCA)/gold nanoparticles (AuNPs) modified with ssDNA. The binding event can be detected by a decrease in the integrated charge of the surface-bound [Ru(NH(3))(6)](3+) which electrostatically absorbed onto the negatively charged phosphate backbones of DNA. In the presence of target molecules, a large amount of TCA/AuNP/ssDNA network associated with [Ru(NH(3))(6)](3+) would be removed from the electrode surface, leading to a significant decrease of redox current. Cyclic voltammetry (CV) signals of [Ru(NH(3))(6)](3+) provides quantitative measures of the concentrations of lysozyme, with a linear calibration ranging from 5 pM to 1 nM and a detection limit is 0.1 pM. The detection limit of the proposed sensor is one order of magnitude and three orders of magnitude more sensitive than the detection limits in the absence of TCA (5 pM) and in the absence of TCA/AuNP/ssDNA network (0.5 nM). This amplification method is promising for broad potential application in clinic assay and various protein analysis.

  7. Multivalent bifunctional chelator scaffolds for gallium-68 based positron emission tomography imaging probe design: signal amplification via multivalency.

    PubMed

    Singh, Ajay N; Liu, Wei; Hao, Guiyang; Kumar, Amit; Gupta, Anjali; Öz, Orhan K; Hsieh, Jer-Tsong; Sun, Xiankai

    2011-08-17

    The role of the multivalent effect has been well recognized in the design of molecular imaging probes toward the desired imaging signal amplification. Recently, we reported a bifunctional chelator (BFC) scaffold design, which provides a simple and versatile approach to impart multivalency to radiometal based nuclear imaging probes. In this work, we report a series of BFC scaffolds ((t)Bu(3)-1-COOH, (t)Bu(3)-2-(COOH)(2), and (t)Bu(3)-3-(COOH)(3)) constructed on the framework of 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA) for (68)Ga-based PET probe design and signal amplification via the multivalent effect. For proof of principle, a known integrin α(v)β(3) specific ligand (c(RGDyK)) was used to build the corresponding NOTA conjugates (H(3)1, H(3)2, and H(3)3), which present 1-3 copies of c(RGDyK) peptide, respectively, in a systematic manner. Using the integrin α(v)β(3) binding affinities (IC(50) values), enhanced specific binding was observed for multivalent conjugates (H(3)2: 43.9 ± 16.1 nM; H(3)3: 14.7 ± 5.0 nM) as compared to their monovalent counterpart (H(3)1: 171 ± 60 nM) and the intact c(RGDyK) peptide (204 ± 76 nM). The obtained conjugates were efficiently labeled with (68)Ga(3+) within 30 min at room temperature in high radiochemical yields (>95%). The in vivo evaluation of the labeled conjugates, (68)Ga-1, (68)Ga-2, and (68)Ga-3, was performed using male severe combined immunodeficiency (SCID) mice bearing integrin α(v)β(3) positive PC-3 tumor xenografts (n = 3). All (68)Ga-labeled conjugates showed high in vivo stability with no detectable metabolites found by radio-HPLC within 2 h postinjection (p.i.). The PET signal amplification in PC-3 tumor by the multivalent effect was clearly displayed by the tumor uptake of the (68)Ga-labeled conjugates ((68)Ga-3: 2.55 ± 0.50%ID/g; (68)Ga-2: 1.90 ± 0.10%ID/g; (68)Ga-1: 1.66 ± 0.15%ID/g) at 2 h p.i. In summary, we have designed and synthesized a series of NOTA-based BFC scaffolds with signal

  8. 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. PMID:24534575

  9. Linear light-scattering of gold nanostars for versatile biosensing of nucleic acids and proteins using exonuclease III as biocatalyst to signal amplification.

    PubMed

    Bi, Sai; Jia, Xiaoqiang; Ye, Jiayan; Dong, Ying

    2015-09-15

    Gold nanomaterials promise a wide range of potential applications in chemical and biological sensing, imaging, and catalysis. In this paper, we demonstrate a facile method for room-temperature synthesis of gold nanostars (AuNSs) with a size of ~50 nm via seeded growth. Significantly, the AuNSs are found to have high light-scattering properties, which are successfully used as labels for sensitive and selective detection of nucleic acids and proteins by using exonuclease III (Exo III) as a biocatalyst. For DNA detection, the binding of targets to the functionalized AuNS probes leads to the Exo III-stimulated cascade recycling amplification. As a result, a large amount of AuNSs are released from magnetic nanoparticles (MNPs) into solution, providing a greatly enhanced light-scattering signal for amplified sensing process. Moreover, a binding-induced DNA three-way junction (DNA TWJ) is introduced to thrombin detection, in which the binding of two aptamers to thrombin triggers assembly of the DNA motifs and initiates the subsequent DNA strand displacement reaction (SDR) and Exo III-assisted cascade recycling amplification. The detection limits of 89 fM and 5.6 pM are achieved for DNA and thrombin, respectively, which are comparable to or even exceed that of the reported isothermal amplification methods. It is noteworthy that based on the DNA TWJ strategy the sequences are independent on target proteins. Additionally, the employment of MNPs in the assays can not only simplify the operations but also improve the detection sensitivity. Therefore, the proposed amplified light-scattering assay with high sensitivity and selectivity, acceptable accuracy, and satisfactory versatility of analytes provides various applications in bioanalysis.

  10. 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

    2011-12-15

    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.

  11. Tuning backbones and side-chains of cationic conjugated polymers for optical signal amplification of fluorescent DNA detection.

    PubMed

    Huang, Yan-Qin; Liu, Xing-Fen; Fan, Qu-Li; Wang, Lihua; Song, Shiping; Wang, Lian-Hui; Fan, Chunhai; Huang, Wei

    2009-06-15

    Three cationic conjugated polymers (CCPs) exhibiting different backbone geometries and charge densities were used to investigate how their conjugated backbone and side chain properties, together with the transitions of DNA amphiphilic properties, interplay in the CCP/DNA-C* (DNA-C*: fluorophore-labeled DNA) complexes to influence the optical signal amplification of fluorescent DNA detection based on Förster resonance energy transfer (FRET). By examining the FRET efficiencies to dsDNA-C* (dsDNA: double-stranded DNA) and ssDNA-C* (ssDNA: single-stranded DNA) for each CCP, twisted conjugated backbones and higher charge densities were proved to facilitate electrostatic attraction in CCP/dsDNA-C* complexes, and induced improved sensitivity to DNA hybridization. Especially, by using the CCP with twisted conjugated backbone and the highest charge density, a more than 7-fold higher efficiency of FRET to dsDNA-C* was found than to ssDNA-C*, indicating a high signal amplification for discriminating between dsDNA and ssDNA. By contrast, linear conjugated backbones and lower charge density were demonstrated to favor hydrophobic interactions in CCP/ssDNA-C* complexes. These findings provided guidelines for the design of novel sensitive CCP, which can be useful to recognize many other important DNA activities involving transitions of DNA amphiphilic properties like DNA hybridization, such as specific DNA binding with ions, some secondary or tertiary structural changes of DNA, and so forth.

  12. 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.

  13. Sensitive and fast detection of fructose in complex media via symmetry breaking and signal amplification using surface-enhanced Raman spectroscopy.

    PubMed

    Sun, Fang; Bai, Tao; Zhang, Lei; Ella-Menye, Jean-Rene; Liu, Sijun; Nowinski, Ann K; Jiang, Shaoyi; Yu, Qiuming

    2014-03-01

    A new strategy is proposed to sensitively and rapidly detect analytes with weak Raman signals in complex media using surface-enhanced Raman spectroscopy (SERS) via detecting the SERS signal changes of the immobilized probe molecules on SERS-active substrates upon binding of the analytes. In this work, 4-mercaptophenylboronic acid (4-MPBA) was selected as the probe molecule which was immobilized on the gold surface of a quasi-three-dimensional plasmonic nanostructure array (Q3D-PNA) SERS substrate to detect fructose. The molecule of 4-MPBA possesses three key functions: molecule recognition and reversible binding of the analyte via the boronic acid group, amplification of SERS signals by the phenyl group and thus shielding of the background noise of complex media, and immobilization on the surface of SERS-active substrates via the thiol group. Most importantly, the symmetry breaking of the 4-MPBA molecule upon fructose binding leads to the change of area ratio between totally symmetric 8a ring mode and nontotally symmetric 8b ring mode, which enables the detection. The detection curves were obtained in phosphate-buffered saline (PBS) and in undiluted artificial urine at clinically relevant concentrations, and the limit of detection of 0.05 mM was achieved.

  14. Ultrasensitive electrochemical immunoassay for DNA methyltransferase activity and inhibitor screening based on methyl binding domain protein of MeCP2 and enzymatic signal amplification.

    PubMed

    Yin, Huanshun; Zhou, Yunlei; Xu, Zhenning; Wang, Mo; Ai, Shiyun

    2013-11-15

    In this work, we fabricated a novel electrochemical immunosensor for detection of DNA methylation, analysis of DNA MTase activity and screening of MTase inhibitor. The immunosensor was on the basis of methyl binding domain protein of MeCP2 as DNA CpG methylation recognization unit, anti-His tag antibody as "immuno-bridge" and horseradish peroxidase labeled immuneglobulin G functionalized gold nanoparticles (AuNPs-IgG-HRP) as signal amplification unit. In the presence of M. SssI MTase, the symmetrical sequence of 5'-CCGG-3' was methylated and then recognized by MeCP2 protein. By the immunoreactions, anti-His tag antibody and AuNPs-IgG-HRP was captured on the electrode surface successively. Under the catalysis effect of HRP towards hydroquinone oxidized by H2O2, the electrochemical reduction signal of benzoquinone was used to analyze M. SssI MTase activity. The electrochemical reduction signal demonstrated a wide linear relationship with M. SssI concentration ranging from 0.05 unit/mL to 90 unit/mL, achieving a detection limit of 0.017 unit/mL (S/N=3). The most important advantages of this method were its high sensitivity and good selectivity, which enabled the detection of even one-base mismatched sequence. In addition, we also verified that the developed method could be applied for screening the inhibitors of DNA MTase and for developing new anticancer drugs.

  15. 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.

  16. Nanogold-functionalized DNAzyme concatamers with redox-active intercalators for quadruple signal amplification of electrochemical immunoassay.

    PubMed

    Zhou, Jun; Lai, Wenqiang; Zhuang, Junyang; Tang, Juan; Tang, Dianping

    2013-04-10

    A novel and in situ amplified immunoassay strategy with quadruple signal amplification was designed for highly efficient electrochemical detection of low-abundance proteins (carcinoembryonic antigen, CEA, as a model) by using nanogold-functionalized DNAzyme concatamers with redox-active intercalators. To construct such an in situ amplification system, streptavidin-labeled gold nanoparticles (AuNP-SA) were initially used for the labelling of initiator strands (S0) and detection antibody (mAb2) with a large ratio (mAb2-AuNP-S0), and then two auxiliary DNA strands S1 and S2 were designed for in situ propagation of DNAzyme concatamers with the hemin/G-quadruplex format. The quadruple signal amplification was implemented by using the avidin-biotin chemistry, nanogold labels, DNA concatamers, and DNAzymes. In the presence of target CEA, the sandwiched immunocomplex was formed between the immobilized primary antibodies on the electrode and the conjugated detection antibodies on the mAb2-AuNP-S0. The carried S0 initiator strands could progress a chain reaction of hybridization events between alternating S1/S2 DNA strands to form a nicked double-helix. Upon addition of hemin, the hemin-binding aptamers could be bound to form the hemin/G-quadruplex-based DNAzymes. The formed double-helix DNA polymers could cause the intercalation of numerous electroactive methylene blue molecules. During the electrochemical measurement, the formed DNAzymes could catalyze the reduction of H2O2 in the solution to amplify the electrochemical signal of the intercalated methylene blue. Under optimal conditions, the electrochemical immunoassay exhibited a wide dynamic range of 1.0 fg mL(-1) to 20 ng mL(-1) toward CEA standards with a low detection limit of 0.5 fg mL(-1). Intra-assay and inter-assay coefficients of variation (CV) were less than 8.5% and 11.5%, respectively. No significant differences at the 0.05 significance level were encountered in the analysis of 14 clinical serum specimens

  17. 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.

  18. Highly sensitive fluorescence assay of T4 polynucleotide kinase activity and inhibition via enzyme-assisted signal amplification.

    PubMed

    Tao, Mangjuan; Zhang, Jing; Jin, Yan; Li, Baoxin

    2014-11-01

    DNA phosphorylation catalyzed by polynucleotide kinase (PNK) is an indispensable process in the repair, replication, and recombination of nucleic acids. Here, an enzyme-assisted amplification strategy was developed for the ultrasensitive monitoring activity and inhibition of T4 PNK. A hairpin oligonucleotide (hpDNA) was designed as a probe whose stem can be degraded from the 5' to 3' direction by lambda exonuclease (λ exo) when its 5' end is phosphorylated by PNK. So, the 3' stem and loop part of hpDNA was released as an initiator strand to open a molecular beacon (MB) that was designed as a fluorescence reporter, leading to a fluorescence restoration. Then, the initiator strand was released again by the nicking endonuclease (Nt.BbvCI) to hybridize with another MB, resulting in a cyclic reaction and accumulation of fluorescence signal. Based on enzyme-assisted amplification, PNK activity can be sensitively and rapidly detected with a detection limit of 1.0×10(-4)U/ml, which is superior to those of most existing approaches. Furthermore, the application of the proposed strategy for screening PNK inhibitors also demonstrated satisfactory results. Therefore, it provided a promising platform for monitoring activity and inhibition of PNK as well as for studying the activity of other nucleases.

  19. 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.

  20. 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.

  1. 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

  2. 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.

  3. Linewidth Enhancement Factor and Amplification of Angle-Modulated Optical Signals in Injection-Locked Quantum Cascade Lasers

    NASA Astrophysics Data System (ADS)

    Chattopadhyay, Taraprasad; Bhattacharyya, Prosenjit

    2016-03-01

    This paper presents a nonlinear analysis of the effect of linewidth enhancement factor (LEF) on the amplification of angle-modulated optical signals in injection-locked mid-infrared (IR) quantum cascade lasers (QCLs). A higher value of LEF tends to conserve the output angle modulation index of the amplified mid-IR signal particularly in the low-modulation frequency region. Further, a higher value of signal injection ratio produces a wider bandwidth of the locked QCL amplifier. The LEF introduces asymmetry in the lockband (LB) of the injection-locked QCL and this asymmetry increases with the increase in the value of LEF. Typically ratio of calculated lower-side LB to upper-side LB for an injection power level of - 20 dB and a LEF of unity is 1.67. The electron relaxation time in the uppermost subband lasing level in a three-level system has a profound effect on the LB asymmetry in a QCL.

  4. Resonance light scattering determination of trace bisphenol A with signal amplification by aptamer-nanogold catalysis.

    PubMed

    Yao, Dongmei; Liang, Aihui; Yin, Wenqing; Jiang, Zhiliang

    2014-08-01

    HAuCl4 was reduced by sodium citrate to prepare 10 nm gold nanoparticles (AuNPs) that were modified by the bisphenol A aptamer (Apt) to obtain an aptamer-nanogold probe (Apt-AuNP) for bisphenol A (BPA). The probes were aggregated nonspecifically to form large clusters, which showed a strong resonance light scattering (RLS) peak at 520 nm, under preparation conditions (pH 7.6 Na2HPO4-NaH2PO4 buffer and ultrasonication). Upon addition of BPA, the probe reacted specifically to form dispersed BPA-Apt-AuNP conjugates that exhibited strong catalysis of the two particle reactions of glucose-Cu(II) and hydrazine hydrochloride-Cu(II) with a strong RLS peak at 360 nm and 510 nm respectively. When the BPA concentration increased, the RLS intensity at 360 nm and 510 nm increased respectively. Accordingly, two new and highly-sensitive RLS methods were established for the detection of BPA, using the Apt-AuNP catalytic amplification.

  5. 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.

  6. 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.

  7. Significant Chiral Signal Amplification of Langmuir Monolayers Probed by Second Harmonic Generation.

    PubMed

    Lv, Kai; Lin, Lu; Wang, Xiaoyu; Zhang, Li; Guo, Yuan; Lu, Zhou; Liu, Minghua

    2015-05-01

    With the development of the nonlinear optical technique such as SHG (second harmonic generation), the in situ measurements of the chirality in the monolayers at the air/water interface have become possible. However, when performing the SHG measurement of the chirality in a monolayer, it is still a great challenge to obtain the chiral signals with a good S/N (signal-to-noise) ratio. In this Letter, interfacial assemblies with induced supramolecular chirality were used to amplify the weak chiral SHG signals from the monolayers at the air/water interface. Tetrakis(4-sulfonatophenyl) porphyrin (TPPS) J aggregates were used as the subphase, and when chiral amphiphilic molecules were spread on it, chiral domains of the amphiphile/TPPS J aggregates were formed and then significantly amplified chiral signals that otherwise could not be detected. Moreover, the sign of the DCE (degree of chiral excess) changed with the chirality of the amphiphilic molecules, thus providing a possible way to obtain the absolute chiral information in situ in the monolayers.

  8. Significant Chiral Signal Amplification of Langmuir Monolayers Probed by Second Harmonic Generation.

    PubMed

    Lv, Kai; Lin, Lu; Wang, Xiaoyu; Zhang, Li; Guo, Yuan; Lu, Zhou; Liu, Minghua

    2015-05-01

    With the development of the nonlinear optical technique such as SHG (second harmonic generation), the in situ measurements of the chirality in the monolayers at the air/water interface have become possible. However, when performing the SHG measurement of the chirality in a monolayer, it is still a great challenge to obtain the chiral signals with a good S/N (signal-to-noise) ratio. In this Letter, interfacial assemblies with induced supramolecular chirality were used to amplify the weak chiral SHG signals from the monolayers at the air/water interface. Tetrakis(4-sulfonatophenyl) porphyrin (TPPS) J aggregates were used as the subphase, and when chiral amphiphilic molecules were spread on it, chiral domains of the amphiphile/TPPS J aggregates were formed and then significantly amplified chiral signals that otherwise could not be detected. Moreover, the sign of the DCE (degree of chiral excess) changed with the chirality of the amphiphilic molecules, thus providing a possible way to obtain the absolute chiral information in situ in the monolayers. PMID:26263339

  9. Amplification of phytochrome induced morphogenesis in plants by the cryptic red light signal (CRS)

    PubMed Central

    Narayanaswamy, Guruprasad Kadur

    2009-01-01

    The regulation of endogenous levels of ascorbic acid in soybean by far-red absorbing form of phytochrome (Pfr) and by cryptic red light signal (CRS) was studied. Cryptic red light signal is produced by red light pre-irradiation of a photoreceptor other than far-red absorbing form of phytochrome (Pfr) and CRS amplifies the action of phytochrome. The endogenous level of ascorbic acid levels enhanced by phytochrome was amplified by CRS. The lifetime of CRS was from 0 to 2 h and the peak of enhancement of ascorbic acid due to CRS was between 16 to 24 h of dark incubation after the end of the treatment. CRS was found to be ineffective on UV-B enhanced endogenous levels of ascorbic acid. PMID:19816105

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

    PubMed

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

    2016-07-01

    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

  11. Signal amplification strategy for sensitive immunoassay of prostate specific antigen (PSA) based on ferrocene incorporated polystyrene spheres.

    PubMed

    Ding, Lu; You, Jinmao; Kong, Rongmei; Qu, Fengli

    2013-09-01

    A new kind of signal amplification strategy based on ferrocene (Fc) incorporated polystyrene spheres (PS-Fc) was proposed. The synthesized PS-Fc displayed narrow size distribution and good stability. PS-Fc was applied as label to develop immunosensors for prostate specific antigen (PSA) after the typical sandwich immunoreaction by linking anti-PSA antibody (Ab2) onto PS-Fc. After the fabrication of the immunosensor, tetrahydrofuran (THF) was dropped to dissolve PS and release the contained Fc for the following stripping voltammetric detection. PS-Fc as a new electrochemical label prevented the leakage of Fc and greatly amplified the immunosensor signal. In addition, the good biocompatibility of PS could maintain the bioactivity of the antibodies. The response current was linear to the logarithm of PSA concentration in the range from 0.01 ng mL(-1) to 20 ng mL(-1) with a detection limit of 1 pg mL(-1). The immunosensor results were validated through the detection of PSA in serum samples with satisfactory results.

  12. 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

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

    PubMed

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

    2016-07-01

    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.

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

    PubMed

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

    2014-11-01

    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.

  15. Amplification of JNK signaling is necessary to complete the murine gammaherpesvirus 68 lytic replication cycle.

    PubMed

    Stahl, James A; Paden, Clinton R; Chavan, Shweta S; MacLeod, Veronica; Edmondson, Ricky D; Speck, Samuel H; Forrest, J Craig

    2012-12-01

    Several studies have previously defined host-derived signaling events capable of driving lytic gammaherpesvirus replication or enhancing immediate-early viral gene expression. Yet signaling pathways that regulate later stages of the productive gammaherpesvirus replication cycle are still poorly defined. In this study, we utilized a mass spectrometric approach to identify c-Jun as an abundant cellular phosphoprotein present in late stages of lytic murine gammaherpesvirus 68 (MHV68) infection. Kinetically, c-Jun phosphorylation was enhanced as infection progressed, and this correlated with enhanced phosphorylation of the c-Jun amino-terminal kinases JNK1 and JNK2 and activation of AP-1 transcription. These events were dependent on progression beyond viral immediate-early gene expression, but not dependent on viral DNA replication. Both pharmacologic and dominant-negative blockade of JNK1/2 activity inhibited viral replication, and this correlated with inhibition of viral DNA synthesis and reduced viral gene expression. These data suggest a model in which MHV68 by necessity amplifies and usurps JNK/c-Jun signaling as infection progresses in order to facilitate late stages of the MHV68 lytic infection cycle.

  16. 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

  17. An electrochemical amplification immunoassay using bi-electrode signal transduction system.

    PubMed

    Chen, Zhao-Peng; Jiang, Jian-Hui; Zhang, Xiao-Bing; Shen, Guo-Li; Yu, Ru-Qin

    2007-03-30

    An electrochemical immunoassay technique has been developed based on the sensitive detection of the enzyme-generated product with a bi-electrode signal transduction system. The system uses two separate electrodes, an immunoelectrode and a detection electrode to form a galvanic cell to implement the redox reactions on two different electrodes, that is the enzyme-generated reductant in the anode region is electrochemically oxidized by an oxidant (silver ions) in the cathode apartment. Based on a sandwich procedure, after immunoelectrode with antibody immobilized on its surface bound with the corresponding antigen and alkaline phosphatase conjugated antibody successively, the immunoelectrode was placed in enzyme reaction solution and wired to the detection electrode which was immerged into a silver deposition solution. These two solutions are connected with a salt bridge. Thus a bi-electrode signal transduction system device is constructed in which the immunoelectrode acts as anode and the detection electrode serves as cathode. The enzyme bound on the anode surface initiates the hydrolysis of ascorbic acid 2-phosphate to produce ascorbic acid in the anode region. The ascorbic acid produced in the anodic apartment is electrochemically oxidized by silver ions coupled with the deposition of silver metal on the cathode. Via a period of 30min deposition, silver will deposited on the detection electrode in an amount corresponding to the quantity of ascorbic acid produced, leading to a great enhancement in the electrochemical stripping signal due to the accumulation of metallic silver by enzyme-generated product. Compared with the method using chemical deposition of silver, the electrochemical deposition of silver on a separate detection electrode apartment avoids the possible influence of silver deposition on the enzyme activity.

  18. Highly sensitive fluorescence assay of DNA methyltransferase activity via methylation-sensitive cleavage coupled with nicking enzyme-assisted signal amplification.

    PubMed

    Zhao, Yongxi; Chen, Feng; Wu, Yayan; Dong, Yanhua; Fan, Chunhai

    2013-04-15

    Herein, using DNA adenine methylation (Dam) methyltransferase (MTase) as a model analyte, a simple, rapid, and highly sensitive fluorescence sensing platform for monitoring the activity and inhibition of DNA MTase was developed on the basis of methylation-sensitive cleavage and nicking enzyme-assisted signal amplification. In the presence of Dam MTase, an elaborately designed hairpin probe was methylated. With the help of methylation-sensitive restriction endonuclease DpnI, the methylated hairpin probe could be cleaved to release a single-stranded DNA (ssDNA). Subsequently, this released ssDNA would hybridize with the molecular beacon (MB) to open its hairpin structure, resulting in the restoration of fluorescence signal as well as formation of the double-stranded recognition site for nicking enzyme Nt.BbvCI. Eventually, an amplified fluorescence signal was observed through the enzymatic recycling cleavage of MBs. Based on this unique strategy, a very low detection limit down to 0.06 U/mL was achieved within a short assay time (60 min) in one step, which is superior to those of most existing approaches. Owing to the specific site recognition of MTase toward its substrate, the proposed sensing system was able to readily discriminate Dam MTase from other MTase such as M.SssI and even detect the target in complex biological matrix. Furthermore, the application of the proposed sensing strategy for screening Dam MTase inhibitors was also demonstrated with satisfactory results. This novel method not only provides a promising platform for monitoring activity and inhibition of DNA MTases, but also shows great potentials in biological process researches, drugs discovery and clinical diagnostics.

  19. 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

    2016-04-01

    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.

  20. 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

    2016-04-01

    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. PMID:26980448

  1. Vesicles for Signal Amplification in a Biosensor for the Detection of Low Antigen Concentrations

    PubMed Central

    Grieshaber, Dorothee; de Lange, Victoria; Hirt, Thomas; Lu, Zhihua; Vörös, Janos

    2008-01-01

    The sensitivity of biosensors is often not sufficient to detect diagnostically relevant biomarker concentrations. In this paper we have utilized a Quartz Crystal Microbalance with Dissipation monitoring (QCM-D) to detect dissipative losses induced by the attachment of intact vesicles. We modified a sandwich assay by coupling the secondary antibodies to vesi-cles. This resulted in an increase of detection sensitivity, achieving a diagnostically relevant detection limit of 5 ng/ml or 30 pM antigens. In addition, we could combine the individual assay steps to decrease the total time to result in about 30 minutes.

  2. A protein tagging system for signal amplification in gene expression and fluorescence imaging

    PubMed Central

    Tanenbaum, Marvin E.; Gilbert, Luke A.; Qi, Lei S.; Weissman, Jonathan S.; Vale, Ronald D.

    2014-01-01

    Summary Signals in many biological processes can be amplified by recruiting multiple copies of regulatory proteins to a site of action. Harnessing this principle, we have developed a novel protein scaffold, a repeating peptide array termed SunTag, which can recruit multiple copies of an antibody-fusion protein. We show that the SunTag can recruit up to 24 copies of GFP, thereby enabling long-term imaging of single protein molecules in living cells. We also use the SunTag to create a potent synthetic transcription factor by recruiting multiple copies of a transcriptional activation domain to a nuclease-deficient CRISPR/Cas9 protein and demonstrate strong activation of endogenous gene expression and re-engineered cell behavior with this system. Thus, the SunTag provides a versatile platform for multimerizing proteins on a target protein scaffold and is likely to have many applications in imaging and in controlling biological outputs. PMID:25307933

  3. 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.

  4. Highly sensitive and selective microRNA detection based on DNA-bio-bar-code and enzyme-assisted strand cycle exponential signal amplification.

    PubMed

    Dong, Haifeng; Meng, Xiangdan; Dai, Wenhao; Cao, Yu; Lu, Huiting; Zhou, Shufeng; Zhang, Xueji

    2015-04-21

    Herein, a highly sensitive and selective microRNA (miRNA) detection strategy using DNA-bio-bar-code amplification (BCA) and Nb·BbvCI nicking enzyme-assisted strand cycle for exponential signal amplification was designed. The DNA-BCA system contains a locked nucleic acid (LNA) modified DNA probe for improving hybridization efficiency, while a signal reported molecular beacon (MB) with an endonuclease recognition site was designed for strand cycle amplification. In the presence of target miRNA, the oligonucleotides functionalized magnetic nanoprobe (MNP-DNA) and gold nanoprobe (AuNP-DNA) with numerous reported probes (RP) can hybridize with target miRNA, respectively, to form a sandwich structure. After sandwich structures were separated from the solution by the magnetic field, the RP were released under high temperature to recognize the MB and cleaved the hairpin DNA to induce the dissociation of RP. The dissociated RP then triggered the next strand cycle to produce exponential fluorescent signal amplification for miRNA detection. Under optimized conditions, the exponential signal amplification system shows a good linear range of 6 orders of magnitude (from 0.3 pM to 3 aM) with limit of detection (LOD) down to 52.5 zM, while the sandwich structure renders the system with high selectivity. Meanwhile, the feasibility of the proposed strategy for cell miRNA detection was confirmed by analyzing miRNA-21 in HeLa lysates. Given the high-performance for miRNA analysis, the strategy has a promising application in biological detection and in clinical diagnosis.

  5. 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.

  6. Detection of Q Fever Specific Antibodies Using Recombinant Antigen in ELISA with Peroxidase Based Signal Amplification

    PubMed Central

    Chen, Hua-Wei; Zhang, Zhiwen; Glennon, Erin; Ching, Wei-Mei

    2014-01-01

    Currently, the accepted method for Q fever serodiagnosis is indirect immunofluorescent antibody assay (IFA) using the whole cell antigen. In this study, we prepared the recombinant antigen of the 27-kDa outer membrane protein (Com1) which has been shown to be recognized by Q fever patient sera. The performance of recombinant Com1 was evaluated in ELISA by IFA confirmed serum samples. Due to the low titers of IgG and IgM in Q fever patients, the standard ELISA signals were further amplified by using biotinylated anti-human IgG or IgM plus streptavidin-HRP polymer. The modified ELISA can detect 88% (29 out of 33) of Q fever patient sera collected from Marines deployed to Iraq. Less than 5% (5 out of 156) of the sera from patients with other febrile diseases reacted with the Com1. These results suggest that the modified ELISA using Com1 may have the potential to improve the detection of Q fever specific antibodies. PMID:26904739

  7. Spontaneous oscillations, signal amplification and synchronization in a model of active hair bundle mechanics

    PubMed Central

    Han, Lijuan; Neiman, Alexander B.

    2010-01-01

    We study spontaneous dynamics and signal transduction in a model of active hair bundle mechanics of sensory hair cells. The hair bundle motion is subjected to internal noise resulted from thermal fluctuations and stochastic dynamics of mechano-electrical transduction ion channels. Similar to other studies we found that in the presence of noise the coherence of stochastic oscillations is maximal at a point on the bifurcation diagram away from the Andronov-Hopf bifurcation and is close to the point of maximum sensitivity of the system to weak periodic mechanical perturbations. Despite decoherent effect of noise the stochastic hair bundle oscillations can be synchronized by external periodic force of few pN amplitude in a finite range of control parameters. We then study effects of receptor potential oscillations on mechanics of the hair bundle and show that the hair bundle oscillations can be synchronized by oscillating receptor voltage. Moreover, using a linear model for the receptor potential we show that bi-directional coupling of the hair bundle and the receptor potential results in significant enhancement of the coherence of spontaneous oscillations and of the sensitivity to the external mechanical perturbations. PMID:20481759

  8. 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.

  9. Spontaneous oscillations, signal amplification, and synchronization in a model of active hair bundle mechanics

    NASA Astrophysics Data System (ADS)

    Han, Lijuan; Neiman, Alexander B.

    2010-04-01

    We study spontaneous dynamics and signal transduction in a model of active hair bundle mechanics of sensory hair cells. The hair bundle motion is subjected to internal noise resulted from thermal fluctuations and stochastic dynamics of mechanoelectrical transduction ion channels. Similar to other studies we found that in the presence of noise the coherence of stochastic oscillations is maximal at a point on the bifurcation diagram away from the Andronov-Hopf bifurcation and is close to the point of maximum sensitivity of the system to weak periodic mechanical perturbations. Despite decoherent effect of noise the stochastic hair bundle oscillations can be synchronized by external periodic force of few pN amplitude in a finite range of control parameters. We then study effects of receptor potential oscillations on mechanics of the hair bundle and show that the hair bundle oscillations can be synchronized by oscillating receptor voltage. Moreover, using a linear model for the receptor potential we show that bidirectional coupling of the hair bundle and the receptor potential results in significant enhancement of the coherence of spontaneous oscillations and of the sensitivity to the external mechanical perturbations.

  10. The spatial pattern of cochlear amplification.

    PubMed

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

    2012-12-01

    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.

  11. 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

  12. 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. PMID:26686924

  13. 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.

  14. The Ras oncogene signals centrosome amplification in mammary epithelial cells through cyclin D1/Cdk4 and Nek2

    PubMed Central

    Zeng, X; Shaikh, FY; Harrison, MK; Adon, AM; Trimboli, AJ; Carroll, KA; Sharma, N; Timmers, C; Chodosh, LA; Leone, G; Saavedra, HI

    2010-01-01

    Centrosome amplification (CA) contributes to carcinogenesis by generating aneuploidy. Elevated frequencies of CA in most benign breast lesions and primary tumors suggest a causative role for CA in breast cancers. Clearly, identifying which and how altered signal transduction pathways contribute to CA is crucial to breast cancer control. Although a causative and cooperative role for c-Myc and Ras in mammary tumorigenesis is well documented, their ability to generate CA during mammary tumor initiation remains unexplored. To answer that question, K-RasG12D and c-Myc were induced in mouse mammary glands. Although CA was observed in mammary tumors initiated by c-Myc or K-RasG12D, it was detected only in premalignant mammary lesions expressing K-RasG12D. CA, both in vivo and in vitro, was associated with increased expression of the centrosome-regulatory proteins, cyclin D1 and Nek2. Abolishing the expression of cyclin D1, Cdk4 or Nek2 in MCF10A human mammary epithelial cells expressing H-RasG12V abrogated Ras-induced CA, whereas silencing cyclin E1 or B2 had no effect. Thus, we conclude that CA precedes mammary tumorigenesis, and interfering with centrosome-regulatory targets suppresses CA. PMID:20581865

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

    PubMed

    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

  16. Fluorescence detection of telomerase activity in cancer cell extracts based on autonomous exonuclease III-assisted isothermal cycling signal amplification.

    PubMed

    Ding, Caifeng; Li, Xiaoqian; Wang, Wei; Chen, Yaoyao

    2016-09-15

    Based on the extension reaction of a telomerase substrate (TS) primer in the presence of the telomerase, strand-displacement process to perform more stable longer duplex chain, and stepwise hydrolysis of mononucleotides from the blunt or the recessed 3'-hydroxyl termini of duplex DNA in the presence of Exonuclease III (Exo III), an amplified fluorescence detection of telomerase activity in the cancer cells was described in this manuscript. A fluorescence probe DNA, a quencher DNA, and a TS primer were mixed to construct a three-chain DNA structure and a two-chain DNA structure because the amount of the TS primer was less than the other two DNA. In the presence of the telomerase, the quencher DNA was replaced from the probe DNA and the telomerase activity could be determined with the fluorescence enhancement. The telomerase activity in HeLa extracts equivalent to 6-2000 cells was detected by this method. Moreover, the strategy was further proved by using telomerase extracted from Romas cells. With the multiple rounds of isothermal strand displacement and the hydrolysis process, constituted consecutive of signal amplification for the novel detection paradigm that allowed measuring of telomerase activity in crude cancer cell extracts confirmed the reliability and practicality of the protocol, which reveal this platform holds great promise in the biochemical assay for the telomerase activity in early diagnosis for cancers.

  17. Signal Amplification Strategy Based on TiO2-Nanotube Layers and Nanobeads Carrying Quantum Dots for Electrochemiluminescent Immunosensors

    PubMed Central

    Gao, Zhi-Da; Zhuang, Qian-Lan; Song, Yan-Yan; Lee, Kiyoung; Schmuki, Patrik

    2013-01-01

    Self-organized TiO2-nanotube layers can be used for immunoassay-type sensing in combination with amplifying CdTe labels in a direct and very sensitive electrochemiluminescent (ECL) configuration. Key properties for this method are the conductivity of the TiO2 nanotubes, and their transparency for light emitted from the CdTe labels at approximately 2.4 eV. To demonstrate the potential of this platform, we constructed a sandwich-type immunoassay onto the TiO2-nanotube wall with a layer of (3-aminopropyl)triethoxysilane as the cross-linker for antibody immobilization. For the counter part of the sandwich, we created an amplification system consisting of TiO2 nanobeads carrying the secondary antibody and multiple CdTe quantum dots (multiQD). For antigen (IgG) detection, we find that this combination of 3D transparent electrode with multiQD labels allows for an ECL detection limit of 0.05 pg mL−1 and a linearity of the signal in the range of 0.1–108 pg mL−1. PMID:24551545

  18. 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

  19. Ultrasensitive electrochemical immunoassay for squamous cell carcinoma antigen using dumbbell-like Pt-Fe₃O₄ nanoparticles as signal amplification.

    PubMed

    Wu, Dan; Fan, Haixia; Li, Yuyang; Zhang, Yong; Liang, Huixin; Wei, Qin

    2013-08-15

    Dumbbell-like Pt-Fe₃O₄ nanoparticles (NPs) were synthesized and used as a novel kind of label for the preparation of electrochemical immunosensor, which is applied to the detection of cancer biomarker squamous cell carcinoma antigen (SCC-Ag). The signal amplification strategy, using the synergetic effect present in Pt-Fe₃O₄ to increase the reduction ability of the NPs toward H₂O₂, improved the sensitivity of the immunosensor. Nitrogen-doped graphene sheets (N-GS) were synthesized from graphite oxides through thermal annealing of graphite oxides in ammonia, which was used to immobilize primary anti-SCC antibody (Ab₁). Secondary anti-SCC antibody (Ab₂) was adsorbed onto the Pt-Fe₃O₄ NPs. The immunosensor was prepared through a sandwich structure and displayed a wide linear range (0.05-18 ng/mL), low detection limit (15.3 pg/mL), good reproducibility and stability. The method has been applied to the analysis of clinical serum samples with satisfactory results. These labels for immunosensors can provide many potential applications for the detection of different biomolecules.

  20. Ultra-sensitive biosensor for K-ras gene detection using enzyme capped gold nanoparticles conjugates for signal amplification.

    PubMed

    Fang, Xian; Bai, Lijuan; Han, Xiaowei; Wang, Jiao; Shi, Anqi; Zhang, Yuzhong

    2014-09-01

    In this study, an ultra-sensitive hairpin DNA-based electrochemical DNA biosensor for K-ras gene detection is described. Gold nanoparticles (Au-NPs) and horseradish peroxidase (HRP)-streptavidin capped Au-NPs (HAS) conjugates are used for signal amplification. Initially, hairpin DNA dually labeled with thiol at its 5' end and with biotin at its 3' end is immobilized on the surface of Au-NPs modified electrode, and the hairpin DNA is in a "closed" state; hence, the HAS conjugates are shielded from being approached by the biotin due to steric hindrance. However, in the presence of target DNA, the target DNA hybridizes with the loop structure of hairpin DNA and causes conformational change, resulting in biotin forced away from the electrode surface, thereby becoming accessible for the HAS conjugates. Thus, the HAS conjugates are linked to the electrode surface via the specific interaction between biotin and streptavidin. Electrochemical detection was performed in phosphate-buffered saline (PBS) containing tetramethylbenzidine (TMB) and H2O2. Under optimal conditions, the peak current differences (ΔI) are linear with the target DNA in the range from 0.1 fM to 1 nM with a detection limit of 0.035 fM. Furthermore, this biosensor also demonstrates its excellent specificity for single-base mismatched DNA. PMID:24939462

  1. A sensitive electrochemical DNA biosensor for specific detection of Enterobacteriaceae bacteria by Exonuclease III-assisted signal amplification.

    PubMed

    Luo, Caihui; Tang, Hua; Cheng, Wei; Yan, Li; Zhang, Decai; Ju, Huangxian; Ding, Shijia

    2013-10-15

    A specific and sensitive methodology was developed successfully for quantitative detection of Enterobacteriaceae bacteria by integrating Exonuclease III-assisted target recycling amplification with a simple electrochemical DNA biosensor. After target DNA hybridizes with capture DNA, Exonuclease III can selectively digest the capture DNA, which releases the target to undergo a new hybridization and cleavage cycle on sensor surface, leading to a successful target recycling. Finally, the left capture DNA is recognized by detection probe to produce the detectable signal, which decreases with the increasing target DNA concentration. Under the optimal conditions, the proposed strategy could detect target DNA down to 8.7 fM with a linear range from 0.01 pM to 1 nM, showing high sensitivity. Meanwhile, the sensing strategy was successfully used for detection of Enterobacteriaceae bacteria down to 40 CFU mL⁻¹ in milk samples. This strategy presented a simple, rapid and sensitive platform for Enterobacteriaceae bacteria detection and would become a versatile and powerful tool for food safety, biothreat detection and environmental monitoring.

  2. 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

  3. Electrochemical immunoassay for procalcitonin antigen detection based on signal amplification strategy of multiple nanocomposites.

    PubMed

    Fang, Yi-Shan; Wang, Hai-Ying; Wang, Li-Shi; Wang, Ju-Fang

    2014-01-15

    Procalcitonin, as a medium of inflammation, has become a new marker of the identification of severe bacterial infections in recent years and has received high attention due to its most ideal diagnostic indicators of specificity with major types of organism systemic inflammation of bacterial infection in the early stages. Thus, a novel method for the determination of procalcitonin (PCT) was developed based on a sandwich-type electrochemical immunosensor, which combined a simple immunosensor array as well as an effectively designed trace tag. The immunosensor was fabricated by layer-by-layer coating graphene (GC), carbon nanotubes (MWCNTs), chitosan (CS), glutaraldehyde (GA) composite on the working electrode, which can increase the electronic transfer rate and improve the surface area to capture a large number of primary antibodies (Ab1). The trace tag was prepared by loading high-content signal horseradish peroxidase labeled secondary PCT antibody (HRP-Ab2) with AuNPs, which were coated with mesoporous silica nanoparticles (MCM-41) through thionine linking. In comparison with conventional methods, the proposed immunosensor for PCT provided a better linear response range from 0.01 to 350 ng/mL and a lower limit of detection (LOD) of 0.5 pg/mL under optimal experimental conditions. In addition, the immunosensor exhibited convenience, low cost, rapidity, good specificity, acceptable stability and reproducibility. Moreover, satisfactory results were obtained for the determination of PCT in real human serum samples, indicating that the developed immunoassay has the potential to find application in clinical detection of PCT and other tumor markers as an alternative approach.

  4. Glucose encapsulating liposome for signal amplification for quantitative detection of biomarkers with glucometer readout.

    PubMed

    Zhao, Yuting; Du, Dan; Lin, Yuehe

    2015-10-15

    A new technology was developed to quantitatively detect a broad range of disease biomarkers and proven to be portable, economical, and conveniently accessible. Measurements were performed based on releasing encapsulated glucose from antibody-tagged liposomes and subsequently detecting the released glucose using a commercial personal glucose meter (GM). The innovative aspect of this approach lies in the quantification of target biomarkers through the detection of glucose, thus expanding the applicability of the GM by broadening the range of target biomarkers instead of detecting only one analyte, glucose. Because of the bilayer membrane of liposomes, which can accommodate tens of thousands of glucose molecules, the sensitivity was greatly enhanced by using glucose encapsulating liposomes as a signal output and an amplifier. Here, the model analyte, protein 53 phosphorylated on Serine 15 (phospho-p53(15)), was captured by primary antibodies bound on magnetic Fe3O4 nanoparticles and then recognized by reporting antibodies conjugated to glucose encapsulating liposomes. Finally, the target phospho-p53(15) was detected by lysing the bound liposomes to release the encapsulated glucose (4 × 10(5) glucose molecules per liposome), which is detected with the GM. This approach was demonstrated to be a universal technology that can be easily produced to quantify a wide variety of biomarkers in medical diagnostics, food safety, public health, and environmental monitoring. In the near future, it is expected that these sensors, in combination with a portable GM, can be used in many fields such as physicians' laboratories, hospitals and the common household. PMID:26005847

  5. 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.

  6. Novel One-Tube-One-Step Real-Time Methodology for Rapid Transcriptomic Biomarker Detection: Signal Amplification by Ternary Initiation Complexes.

    PubMed

    Fujita, Hiroto; Kataoka, Yuka; Tobita, Seiji; Kuwahara, Masayasu; Sugimoto, Naoki

    2016-07-19

    We have developed a novel RNA detection method, termed signal amplification by ternary initiation complexes (SATIC), in which an analyte sample is simply mixed with the relevant reagents and allowed to stand for a short time under isothermal conditions (37 °C). The advantage of the technique is that there is no requirement for (i) heat annealing, (ii) thermal cycling during the reaction, (iii) a reverse transcription step, or (iv) enzymatic or mechanical fragmentation of the target RNA. SATIC involves the formation of a ternary initiation complex between the target RNA, a circular DNA template, and a DNA primer, followed by rolling circle amplification (RCA) to generate multiple copies of G-quadruplex (G4) on a long DNA strand like beads on a string. The G4s can be specifically fluorescence-stained with N(3)-hydroxyethyl thioflavin T (ThT-HE), which emits weakly with single- and double-stranded RNA/DNA but strongly with parallel G4s. An improved dual SATIC system, which involves the formation of two different ternary initiation complexes in the RCA process, exhibited a wide quantitative detection range of 1-5000 pM. Furthermore, this enabled visual observation-based RNA detection, which is more rapid and convenient than conventional isothermal methods, such as reverse transcription-loop-mediated isothermal amplification, signal mediated amplification of RNA technology, and RNA-primed rolling circle amplification. Thus, SATIC methodology may serve as an on-site and real-time measurement technique for transcriptomic biomarkers for various diseases.

  7. Novel One-Tube-One-Step Real-Time Methodology for Rapid Transcriptomic Biomarker Detection: Signal Amplification by Ternary Initiation Complexes.

    PubMed

    Fujita, Hiroto; Kataoka, Yuka; Tobita, Seiji; Kuwahara, Masayasu; Sugimoto, Naoki

    2016-07-19

    We have developed a novel RNA detection method, termed signal amplification by ternary initiation complexes (SATIC), in which an analyte sample is simply mixed with the relevant reagents and allowed to stand for a short time under isothermal conditions (37 °C). The advantage of the technique is that there is no requirement for (i) heat annealing, (ii) thermal cycling during the reaction, (iii) a reverse transcription step, or (iv) enzymatic or mechanical fragmentation of the target RNA. SATIC involves the formation of a ternary initiation complex between the target RNA, a circular DNA template, and a DNA primer, followed by rolling circle amplification (RCA) to generate multiple copies of G-quadruplex (G4) on a long DNA strand like beads on a string. The G4s can be specifically fluorescence-stained with N(3)-hydroxyethyl thioflavin T (ThT-HE), which emits weakly with single- and double-stranded RNA/DNA but strongly with parallel G4s. An improved dual SATIC system, which involves the formation of two different ternary initiation complexes in the RCA process, exhibited a wide quantitative detection range of 1-5000 pM. Furthermore, this enabled visual observation-based RNA detection, which is more rapid and convenient than conventional isothermal methods, such as reverse transcription-loop-mediated isothermal amplification, signal mediated amplification of RNA technology, and RNA-primed rolling circle amplification. Thus, SATIC methodology may serve as an on-site and real-time measurement technique for transcriptomic biomarkers for various diseases. PMID:27347743

  8. Amplification of stimulated Brillouin scattering of two collinear pulsed laser beams with orthogonal polarizations.

    PubMed

    Shi, Jinwei; Chen, Xudong; Ouyang, Min; Liu, Juan; Liu, Dahe

    2009-06-10

    A polarization-controlling device was developed based on the fact that there can be a time delay between the seeder and the pumping beams during the amplification of a stimulated Brillouin scattering signal. The device causes two coaxially transmitted pulsed beams with orthogonal polarizations to have the same polarization in order to implement amplification by the pumping effect. An experiment showed that good pumping amplification can be achieved by using this technique. PMID:19516374

  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.

    PubMed

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

    2014-03-01

    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

    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.

  12. 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

    2014-03-01

    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. PMID:24477782

  13. Prolactin-induced prostate tumorigenesis links sustained Stat5 signaling with the amplification of basal/stem cells and emergence of putative luminal progenitors.

    PubMed

    Sackmann-Sala, Lucila; Chiche, Aurélie; Mosquera-Garrote, Nerea; Boutillon, Florence; Cordier, Corinne; Pourmir, Ivan; Pascual-Mathey, Luz; Kessal, Karima; Pigat, Natascha; Camparo, Philippe; Goffin, Vincent

    2014-11-01

    Current androgen ablation therapies for prostate cancer are initially successful, but the frequent development of castration resistance urges the generation of alternative therapies and represents an important health concern. Prolactin/signal transducer and activator of transcription 5 (STAT5) signaling is emerging as a putative target for alternative treatment for prostate cancer. However, mechanistic data for its role in development or progression of prostate tumors are scarce. In vivo mouse studies found that local prolactin induced the amplification of prostate epithelial basal/stem cells. Because these cells are proposed cells of origin for prostate cancer and disease recurrence, we looked further into this amplification. Our results indicated that sustained Stat5 activation was associated with the occurrence of abnormal basal/stem cell clusters in prostate epithelium of prostate-specific prolactin-transgenic mice. Analysis of epithelial areas containing these clusters found high proliferation, Stat5 activation, and expression of stem cell antigen 1. Furthermore, enhanced prolactin signaling also led to amplification of a luminal cell population that was positive for stem cell antigen 1. These cells may originate from amplified basal/stem cells and might represent important progenitors for tumor development in prostate epithelium. These data provide a deeper understanding of the initial stages of prostate tumorigenesis induced by prolactin to help determine whether this hormone or its downstream messengers could be useful targets for prostate cancer treatment in the future.

  14. 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.

  15. Membrane conductances involved in amplification of small signals by sodium channels in photoreceptors of drone honey bee.

    PubMed Central

    Vallet, A M; Coles, J A; Eilbeck, J C; Scott, A C

    1992-01-01

    1. Voltage signals of about 1 mV evoked in photoreceptors of the drone honey bee by shallow modulation of a background illumination of an intensity useful for behaviour are thought to be amplified by voltage-dependent Na+ channels. To elucidate the roles of the various membrane conductances in this amplification we have studied the effects of the Na+ channel blocker tetrodotoxin (TTX) and various putative K+ channel blockers on the membrane potential, Vm. 2. Superfusion of a slice of retina with 0.5-10 mM-4-aminopyridine (4-AP) depolarized the membrane and, in fifty of sixty-three cells induced repetitive action potentials. Ionophoretic injection of tetraethylammonium produced similar effects. 3. In order to measure the depolarization caused by 4-AP, action potentials were prevented by application of TTX: 4-AP was applied when the membrane was depolarized to different levels by light. 4-AP induced an additional depolarization at all membrane potentials tested (-64 to -27 mV). We conclude that there are 4-AP-sensitive K+ channels that are open at constant voltage over this range. 4. 4-AP slowed down the recovery phase of the action potential induced by a light flash by a factor that ranged from 0.51 to 0.16. This reduction could be accounted for by the reduction in a voltage-independent K+ conductance estimated from the steady-state depolarization. 5. After the voltage-gated Na+ channels had been blocked by TTX, exposure to 4-AP further changed the amplitude of the response to a small (approximately 10%) decremental light stimulus. The change was an increase when the background illumination brought Vm to potentials more negative than about -40 mV; it was a decrease when Vm > -40 mV. The data could be fitted by a circuit representation of the membrane with a light-activated conductance and a K+ conductance (EK = -66 mV) that was partly blocked by 4-AP. The voltage range studied was from -52 to -27 mV; neither conductance in the model was voltage dependent. 6. The

  16. Membrane conductances involved in amplification of small signals by sodium channels in photoreceptors of drone honey bee.

    PubMed

    Vallet, A M; Coles, J A; Eilbeck, J C; Scott, A C

    1992-10-01

    1. Voltage signals of about 1 mV evoked in photoreceptors of the drone honey bee by shallow modulation of a background illumination of an intensity useful for behaviour are thought to be amplified by voltage-dependent Na+ channels. To elucidate the roles of the various membrane conductances in this amplification we have studied the effects of the Na+ channel blocker tetrodotoxin (TTX) and various putative K+ channel blockers on the membrane potential, Vm. 2. Superfusion of a slice of retina with 0.5-10 mM-4-aminopyridine (4-AP) depolarized the membrane and, in fifty of sixty-three cells induced repetitive action potentials. Ionophoretic injection of tetraethylammonium produced similar effects. 3. In order to measure the depolarization caused by 4-AP, action potentials were prevented by application of TTX: 4-AP was applied when the membrane was depolarized to different levels by light. 4-AP induced an additional depolarization at all membrane potentials tested (-64 to -27 mV). We conclude that there are 4-AP-sensitive K+ channels that are open at constant voltage over this range. 4. 4-AP slowed down the recovery phase of the action potential induced by a light flash by a factor that ranged from 0.51 to 0.16. This reduction could be accounted for by the reduction in a voltage-independent K+ conductance estimated from the steady-state depolarization. 5. After the voltage-gated Na+ channels had been blocked by TTX, exposure to 4-AP further changed the amplitude of the response to a small (approximately 10%) decremental light stimulus. The change was an increase when the background illumination brought Vm to potentials more negative than about -40 mV; it was a decrease when Vm > -40 mV. The data could be fitted by a circuit representation of the membrane with a light-activated conductance and a K+ conductance (EK = -66 mV) that was partly blocked by 4-AP. The voltage range studied was from -52 to -27 mV; neither conductance in the model was voltage dependent. 6. The

  17. Membrane conductances involved in amplification of small signals by sodium channels in photoreceptors of drone honey bee.

    PubMed

    Vallet, A M; Coles, J A; Eilbeck, J C; Scott, A C

    1992-10-01

    1. Voltage signals of about 1 mV evoked in photoreceptors of the drone honey bee by shallow modulation of a background illumination of an intensity useful for behaviour are thought to be amplified by voltage-dependent Na+ channels. To elucidate the roles of the various membrane conductances in this amplification we have studied the effects of the Na+ channel blocker tetrodotoxin (TTX) and various putative K+ channel blockers on the membrane potential, Vm. 2. Superfusion of a slice of retina with 0.5-10 mM-4-aminopyridine (4-AP) depolarized the membrane and, in fifty of sixty-three cells induced repetitive action potentials. Ionophoretic injection of tetraethylammonium produced similar effects. 3. In order to measure the depolarization caused by 4-AP, action potentials were prevented by application of TTX: 4-AP was applied when the membrane was depolarized to different levels by light. 4-AP induced an additional depolarization at all membrane potentials tested (-64 to -27 mV). We conclude that there are 4-AP-sensitive K+ channels that are open at constant voltage over this range. 4. 4-AP slowed down the recovery phase of the action potential induced by a light flash by a factor that ranged from 0.51 to 0.16. This reduction could be accounted for by the reduction in a voltage-independent K+ conductance estimated from the steady-state depolarization. 5. After the voltage-gated Na+ channels had been blocked by TTX, exposure to 4-AP further changed the amplitude of the response to a small (approximately 10%) decremental light stimulus. The change was an increase when the background illumination brought Vm to potentials more negative than about -40 mV; it was a decrease when Vm > -40 mV. The data could be fitted by a circuit representation of the membrane with a light-activated conductance and a K+ conductance (EK = -66 mV) that was partly blocked by 4-AP. The voltage range studied was from -52 to -27 mV; neither conductance in the model was voltage dependent. 6. The

  18. 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

    2016-04-01

    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.

  19. 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 Central

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

    2016-01-01

    We developed a new magnetic nanoparticles sandwich-like immunoassay using protein cage nanoparticles (PCN) for signal amplification together with graphite furnace atomic absorption spectrometry (GFAAS) for quantification of 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 TiO2-MNPs/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 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. PMID:26953358

  20. 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

    2016-04-01

    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. PMID:26953358

  1. Hybrid chirped-pulse amplification.

    PubMed

    Jovanovic, Igor; Ebbers, Christopher A; Barty, C P J

    2002-09-15

    Conversion efficiency in optical parametric chirped-pulse amplification is limited by spatiotemporal characteristics of the pump pulse. We have demonstrated a novel hybrid chirped-pulse amplification scheme that uses a single pump pulse and combines optical parametric amplification and laser amplification to achieve high gain, high conversion efficiency, and high prepulse contrast without utilization of electro-optic modulators. We achieved an overall conversion efficiency of 37% from the hybrid amplification system at a center wavelength of 820nm. Generation of multiterawatt pulses is possible by use of this simple method and commercial Q -switched pump lasers.

  2. Nanocrystal-based electrochemiluminescence sensor for cell detection with Au nanoparticles and isothermal circular double-assisted signal amplification.

    PubMed

    Dai, Pan-Pan; Li, Jin-Yi; Yu, Tao; Xu, Jing-Juan; Chen, Hong-Yuan

    2015-08-15

    Here we have developed a sensitive cancer cell amplified detection method which combined Au NPs enhanced electrochemiluminescence (ECL) of CdS nanocrystals (NCs) film, with isothermal circular amplification reaction of polymerase. In DNA circular amplification detection system, hairpin DNA beacon/Au NPs composite modified CdS NCs film was used as an ECL emitter. Messenger DNA is hybridized with the aptamer modified on magnetic beads (MBs) to form MB-Au bioconjugates. In the presence of HL-60 cell, the aptamer would conjugate with the glycoprotein at cell surface and messenger DNA sequence would be released. The released messenger DNA sequence was then introduced into the cycle amplification system to trigger circular polymerizations. This assay allows us to determine the released messenger DNA equivalent to 10 cells and exhibits a significant specificity for HL-60 cells.

  3. 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.

  4. Sensitive detection of microRNA in complex biological samples via enzymatic signal amplification using DNA polymerase coupled with nicking endonuclease.

    PubMed

    Yin, Bin-Cheng; Liu, Yu-Qiang; Ye, Bang-Ce

    2013-12-01

    MicroRNA (miRNA) has become an ideal biomarker candidate for cancer diagnosis, prognosis, and therapy. In this study, we have developed a novel one-step method for sensitive and specific miRNA detection via enzymatic signal amplification and demonstrated its practical application in biological samples. The proposed signal amplification strategy is an integrated "biological circuit" designed to initiate a cascade of enzymatic polymerization reactions in order to detect, amplify, and measure a specific miRNA sequence by using the isothermal strand-displacement property of a mesophilic DNA polymerase together with the nicking activity of a restriction endonuclease. The circuit is composed of two molecular switches operating in series: the nicking endonuclease-assisted isothermal polymerization reaction activated by a specific miRNA and the strand-displacement polymerization reaction designed to initiate molecular beacon-assisted amplification and signal transduction. The hsa-miR-141 (miR-141) was chosen as a target miRNA because its level specifically elevates in prostate cancer. The proposed method allowed quantitative sequence-specific detection of miR-141 in a dynamic range from 1 fM to 100 nM, with an excellent ability to discriminate differences among miR-200 family members. Moreover, the detection assay was applied to quantify miR-141 in cancerous cell lysates. The results are in excellent agreement with those from the reverse transcription polymerase chain reaction method. On the basis of these findings, we believe that this proposed sensitive and specific assay has great potential as a miRNA quantification method for use in biomedical research and clinical diagnosis.

  5. Ultrasensitive detection of mRNA extracted from cancerous cells achieved by DNA rotaxane-based cross-rolling circle amplification.

    PubMed

    Bi, Sai; Cui, Yangyang; Li, Li

    2013-01-01

    An ultrasensitive and highly selective method for polymerase chain reaction-free (PCR-free) messenger RNA (mRNA) expression profiling is developed through a novel cross-rolling circle amplification (C-RCA) process based on DNA-rotaxane nanostructures. Two species of DNA pseudorotaxane (DPR) superstructures (DPR-I and DPR-II) are assembled by threading a linear DNA rod through a double-stranded DNA (dsDNA) ring containing two single-stranded gaps. In this assay, cDNA that is specific for β-actin (ACTB) mRNA is taken as a model analyte. Upon the introduction of the target cDNA, the cDNA and the biotin-modified primer are hybridized to the single-stranded regions of the DNA rod and the gap-ring, respectively. As a result, the DPR-I dethreads into free DNA macrocycle and a dumbbell-shaped DNA nanostructure. In the presence of DNA polymerase/dNTPs, two release-DNA on the DPR-I are replaced by polymerase with strand-displacement activity, which can act as the input of the DPR-II to trigger the dethreading of DPR-II and the RCA reaction, releasing another two specified release-DNA strands those in turn serve as the "mimic cDNA" for DPR-I. The C-RCA reaction then proceeds autonomously. To overcome the high background induced by hemin itself, the biotinylated rolling circle products are captured by streptavidin-coated MNPs, achieving a detection limit as low as 0.1 zmol cDNA. The assay also exhibits an excellent selectivity due to its unique DNA nanostructure fabricated through base pairing hybridization. The ACTB mRNA expression in mammary cancer cells (MCF-7) is successfully detected.

  6. Target-induced formation of gold amalgamation on DNA-based sensing platform for electrochemical monitoring of mercury ion coupling with cycling signal amplification strategy.

    PubMed

    Chen, Jinfeng; Tang, Juan; Zhou, Jun; Zhang, Lan; Chen, Guonan; Tang, Dianping

    2014-01-31

    Heavy metal ion pollution poses severe risks in human health and environmental pollutant, because of the likelihood of bioaccumulation and toxicity. Driven by the requirement to monitor trace-level mercury ion (Hg(2+)), herein we construct a new DNA-based sensor for sensitive electrochemical monitoring of Hg(2+) by coupling target-induced formation of gold amalgamation on DNA-based sensing platform with gold amalgamation-catalyzed cycling signal amplification strategy. The sensor was simply prepared by covalent conjugation of aminated poly-T(25) oligonucleotide onto the glassy carbon electrode by typical carbodiimide coupling. Upon introduction of target analyte, Hg(2+) ion was intercalated into the DNA polyion complex membrane based on T-Hg(2+)-T coordination chemistry. The chelated Hg(2+) ion could induce the formation of gold amalgamation, which could catalyze the p-nitrophenol with the aid of NaBH4 and Ru(NH3)6(3+) for cycling signal amplification. Experimental results indicated that the electronic signal of our system increased with the increasing Hg(2+) level in the sample, and has a detection limit of 0.02nM with a dynamic range of up to 1000nM Hg(2+). The strategy afforded exquisite selectivity for Hg(2+) against other environmentally related metal ions. In addition, the methodology was evaluated for the analysis of Hg(2+) in spiked tap-water samples, and the recovery was 87.9-113.8%. PMID:24439499

  7. 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.

  8. 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.

  9. 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.

  10. 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.

  11. 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.

  12. 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

    2015-12-01

    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.

  13. 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.

  14. One-step highly sensitive florescence detection of T4 polynucleotide kinase activity and biological small molecules by ligation-nicking coupled reaction-mediated signal amplification.

    PubMed

    Chen, Feng; Zhao, Yongxi; Qi, Lin; Fan, Chunhai

    2013-09-15

    DNA phosphorylation, catalyzed by polynucleotide kinase (PNK), plays significant regulatory roles in many biological events. Herein, using T4 PNK as a model target, we describe a one-step, highly sensitive, simple and rapid fluorescence approach for monitoring its activity and inhibition. This innovative strategy is inspired by the great amplification capability of ligation-nicking coupled reaction-mediated signal amplification. In the presence of T4 PNK, one of two short oligonucleotides complementary to the loop sequence of molecular beacon (MB) are phosphorylated, and then ligated with the other by DNA ligase. Upon formation of the stable duplex between the ligated DNA and MB, the fluorescence is restored and further significantly amplified through nicking endonuclease assisted cleavage of multiple MBs. Meanwhile, the cleavage of MBs will also generate new nicks to initiate the ligation reaction. Eventually, a maximum fluorescence enhancement is obtained when the ligation and nicking process reached a dynamic equilibrium. As compared to those of the existing approaches except for the assay based on single nanoparticle counting, all limited to 1:1 signal transduction function, the sensitivity (0.00001U/mL) of the proposed strategy is 100-1700 times higher. The application of the sensing system in complex biological matrix and screening of T4 PNK inhibition are demonstrated with satisfactory results. Moreover, this approach is also successfully used to detect biological small molecules such as adenosine triphosphate (ATP), and can be further extended for nicotinamide adenine dinucleotide (NAD(+)) detection.

  15. 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.

  16. Assistant DNA recycling with nicking endonuclease and molecular beacon for signal amplification using a target-complementary arched structure.

    PubMed

    Gao, Fenglei; Lei, Jianping; Ju, Huangxian

    2013-05-11

    A simple and universal method for ultrasensitive "signal on" detection of DNA was developed with a target-complementary arched structure to release assistant DNA, which was recycled with nicking endonuclease to amplify the detectable fluorescent signal of molecular beacons.

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

    PubMed

    Dutta, Anindita; Bahar, Ivet

    2010-09-01

    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 data set 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.

  18. Quantum Feedback Amplification

    NASA Astrophysics Data System (ADS)

    Yamamoto, Naoki

    2016-04-01

    Quantum amplification is essential for various quantum technologies such as communication and weak-signal detection. However, its practical use is still limited due to inevitable device fragility that brings about distortion in the output signal or state. This paper presents a general theory that solves this critical issue. The key idea is simple and easy to implement: just a passive feedback of the amplifier's auxiliary mode, which is usually thrown away. In fact, this scheme makes the controlled amplifier significantly robust, and furthermore it realizes the minimum-noise amplification even under realistic imperfections. Hence, the presented theory enables the quantum amplification to be implemented at a practical level. Also, a nondegenerate parametric amplifier subjected to a special detuning is proposed to show that, additionally, it has a broadband nature.

  19. 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.

  20. 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. PMID:26706801

  1. 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

    2014-06-01

    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.

  2. G-quadruplex DNAzyme-based chemiluminescence biosensing platform based on dual signal amplification for label-free and sensitive detection of protein.

    PubMed

    Zou, Pei; Liu, Yaling; Wang, Hongyong; Wu, Jun; Zhu, Feifan; Wu, Hao

    2016-05-15

    Detection of ultralow concentration of specific protein plays a key role in biotechnological applications and medical diagnostics. In this study, we demonstrate an amplified chemiluminescence biosensing platform for sensitive detection of protein. The biosensing platform ingeniously combines target-catalyzed hairpin assembly and Exo III-assisted signal amplification. A catalytic G-quadruplex-hemin DNAzyme is further employed to stimulate the generation of chemiluminescence in the presence of luminol and H2O2. These designs together allow a high sensitivity for the biotarget, human α-thrombin, resulting in a detection limit of 0.92 pM, and it is much lower than previous reported studies. In addition, the proposed biosensing platform is versatile. By conjugating with various recognition units, it could be employed to sensitive detect various DNA-binding proteins and might find wide applications in biomedical fields. PMID:26686920

  3. 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.

  4. 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

  5. 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

  6. 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.

  7. The flat fielding and achievable signal-to-noise of the MAMA detectors

    NASA Astrophysics Data System (ADS)

    Kaiser, Mary Elizabeth; Lindler, Don J.; Bohlin, Ralph C.

    1997-01-01

    The Space Telescope Imaging Spectrograph (STIS) was designed to achieve a signal-to-noise (S/N) of at least 100:1 per resolution element. Multi-Anode Microchannel Arrays (MAMA) observations during Servicing Mission Orbital Verification (SMOV) confirm that this specification can be met. From analysis of a single spectrum of GD153, with counting statistics of approximately 165 a S/N of approximately 125 is achieved per spectral resolution element in the far ultraviolet (FUV) over the spectral range of 1280A to 1455A. Co-adding spectra of GRW+7OD5824 to increase the counting statistics to approximately 300 yields a S/N of approximately 190 per spectral resolution element over the region extending from 1347A to 1480A in the FUV. In the near ultraviolet (NUV), a single spectrum of GRW+7OD5824 with counting statistics of approximately 200 yields a S/N of approximately 150 per spectral resolution element over the spectral region extending from 2167 to 2520A. Details of the flat field construction, the spectral extraction, and the definition of a spectral resolution element will be described in the text.

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

    PubMed

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

    2013-08-21

    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 × 10(10). 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.

  9. Ultrasensitive electroanalysis of low-level free microRNAs in blood by maximum signal amplification of catalytic silver deposition using alkaline phosphatase-incorporated gold nanoclusters.

    PubMed

    Si, Yanmei; Sun, Zongzhao; Zhang, Ning; Qi, Wei; Li, Shuying; Chen, Lijun; Wang, Hua

    2014-10-21

    An ultrasensitive sandwich-type analysis method has been initially developed for probing low-level free microRNAs (miRNAs) in blood by a maximal signal amplification protocol of catalytic silver deposition. Gold nanoclusters (AuNCs) were first synthesized and in-site incorporated into alkaline phosphatase (ALP) to form the ALP-AuNCs. Unexpectedly, the so incorporated AuNCs could dramatically enhance the catalysis activities of ALP-AuNCs versus native ALP. A sandwiched hybridization protocol was then proposed using ALP-AuNCs as the catalytic labels of the DNA detection probes for targeting miRNAs that were magnetically caught from blood samples by DNA capture probes, followed by the catalytic ligation of two DNA probes complementary to the targets. Herein, the ALP-AuNC labels could act as the bicatalysts separately in the ALP-catalyzed substrate dephosphorylation reaction and the AuNCs-accelerated silver deposition reaction. The signal amplification of ALP-AuNCs-catalyzed silver deposition was thereby maximized to be measured by the electrochemical outputs. The developed electroanalysis strategy could allow for the ultrasensitive detection of free miRNAs in blood with the detection limit as low as 21.5 aM, including the accurate identification of single-base mutant levels in miRNAs. Such a sandwich-type analysis method may circumvent the bottlenecks of the current detection techniques in probing short-chain miRNAs. It would be tailored as an ultrasensitive detection candidate for low-level free miRNAs in blood toward the diagnosis of cancer and the warning or monitoring of cancer metastasis in the clinical laboratory.

  10. Temporal and Spatial Properties of a Yeast Multi-Cellular Amplification System Based on Signal Molecule Diffusion

    PubMed Central

    Jahn, Michael; Mölle, Annett; Rödel, Gerhard; Ostermann, Kai

    2013-01-01

    We report on the spatial and temporal signaling properties of a yeast pheromone-based cell communication and amplifier system. It utilizes the Saccharomyces cerevisiae mating response pathway and relies on diffusion of the pheromone α–factor as key signaling molecule between two cell types. One cell type represents the α–factor secreting sensor part and the other the reporter part emitting fluorescence upon activation. Although multi-cellular signaling systems promise higher specificity and modularity, the complex interaction of the cells makes prediction of sensor performance difficult. To test the maximum distance and response time between sensor and reporter cells, the two cell types were spatially separated in defined compartments of agarose hydrogel (5 × 5 mm) and reconnected by diffusion of the yeast pheromone. Different ratios of sensor to reporter cells were tested to evaluate the minimum amount of sensor cells required for signal transduction. Even the smallest ratio, one α–factor-secreting cell to twenty reporter cells, generated a distinct fluorescence signal. When using a 1:1 ratio, the secreted pheromone induced fluorescence in a distance of up to four millimeters after six hours. We conclude from both our experimental results and a mathematical diffusion model that in our approach: (1) the maximum dimension of separated compartments should not exceed five millimeters in gradient direction; and (2) the time-limiting step is not diffusion of the signaling molecule but production of the reporter protein. PMID:24233076

  11. An electrochemical microRNAs biosensor with the signal amplification of alkaline phosphatase and electrochemical-chemical-chemical redox cycling.

    PubMed

    Xia, Ning; Zhang, Youjuan; Wei, Xin; Huang, Yaping; Liu, Lin

    2015-06-01

    MicroRNAs (MiRNAs) have been regarded as clinically important biomarkers and drug discovery targets. In this work, we reported a simple and ultrasensitive electrochemical method for miRNAs detection based on single enzyme amplification and electrochemical-chemical-chemical (ECC) redox cycling. Specifically, upon contact with the target miRNAs, the hairpin structure of biotinylated DNA immobilized on gold electrode was destroyed and the biotin group in DNA was forced away from the electrode surface, allowing for the coupling of streptavidin-conjugated alkaline phosphatase (SA-ALP). Then, ascorbic acid (AA, the enzymatic product of ALP) triggered the ECC redox cycling with ferrocene methanol (FcM) and tris(2-carboxyethyl)phosphine (TCEP) as the redox mediator and the chemical reducing reagent, respectively. The method was more sensitive than that with horseradish peroxidase (HRP) or glucose oxidase (GOx) triggered recycling since one ALP molecule captured by one target miRNA molecule promoted the production of thousands of AA. Analytical merits (e.g., detection limit, dynamic range, specificity, regeneration and reproducibility) were evaluated. The feasibility of the method for analysis of miRNA-21 in human serum has also been demonstrated.

  12. MeV-GeV neutrino propagation as a signal of magnetic field amplification in neutron star merger

    NASA Astrophysics Data System (ADS)

    Fraija, N.

    2016-09-01

    Short gamma-ray bursts (sGRBs) have widely been accepted to arise from a compact object binary merger; neutron star-neutron star or neutron star-black hole. During the merger of a binary neutron star system, magnetic field can be amplified beyond magnetar field strength (∼1015-1016 G) by Kelvin-Helmholtz instabilities. Considering this effect on the GRB "fireball" dynamics, we study the emission, propagation and oscillation of multi MeV-GeV neutrinos through their self-energies and using these we compute the neutrino effective potential up to order MW-4. Additionally, we calculate the number of neutrino events and neutrino flavor ratios that we would expect on Hyper-Kamiokande and DeepCore experiments. We found that MeV neutrinos in a strong magnetic field could provide information of the topology of the field, and that the number of GeV neutrinos expected in DeepCore detector would be directly affected by the strength of the field. It is worth noting that our estimates correspond to the only trustworthy method for verifying the effect of the magnetic field amplification.

  13. An electrochemical microRNAs biosensor with the signal amplification of alkaline phosphatase and electrochemical-chemical-chemical redox cycling.

    PubMed

    Xia, Ning; Zhang, Youjuan; Wei, Xin; Huang, Yaping; Liu, Lin

    2015-06-01

    MicroRNAs (MiRNAs) have been regarded as clinically important biomarkers and drug discovery targets. In this work, we reported a simple and ultrasensitive electrochemical method for miRNAs detection based on single enzyme amplification and electrochemical-chemical-chemical (ECC) redox cycling. Specifically, upon contact with the target miRNAs, the hairpin structure of biotinylated DNA immobilized on gold electrode was destroyed and the biotin group in DNA was forced away from the electrode surface, allowing for the coupling of streptavidin-conjugated alkaline phosphatase (SA-ALP). Then, ascorbic acid (AA, the enzymatic product of ALP) triggered the ECC redox cycling with ferrocene methanol (FcM) and tris(2-carboxyethyl)phosphine (TCEP) as the redox mediator and the chemical reducing reagent, respectively. The method was more sensitive than that with horseradish peroxidase (HRP) or glucose oxidase (GOx) triggered recycling since one ALP molecule captured by one target miRNA molecule promoted the production of thousands of AA. Analytical merits (e.g., detection limit, dynamic range, specificity, regeneration and reproducibility) were evaluated. The feasibility of the method for analysis of miRNA-21 in human serum has also been demonstrated. PMID:26002330

  14. 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.

  15. 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. PMID:26386905

  16. Amplification of the signal intensity of fluorescence-based fiber-optic biosensors using a Fabry-Perot resonator structure.

    PubMed

    Hsieh, Meng-Chang; Chiu, Yi-Hsin; Lin, Sheng-Fu; Chang, Jenq-Yang; Chang, Chia-Ou; Chiang, Huihua Kenny

    2015-01-01

    Fluorescent biosensors have been widely used in biomedical applications. To amplify the intensity of fluorescence signals, this study developed a novel structure for an evanescent wave fiber-optic biosensor by using a Fabry-Perot resonator structure. An excitation light was coupled into the optical fiber through a laser-drilled hole on the proximal end of the resonator. After entering the resonator, the excitation light was reflected back and forth inside the resonator, thereby amplifying the intensity of the light in the fiber. Subsequently, the light was used to excite the fluorescent molecules in the reactive region of the sensor. The experimental results showed that the biosensor signal was amplified eight-fold when the resonator reflector was formed using a 92% reflective coating. Furthermore, in a simulation, the biosensor signal could be amplified 20-fold by using a 99% reflector. PMID:25690548

  17. Graphene Ambipolar Nanoelectronics for High Noise Rejection Amplification.

    PubMed

    Liu, Che-Hung; Chen, Qi; Liu, Chang-Hua; Zhong, Zhaohui

    2016-02-10

    In a modern wireless communication system, signal amplification is critical for overcoming losses during multiple data transformations/processes and long-distance transmission. Common mode and differential mode are two fundamental amplification mechanisms, and they utilize totally different circuit configurations. In this paper, we report a new type of dual-gate graphene ambipolar device with capability of operating under both common and differential modes to realize signal amplification. The signal goes through two stages of modulation where the phase of signal can be individually modulated to be either in-phase or out-of-phase at two stages by exploiting the ambipolarity of graphene. As a result, both common and differential mode amplifications can be achieved within one single device, which is not possible in the conventional circuit configuration. In addition, a common-mode rejection ratio as high as 80 dB can be achieved, making it possible for low noise circuit application. These results open up new directions of graphene-based ambipolar electronics that greatly simplify the RF circuit complexity and the design of multifunction device operation. PMID:26808093

  18. 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…

  19. 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

  20. 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.

  1. 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.

  2. 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.

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

    PubMed Central

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

    2015-01-01

    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 102 to 107 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. PMID:26251911

  4. A simple and rapid detection assay for peptides based on the specific recognition of aptamer and signal amplification of hybridization chain reaction.

    PubMed

    Ma, Chao; Liu, Haiyun; Tian, Tian; Song, Xianrang; Yu, Jinghua; Yan, Mei

    2016-09-15

    A simple and rapid assay for the detection of peptides is designed based on the specific recognition of aptamer, the quenching effect of graphene oxide (GO) and the efficient signal amplification of hybrid chain reaction (HCR). In this assay, the hairpin structure of aptamer is opened after binding with targets, and the initiation sequence could be exposed to hairpin probe 1 (H1) to open its hairpin structure. Then the opened H1 will open the hairpin structure of hairpin probe 2 (H2), and in turn, the opened initiation sequence of H2 continues to open H1. As a result, the specific recognition of target and fluorescent signals are accumulated through the process in short 1h. Attentively, the aptamer can not only identify target peptides, but also initiate the HCR between H1 and H2. More importantly, the HCR is initiated only after the target recognition of aptamer. After HCR, the excess hairpin probes will be anchored on the GO surface, and the background is greatly reduced due to the quenching effect of GO. By using Mucin-1(MUC1) as a model peptide, the assay has a wide linear range as two orders of magnitude and the detection range is from 0.01 to 5nM with low detection limit of 3.33pM. Therefore, the simple and rapid detection of the target can be realized, and the novel assay has great potential in detecting various peptides and even cancer cells.

  5. Detection of vascular endothelial growth factor based on rolling circle amplification as a means of signal enhancement in surface plasmon resonance.

    PubMed

    Chen, Hongxia; Hou, Yafei; Qi, Fangjie; Zhang, Jiangjiang; Koh, Kwangnak; Shen, Zhongming; Li, Genxi

    2014-11-15

    Vascular endothelial growth factor (VEGF) is a major regulator of angiogenesis. It has been identified as an ideal biomarker for staging of many kinds of cancers, so more specific and intense signal is desirable for VEGF biosensors so that the sensors may have more valuable clinical application. Herein, we report a highly sensitive and selective surface plasmon resonance (SPR) sensor for VEGF detection by using two DNA aptamers which target different VEGF domains used as the capture and detection probe, respectively. Moreover, by making use of carboxyl-coated polystyrene microspheres, 3'-NH2 immobilized aptamer and 3'-NH2 modified primer DNA are loaded through amidation onto the sensing layer for further rolling circle amplification (RCA) process to amplify the SPR signal. With the well-designed sensing platform, VEGF can be determined in a linear range from 100 pg mL(-1) to 1 μg mL(-1) with a detection limit of 100 pg mL(-1). Due to its high specificity and desirable sensitivity, this RCA assisted SPR method may be a useful tool for the assay of VEGF in the future. What is more, by replacing the sensing element, i.e., the aptamer of VEGF used in this work, more biosensors for sensitive detection of other biomarkers proteins can be fabricated based on the strategy proposed in this study.

  6. Sensitive pseudobienzyme electrocatalytic DNA biosensor for mercury(II) ion by using the autonomously assembled hemin/G-quadruplex DNAzyme nanowires for signal amplification.

    PubMed

    Yuan, Yali; Gao, Min; Liu, Guangpeng; Chai, Yaqin; Wei, Shiqing; Yuan, Ruo

    2014-02-01

    Herein, a novel sensitive pseudobienzyme electrocatalytic DNA biosensor was proposed for mercury ion (Hg(2+)) detection by using autonomously assembled hemin/G-quadruplex DNAzyme nanowires for signal amplification. Thiol functionalized capture DNA was firstly immobilized on a nano-Au modified glass carbon electrode (GCE). In presence of Hg(2+), the specific coordination between Hg(2+) and T could result in the assembly of primer DNA on the electrode, which successfully triggered the HCR to form the hemin/G-quadruplex DNAzyme nanowires with substantial redox probe thionine (Thi). In the electrolyte of PBS containing NADH, the hemin/G-quadruplex nanowires firstly acted as an NADH oxidase to assist the concomitant formation of H2O2 in the presence of dissolved O2. Then, with the redox probe Thi as electron mediator, the hemin/G-quadruplex nanowires acted as an HRP-mimicking DNAzyme that quickly bioelectrocatalyzed the reduction of produced H2O2, which finally led to a dramatically amplified electrochemical signal. This method has demonstrated a high sensitivity of Hg(2+) detection with the dynamic concentration range spanning from 1.0 ng L(-1) to 10 mg L(-1) Hg(2+) and a detection limit of 0.5 ng L(-1) (2.5 pM) at the 3Sblank level, and it also demonstrated excellent selectivity against other interferential metal ions.

  7. 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.

  8. A simple and rapid detection assay for peptides based on the specific recognition of aptamer and signal amplification of hybridization chain reaction.

    PubMed

    Ma, Chao; Liu, Haiyun; Tian, Tian; Song, Xianrang; Yu, Jinghua; Yan, Mei

    2016-09-15

    A simple and rapid assay for the detection of peptides is designed based on the specific recognition of aptamer, the quenching effect of graphene oxide (GO) and the efficient signal amplification of hybrid chain reaction (HCR). In this assay, the hairpin structure of aptamer is opened after binding with targets, and the initiation sequence could be exposed to hairpin probe 1 (H1) to open its hairpin structure. Then the opened H1 will open the hairpin structure of hairpin probe 2 (H2), and in turn, the opened initiation sequence of H2 continues to open H1. As a result, the specific recognition of target and fluorescent signals are accumulated through the process in short 1h. Attentively, the aptamer can not only identify target peptides, but also initiate the HCR between H1 and H2. More importantly, the HCR is initiated only after the target recognition of aptamer. After HCR, the excess hairpin probes will be anchored on the GO surface, and the background is greatly reduced due to the quenching effect of GO. By using Mucin-1(MUC1) as a model peptide, the assay has a wide linear range as two orders of magnitude and the detection range is from 0.01 to 5nM with low detection limit of 3.33pM. Therefore, the simple and rapid detection of the target can be realized, and the novel assay has great potential in detecting various peptides and even cancer cells. PMID:27093485

  9. 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

    2015-01-01

    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. PMID:26251911

  10. Specific sorting of single bacterial cells with microfabricated fluorescence-activated cell sorting and tyramide signal amplification fluorescence in situ hybridization.

    PubMed

    Chen, Chun H; Cho, Sung H; Chiang, Hsin-I; Tsai, Frank; Zhang, Kun; Lo, Yu-Hwa

    2011-10-01

    When attempting to probe the genetic makeup of diverse bacterial communities that elude cell culturing, researchers face two primary challenges: isolation of rare bacteria from microbial samples and removal of contaminating cell-free DNA. We report a compact, low-cost, and high-performance microfabricated fluorescence-activated cell sorting (μFACS) technology in combination with a tyramide signal amplification fluorescence in situ hybridization (TSA-FISH) to address these two challenges. The TSA-FISH protocol that was adapted for flow cytometry yields a 10-30-fold enhancement in fluorescence intensity over standard FISH methods. The μFACS technology, capable of enhancing its sensitivity by ~18 dB through signal processing, was able to enrich TSA-FISH-labeled E. coli cells by 223-fold. The μFACS technology was also used to remove contaminating cell-free DNA. After two rounds of sorting on E. coli mixed with λ-phage DNA (10 ng/μL), we demonstrated over 100,000-fold reduction in λ-DNA concentration. The integrated μFACS and TSA-FISH technologies provide a highly effective and low-cost solution for research on the genomic complexity of bacteria as well as single-cell genomic analysis of other sample types. PMID:21809842

  11. 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

    2016-08-01

    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.

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

    PubMed

    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 Mn(2+) 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 Mn(2+) ions. Binding to proteins increases the relaxivity of Mn(2+) 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. PMID:27183055

  13. 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.

  14. 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.

  15. Site-selective in situ growth of fluorescent polymer-antibody conjugates with enhanced antigen detection by signal amplification.

    PubMed

    Zhang, Libin; Zhao, Wenguo; Liu, Xinyu; Wang, Guilin; Wang, Yang; Li, Dong; Xie, Liangzhi; Gao, Yan; Deng, Haiteng; Gao, Weiping

    2015-09-01

    This paper reports a new and general in situ methodology to grow fluorescent polymer conjugates from the interchain disulfide bridging sites of a monoclonal antibody. Atom transfer radical polymerization (ATRP) initiators were attached to a monoclonal antibody at its interchain disulfide bridging sites by disulfide re-bridging to yield a macroinitiator. Subsequent in situ ATRP of PEG-like monomers with dye-functionalized monomers from the macroinitiator formed antibody-polymer-dye conjugates with site-selectivity and tunable dye-to-antibody ratios. Notably, these conjugates can amplify antigen detection signal by reducing label-density dependent fluorescence quenching and by increasing dye-to-antibody ratios. The method developed may be applicable to a variety of antibodies, dyes and drugs to create a number of antibody-polymer-dye/drug conjugates for advanced diagnosis and therapy of diseases. PMID:26102329

  16. Development of silicon matrixes and channels of amplification of signals for a telescope-spectrometer of charged particles

    NASA Astrophysics Data System (ADS)

    Frolov, O. S.; Dudnik, A. V.; Sadovnichiy, A. A.; Zaitchevskiy, I. L.; Shevchenko, V. A.; Rysin, V.S.; Frolov, D. O.

    2001-08-01

    The silicon detectors will widely be used at registration of cosmic rays. If it is required to determine angular distributions of particles, will be used multi-element (for example, matrix) receivers. In the report the outcomes of development of the silicon matrix detector of the format 6x6 units and multichannel systems of preamplification formation of signals intended for a satellite telescope of charged particles are adduced. The area of a unit of the detector is 0,5 m2 , depth of a slice is 300-400 microns, concentration of an impurity in a substrate 1012 cm 3 . The role of effects of ionic drift on a surface of the detector inverse channels, initiation derivation of leakage is analysed. For their supression will be used of a protective dielectric film and guard rings of a special design. In outcome the level of an electrical noise of a unit matrix makes 5 keV at Т=25o С and 10-16 keV at Т=40o С. It provides the sure registration of electrons of high energy, that confirms also by analysis of β-spectra. The energy resolution of a unit matrix on a-particles makes about 10 keV. It is possible to lower a degree of influence of a γbackground up to 10-5 . At mining the multiway unit of preamplifiers shapers of signals it is necessary to take into account as the customary requirements (low noise, fast response time, high volume range etc.), and feature of operation of satellite instrumentation (temperature range, interference on the power supply circuit, low power consumption, high level of a radiation dose). The route of mining of amplifying channels with the purpose are analysed to satisfy with all these requirements. On samples are obtained a input noise of a channel an 10 keV, displacement of a stationary value of component on an output at a temperature variation and loading of a channel < 1 mV, volume range more than 60 dB, power consumption in one channel less than 150 mW. Correspondence to: Shevchenko Valeriy, Turgenevskaia 2, r.11, Kiev, Ukraine, 01054

  17. 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.

  18. rhEPO/EPO discrimination with ultrasensitive electrochemical biosensor based on sandwich-type nano-Au/ZnO sol-gel/nano-Au signal amplification.

    PubMed

    Zhang, Liqun; Wang, Yunxia; Wang, Jingjing; Shi, Jianfeng; Deng, Kun; Fu, Weiling

    2013-12-15

    This research established a non-labeled electrochemical biosensor for discrimination of recombinant human erythropoietin (rhEPO) and endogenous erythropoietin (EPO). We prepared a glassy carbon electrode (GCE) modified by a unique sandwich-like nano-Au/ZnO sol-gel/nano-Au compound membrane for signal amplification. The porous sol-gel structure facilitates protein activity maintenance and thermostability. Nano-Au is characterized by a large specific surface area, high surface activity, high absorbability, and good electro-conductivity and biocompatibility. By combining the advantages of both ZnO sol-gel and nano-Au, the amount of erythropoietin receptor (EPOR) increased substantially, and electron transfer of EPOR protein and electrode surface increased accordingly. In the present study, the effects of experimental conditions such as nano-Au electrodeposition time and nano-Au concentration were investigated by cyclic voltammetry, and the process of GCE modification was characterized electrochemically. We successfully developed a new method for electrochemical detection of trace rhEPO/EPO. More importantly, the response current change (ΔI) of the nano-Au/ZnO sol-gel/nano-Au modified GCE increases 3-fold when compared with that of the unmodified electrode and the sensor detection sensitivity increases significantly. In conclusion, this electrochemical biosensor is simple to prepare and allows fast, accurate, and specific detection of trace rhEPO in clinical monitoring and stimulant discrimination.

  19. 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.

  20. Target-induced reconfiguration of DNA probes for recycling amplification and signal-on electrochemical detection of hereditary tyrosinemia type I gene.

    PubMed

    Dou, Baoting; Yang, Cuiyun; Chai, Yaqin; Yuan, Ruo; Xiang, Yun

    2015-09-01

    By coupling target DNA-induced reconfiguration of the dsDNA probes with enzyme-assisted target recycling amplification, we describe the development of a signal-on electrochemical sensing approach for sensitive detection of hereditary tyrosinemia type I gene. The dsDNA probes are self-assembled on the sensing electrode, and the addition of the target DNA reconfigures and switches the dsDNA probes into active substrates for exonuclease III, which catalytically digests the probes and leads to cyclic reuse of the target DNA. The target DNA recycling and the removal of one of the ssDNA from the dsDNA probes by exonuclease III result in the formation of many hairpin structures on the sensor surface, which brings the electroactive methylene blue labels into proximity with the electrode and produces a significantly amplified current response for sensitive detection of the target gene down to 0.24 pM. This method is also selective to discriminate single-base mismatch and can be employed to detect the target gene in human serum samples. With the demonstration for the detection of the target gene, we expect the developed method to be a universal sensitive sensing platform for the detection of different nucleic acid sequences.

  1. 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.

  2. An electrochemical aptasensor for thrombin detection based on the recycling of exonuclease III and double-stranded DNA-templated copper nanoparticles assisted signal amplification.

    PubMed

    Zhao, Jing; Xin, Meiling; Cao, Ya; Yin, Yongmei; Shu, Yongqian; Ma, Wenli

    2015-02-20

    In this paper, we report an improved electrochemical aptasensor based on exonuclease III and double-stranded DNA (dsDNA)-templated copper nanoparticles (CuNPs) assisted signal amplification. In this sensor, duplex DNA from the hybridization of ligated thrombin-binding aptamer (TBA) subunits and probe DNA can act as an effective template for the formation of CuNPs on the electrode surface, so copper ions released from acid-dissolution of CuNPs may catalyze the oxidation of ο-phenylenediamine to produce an amplified electrochemical response. In the presence of thrombin, a short duplex domain with four complementary base pairs can be stabilized by the binding of TBA subunits with thrombin, in which TBA subunit 2 can be partially digested from 3' terminal with the cycle of exonuclease III, so the ligation of TBA subunits and the subsequent formation of CuNPs can be inhibited. By electrochemical characterization of dsDNA-templated CuNPs on the electrode surface, our aptasensor can display excellent performances for the detection of thrombin in a broad linear range from 100 fM to 1 nM with a low detection limit of 20.3 fM, which can also specially distinguish thrombin in both PBS and serum samples. Therefore, our aptasensor might have great potential for clinical diagnosis of biomarkers in the future.

  3. Ultrasensitive strategy based on PtPd nanodendrite/nano-flower-like@GO signal amplification for the detection of long non-coding RNA.

    PubMed

    Liu, Fei; Xiang, Guiming; Jiang, Dongneng; Zhang, Liqun; Chen, Xuemei; Liu, Linlin; Luo, Fukang; Li, Yi; Liu, Chang; Pu, Xiaoyun

    2015-12-15

    Highly up-regulated in liver cancer (HULC) is a novel promising noninvasive biomarker for hepatocellular carcinoma (HCC), which is a kind of long non-coding RNAs (lncRNAs). But traditional methods limited HULC clinical detection for ownself drawbacks. Development a new HULC detection approach is urgent and necessary. Electrochemical nucleic acid sensor based on different signal amplification strategies with high sensitivity, fast, simple, and convenient, may solve this problem. Herein, we propose a novel strategy based on Pt-Pd bimetallic nanodendrites/nanoflower-like clusters on graphene oxide/Au/horseradish peroxidase (PtPd BND/BNF@GO/Au/HRP) to enhance the catalytic efficiency and sensitivity. And Au particles were simultaneously and separately capped with thionine or detection probe, which increase the binding amount of detection probe and decrease the electronic background. The results indicated that the catalytic effect was noticeably elevated and that the biosensor provides ultrasensitive detection for the lncRNA HULC. The linear calibration of the biosensor ranged from 1.00×10(-3) to 1.00×10(3) pM/mL, and the limit of detection was 0.247 fM/mL. The lncRNA biosensor based on the PtPd BND/BNF@GO/Au/HRP/Au/thionine exhibited acceptable reproducibility and clear selectivity. This strategy may provide a new alternative for clinical HCC diagnosis through the detection of HULC.

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

    PubMed

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

    2015-12-09

    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.

  5. 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

  6. 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. PMID:27392235

  7. 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.

  8. 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.

  9. Ultrasensitive electrochemical immunosensor based on dual signal amplification process for p16(INK4a) cervical cancer detection in clinical samples.

    PubMed

    Duangkaew, Pattasuda; Tapaneeyakorn, Satita; Apiwat, Chayachon; Dharakul, Tararaj; Laiwejpithaya, Somsak; Kanatharana, Proespichaya; Laocharoensuk, Rawiwan

    2015-12-15

    The p16(INK4a) (p16) is a cyclin-dependent kinase inhibitor, which has been evaluated in several studies as a diagnostic marker of cervical cancer. Immunostaining using p16 specific antibody has confirmed an over-expression of p16 protein in cervical cancer cells and its association with disease progression. This article reports an ultrasensitive electrochemical immunosensor for specific detection of p16 and demonstrates its performance for detection of solubilized p16 protein in cell lysates obtained from patients. Sandwich-based immunoreaction couple with double signal amplification strategy based on catalytic enlargement of particle tag was used for high sensitivity and specificity. The conditions were optimized to create an immunoassay protocol. Disposable screen-printed electrode modified with capture antibodies (Ab1) was selected for further implementation towards point-of-care diagnostics. Small gold nanoparticles (15 nm diameter) conjugated with detection antibodies (Ab2) were found to better serve as a detection label due to limited interference with antigen-antibody interaction. Double signal enhancement was performed by sequential depositions of gold and silver layers. This gave the sensitivity of 1.78 μA mL(ng GST-p16)(-1) cm(-2) and detection limit of 1.3 ng mL(-1) for GST-p16 protein which is equivalent to 0.49 ng mL(-1) for p16 protein and 28 cells for HeLa cervical cancer cells. In addition to purified protein, the proposed immunosensor effectively detected elevated p16 level in cervical swab samples obtained from 10 patients with positive result from standard Pap smear test, indicating that an electrochemical immunosensors hold an excellent promise for detection of cervical cancer in clinical setting. PMID:26201985

  10. A highly sensitive homogeneous electrochemical assay for alkaline phosphatase activity based on single molecular beacon-initiated T7 exonuclease-mediated signal amplification.

    PubMed

    Zhang, Lianfang; Hou, Ting; Li, Haiyin; Li, Feng

    2015-06-21

    Alkaline phosphatase (ALP), a class of enzymes that catalyzes the dephosphorylation of a variety of substrates, is one of the most commonly assayed enzymes in routine clinical practice, and an important biomarker related to many human diseases. Herein, a facile and highly sensitive homogeneous electrochemical biosensing strategy was proposed for the ALP activity detection based on single molecular beacon-initiated T7 exonuclease-assisted signal amplification. One 3'-phosphorylated and 5'-methylene blue (MB) labeled hairpin probe (HP) is ingeniously designed. In the presence of ALP, the dephosphorylation of HP, the subsequent Klenow fragment (KF) polymerase-catalyzed elongation and T7 exonuclease-catalyzed digestion of the duplex stem of HP take place, releasing MB-labeled mononucleotides and the trigger DNA (tDNA). tDNA then hybridizes with another HP and initiates the subsequent cycling cleavage process. As a result, a large amount of MB-labeled mononucleotides are released, generating a significantly amplified electrochemical signal toward the ALP activity assay. A directly measured detection limit as low as 0.1 U L(-1) is obtained, which is comparable to that of the fluorescence method and up to three orders of magnitude lower than that of the immobilization-based electrochemical strategy previously reported. In addition to high sensitivity and good selectivity, the as-proposed strategy also exhibits the advantages of simplicity and convenience, because the assay is carried out in the homogeneous solution phase and sophisticated electrode modification processes are avoided. Therefore, the homogeneous electrochemical method we proposed here is an ideal candidate for ALP activity detection in biochemical research and clinical practices. PMID:25924941

  11. A highly sensitive homogeneous electrochemical assay for alkaline phosphatase activity based on single molecular beacon-initiated T7 exonuclease-mediated signal amplification.

    PubMed

    Zhang, Lianfang; Hou, Ting; Li, Haiyin; Li, Feng

    2015-06-21

    Alkaline phosphatase (ALP), a class of enzymes that catalyzes the dephosphorylation of a variety of substrates, is one of the most commonly assayed enzymes in routine clinical practice, and an important biomarker related to many human diseases. Herein, a facile and highly sensitive homogeneous electrochemical biosensing strategy was proposed for the ALP activity detection based on single molecular beacon-initiated T7 exonuclease-assisted signal amplification. One 3'-phosphorylated and 5'-methylene blue (MB) labeled hairpin probe (HP) is ingeniously designed. In the presence of ALP, the dephosphorylation of HP, the subsequent Klenow fragment (KF) polymerase-catalyzed elongation and T7 exonuclease-catalyzed digestion of the duplex stem of HP take place, releasing MB-labeled mononucleotides and the trigger DNA (tDNA). tDNA then hybridizes with another HP and initiates the subsequent cycling cleavage process. As a result, a large amount of MB-labeled mononucleotides are released, generating a significantly amplified electrochemical signal toward the ALP activity assay. A directly measured detection limit as low as 0.1 U L(-1) is obtained, which is comparable to that of the fluorescence method and up to three orders of magnitude lower than that of the immobilization-based electrochemical strategy previously reported. In addition to high sensitivity and good selectivity, the as-proposed strategy also exhibits the advantages of simplicity and convenience, because the assay is carried out in the homogeneous solution phase and sophisticated electrode modification processes are avoided. Therefore, the homogeneous electrochemical method we proposed here is an ideal candidate for ALP activity detection in biochemical research and clinical practices.

  12. 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

    2016-06-01

    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. PMID:27142084

  13. Pulse Recycling and Weak Value Amplification for Precision Metrology

    NASA Astrophysics Data System (ADS)

    Graham, Trent; Byard, Courtney; Kwiat, Paul; Jordan, Andrew

    2015-05-01

    Weak-value measurements have been shown to be useful for making precision optical measurements, owing to the huge amplification of tiny effects which is achievable with the technique (Hosten 2008, Dixon 2009, Egan 2012, Viza 2013). This amplification is especially helpful in the case where technical noise limits the resolution. However, if the intrinsic shot noise limits the resolution, weak-value measurements offer no advantage because the amplification is achieved via a postselection which discards most of the photons input into the measuring system. The reduction in photon number cancels the increase in signal from the amplification, and the resolution is not increased. To overcome this, we implement a method for recycling the discarded photons. We show that, for a given number of photons input to the system, recycling gives an improvement over the resolution of a conventional measurement. Our work with a simple double-pass recycling system demonstrated a 1.4x improvement over the standard shot-noise limit. We also present our work toward achieving a many-pass recycling system, for which we expect a five-fold improvement over the shot-noise limit. Such a weak-measurement recycling system could be combined with quantum states to further enhance the achievable resolution.

  14. NASBA: A detection and amplification system uniquely suited for RNA

    SciTech Connect

    Sooknanan, R.; Malek, L.T.

    1995-06-01

    The invention of PCR (polymerase chain reaction) has revolutionized our ability to amplify and manipulate a nucleic acid sequence in vitro. The commercial rewards of this revolution have driven the development of other nuclei acid amplification and detection methodologies. This has created an alphabet soup of technologies that use different amplification methods, including NASBA (nucleic acid sequence-based amplification), LCR (ligase chain reaction), SDA (strand displacement amplification), QBR (Q-beta replicase), CPR (cycling probe reaction), and bDNA (branched DNA). Despite the differences in their processes, these amplification systems can be separated into two broad categories based on how they achieve their goal: sequence-based amplification systems, such as PCR, NASBA, and SDA, amplify a target nucleic acid sequence. Signal-based amplification systems, such as LCR, QBR, CPR and bDNA, amplify or alter a signal from a detection reaction that is target-dependent. While the various methods have relative strengths and weaknesses, only NASBA offers the unique ability to homogeneously amplify an RNA analyte in the presence of homologous genomic DNA under isothermal conditions. Since the detection of RNA sequences almost invariably measures biological activity, it is an excellent prognostic indicator of activities as diverse as virus production, gene expression, and cell viability. The isothermal nature of the reaction makes NASBA especially suitable for large-scale manual screening. These features extend NASBA`s application range from research to commercial diagnostic applications. Field test kits are presently under development for human diagnostics as well as the burgeoning fields of food and environmental diagnostic testing. These developments suggest future integration of NASBA into robotic workstations for high-throughput screening as well. 17 refs., 1 tab.

  15. 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.

  16. 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. PMID:26451956

  17. 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.

  18. 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.

  19. 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. PMID:27498326

  20. 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

  1. Stacking nonenzymatic circuits for high signal gain

    PubMed Central

    Chen, Xi; Briggs, Neima; McLain, Jeremy R.; Ellington, Andrew D.

    2013-01-01

    Signal amplification schemes that do not rely on protein enzymes show great potential in areas as abstruse as DNA computation and as applied as point-of-care molecular diagnostics. Toehold-mediated strand displacement, a programmable form of dynamic DNA hybridization, can be used to design powerful amplification cascades that can achieve polynomial or exponential amplification of input signals. However, experimental implementation of such amplification cascades has been severely hindered by circuit leakage due to catalyst-independent side reactions. In this study, we systematically analyzed the origins, characteristics, and outcomes of circuit leakage in amplification cascades and devised unique methods to obtain high-quality DNA circuits that exhibit minimal leakage. We successfully implemented a two-layer cascade that yielded 7,000-fold signal amplification and a two-stage, four-layer cascade that yielded upward of 600,000-fold signal amplification. Implementation of these unique methods and design principles should greatly empower molecular programming in general and DNA-based molecular diagnostics in particular. PMID:23509255

  2. Limitation of the achievable signal-to-noise ratio in optical coherence tomography due to mismatch of the balanced receiver.

    PubMed

    Rosa, Carla Carmelo; Podoleanu, Adrian Gh

    2004-09-01

    Owing to the limited spectral response of the fiber directional coupler used in a balanced optical coherence tomography configuration, the spectra are different in the two outputs. This affects unfavorably operation of the balanced photodetector unit. Excess photon noise makes a larger contribution than a directional coupler with a flat spectral response. A theoretical model is developed that shows that an optimum set of parameters may be defined to maximize the achievable signal-to-noise ratio. The model leads to a redefinition of the effective noise bandwidth, which takes into account the nonflat response of the directional coupler used. The model also predicts a limitation on the signal-to-noise ratio even when the stray reflectances in the interferometer are brought to zero. PMID:15449466

  3. Single Cell Transcriptome Amplification with MALBAC

    PubMed Central

    Tan, Longzhi; Tang, Fuchou; Xie, X. Sunney

    2015-01-01

    Recently, Multiple Annealing and Looping-Based Amplification Cycles (MALBAC) has been developed for whole genome amplification of an individual cell, relying on quasilinear instead of exponential amplification to achieve high coverage. Here we adapt MALBAC for single-cell transcriptome amplification, which gives consistently high detection efficiency, accuracy and reproducibility. With this newly developed technique, we successfully amplified and sequenced single cells from 3 germ layers from mouse embryos in the early gastrulation stage, and examined the epithelial-mesenchymal transition (EMT) program among cells in the mesoderm layer on a single-cell level. PMID:25822772

  4. Ultrasensitive electrochemical aptasensor for the detection of thrombin based on dual signal amplification strategy of Au@GS and DNA-CoPd NPs conjugates.

    PubMed

    Wang, Yaoguang; Zhang, Yong; Yan, Tao; Fan, Dawei; Du, Bin; Ma, Hongmin; Wei, Qin

    2016-06-15

    In this work, an ultrasensitive electrochemical aptasensor for the detection of thrombin was developed based on Au nanoparticles decorated graphene sheet (Au@GS) and CoPd binary nanoparticles (CoPd NPs). A sulfydryl-labeled thrombin capture probe (Apt1) and a biotin-labeled thrombin reporter probe (Apt2) were designed to achieve a sandwich-type strategy. Au@GS was used as a sensing platform for the facile immobilization of Apt1 through Au-S bond, forming a sensing interface for thrombin. The specific recognition of thrombin induced the attachment of Apt2-CoPd NPs to the electrode. The labeled CoPd NPs showed good catalytic properties toward the reduction of H2O2, resulting in an amperometric signal. The amperometric response was correlated to the thrombin concentration in sample solutions. Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) confirmed the successful fabrication of the aptasensor. A linear response to thrombin in the range of 0.01-2.00 ng mL(-1) with a low detection limit (5 pg mL(-1)) was achieved. The proposed aptasensor showed good selectivity, good reproducibility and acceptable stability. This proposed strategy may find many potential applications in the detection of other biomolecules.

  5. Theoretical limit of repeater spacing in an optical transmission line utilizing Raman amplification

    NASA Astrophysics Data System (ADS)

    Nakashima, T.; Seikai, S.; Nakazawa, M.; Negishi, Y.

    1986-08-01

    The limit of repeater spacing is investigated theoretically for optical transmission lines utilizing stimulated Raman scattering to amplify the signal light. Achievable repeater spacing is numerically estimated on the basis of coupled power equations and measured fiber characteristics for various signal wavelengths and relative index differences. Three types of transmission line configurations are considered, i.e., utilization of forward only, backward only, and bidirectional amplification. In the third case, a transmission distance of more than 400 km is predicted for an input signal light power of 100 microW, a signal wavelength of 1.57 microns, and a pump power of 0.5 W using a fiber with a relative index difference of 1 percent for signal light amplification.

  6. 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.

  7. 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.

  8. Highly Robust Nanopore-Based Dual-Signal-Output Ion Detection System for Achieving Three Successive Calibration Curves.

    PubMed

    Xu, Xuemei; Hou, Ruizuo; Gao, Pengcheng; Miao, Mao; Lou, Xiaoding; Liu, Bifeng; Xia, Fan

    2016-02-16

    In recent years, artificial stimuli-responsive bioinspired nanopores have attracted a lot of attention due to their unique property of confined spaces and flexibility in terms of shapes and sizes. Most of the nanopore systems demonstrated their transmembrane properties and applications in target detections. However, almost all of the nanopores can be used only once due to either the irreversible reactions between targets and probes or the plugged nanopores not easily being unplugged again. In this work, we propose a dual-signal-output nanopore system that could detect the cations (Hg(2+)) inducing the plugged nanopores. The detection system is highly recoverable by the anions (S(2-)) inducing the unplugged nanopores. More importantly, as far as we know, it is seldom reported for the same nanopores to achieve successive calibration curves for three times by subsequent reversible plug-unplug processes, which strongly demonstrates the high robustness of this novel nanopore-detection system. In addition, unlike monitoring the plug-unplug phenomena by only one type of signal, we combined the ionic current signal with the fluorescence output and could directly observe that the change of ionic current does in fact correspond to the plug-unplug of the nanopores by the target stimuli. PMID:26754059

  9. Phase matched parametric amplification via four-wave mixing in optical microfibers.

    PubMed

    Abdul Khudus, Muhammad I M; De Lucia, Francesco; Corbari, Costantino; Lee, Timothy; Horak, Peter; Sazio, Pier; Brambilla, Gilberto

    2016-02-15

    Four-wave mixing (FWM) based parametric amplification in optical microfibers (OMFs) is demonstrated over a wavelength range of over 1000 nm by exploiting their tailorable dispersion characteristics to achieve phase matching. Simulations indicate that for any set of wavelengths satisfying the FWM energy conservation condition there are two diameters at which phase matching in the fundamental mode can occur. Experiments with a high-power pulsed source working in conjunction with a periodically poled silica fiber (PPSF), producing both fundamental and second harmonic signals, are undertaken to investigate the possibility of FWM parametric amplification in OMFs. Large increases of idler output power at the third harmonic wavelength were recorded for diameters close to the two phase matching diameters. A total amplification of more than 25 dB from the initial signal was observed in a 6 mm long optical microfiber, after accounting for the thermal drift of the PPSF and other losses in the system. PMID:26872182

  10. 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.

  11. 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).

  12. 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

  13. Hybrid chirped pulse amplification system

    DOEpatents

    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.

  14. Intermolecular communication on a liposomal membrane: enzymatic amplification of a photonic signal with a gemini peptide lipid as a membrane-bound artificial receptor.

    PubMed

    Mukai, Masaru; Maruo, Kohei; Sasaki, Yoshihiro; Kikuchi, Jun-ichi

    2012-03-12

    A supramolecular system that can activate an enzyme through photo-isomerization was constructed by using a liposomal membrane scaffold. The design of the system was inspired by natural signal transduction systems, in which enzymes amplify external signals to control signal transduction pathways. The liposomal membrane, which provided a scaffold for the system, was prepared by self-assembly of a photoresponsive receptor and a cationic synthetic lipid. NADH-dependent L-lactate dehydrogenase, the signal amplifier, was immobilized on the liposomal surface by electrostatic interactions. Recognition of photonic signals by the membrane-bound receptor induced photo-isomerization, which significantly altered the receptor's metal-binding affinity. The response to the photonic signal was transmitted to the enzyme by Cu(2+) ions. The enzyme amplified the chemical information through a catalytic reaction to generate the intended output signal.

  15. MYCN Gene Amplification

    PubMed Central

    Yoshimoto, Maisa; Caminada de Toledo, Silvia Regina; Monteiro Caran, Eliana Maria; de Seixas, Maria Teresa; de Martino Lee, Maria Lucia; de Campos Vieira Abib, Simone; Vianna, Sonia Maria Rossi; Schettini, Sergio Thomaz; Anderson Duffles Andrade, Joyce

    1999-01-01

    Neuroblastoma is the second most common solid tumor occurring in children. Amplification of the MYCN oncogene is associated with poor prognosis. To identify neuroblastoma tumors with MYCN amplification, we studied the number of copies of MYCN in interphase cells by fluorescence in situ hybridization in 20 neuroblastoma patients. MYCN amplification appeared in 7 tumor specimens. Interphase and metaphase studies showed a tumor cell population with both forms of amplification, double minutes and homogeneously staining regions, in two patients. These patients showed a smaller tumor cell subpopulation with the presence of more than one homogeneously staining region, suggesting that gene amplification was undergoing karyotype evolution. PMID:10550298

  16. 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.

  17. Backaction amplification and quantum limits in optomechanical measurements.

    PubMed

    Verlot, P; Tavernarakis, A; Briant, T; Cohadon, P-F; Heidmann, A

    2010-04-01

    Optical interferometry is by far the most sensitive displacement measurement technique available, with sensitivities at the 10(-20) m/square root(Hz) level in the large-scale gravitational-wave interferometers currently in operation. Second-generation interferometers will experience a tenfold improvement in sensitivity and be mainly limited by quantum noise, close to the standard quantum limit (SQL), once considered as the ultimate displacement sensitivity achievable by interferometry. In this Letter, we experimentally demonstrate one of the techniques envisioned to go beyond the SQL: amplification of a signal by radiation-pressure backaction in a detuned cavity.

  18. Can Anomalous Amplification be Attained without Postselection?

    NASA Astrophysics Data System (ADS)

    Martínez-Rincón, Julián; Liu, Wei-Tao; Viza, Gerardo I.; Howell, John C.

    2016-03-01

    We present a parameter estimation technique based on performing joint measurements of a weak interaction away from the weak-value-amplification approximation. Two detectors are used to collect full statistics of the correlations between two weakly entangled degrees of freedom. Without discarding of data, the protocol resembles the anomalous amplification of an imaginary-weak-value-like response. The amplification is induced in the difference signal of both detectors allowing robustness to different sources of technical noise, and offering in addition the advantages of balanced signals for precision metrology. All of the Fisher information about the parameter of interest is collected. A tunable phase controls the strength of the amplification response. We experimentally demonstrate the proposed technique by measuring polarization rotations in a linearly polarized laser pulse. We show that in the presence of technical noise the effective sensitivity and precision of a split detector is increased when compared to a conventional continuous-wave balanced detection technique.

  19. Cochlear amplification, outer hair cells and prestin

    PubMed Central

    Dallos, Peter

    2008-01-01

    Mechanical amplification of acoustic signals is apparently a common feature of vertebrate auditory organs. In non-mammalian vertebrates amplification is produced by stereociliary processes, related to the mechanotransducer channel complex and probably to the phenomenon of fast adaptation. The extended frequency range of the mammalian cochlea has likely co-evolved with a novel hair cell type, the outer hair cell and its constituent membrane protein, prestin. Cylindrical outer hair cells are motile and their somatic length changes are voltage driven and powered by prestin. One of the central outstanding problems in mammalian cochlear neurobiology is the relation between the two amplification processes. PMID:18809494

  20. In vivo and in vitro glucose-induced biphasic insulin secretion in the mouse: pattern and role of cytoplasmic Ca2+ and amplification signals in beta-cells.

    PubMed

    Henquin, Jean-Claude; Nenquin, Myriam; Stiernet, Patrick; Ahren, Bo

    2006-02-01

    The mechanisms underlying biphasic insulin secretion have not been completely elucidated. We compared the pattern of plasma insulin changes during hyperglycemic clamps in mice to that of glucose-induced insulin secretion and cytosolic calcium concentration ([Ca(2+)](c)) changes in perifused mouse islets. Anesthetized mice were infused with glucose to clamp blood glucose at 8.5 (baseline), 11.1, 16.7, or 30 mmol/l. A first-phase insulin response consistently peaked at 1 min, and a slowly ascending second phase occurred at 16.7 and 30 mmol/l glucose. Glucose-induced insulin secretion in vivo is thus biphasic, with a similarly increasing second phase in the mouse as in humans. In vitro, square-wave stimulation from a baseline of 3 mmol/l glucose induced similar biphasic insulin secretion and [Ca(2+)](c) increases, with sustained and flat second phases. The glucose dependency (3-30 mmol/l) of both changes was sigmoidal with, however, a shift to the right of the relation for insulin secretion compared with that for [Ca(2+)](c). The maximum [Ca(2+)](c) increase was achieved by glucose concentrations, causing half-maximum insulin secretion. Because this was true for both phases, we propose that contrary to current concepts, amplifying signals are also implicated in first-phase glucose-induced insulin secretion. To mimic in vivo conditions, islets were stimulated with high glucose after being initially perifused with 8.5 instead of 3.0 mmol/l glucose. First-phase insulin secretion induced by glucose at 11.1, 16.7, and 30 mmol/l was decreased by approximately 50%, an inhibition that could not be explained by commensurate decreases in [Ca(2+)](c) or in the pool of readily releasable granules. Also unexpected was the gradually ascending pattern of the second phase, now similar to that in vivo. These observations indicated that variations in prestimulatory glucose can secondarily affect the magnitude and pattern of subsequent glucose-induced insulin secretion.

  1. 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.

  2. 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.

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

    PubMed

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

    2016-08-19

    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. PMID:27378514

  4. A new signal amplification strategy of photoelectrochemical immunoassay for highly sensitive interleukin-6 detection based on TiO2/CdS/CdSe dual co-sensitized structure.

    PubMed

    Fan, Gao-Chao; Ren, Xiao-Lin; Zhu, Cheng; Zhang, Jian-Rong; Zhu, Jun-Jie

    2014-09-15

    Dual co-sensitized structure of TiO2/CdS/CdSe was designed to develop a novel photoelectrochemical immunoassay for highly sensitive detection of human interleukin-6 (IL-6). To construct a sensing electrode, TiO2/CdS hybrid was prepared by successive adsorption and reaction of Cd(2+) and S(2-) ions on the surface of TiO2 and then was employed as matrix for immobilization of anti-IL-6 antibody, whereas CdSe QDs linked to IL-6 were used for signal amplification via the specific antibody-antigen immunoreaction between anti-IL-6 and IL-6-CdSe bioconjugate. Greatly enhanced sensitivity for IL-6 detection was derived from the new co-sensitization signal amplification strategy. First, the TiO2/CdS/CdSe co-sensitized structure extended the absorption range to long wavelength of white light, which adequately utilized the light energy. Second, the TiO2/CdS/CdSe co-sensitized structure possessed stepwise band-edge levels favoring ultrafast transfer of photogenerated electrons and significantly prompted the photoelectrochemical performance. Besides, the introduction of CdSe effectively prevented the recombination of photogenerated electrons in the conduction band of CdS, further causing an enhanced photocurrent. Accordingly, upon the co-sensitization strategy, a novel immunoassay based on the competitive binding of anti-IL-6 antibody with IL-6 antigen and IL-6-CdSe bioconjugate was developed, and it exhibited a wide linear range from 1.0 pg/mL to 100 ng/mL with a low detection limit of 0.38 pg/mL for IL-6 detection. The proposed co-sensitization strategy presented high sensitivity, reproducibility, specificity and stability, and also opened up a new promising platform for detection of other biomarkers.

  5. Ultrasensitive solution-phase electrochemical molecular beacon-based DNA detection with signal amplification by exonuclease III-assisted target recycling.

    PubMed

    Xuan, Feng; Luo, Xiaoteng; Hsing, I-Ming

    2012-06-19

    Taking advantage of the preferential exodeoxyribonuclease activity of exonuclease III in combination with the difference in diffusivity between an oligonucleotide and a mononucleotide toward a negatively charged ITO electrode, a highly sensitive and selective electrochemical molecular beacon (eMB)-based DNA sensor has been developed. This sensor realizes electrochemical detection of DNA in a homogeneous solution, with sensing signals amplified by an exonuclease III-based target recycling strategy. A hairpin-shaped oligonucleotide containing the target DNA recognition sequence, with a methylene blue tag close to the 3' terminus, is designed as the signaling probe. Hybridization with the target DNA transforms the probe's exonuclease III-inactive protruding 3' terminus into an exonuclease III-active blunt end, triggering the digestion of the probe into mononucleotides including a methylene blue-labeled electro-active mononucleotide (eNT). The released eNT, due to its less negative charge and small size, diffuses easily to the negative ITO electrode, resulting in an increased electrochemical signal. Meanwhile, the intact target DNA returns freely to the solution and hybridizes with other probes, releasing multiple eNTs and thereby further amplifies the electrochemical signal. This new immobilization-free, signal-amplified electrochemical DNA detection strategy shows great potential to be integrated in portable and cost-effective DNA sensing devices.

  6. Spheromak Impedance and Current Amplification

    SciTech Connect

    Fowler, T K; Hua, D D; Stallard, B W

    2002-01-31

    It is shown that high current amplification can be achieved only by injecting helicity on the timescale for reconnection, {tau}{sub REC}, which determines the effective impedance of the spheromak. An approximate equation for current amplification is: dI{sub TOR}{sup 2}/dt {approx} I{sup 2}/{tau}{sub REC} - I{sub TOR}{sup 2}/{tau}{sub closed} where I is the gun current, I{sub TOR} is the spheromak toroidal current and {tau}{sub CLOSED} is the ohmic decay time of the spheromak. Achieving high current amplification, I{sub TOR} >> I, requires {tau}{sub REC} <<{tau}{sub CLOSED}. For resistive reconnection, this requires reconnection in a cold zone feeding helicity into a hot zone. Here we propose an impedance model based on these ideas in a form that can be implemented in the Corsica-based helicity transport code. The most important feature of the model is the possibility that {tau}{sub REC} actually increases as the spheromak temperature increases, perhaps accounting for the ''voltage sag'' observed in some experiments, and a tendency toward a constant ratio of field to current, B {proportional_to} I, or I{sub TOR} {approx} I. Program implications are discussed.

  7. Multiplexed protein profiling on microarrays by rolling-circle amplification

    PubMed Central

    Schweitzer, Barry; Roberts, Scott; Grimwade, Brian; Shao, Weiping; Wang, Minjuan; Fu, Qin; Shu, Quiping; Laroche, Isabelle; Zhou, Zhimin; Tchernev, Velizar T.; Christiansen, Jason; Velleca, Mark; Kingsmore, Stephen F.

    2010-01-01

    Fluorescent-sandwich immunoassays on microarrays hold appeal for proteomics studies, because equipment and antibodies are readily available, and assays are simple, scalable, and reproducible. The achievement of adequate sensitivity and specificity, however, requires a general method of immunoassay amplification. We describe coupling of isothermal rolling-circle amplification (RCA) to universal antibodies for this purpose. A total of 75 cytokines were measured simultaneously on glass arrays with signal amplification by RCA with high specificity, femtomolar sensitivity, 3 log quantitative range, and economy of sample consumption. A 51-feature RCA cytokine glass array was used to measure secretion from human dendritic cells (DCs) induced by lipopolysaccharide (LPS) or tumor necrosis factor-α (TNF-α). As expected, LPS induced rapid secretion of inflammatory cytokines such as macrophage inflammatory protein (MIP)-1β, interleukin (IL)-8, and interferon-inducible protein (IP)-10. We found that eotaxin-2 and I-309 were induced by LPS; in addition, macrophage-derived chemokine (MDC), thymus and activation-regulated chemokine (TARC), soluble interleukin 6 receptor (sIL-6R), and soluble tumor necrosis factor receptor I (sTNF-RI) were induced by TNF-α treatment. Because microarrays can accommodat ~1,000 sandwich immunoassays of this type, a relatively small number of RCA microarrays seem to offer a tractable approach for proteomic surveys. PMID:11923841

  8. 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.

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

    DOE PAGES

    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.

  10. 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. PMID:18710880

  11. 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.

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

    PubMed Central

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

    2008-01-01

    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. PMID:18710880

  13. 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.

  14. 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

  15. Hydrogen peroxide stimulation of CFTR reveals an Epac-mediated, soluble AC-dependent cAMP amplification pathway common to GPCR signalling

    PubMed Central

    Ivonnet, P; Salathe, M; Conner, G E

    2015-01-01

    BACKGROUND AND PURPOSE H2O2 is widely understood to regulate intracellular signalling. In airway epithelia, H2O2 stimulates anion secretion primarily by activating an autocrine PGE2 signalling pathway via EP4 and EP1 receptors to initiate cytic fibrosis transmembrane regulator (CFTR)-mediated Cl− secretion. This study investigated signalling downstream of the receptors activated by H2O2. EXPERIMENTAL APPROACH Anion secretion by differentiated bronchial epithelial cells was measured in Ussing chambers during stimulation with H2O2, an EP4 receptor agonist or β2-adrenoceptor agonist in the presence and absence of inhibitors of ACs and downstream effectors. Intracellular calcium ([Ca2+]I) changes were followed by microscopy using fura–2-loaded cells and PKA activation followed by FRET microscopy. KEY RESULTS Transmembrane adenylyl cyclase (tmAC) and soluble AC (sAC) were both necessary for H2O2 and EP4 receptor-mediated CFTR activation in bronchial epithelia. H2O2 and EP4 receptor agonist stimulated tmAC to increase exchange protein activated by cAMP (Epac) activity that drives PLC activation to raise [Ca2+]i via Ca2+ store release (and not entry). Increased [Ca2+]i led to sAC activation and further increases in CFTR activity. Stimulation of sAC did not depend on changes in [HCO3−]. Ca2+-activated apical KCa1.1 channels and cAMP-activated basolateral KV7.1 channels contributed to H2O2-stimulated anion currents. A similar Epac-mediated pathway was seen following β2-adrenoceptor or forskolin stimulation. CONCLUSIONS AND IMPLICATIONS H2O2 initiated a complex signalling cascade that used direct stimulation of tmACs by Gαs followed by Epac-mediated Ca2+ crosstalk to activate sAC. The Epac-mediated Ca2+ signal constituted a positive feedback loop that amplified CFTR anion secretion following stimulation of tmAC by a variety of stimuli. PMID:25220136

  16. Weak value amplification considered harmful

    NASA Astrophysics Data System (ADS)

    Ferrie, Christopher; Combes, Joshua

    2014-03-01

    We show using statistically rigorous arguments that the technique of weak value amplification does not perform better than standard statistical techniques for the tasks of parameter estimation and signal detection. We show that using all data and considering the joint distribution of all measurement outcomes yields the optimal estimator. Moreover, we show estimation using the maximum likelihood technique with weak values as small as possible produces better performance for quantum metrology. In doing so, we identify the optimal experimental arrangement to be the one which reveals the maximal eigenvalue of the square of system observables. We also show these conclusions do not change in the presence of technical noise.

  17. 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%.

  18. 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.

  19. 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%.

  20. 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.

  1. 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.

  2. 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. PMID:27452670

  3. Short-pulse amplification by strongly coupled stimulated Brillouin scattering

    NASA Astrophysics Data System (ADS)

    Edwards, Matthew R.; Jia, Qing; Mikhailova, Julia M.; Fisch, Nathaniel J.

    2016-08-01

    We examine the feasibility of strongly coupled stimulated Brillouin scattering as a mechanism for the plasma-based amplification of sub-picosecond pulses. In particular, we use fluid theory and particle-in-cell simulations to compare the relative advantages of Raman and Brillouin amplification over a broad range of achievable parameters.

  4. 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. PMID:26164013

  5. 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.

  6. 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.}

  7. Remote fiber sensors and optical amplification

    NASA Astrophysics Data System (ADS)

    Pontes, M. J.; Coelho, Thiago V. N.; Carvalho, Joel P.; Santos, J. L.; Guerreiro, A.

    2013-11-01

    This work discusses remote fiber sensors enabled by optical amplification. Continuous wave numerical modeling based on the propagation of pumps and signal lasers coupled to optical fibers explores Raman amplification schemes to predict the sensor's behavior. Experimental analyses report the results to a temperature remote optical sensor with 50 km distance between the central unit and the sensor head. An electrical interrogation scheme is used due to their low cost and good time response. Different architectures in remote sensor systems are evaluated, where diffraction gratings are the sensor element. A validation of calculated results is performed by experimental analyses and, as an application, the noise generated by Raman amplification in the remote sensors systems is simulated applying such numerical modeling. The analyses of sensors systems based on diffraction gratings requires optical broadband sources to interrogate the optical sensor unit, mainly in long period gratings that shows a characteristic rejection band. Therefore, the sensor distance is limited to a few kilometers due to the attenuation in optical fibers. Additional attenuation is introduced by the sensor element. Hence, to extend the distance in the optical sensor system, the optical amplification system is needed to compensate the losses in the optical fibers. The Raman amplification technology was selected mainly due to the flexibility in the gain bandwidth. The modeling can be applied to sensor systems that monitor sites located at long distances, or in places that the access is restricted due to harsh environment conditions in such cases conventional sensors are relatively fast deteriorated.

  8. Quantitative detection of hepatitis C virus RNA with a solid-phase signal amplification method: definition of optimal conditions for specimen collection and clinical application in interferon-treated patients.

    PubMed

    Davis, G L; Lau, J Y; Urdea, M S; Neuwald, P D; Wilber, J C; Lindsay, K; Perrillo, R P; Albrecht, J

    1994-06-01

    To determine the optimal conditions for preparation of serum specimens for quantitative hepatitis C virus RNA determination, patient samples were processed such that differences in time from clot formation to centrifugation, centrifugation to separation of serum and collection of serum until freezing could be independently assessed. The effects of multiple cycles of freezing and thawing were also determined. There was progressive and significant loss of hepatitis C virus RNA activity when the time from the formation of the clot until centrifugation was longer than 2 hr. This reduction reached 32% after 6 hr and 49% after 24 hr. If centrifugation was performed immediately after formation of the clot, loss of hepatitis C virus RNA activity was reduced to less than 10% even though the serum remained unseparated from the clot for up to 6 hr. Centrifugation of blood through a paraffin plug (serum separator tube) prevented loss of hepatitis C virus RNA activity for up to 24 hr. There was no loss of hepatitis C virus RNA activity with up to three freeze-thaw cycles. When patient specimens were prepared under these optimal conditions, the sensitivity of the quantitative branched DNA signal amplification assay in patients with hepatitis C virus infection was 83% and the specificity in patients with liver disease was 100%. Fluctuations in hepatitis C virus RNA levels were shown to correlate with biochemical changes observed in patients treated with recombinant interferon-alpha 2b. These data demonstrate that improper or inconsistent methods of serum preparation may result in falsely low and unreliable levels of hepatitis C virus RNA.(ABSTRACT TRUNCATED AT 250 WORDS)

  9. Improved PCR Amplification of Broad Spectrum GC DNA Templates

    PubMed Central

    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. PMID:27271574

  10. 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.

  11. 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.)

  12. On soliton amplification

    NASA Technical Reports Server (NTRS)

    Leibovich, S.; Randall, J. D.

    1979-01-01

    The paper considers a modified Korteweg-de Vries equation that permits wave amplification or damping. A 'terminal similarity' solution is identified for large times in amplified systems. Numerical results are given which confirm that the terminal similarity solution is a valid local approximation for mu t sufficiently large and positive, even though the approximation is not uniformly valid in space.

  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. Weak-value amplification of the fast-light effect in rubidium vapor

    NASA Astrophysics Data System (ADS)

    Mirhosseini, Mohammad; Viza, Gerardo I.; Magaña-Loaiza, Omar S.; Malik, Mehul; Howell, John C.; Boyd, Robert W.

    2016-05-01

    We use weak-value amplification to enhance the polarization-sensitive fast-light effect from induced Raman absorption in hot rubidium vapor. We experimentally demonstrate that projecting the output signal into an appropriate polarization state enables a pulse advancement of 4.2 μ s , which is more than 15 times larger than that naturally caused by dispersion. More significantly, we show that combining weak-value amplification with the dispersive response of an atomic system provides a clear advantage in terms of the maximum pulse advance achievable for a given value of loss. This technique has potential applications for designing novel quantum-information-processing gates and optical buffers for telecommunication systems.

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

    SciTech Connect

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

    2009-07-15

    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 {mu}m. 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 {mu}m pitch arrays employing discrete photodiodes culminated in a novel design providing an optical fill factor of {approx}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 {mu}m 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

  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. PMID:19736969

  18. 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.

  19. Enzyme-free amplification for sensitive electrochemical detection of DNA via target-catalyzed hairpin assembly assisted current change.

    PubMed

    Qian, Yong; Wang, Chunyan; Gao, Fenglei

    2014-12-01

    An isothermal, enzyme-free and sensitive method for electrochemical detection of DNA is proposed based on target catalyzed hairpin assembly and for signal amplification. Molecular beacon 1 (MB1) contains a ferrocene (Fc) tag, which was immobilized on the gold electrode as recognition probe to hybridize with target DNA. Then, molecular beacon 2 hybridized with the opened MB1, allowing the target to be displaced. The displaced target again triggered the next round of strand exchange reaction resulting in many Fc far away from the GE to achieve signal amplification for sensitive DNA detection. The current signal amplification strategy is relatively simple and inexpensive owing to avoid the use of any kind of enzyme or sophisticated equipment. It can achieve a sensitivity of 42 fM with a wide linear dynamic range from 10(-13) to 10(-9)M and discriminate mismatched DNA from perfect matched target DNA with a high selectivity. The proposed method showed excellent specificity, high sensitivity and low detection limit, and could be applied in analysis of real samples.

  20. 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).

  1. Light amplification using semiconductors

    SciTech Connect

    Dupuis, R.D.

    1987-06-01

    During the summer of 1953, John von Neumann discussed his ideas concerning light amplification using semiconductors with Edward Teller. In September of that year, von Neumann sent a manuscript containing his ideas and calculations on this subject to Teller for his comments. To the best of our knowledge, von Neumann did not take time to work further on these ideas, and the manuscript remained unpublished. These previously unpublished writings of John von Neumann on the subject of light amplification in semiconductors are printed as a service to the laser community. While von Neumann's original manuscript and his letter to Teller are available to anyone who visits the Library of Congress, it is much more convenient to have this paper appear in an archival journal.

  2. Flux amplification in SSPX

    NASA Astrophysics Data System (ADS)

    Lodestro, Lynda; Hooper, E. B.; Jayakumar, R. J.; Pearlstein, L. D.; Wood, R. D.; McLean, H. S.

    2007-11-01

    Flux amplification---the ratio of poloidal flux enclosed between the magnetic and geometric axes to that between the separatrix and the geometric axis---is a key measure of efficiency for edge-current-driven spheromaks. With the new, modular capacitor bank, permitting flexible programming of the gun current, studies of flux amplification under various drive scenarios can be performed. Analysis of recent results of pulsed operation with the new bank finds an efficiency ˜ 0.2, in selected shots, of the conversion of gun energy to confined magnetic energy during the pulses, and suggests a route toward sustained efficiency at 0.2. Results of experiments, a model calculation of field build-up, and NIMROD simulations exploring this newly suggested scenario will be presented.

  3. 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.

  4. 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".

  5. Prevention effect of rare ginsenosides against stress-hormone induced MTOC amplification

    PubMed Central

    Lee, Jee-Hyun; Cheong, Kyu Jin; Jung, Youn-Sang; Woo, Tae-Gyun; Yoon, Min-Ho; Oh, Ah-Young; Kang, So-Mi; Lee, Chunghui; Sun, Hokeun; Hwang, Jihwan; Song, Gyu-Yong; Park, Bum-Joon

    2016-01-01

    Stress has been suggested as one of important cause of human cancer without molecular biological evidence. Thus, we test the effect of stress-related hormones on cell viability and mitotic fidelity. Similarly to estrogen, stress hormone cortisol and its relative cortisone increase microtubule organizing center (MTOC) number through elevated expression of γ-tubulin and provide the Taxol resistance to human cancer cell lines. However, these effects are achieved by glucocorticoid hormone receptor (GR) but not by estrogen receptor (ER). Since ginsenosides possess steroid-like structure, we hypothesized that it would block the stress or estrogen-induced MTOC amplification and Taxol resistance. Among tested chemicals, rare ginsenoside, CSH1 (Rg6) shows obvious effect on inhibition of MTOC amplification, γ-tubulin induction and Taxol resistance. Comparing to Fulvestant (FST), ER-α specific inhibitor, this chemical can block the cortisol/cortisone-induced MTOC deregulation as well as ER-α signaling. Our results suggest that stress hormone induced tumorigenesis would be achieved by MTOC amplification, and CSH1 would be useful for prevention of stress-hormone or steroid hormone-induced chromosomal instability. PMID:27147573

  6. A versatile immobilization-free photoelectrochemical biosensor for ultrasensitive detection of cancer biomarker based on enzyme-free cascaded quadratic amplification strategy.

    PubMed

    Ge, Lei; Wang, Wenxiao; Hou, Ting; Li, Feng

    2016-03-15

    In this work, an ultrasensitive immobilization-free photoelectrochemical (PEC) biosensor was successfully developed for the first time based on a novel enzyme-free cascaded quadratic signal amplification strategy. This rationally designed homogeneous dual amplification strategy consists of a target-analog recycling circuit based on catalytic hairpin assembly (CHA) and a hybridization chain reaction (HCR) mediated amplification circuit. In the presence of carcinoembryonic antigen (CEA), a proof-of-concept target, target-analog is released to trigger the upstream CHA recycling circuit. The generated dsDNA complexes from CHA recycling could further induce the downstream HCR amplification, leading to the formation of numerous hemin/G-quadruplex DNAzymes. This would accordingly stimulate the biocatalytic precipitation of 4-chloro-1-naphthol, inducing a distinct decrease in the photocurrent signal due to the formed insoluble/insulating products on electrode surface. Under the optimal conditions, this PEC biosensor achieved ultrasensitive detection of CEA down to the atto-gram level. The introduction of this aptamer-based cascaded quadratic amplification strategy not only remarkably improves the selectivity and sensitivity of CEA assay, but also allows the ultrasensitive detection of other proteins by designing specific aptamers, providing a universal, isothermal and label-free PEC biosensing platform for ultrasensitive detection of different kinds of cancer biomarkers and holding a great potential for early-diagnosis of cancer.

  7. Quantum Amplification by Superradiant Emission of Radiation

    NASA Astrophysics Data System (ADS)

    Svidzinsky, Anatoly A.; Yuan, Luqi; Scully, Marlan O.

    2013-10-01

    A laser generates light through stimulated emission of radiation and requires population inversion. Quantum interference can yield lasing without inversion. However, such phase-sensitive quantum amplification still requires some atomic population in the excited state. Here, we present a new kind of quantum amplifier based on collective superradiant emission which does not need any population in the excited state. We show that parametric resonance between the driving (e.g., infrared) field and collective superradiant oscillations of the atomic polarization can yield light amplification at high (e.g., XUV) frequencies. To achieve gain, one must suppress a time-dependent Stark shift caused by the driving field. The resulting superradiant amplifier is many orders of magnitude more efficient than the usual nonlinear multiphoton excitation and holds promise for a new kind of generator of high-frequency coherent radiation. In addition to a detailed analytical analysis, confirmed by numerical simulations, we provide a physically appealing explanation of the quantum amplification by superradiant emission of radiation (QASER) operation in terms of coupled classical oscillators. We also present an experiment that demonstrates the QASER amplification mechanism in an electronic circuit, which, to the best of our knowledge, is the first experimental demonstration of the difference combination resonance.

  8. Modeling of the Princeton Raman Amplification Experiment

    NASA Astrophysics Data System (ADS)

    Hur, Min Sup; Lindberg, Ryan; Wurtele, Jonathan; Cheng, W.; Avitzour, Y.; Ping, Y.; Suckewer, S.; Fisch, N. J.

    2004-11-01

    We numerically model the Princeton experiments on Raman amplification [1] using averaged-PIC (aPIC) [2] and 3-wave codes. Recently, there has been a series of experimental results performed in Princeton University [3]. Amplification factors up to 500 in intensity were obtained using a subpicosecond pulse propagating in a 3mm plasma. The plasma was created using a gas jet. The intensity of the amplified pulse exceeds the intensity of the pump pulse, indicating that the process of Raman amplification is in the nonlinear regime. Comparisons are made between 3-wave models, kinetic models and the experimental data. It is found that better agreement is achieved when kinetic effects are included. Discussion of the influence of a range of potentially deleterious phenomena such as density inhomogeneity, particle trapping, ionization, and pump depletion by noise amplification will be examined. References [1] V.M. Malkin, G. Shvets, and N.J. Fisch, Phys. Rev. Lett. vol.82, 1999. [2] M.S. Hur, G. Penn, J.S. Wurtele, R. Lindberg, to appear in Phys. Plasmas. [3] Y. Ping, et al., Phys. Rev. E vol. 66, 2002; Phys. Rev. E vol. 67, 2003;

  9. Ultrabroadband noncollinear optical parametric amplification with LBO crystal.

    PubMed

    Zhao, Baozhen; Jiang, Yongliang; Sueda, Keiich; Miyanaga, Noriaki; Kobayashi, Takayoshi

    2008-11-10

    Ultrabroadband visible noncollinear optical parametric amplification (NOPA) was achieved in an LBO crystal, with a continuum seed pulse generated from a sapphire plate. The spectral bandwidth of the amplified visible pulse was about 200 nm, which can support sub-5 fs pulse amplification. An amplified output of 0.21 microJ with an average gain of about 210 was achieved. This provides, to the best of our knowledge, the first-time demonstration of such broadband amplification with a biaxial nonlinear optical crystal. Both the simulation and experimental results indicate that the LBO has a great potential as nonlinear medium in power amplifier for TW to PW noncollinear optical parametric chirped pulse amplification (NOPCPA) systems. PMID:19581976

  10. 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.

  11. 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.

  12. Self-locked aptamer probe mediated cascade amplification strategy for highly sensitive and selective detection of protein and small molecule.

    PubMed

    Li, Wei; Jiang, Wei; Wang, Lei

    2016-10-12

    In this work, a novel self-locked aptamer probe mediated cascade amplification strategy has been constructed for highly sensitive and specific detection of protein. First, the self-locked aptamer probe was designed with three functions: one was specific molecular recognition attributed to the aptamer sequence, the second was signal transduction owing to the transduction sequence, and the third was self-locking through the hybridization of the transduction sequence and part of the aptamer sequence. Then, the aptamer sequence specific recognized the target and folded into a three-way helix junction, leading to the release of the transduction sequence. Next, the 3'-end of this three-way junction acted as primer to trigger the strand displacement amplification (SDA), yielding a large amount of primers. Finally, the primers initiated the dual-exponential rolling circle amplification (DE-RCA) and generated numerous G-quadruples sequences. By inserting the fluorescent dye N-methyl mesoporphyrin IX (NMM), enhanced fluorescence signal was achieved. In this strategy, the self-locked aptamer probe was more stable to reduce the interference signals generated by the uncontrollable folding in unbounded state. Through the cascade amplification of SDA and DE-RCA, the sensitivity was further improved with a detection limit of 3.8 × 10(-16) mol/L for protein detection. Furthermore, by changing the aptamer sequence of the probe, sensitive and selective detection of adenosine has been also achieved, suggesting that the proposed strategy has good versatility and can be widely used in sensitive and selective detection of biomolecules.

  13. Self-locked aptamer probe mediated cascade amplification strategy for highly sensitive and selective detection of protein and small molecule.

    PubMed

    Li, Wei; Jiang, Wei; Wang, Lei

    2016-10-12

    In this work, a novel self-locked aptamer probe mediated cascade amplification strategy has been constructed for highly sensitive and specific detection of protein. First, the self-locked aptamer probe was designed with three functions: one was specific molecular recognition attributed to the aptamer sequence, the second was signal transduction owing to the transduction sequence, and the third was self-locking through the hybridization of the transduction sequence and part of the aptamer sequence. Then, the aptamer sequence specific recognized the target and folded into a three-way helix junction, leading to the release of the transduction sequence. Next, the 3'-end of this three-way junction acted as primer to trigger the strand displacement amplification (SDA), yielding a large amount of primers. Finally, the primers initiated the dual-exponential rolling circle amplification (DE-RCA) and generated numerous G-quadruples sequences. By inserting the fluorescent dye N-methyl mesoporphyrin IX (NMM), enhanced fluorescence signal was achieved. In this strategy, the self-locked aptamer probe was more stable to reduce the interference signals generated by the uncontrollable folding in unbounded state. Through the cascade amplification of SDA and DE-RCA, the sensitivity was further improved with a detection limit of 3.8 × 10(-16) mol/L for protein detection. Furthermore, by changing the aptamer sequence of the probe, sensitive and selective detection of adenosine has been also achieved, suggesting that the proposed strategy has good versatility and can be widely used in sensitive and selective detection of biomolecules. PMID:27662754

  14. Evidence of high-elevation amplification versus Arctic amplification.

    PubMed

    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

  15. Graphene fluorescence switch-based cooperative amplification: a sensitive and accurate method to detection microRNA.

    PubMed

    Liu, Haiyun; Li, Lu; Wang, Qian; Duan, Lili; Tang, Bo

    2014-06-01

    MicroRNAs (miRNAs) play significant roles in a diverse range of biological progress and have been regarded as biomarkers and therapeutic targets in cancer treatment. Sensitive and accurate detection of miRNAs is crucial for better understanding their roles in cancer cells and further validating their function in clinical diagnosis. Here, we developed a stable, sensitive, and specific miRNAs detection method on the basis of cooperative amplification combining with the graphene oxide (GO) fluorescence switch-based circular exponential amplification and the multimolecules labeling of SYBR Green I (SG). First, the target miRNA is adsorbed on the surface of GO, which can protect the miRNA from enzyme digest. Next, the miRNA hybridizes with a partial hairpin probe and then acts as a primer to initiate a strand displacement reaction to form a complete duplex. Finally, under the action of nicking enzyme, universal DNA fragments are released and used as triggers to initiate next reaction cycle, constituting a new circular exponential amplification. In the proposed strategy, a small amount of target miRNA can be converted to a large number of stable DNA triggers, leading to a remarkable amplification for the target. Moreover, compared with labeling with a 1:1 stoichiometric ratio, multimolecules binding of intercalating dye SG to double-stranded DNA (dsDNA) can induce significant enhancement of fluorescence signal and further improve the detection sensitivity. The extraordinary fluorescence quenching of GO used here guarantees the high signal-to-noise ratio. Due to the protection for target miRNA by GO, the cooperative amplification, and low fluorescence background, sensitive and accurate detection of miRNAs has been achieved. The strategy proposed here will offer a new approach for reliable quantification of miRNAs in medical research and early clinical diagnostics. PMID:24823448

  16. A novel power amplification scheme for nuclear magnetic resonance/nuclear quadrupole resonance systems.

    PubMed

    Zhang, Xinwang; Schemm, Nathan; Balkır, Sina

    2011-03-01

    Nuclear magnetic resonance (NMR) and nuclear quadrupole resonance (NQR)-based chemical analysis systems have been widely utilized in various areas such as medicine, security, and academic research. In these applications, the power amplifier stage plays a key role in generating the required oscillating magnetic fields within a radio frequency coil that serves as the probe. However, the bulky size and relatively low efficiency of the traditional power amplification schemes employed present a bottleneck for the realization of compact sized and portable NMR and NQR systems. To address this problem, this work presents a class D voltage-switching power amplification scheme with novel fast-start and fast-stop functions that are suitable for generating ideal NMR and NQR excitation signals. Compared to the traditional analog power amplifiers (PAs), the proposed switched-mode PA can achieve significant improvement on the power efficiency as well as the physical volume. A PA circuit for portable NQR-based explosive detection systems has been designed and built using the proposed scheme with 1 kW possible maximum output power and 10 MHz maximum operating frequency. Test results show that the presented PA achieves more than 60% measured efficiency within a highly compact volume while sustaining fast start and stop of excitation signals in the order of microseconds.

  17. How do dynamic cellular signals travel long distances?

    PubMed

    Nussinov, Ruth

    2012-01-01

    Communication is essential. It is vital between cells in multi-cellular organisms, and within cells. A signaling molecule binds to a receptor protein, and initiates a cascade of dynamic events. Signaling is a multistep pathway, which allows signal amplification: if some of the molecules in a pathway transmit the signal to multiple molecules, the result can be a large number of activated molecules across the cell and multiple reactions. That is how a small number of extracellular signaling molecules can produce a major cellular response. The pathway can relay signals from the extracellular space to the nucleus. How do signals travel efficiently over long-distances across the cell? Here we argue that evolution has utilized three properties: a modular functional organization of the cellular network; sequences in some key regions of proteins, such as linkers or loops, which were pre-encoded by evolution to facilitate signaling among domains; and compact interactions between proteins which is achieved via conformational disorder.

  18. 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.

  19. Self-induced parametric amplification arising from nonlinear elastic coupling in a micromechanical resonating disk gyroscope

    PubMed Central

    Nitzan, Sarah H.; Zega, Valentina; Li, Mo; Ahn, Chae H.; Corigliano, Alberto; Kenny, Thomas W.; Horsley, David A.

    2015-01-01

    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. PMID:25762243

  20. Self-induced parametric amplification arising from nonlinear elastic coupling in a micromechanical resonating disk gyroscope

    NASA Astrophysics Data System (ADS)

    Nitzan, Sarah H.; Zega, Valentina; Li, Mo; Ahn, Chae H.; Corigliano, Alberto; Kenny, Thomas W.; Horsley, David A.

    2015-03-01

    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.

  1. 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. PMID:26551336

  2. 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.

  3. 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.

  4. Universal probe amplification: multiplex screening technologies for genetic variations.

    PubMed

    Park, Jung Hun; Park, Ki Soo; Lee, Kyungmee; Jang, Hyowon; Park, Hyun Gyu

    2015-01-01

    In order to achieve multiplex screening of genetic variations, multiplex amplification of target genomic DNA is necessary. Universal amplification technology meets this requirement by simultaneously amplifying a number of different regions within the target genomic DNA using a single pair of universal primers and thus eliminating the limitations associated with the use of multiple pairs of primers. We comprehensively review universal probe amplification and its use with multiplex technologies for the identification of the most representative genetic variation, i. e. single nucleotide polymorphisms. The progress and key issues relating to universal probe amplification are discussed and the representative technologies are summarized with an emphasis on their application for the identification of susceptibility to human diseases.

  5. 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

  6. Laser light triggers increased Raman amplification in the regime of nonlinear Landau damping.

    PubMed

    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

  7. Nonlinearities and Parametric Amplification of Superconducting Coplanar Waveguide Resonators

    NASA Astrophysics Data System (ADS)

    Haviland, David; Tholén, Erik; Ergul, Adem

    2008-03-01

    We have experimentally studied the nonlinear properties of superconducting coplanar stripline resonators fabricated from Al and Nb films with small transverse dimensions (gap size 1μm). Magnetic field penetration into the superconductor causes a current-dependant kinetic inductance, which gives an ideal Kerr nonlinearity. When the nonlinear oscillator is pumped very near its dynamic instability, it can be used to realize parametric amplification. We have achieved a gain of +22.4dB in a 5.8 GHz resonator cooled to 450 mK [E. Thol'en et. al. Appl. Phys. Lett. 90, 253509 (2007)]. Parametric deamplification or squeezing of a signal has also been verified with squeezing of 30 dB. The later effect is interesting because it can be used to generate squeezed vacuum states of the electromagnetic field. We have modeled the data using a theory developed by Yurke and Buks [J. Lightwave Technol. 24, 5054 (2006)]. Excellent fit of the model to the measured data can be achieved over a wide range of pump power, and the strength of the nonlinear terms can be obtained with high accuracy.

  8. 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.

  9. Enhanced acoustic sensing through wave compression and pressure amplification in anisotropic metamaterials

    NASA Astrophysics Data System (ADS)

    Chen, Yongyao; Liu, Haijun; Reilly, Michael; Bae, Hyungdae; Yu, Miao

    2014-10-01

    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.

  10. 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.

  11. 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. PMID:26512524

  12. Highly sensitive fluorescence detection of target DNA by coupling exonuclease-assisted cascade target recycling and DNAzyme amplification.

    PubMed

    Liu, Shufeng; Cheng, Chuanbin; Liu, Tao; Wang, Li; Gong, Hongwei; Li, Feng

    2015-01-15

    Because of the intrinsic importance of nucleic acid as bio-targets, the simple and sensitive detection of nucleic acid is very essential for biological studies and medical diagnostics. Herein, a simple, isothermal and highly sensitive fluorescence detection of target DNA was developed with the combination of exonuclease III (Exo III)-assisted cascade target recycling and DNAzyme amplification. A hairpin DNA probe was designed, which contained the 3'-protruding DNA fragment as target recognition unit, the caged DNA fragment in the stem region as target analogue, and the caged 8-17 DNAzyme sequence in the loop region as signal response unit. Upon sensing of target DNA, the 3'-strand of hairpin DNA probe could be stepwise removed by Exo III, accompanied by the releasing of target DNA and autonomous generation of new target analogues for the successive hybridization and cleavage process. Simultaneously, the 8-17 DNAzyme unit could be exponentially released from this hairpin DNA probe and activated for the cyclic cleavage toward the ribonucleotide-containing molecular beacon substrate, inducing a remarkable fluorescence signal amplification for target detection. A low detection limit of 20 fM with an excellent selectivity toward target DNA could be achieved. The developed cascade amplification strategy may be further extended for the detection of a wide spectrum of analytes including protein and biological small molecules by combining DNA aptamer technology.

  13. A target responsive aptamer machine for label-free and sensitive non-enzymatic recycling amplification detection of ATP.

    PubMed

    Li, Xin; Peng, Ying; Chai, Yaqin; Yuan, Ruo; Xiang, Yun

    2016-03-01

    Based on a new target-triggered aptamer molecular machine, a label-free and non-enzymatic target recycling amplification strategy for sensitive fluorescence detection of ATP in human serums is described. The presence of the target ATP together with the DNA fuel strand initiates the operation of the aptamer machine and leads to cyclic reuse of ATP and the release of many G-quadruplex sequences, which associate with a fluorescent dye to generate significantly amplified fluorescence signals to achieve sensitive detection of ATP. PMID:26853492

  14. Optical parametric chirped pulse amplification and spectral shaping of a continuum generated in a photonic band gap fiber.

    PubMed

    Hugonnot, E; Somekh, M; Villate, D; Salin, F; Freysz, E

    2004-05-31

    A chirped pulse, spectrally broadened in a photonic bandgap optical fiber by 120 fs Ti:Sapphire laser pulses, is parametrically amplified in a BBO crystal pumped by a frequency doubled nanosecond Nd:YAG laser pulse. Without changing the frequency of the Ti:Sapphire, a spectral tunability of the amplified pulses is demonstrated. The possibility to achieve broader spectral range amplification is confirmed for a non-collinear pump-signal interaction geometry. For optimal non-collinear interaction geometry, the pulse duration of the original and amplified pulse are similar. Finally, we demonstrate that the combination of two BBO crystals makes it possible to spectrally shape the amplified pulses.

  15. 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.

  16. New Technologies in Amplification: Applications to the Pediatric Population.

    ERIC Educational Resources Information Center

    Kopun, Judy

    1995-01-01

    Discussion of technological advances in amplification for children with hearing impairments focuses on the advantages and limitations of fitting children with devices that have features such as dynamic-range compression, multiband signal processing, multimemory capability, digital feedback reduction, and frequency transposition. (Author/DB)

  17. 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

  18. Regulation of ribosomal DNA amplification by the TOR pathway.

    PubMed

    Jack, Carmen V; Cruz, Cristina; Hull, Ryan M; Keller, Markus A; Ralser, Markus; Houseley, Jonathan

    2015-08-01

    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.

  19. 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.

  20. 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.

  1. Kinase signaling in the spindle checkpoint.

    PubMed

    Kang, Jungseog; Yu, Hongtao

    2009-06-01

    The spindle checkpoint is a cell cycle surveillance system that ensures the fidelity of chromosome segregation. In mitosis, it elicits the "wait anaphase" signal to inhibit the anaphase-promoting complex or cyclosome until all chromosomes achieve bipolar microtubule attachment and align at the metaphase plate. Because a single kinetochore unattached to microtubules activates the checkpoint, the wait anaphase signal is thought to be generated by this kinetochore and is then amplified and distributed throughout the cell to inhibit the anaphase-promoting complex/cyclosome. Several spindle checkpoint kinases participate in the generation and amplification of this signal. Recent studies have begun to reveal the activation mechanisms of these checkpoint kinases. Increasing evidence also indicates that the checkpoint kinases not only help to generate the wait anaphase signal but also actively correct kinetochore-microtubule attachment defects. PMID:19228686

  2. 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.

  3. Ultra-broad bandwidth parametric amplification at degeneracy.

    PubMed

    Limpert, J; Aguergaray, C; Montant, S; Manek-Hönninger, I; Petit, S; Descamps, D; Cormier, E; Salin, F

    2005-09-19

    We report on a novel approach of ultra-broad bandwidth parametric amplification around degeneracy. A bandwidth of up to 400 nm centered around 800 nm is amplified in a BBO crystal by using chirped pump pulses with a bandwitdth as broad as 10 nm. A supercontinuum signal is generated in a microstructured fiber, having to first order a quadratic chirp, which is necessary to ensure temporal overlap of the interacting waves over this broad bandwidth. Furthermore, we discuss the potential of this approach for an octave-spanning parametric amplification.

  4. 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.

  5. In vitro amplification techniques for the detection of nucleic acids: new tools for the diagnostic laboratory.

    PubMed

    Persing, D H; Landry, M L

    1989-01-01

    The acceptance of nucleic acid probes as diagnostic tools for the clinical laboratory has been hampered by a number of factors, including laborious techniques and limited sensitivity. The focus of this review is on the recent development of amplification techniques to enhance the signal generated by nucleic acid-based detection systems. Three general areas are discussed: (1) amplification of target sequences using the polymerase chain reaction or the transcript amplification system, (2) amplification of the probe sequences using Q beta replicase, and (3) amplification of probe-generated signals with compound or "Christmas tree" probes. The hope of these new technologies is to simplify yet improve on the sensitivity of nucleic acid-based tests to enable them to attain a more prominent place in the diagnostic repertoire of the clinical laboratory.

  6. 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.

  7. 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.

  8. 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.

  9. 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

  10. Tidal amplification of seabed light

    NASA Astrophysics Data System (ADS)

    Bowers, D. G.; Brubaker, J. M.

    2010-09-01

    Because solar irradiance decreases approximately exponentially with depth in the sea, the increase in irradiance at the seabed from mid to low tide is greater than the decrease from mid to high tide. Summed over a day, this can lead to a net amplification of seabed irradiance in tidal waters compared to nontidal waters with the same mean depth and transparency. In this paper, this effect is quantified by numerical and analytical integration of the Lambert-Beer equation to derive the ratio of daily total seabed irradiance with and without a tide. Greatest amplification occurs in turbid water with large tidal range and low tide occurring at noon. The theoretical prediction is tested against observations of seabed irradiance in the coastal waters of North Wales where tidal amplification of seabed light by up to a factor of 7 is both observed and predicted. Increasing the strength of tidal currents tends to increase the turbidity of the water and hence reduce the light reaching the seabed, but this effect is made less by increasing tidal amplification, especially when low water is in the middle of the day. The ecological implications of tidal amplification are discussed. The productivity of benthic algae will be greater than that predicted by simple models which calculate seabed irradiance using the mean depth of water alone. Benthic algae are also able to live at greater depths in tidal waters than in nontidal waters with the same transparency.

  11. 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.

  12. Gene amplification and insecticide resistance.

    PubMed

    Bass, Chris; Field, Linda M

    2011-08-01

    Pesticide resistance in arthropods has been shown to evolve by two main mechanisms, the enhanced production of metabolic enzymes, which bind to and/or detoxify the pesticide, and mutation of the target protein, which makes it less sensitive to the pesticide. One route that leads to enhanced metabolism is the duplication or amplification of the structural gene(s) encoding the detoxifying enzyme, and this has now been described for the three main families (esterases, glutathione S-transferases and cytochrome P450 monooxygenases) implicated in resistance. More recently, a direct or indirect role for gene duplication or amplification has been described for target-site resistance in several arthropod species. This mini-review summarises the involvement of gene duplication/amplification in the insecticide/acaricide resistance of insect and mite pests and highlights recent developments in this area in relation to P450-mediated and target-site resistance.

  13. DNA extraction, preservation, and amplification.

    PubMed

    Knebelsberger, Thomas; Stöger, Isabella

    2012-01-01

    The effectiveness of DNA barcoding as a routine practice in biodiversity research is strongly dependent on the quality of the source material, DNA extraction method, and selection of adequate primers in combination with optimized polymerase chain reaction (PCR) conditions. For the isolation of nucleic acids, silica-gel membrane methods are to be favored because they are easy to handle, applicable for high sample throughput, relatively inexpensive, and provide high DNA quality, quantity, and purity which are pre-requisites for successful PCR amplification and long-term storage of nucleic acids in biorepositories, such as DNA banks. In this section, standard protocols and workflow schemes for sample preparation, DNA isolation, DNA storage, PCR amplification, PCR product quality control, and PCR product cleanup are proposed and described in detail. A PCR troubleshooting and primer design section may help to solve problems that hinder successful amplification of the desired barcoding gene region.

  14. 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.

  15. 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.

  16. Optimization of noncollinear optical parametric amplification

    NASA Astrophysics Data System (ADS)

    Schimpf, D. N.; Rothardt, J.; Limpert, J.; Tünnermann, A.

    2007-02-01

    Noncollinearly phase-matched optical parametric amplifiers (NOPAs) - pumped with the green light of a frequency doubled Yb-doped fiber-amplifier system 1, 2 - permit convenient generation of ultrashort pulses in the visible (VIS) and near infrared (NIR) 3. The broad bandwidth of the parametric gain via the noncollinear pump configuration allows amplification of few-cycle optical pulses when seeded with a spectrally flat, re-compressible signal. The short pulses tunable over a wide region in the visible permit transcend of frontiers in physics and lifescience. For instance, the resulting high temporal resolution is of significance for many spectroscopic techniques. Furthermore, the high magnitudes of the peak-powers of the produced pulses allow research in high-field physics. To understand the demands of noncollinear optical parametric amplification using a fiber pump source, it is important to investigate this configuration in detail 4. An analysis provides not only insight into the parametric process but also determines an optimal choice of experimental parameters for the objective. Here, the intention is to design a configuration which yields the shortest possible temporal pulse. As a consequence of this analysis, the experimental setup could be optimized. A number of aspects of optical parametric amplifier performance have been treated analytically and computationally 5, but these do not fully cover the situation under consideration here.

  17. Detection of viral sequences by internally calibrated gene amplification.

    PubMed

    Cheung, R K; Hui, M F; Dosch, H M; Ewart, T E

    1993-05-01

    Inherent pitfalls of the polymerase chain reaction (PCR) can become serious difficulties when transferring research applications to high-volume routine procedures such as biofermentation process control and clinical diagnostics. Difficulties include 1) the danger of accidental sample contamination with positive control templates; 2) variable amplification due to positional effects in thermocycler blocks and unequal primer efficiency for sense/anti-sense strands; and 3) the need for reliable controls, which provide confidence for reporting negative reactions. Using the PCR detection system for Epstein-Barr virus as a model, we have developed a quick process to generate mutant internal co-amplification templates. These can be used for titration of amplification sensitivity. More importantly, single tube co-amplification without titrations allows determination of the minimum sensitivity achieved in each individual reaction; critical information when reporting negative diagnostic results. Mutant and native fragments are easy to distinguish by size, and sample cross contamination can be readily identified. The system should be easily adaptable to gene amplification procedures, which aim to routinely detect the presence of a given gene fragment in a controlled fashion.

  18. Heralded noiseless amplification of a photon polarization qubit

    NASA Astrophysics Data System (ADS)

    Kocsis, S.; Xiang, G. Y.; Ralph, T. C.; Pryde, G. J.

    2013-01-01

    Photons are the best long-range carriers of quantum information, but the unavoidable absorption and scattering in a transmission channel places a serious limitation on viable communication distances. Signal amplification will therefore be an essential feature of quantum technologies, with direct applications to quantum communication, metrology and fundamental tests of quantum theory. Non-deterministic noiseless amplification of a single mode can circumvent the challenges related to amplifying a quantum signal, such as the no-cloning theorem and the minimum noise cost for deterministic quantum state amplification. However, existing devices are not suitable for amplifying the fundamental optical quantum information carrier: a qubit coherently encoded across two optical modes. Here, we construct a coherent two-mode amplifier to demonstrate the first heralded noiseless linear amplification of a qubit encoded in the polarization state of a single photon. In doing so, we increase the transmission fidelity of a realistic qubit channel by up to a factor of five. Qubit amplifiers promise to extend the range of secure quantum communication and other quantum information science and technology protocols.

  19. Highly sensitive detection of T4 polynucleotide kinase activity by coupling split DNAzyme and ligation-triggered DNAzyme cascade amplification.

    PubMed

    Liu, Shufeng; Ming, Jingjing; Lin, Ying; Wang, Chunfeng; Cheng, Chuanbin; Liu, Tao; Wang, Li

    2014-05-15

    In current study, a dual strategy for sensitive detection of T4 polynucleotide kinase (T4 PNK) activity was proposed, which combined split DNAzyme-based background reduction with ligation-triggered DNAzyme cascade for signal amplification. The 8-17 DNAzyme is split into two separate oligonucleotide fragments, which can be separately hybridized to the template DNA to form a ligatable nick after one of the fragments is phosphorylated at the 5at the yl by T4 PNK. With the further addition of Escherichia coli DNA ligase, the two oligonucleotides can be ligated to produce the activated 8-17 DNAzyme, the amount of which is positively related to the activity of T4 PNK. The signal amplification can be achieved through the cyclic cleavage of 8-17 DNAzyme toward the molecular beacon substrate, resulting in an evident fluorescence signal enhancement. The current dual strategy can significantly improve the detection sensitivity of the sensing systems, resulting in a detection limit of 0.001 U mL(-1) for T4 PNK activity, which is superior or comparable to the reported methods. Furthermore, the inhibition effects of adenosine diphosphate and sodium hydrogen phosphate on T4 PNK activity have also been demonstrated with satisfactory results. The current method may be further developed as a universal protocol for monitoring activity and inhibition of nucleotide kinase, and may show the huge potentials in biological process researches, drug discovery, and clinic diagnostics.

  20. Chronic centrosome amplification without tumorigenesis

    PubMed Central

    Vitre, Benjamin; Holland, Andrew J.; Kulukian, Anita; Shoshani, Ofer; Hirai, Maretoshi; Wang, Yin; Maldonado, Marcus; Cho, Thomas; Boubaker, Jihane; Swing, Deborah A.; Tessarollo, Lino; Evans, Sylvia M.; Fuchs, Elaine; Cleveland, Don W.

    2015-01-01

    Centrosomes are microtubule-organizing centers that facilitate bipolar mitotic spindle assembly and chromosome segregation. Recognizing that centrosome amplification is a common feature of aneuploid cancer cells, we tested whether supernumerary centrosomes are sufficient to drive tumor development. To do this, we constructed and analyzed mice in which centrosome amplification can be induced by a Cre-recombinase–mediated increase in expression of Polo-like kinase 4 (Plk4). Elevated Plk4 in mouse fibroblasts produced supernumerary centrosomes and enhanced the expected mitotic errors, but proliferation continued only after inactivation of the p53 tumor suppressor. Increasing Plk4 levels in mice with functional p53 produced centrosome amplification in liver and skin, but this did not promote spontaneous tumor development in these tissues or enhance the growth of chemically induced skin tumors. In the absence of p53, Plk4 overexpression generated widespread centrosome amplification, but did not drive additional tumors or affect development of the fatal thymic lymphomas that arise in animals lacking p53. We conclude that, independent of p53 status, supernumerary centrosomes are not sufficient to drive tumor formation. PMID:26578792

  1. Boosting riboswitch efficiency by RNA amplification.

    PubMed

    Emadpour, Masoumeh; Karcher, Daniel; Bock, Ralph

    2015-05-26

    Riboswitches are RNA sensors that regulate gene expression in response to binding of small molecules. Although they conceptually represent simple on/off switches and, therefore, hold great promise for biotechnology and future synthetic biology applications, the induction of gene expression by natural riboswitches after ligand addition or removal is often only moderate and, consequently, the achievable expression levels are not very high. Here, we have designed an RNA amplification-based system that strongly improves the efficiency of riboswitches. We have successfully implemented the method in a biological system for which currently no efficient endogenous tools for inducible (trans)gene expression are available: the chloroplasts of higher plants. We further show that an HIV antigen whose constitutive expression from the chloroplast genome is deleterious to the plant can be inducibly expressed under the control of the RNA amplification-enhanced riboswitch (RAmpER) without causing a mutant phenotype, demonstrating the potential of the method for the production of proteins and metabolites that are toxic to the host cell. PMID:25824954

  2. Boosting riboswitch efficiency by RNA amplification

    PubMed Central

    Emadpour, Masoumeh; Karcher, Daniel; Bock, Ralph

    2015-01-01

    Riboswitches are RNA sensors that regulate gene expression in response to binding of small molecules. Although they conceptually represent simple on/off switches and, therefore, hold great promise for biotechnology and future synthetic biology applications, the induction of gene expression by natural riboswitches after ligand addition or removal is often only moderate and, consequently, the achievable expression levels are not very high. Here, we have designed an RNA amplification-based system that strongly improves the efficiency of riboswitches. We have successfully implemented the method in a biological system for which currently no efficient endogenous tools for inducible (trans)gene expression are available: the chloroplasts of higher plants. We further show that an HIV antigen whose constitutive expression from the chloroplast genome is deleterious to the plant can be inducibly expressed under the control of the RNA amplification-enhanced riboswitch (RAmpER) without causing a mutant phenotype, demonstrating the potential of the method for the production of proteins and metabolites that are toxic to the host cell. PMID:25824954

  3. 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.

  4. Highly efficient sample stacking by enhanced field amplification on a simple paper device.

    PubMed

    Ma, Biao; Song, Yi-Zhen; Niu, Ji-Cheng; Wu, Zhi-Yong

    2016-09-21

    We present a novel electrokinetic stacking (ES) method based on field amplification on a simple paper device for sample preconcentration. With voltage application, charged probe ions in a solution of lower conductivity stack and form a narrow band at the boundary between the sample and the background electrolyte of higher conductivity. The stacking band appears quickly and stabilizes in a few minutes. With this ES method, three orders of magnitude signal improvement was successfully achieved for both a fluorescein probe and a double-stranded DNA within 300 s. This enhanced stacking efficiency is attributed to a focusing effect due to the balance between electromigration and counter electroosmotic flow. We also applied this ES method to other low-cost fiber substrates such as cloth and thread. Such a simple and highly efficient ES method will find wide applications in the development of sensitive paper-based analytical devices (PADs), especially for low-cost point-of-care testing (POCT).

  5. 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.

  6. Amplification and noise properties of an erbium-doped multicore fiber amplifier.

    PubMed

    Abedin, K S; Taunay, T F; Fishteyn, M; Yan, M F; Zhu, B; Fini, J M; Monberg, E M; Dimarcello, F V; Wisk, P W

    2011-08-15

    A multicore erbium-doped fiber (MC-EDF) amplifier for simultaneous amplification in the 7-cores has been developed, and the gain and noise properties of individual cores have been studied. The pump and signal radiation were coupled to individual cores of MC-EDF using two tapered fiber bundled (TFB) couplers with low insertion loss. For a pump power of 146 mW, the average gain achieved in the MC-EDF fiber was 30 dB, and noise figure was less than 4 dB. The net useful gain from the multicore-amplifier, after taking into consideration of all the passive losses, was about 23-27 dB. Pump induced ASE noise transfer between the neighboring channel was negligible. PMID:21935033

  7. 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.

  8. 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.

  9. 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.

  10. Ultrasensitive Detection of Low-Abundance Surface-Marker Protein using Isothermal Rolling Circle Amplification in Microfluidic Nano-Liter Platform

    PubMed Central

    Konry, Tania; Yarmush, Joel M.; Irimia, Daniel

    2011-01-01

    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, we have applied a highly sensitive labeling method that translates antigen-antibody recognition processes into DNA detection event that can be greatly amplified via isothermal Rolling Circle Amplification (RCA). By merging the single-molecule detection power of RCA reaction with microfluidic technology we were able to demonstrate that identification of specific protein markers can be achieved on tumor cell surface in miniaturized nano-liter 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. PMID:21294269

  11. 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.

  12. 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.

  13. 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. PMID:26085615

  14. Deterministic amplification of Schrödinger cat states in circuit quantum electrodynamics

    NASA Astrophysics Data System (ADS)

    Joo, Jaewoo; Elliott, Matthew; Oi, Daniel K. L.; Ginossar, Eran; Spiller, Timothy P.

    2016-02-01

    Perfect deterministic amplification of arbitrary quantum states is prohibited by quantum mechanics, but determinism can be achieved by compromising between fidelity and amplification power. We propose a dynamical scheme for deterministically amplifying photonic Schrödinger cat states, which show great promise as a tool for quantum information processing. Our protocol is designed for strongly coupled circuit quantum electrodynamics and utilizes artificial atomic states and external microwave controls to engineer a set of optimal state transfers and achieve high fidelity amplification. We compare analytical results with full simulations of the open, driven Jaynes-Cummings model, using realistic device parameters for state of the art superconducting circuits. Amplification with a fidelity of 0.9 can be achieved for sizable cat states in the presence of cavity and atomic-level decoherence. This tool could be applied to practical continuous-variable information processing for the purification and stabilization of cat states in the presence of photon losses.

  15. 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.

  16. 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. PMID:26999622

  17. 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.

  18. Cross-triggered and cascaded recycling amplification for ultrasensitive electrochemical sensing of the mutant human p53 gene.

    PubMed

    Yang, Cuiyun; Dou, Baoting; Yang, Jianmei; Yuan, Ruo; Xiang, Yun

    2016-07-01

    Based on an endonuclease-assisted, cross-triggered and cascaded recycling amplification strategy, the construction of a simple electrochemical sensing platform for the ultrasensitive detection of the mutant p53 gene in human serum is described. Using this new signal amplification approach, the sub-femtomolar level of the mutant p53 gene can be selectively detected.

  19. Improvement of φ29 DNA polymerase amplification performance by fusion of DNA binding motifs

    PubMed Central

    de Vega, Miguel; Lázaro, José M.; Mencía, Mario; Blanco, Luis; Salas, Margarita

    2010-01-01

    Bacteriophage φ29 DNA polymerase is a unique enzyme endowed with two distinctive properties, high processivity and faithful polymerization coupled to strand displacement, that have led to the development of protocols to achieve isothermal amplification of limiting amounts of both circular plasmids and genomic DNA. To enhance the amplification efficiency of φ29 DNA polymerase, we have constructed chimerical DNA polymerases by fusing DNA binding domains to the C terminus of the polymerase. The results show that the addition of Helix-hairpin-Helix [(HhH)2] domains increases DNA binding of the hybrid polymerases without hindering their replication rate. In addition, the chimerical DNA polymerases display an improved and faithful multiply primed DNA amplification proficiency on both circular plasmids and genomic DNA and are unique φ29 DNA polymerase variants with enhanced amplification performance. The reported chimerical DNA polymerases will contribute to make φ29 DNA polymerase-based amplification technologies one of the most powerful tools for genomics. PMID:20823261

  20. Improvement of φ29 DNA polymerase amplification performance by fusion of DNA binding motifs.

    PubMed

    de Vega, Miguel; Lázaro, José M; Mencía, Mario; Blanco, Luis; Salas, Margarita

    2010-09-21

    Bacteriophage ϕ29 DNA polymerase is a unique enzyme endowed with two distinctive properties, high processivity and faithful polymerization coupled to strand displacement, that have led to the development of protocols to achieve isothermal amplification of limiting amounts of both circular plasmids and genomic DNA. To enhance the amplification efficiency of ϕ29 DNA polymerase, we have constructed chimerical DNA polymerases by fusing DNA binding domains to the C terminus of the polymerase. The results show that the addition of Helix-hairpin-Helix [(HhH)(2)] domains increases DNA binding of the hybrid polymerases without hindering their replication rate. In addition, the chimerical DNA polymerases display an improved and faithful multiply primed DNA amplification proficiency on both circular plasmids and genomic DNA and are unique ϕ29 DNA polymerase variants with enhanced amplification performance. The reported chimerical DNA polymerases will contribute to make ϕ29 DNA polymerase-based amplification technologies one of the most powerful tools for genomics. PMID:20823261

  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

    NASA Astrophysics Data System (ADS)

    Schmidt, Bruno E.; Thiré, Nicolas; Boivin, Maxime; Laramée, Antoine; Poitras, François; Lebrun, Guy; Ozaki, Tsuneyuki; Ibrahim, Heide; Légaré, François

    2014-05-01

    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.

  3. 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

  4. Numerical simulation of regenerative amplification of nanosecond pulses in a CO/sub 2/ laser

    SciTech Connect

    Apollonov, V.V.; Bunkin, F.V.; Sorochenko, V.R.; Firsov, K.N.; Shakir, Y.A.

    1982-04-01

    The results are given of a numerical simulation of regenerative amplification of nanosecond pulses in a high-power electric-discharge CO/sub 2/ laser. The finite contrast of the injected pulse is allowed for in a determination of the ranges of the injection energy and time in which regenerative amplification can be expected. It is shown that the shape of a small-signal pulse has a considerable influence on the energy and contrast of the output pulse. The optimal combinations of the parameters of a regenerative amplification system are determined so as to maximize the energy and contrast of nanosecond pulses.

  5. 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.

  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. 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.

  8. Master curves for gas amplification in low vacuum and environmental scanning electron microscopy.

    PubMed

    Thiel, Bradley L

    2004-02-01

    The concept of universal amplification profiles for gas cascade amplification of signals in low vacuum and environmental scanning electron microscopes is demonstrated both experimentally and theoretically using water vapor. For a given gas, cascade amplification gain profiles can be plotted onto a single master curve where the independent reduced parameter is the ratio of pressure to amplification field strength. When plotted in this fashion, both desired secondary electron and spurious background signal components fall onto respective master curves, with the amplitude being a function of anode bias only. These master curves can be described by simple Townsend Gas Capacitor equations using only two gas-specific parameters. As long as single scattering conditions apply, this approach allows for simplified, direct comparison of the gain characteristics of different gases and allows more intelligent selection of imaging conditions. The utility of treating signal amplification in this manner is demonstrated through a series of images collected under a variety of conditions, but with the ratio of pressure to amplification field strength kept constant. In practice, the range of operational parameter space in which this description can be applied to imaging is limited, as images typically have a mixture of secondary and backscattered contributions.

  9. Ultrasensitive, colorimetric detection of microRNAs based on isothermal exponential amplification reaction-assisted gold nanoparticle amplification.

    PubMed

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

    2016-12-15

    MicroRNAs (miRNAs) play important roles in a wide range of biological processes, and their aberrant expressions are linked to a large number of human diseases and disorders. In this work, we developed a colorimetric method for rapid, ultrasensitive miRNA detection via isothermal exponential amplification reaction (EXPAR)-assisted gold nanoparticle (AuNP) amplification. The sensing probe designed with a tandem phosphorothioate modification in the backbone of the polyadenines at the 5' terminus was employed to directly assemble onto the surface of AuNP with high adsorption affinity. The recognition domain at the 3' terminus of the sensing probe hybridizes with target miRNAs to trigger EXPAR with exponential signal amplification. With the amplification reaction with the action of DNA polymerase, the sensing probe gradually detaches from the AuNP, resulting in the aggregation of bare AuNPs in the high-salt reaction environment due to lack of DNA protection. The presence of AuNP aggregation is conveniently measured by UV-vis spectroscopy. Our proposed method could provide a linear detection range from 50fM to 10nM with a detection limit of ∼46fM within 60min, and also discriminate a single-nucleotide difference between homologous miRNAs. PMID:27498329

  10. 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.

  11. Envelope matching for enhanced backward Raman amplification by using self-ionizing plasmas

    SciTech Connect

    Zhang, Z. M.; Zhang, B.; Hong, W.; Teng, J.; He, S. K.; Gu, Y. Q.; Yu, M. Y.

    2014-12-15

    Backward Raman amplification (BRA) in plasmas has been promoted as a means for generating ultrapowerful laser pulses. For the purpose of achieving the maximum intensities over the shortest distances, an envelope matching between the seed pulse and the amplification gain is required, i.e., the seed pulse propagates at the same velocity with the gain such that the peak of the seed pulse can always enjoy the maximum gain. However, such an envelope matching is absent in traditional BRA because in the latter the amplification gain propagates at superluminous velocity while the seed pulse propagates at the group velocity, which is less than the speed of light. It is shown here that, by using self-ionizing plasmas, the speed of the amplification gain can be well reduced to reach the envelope matching regime. This results in a favorable BRA process, in which higher saturated intensity, shorter interaction length and higher energy-transfer efficiency are achieved.

  12. Dynamic characteristics of a hydraulic amplification mechanism for large displacement actuators systems.

    PubMed

    Arouette, Xavier; Matsumoto, Yasuaki; Ninomiya, Takeshi; Okayama, Yoshiyuki; Miki, Norihisa

    2010-01-01

    We have developed a hydraulic displacement amplification mechanism (HDAM) and studied its dynamic response when combined with a piezoelectric actuator. The HDAM consists of an incompressible fluid sealed in a microcavity by two largely deformable polydimethylsiloxane (PDMS) membranes. The geometry with input and output surfaces having different cross-sectional areas creates amplification. By combining the HDAM with micro-actuators, we can amplify the input displacement generated by the actuators, which is useful for applications requiring large deformation, such as tactile displays. We achieved a mechanism offering up to 18-fold displacement amplification for static actuation and 12-fold for 55 Hz dynamic actuation. PMID:22319281

  13. Label-free electrochemical nucleic acid biosensing by tandem polymerization and cleavage-mediated cascade target recycling and DNAzyme amplification.

    PubMed

    Liu, Shufeng; Gong, Hongwei; Wang, Yanqun; Wang, Li

    2016-03-15

    Owing to the intrinsic importance of nucleic acid as bio-targets, the achievement of its simple and sensitive detection with high confidence is very essential for biological studies and diagnostic purposes. Herein, a label-free, isothermal, and ultrasensitive electrochemical detection of target DNA was developed by using a tandem polymerization and cleavage-mediated cascade target recycling and DNAzyme releasing amplification strategy. Upon sensing of the nucleic acid analyte for the assembled hairpin-like probe DNA on the electrode, the DNA polymerase guided the target recycling and simultaneously triggered the lambda exonuclease cleavage, accompanied by the cascade recycling of the released new complementary strand and the amplified liberation of the G-rich sequence of the HRP-mimicking DNAzyme. The electrocatalytic reduction of H2O2 by the generated hemin/G-quadruplex DNAzyme was used for the signal readout and further amplification toward target response. Such tandem functional operation by DNA polymerase, lambda exonuclease and DNAzyme endows the developed biosensor with a high sensitivity and also a high confidence. A low detection limit of 5 fM with an excellent selectivity toward target DNA could be achieved. It also exhibits the distinct advantages of simplicity in probe design and biosensor fabrication, and label-free electrochemical detection, thus may offer a promising avenue for the applications in disease diagnosis and clinical biomedicine. PMID:26513289

  14. Label-free electrochemical nucleic acid biosensing by tandem polymerization and cleavage-mediated cascade target recycling and DNAzyme amplification.

    PubMed

    Liu, Shufeng; Gong, Hongwei; Wang, Yanqun; Wang, Li

    2016-03-15

    Owing to the intrinsic importance of nucleic acid as bio-targets, the achievement of its simple and sensitive detection with high confidence is very essential for biological studies and diagnostic purposes. Herein, a label-free, isothermal, and ultrasensitive electrochemical detection of target DNA was developed by using a tandem polymerization and cleavage-mediated cascade target recycling and DNAzyme releasing amplification strategy. Upon sensing of the nucleic acid analyte for the assembled hairpin-like probe DNA on the electrode, the DNA polymerase guided the target recycling and simultaneously triggered the lambda exonuclease cleavage, accompanied by the cascade recycling of the released new complementary strand and the amplified liberation of the G-rich sequence of the HRP-mimicking DNAzyme. The electrocatalytic reduction of H2O2 by the generated hemin/G-quadruplex DNAzyme was used for the signal readout and further amplification toward target response. Such tandem functional operation by DNA polymerase, lambda exonuclease and DNAzyme endows the developed biosensor with a high sensitivity and also a high confidence. A low detection limit of 5 fM with an excellent selectivity toward target DNA could be achieved. It also exhibits the distinct advantages of simplicity in probe design and biosensor fabrication, and label-free electrochemical detection, thus may offer a promising avenue for the applications in disease diagnosis and clinical biomedicine.

  15. Corruption of ant acoustical signals by mimetic social parasites: Maculinea butterflies achieve elevated status in host societies by mimicking the acoustics of queen ants.

    PubMed

    Thomas, Jeremy A; Schönrogge, Karsten; Bonelli, Simona; Barbero, Francesca; Balletto, Emilio

    2010-03-01

    Recent recordings of the stridulations of Myrmica ants revealed that their queens made distinctive sounds from their workers, although the acoustics of queens and workers, respectively, were the same in different species of Myrmica. Queen recordings induced enhanced protective behavior when played to workers in the one species tested. Larvae and pupae of the butterfly genus Maculinea inhabit Myrmica colonies as social parasites, and both stages generate sounds that mimic those of a Myrmica queen, inducing similar superior treatments from workers as their model. We discuss how initial penetration and acceptance as a colony member is achieved by Maculinea through mimicking the species-specific semio-chemicals of their hosts, and how acoustical mimicry is then employed to elevate the parasite's membership of that society towards the highest attainable level in their host's hierarchy. We postulate that, if acoustics is as well developed a means of communication in certain ants as these studies suggest, then others among an estimated 10,000 species of ant social parasite may supplement their well-known use of chemical and tactile mimicry to trick host ants with mimicry of host acoustical systems.

  16. Brief review of monitoring methods for loop-mediated isothermal amplification (LAMP).

    PubMed

    Zhang, Xuzhi; Lowe, Stuart B; Gooding, John Justin

    2014-11-15

    The loop-mediated isothermal amplification (LAMP) technique has the potential to revolutionize molecular biology because it allows DNA amplification under isothermal conditions and is highly compatible with point-of-care analysis. To achieve efficient genetic analysis of samples, the method of real-time or endpoint determination selected to monitor the biochemical reaction is of great importance. In this paper we briefly review progress in the development of monitoring methods for LAMP.

  17. Universal nucleic acid sequence-based amplification for simultaneous amplification of messengerRNAs and microRNAs.

    PubMed

    Mader, Andreas; Riehle, Ulrike; Brandstetter, Thomas; Stickeler, Elmar; Ruehe, Juergen

    2012-11-19

    A universal NASBA assay is presented for simultaneous amplification of multiple microRNA (miRNA) and messengerRNA (mRNA) sequences. First, miRNA and mRNA sequences are reverse transcribed using tailed reverse transcription primer pairs containing a gene-specific and an non-specific region. For reverse transcription of small miRNA molecules a non-specific region is incorporated into a structured stem-loop reverse transcription primer. Second, a universal NASBA primer pair that recognizes the tagged cDNA molecules enables a simultaneous, transcription-based amplification reaction (NASBA) of all different cDNA molecules in one reaction. The NASBA products (RNA copies) are detected by gene-specific DNA probes immobilized on a biochip. By using the multiplex reverse transcription combined with the universal NASBA amplification up to 14 different mRNA and miRNA sequences can be specifically amplified and detected in parallel. In comparison with standard multiplex NASBA assays this approach strongly enhances the multiplex capacity of NASBA-based amplification reactions. Furthermore simultaneous assaying of different RNA classes can be achieved that might be beneficial for studying miRNA-based regulation of gene expression or for RNA-based tumor diagnostics. PMID:23140948

  18. 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.

  19. Increasingly branched rolling circle amplification for the cancer gene detection.

    PubMed

    Li, Hongling; Xu, Jianguo; Wang, Zhenmeng; Wu, Zai-Sheng; Jia, Lee

    2016-12-15

    An increasingly branched rolling circle amplification (IB-RCA) which contains a padlock probe (PP) and a structurally tailored molecular beacon (MB) was innovatively developed for highly sensitive detection of cancer gene, Kras gene codon 12. In this system, the PP can be circularized after hybridization with the precisely-matched target DNA, while the stem of MB can be also opened by target DNA, resulting in hybridization with the circularized PP to generate a long tandem single-stranded DNA (ssDNA) product. Since the MB is also designed to hybridize with ssDNA product, the newly-opened MBs are able to trigger the next RCA reactions, therapy producing branched rolling circle amplification (RCA) products and in turn leading to the increasingly branched RCA (IB-RCA). This alternately and continuously operates hybridization-based MB opening and opened MBs-triggered RCA. As a result, a great number of MBs are opened that is associated with a dramatically amplified fluorescent signal, enabling to quantify target DNA down to 100 fM. This sensing method demonstrates a new concept of IB-RCA amplification even in a simple way to efficiently transduce the fluorescence signal, accomplishing the highly sensitive and selective detection of cancer gene. PMID:27569300

  20. Increasingly branched rolling circle amplification for the cancer gene detection.

    PubMed

    Li, Hongling; Xu, Jianguo; Wang, Zhenmeng; Wu, Zai-Sheng; Jia, Lee

    2016-12-15

    An increasingly branched rolling circle amplification (IB-RCA) which contains a padlock probe (PP) and a structurally tailored molecular beacon (MB) was innovatively developed for highly sensitive detection of cancer gene, Kras gene codon 12. In this system, the PP can be circularized after hybridization with the precisely-matched target DNA, while the stem of MB can be also opened by target DNA, resulting in hybridization with the circularized PP to generate a long tandem single-stranded DNA (ssDNA) product. Since the MB is also designed to hybridize with ssDNA product, the newly-opened MBs are able to trigger the next RCA reactions, therapy producing branched rolling circle amplification (RCA) products and in turn leading to the increasingly branched RCA (IB-RCA). This alternately and continuously operates hybridization-based MB opening and opened MBs-triggered RCA. As a result, a great number of MBs are opened that is associated with a dramatically amplified fluorescent signal, enabling to quantify target DNA down to 100 fM. This sensing method demonstrates a new concept of IB-RCA amplification even in a simple way to efficiently transduce the fluorescence signal, accomplishing the highly sensitive and selective detection of cancer gene.

  1. 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.

  2. Lidar measurements of atmospheric backscattering amplification

    NASA Astrophysics Data System (ADS)

    Banakh, V. A.; Razenkov, I. A.

    2016-02-01

    Results of long-term continuous measurements of the atmospheric backscattering amplification coefficient on a 2-km-long near-ground path with the use of a two-channel micropulse lidar based on a waveguide laser are presented. It is shown that the backscattering amplification coefficient has a pronounced daily variation. In the night and in the afternoon, atmospheric backscattering amplification is maximal and the amplification coefficient can exceed 2. The amplification is low or absent in morning and evening hours at neutral temperature stratification in the near-ground layer of the atmosphere. The backscattering amplification coefficient increases with an increase in the structure constant of the air refracting index and variance of the image jitter of the illumination spot created by the probing laser beam on the wall of a 2-km-distant building.

  3. 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.

  4. Dynamics and control of DNA sequence amplification.

    PubMed

    Marimuthu, Karthikeyan; Chakrabarti, Raj

    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. PMID:25362284

  5. 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.

  6. 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.

  7. Weak value amplification is suboptimal for estimation and detection.

    PubMed

    Ferrie, Christopher; Combes, Joshua

    2014-01-31

    We show by using statistically rigorous arguments that the technique of weak value amplification does not perform better than standard statistical techniques for the tasks of single parameter estimation and signal detection. Specifically, we prove that postselection, a necessary ingredient for weak value amplification, decreases estimation accuracy and, moreover, arranging for anomalously large weak values is a suboptimal strategy. In doing so, we explicitly provide the optimal estimator, which in turn allows us to identify the optimal experimental arrangement to be the one in which all outcomes have equal weak values (all as small as possible) and the initial state of the meter is the maximal eigenvalue of the square of the system observable. Finally, we give precise quantitative conditions for when weak measurement (measurements without postselection or anomalously large weak values) can mitigate the effect of uncharacterized technical noise in estimation.

  8. 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

  9. Trophic amplification of climate warming.

    PubMed

    Kirby, Richard R; Beaugrand, Gregory

    2009-12-01

    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.

  10. 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.

  11. The MDM2 gene amplification database.

    PubMed Central

    Momand, J; Jung, D; Wilczynski, S; Niland, J

    1998-01-01

    The p53 tumor suppressor gene is inactivated in human tumors by several distinct mechanisms. The best characterized inactivation mechanisms are: (i) gene mutation; (ii) p53 protein association with viral proteins; (iii) p53 protein association with the MDM2 cellular oncoprotein. The MDM2 gene has been shown to be abnormally up-regulated in human tumors and tumor cell lines by gene amplification, increased transcript levels and enhanced translation. This communication presents a brief review of the spectrum of MDM2 abnormalities in human tumors and compares the tissue distribution of MDM2 amplification and p53 mutation frequencies. In this study, 3889 samples from tumors or xenografts from 28 tumor types were examined for MDM2 amplification from previously published sources. The overall frequency of MDM2 amplification in these human tumors was 7%. Gene amplification was observed in 19 tumor types, with the highest frequency observed in soft tissue tumors (20%), osteosarcomas (16%) and esophageal carcinomas (13%). Tumors which showed a higher incidence of MDM2 amplification than p53 mutation were soft tissue tumors, testicular germ cell cancers and neuro-blastomas. Data from studies where both MDM2 amplification and p53 mutations were analyzed within the same samples showed that mutations in these two genes do not generally occur within the same tumor. In these studies, 29 out of a total of 33 MDM2 amplification-positive tumors had wild-type p53. We hypothesize that heretofore uncharacterized carcinogens favor MDM2 amplification over p53 mutations in certain tumor types. A database listing the MDM2 gene amplifications is available on the World Wide Web at http://www. infosci.coh.org/mdm2 . Charts of MDM2 amplification frequencies and comparisons with p53 genetic alterations are also available at this Web site. PMID:9671804

  12. 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.

  13. Parametric amplification in single-walled carbon nanotube nanoelectromechanical resonators

    NASA Astrophysics Data System (ADS)

    Wu, Chung-Chiang; Zhong, Zhaohui

    2011-08-01

    The low quality factor (Q) of single-walled carbon nanotube (SWNT) resonators has limited their sensitivity in sensing application. To this end, we employ the technique of parametric amplification by modulating the spring constant of SWNT resonators at twice the resonant frequency and achieve 10 times Q enhancement. The highest Q obtained at room temperature is around ˜700, which is 3-4 times better than previous Q record reported for doubly clamped SWNT resonators. Furthermore, efficient parametric amplification is found to only occur in the catenary vibration regime. Our results open up the possibility to employ light-weight and high-Q carbon nanotube resonators in single molecule and atomic mass sensing.

  14. Digital droplet multiple displacement amplification (ddMDA) for whole genome sequencing of limited DNA samples

    DOE PAGES

    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,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

  15. 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

  16. Digital Droplet Multiple Displacement Amplification (ddMDA) for Whole Genome Sequencing of Limited DNA Samples.

    PubMed

    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

  17. 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.

  18. 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.

  19. Active amplification by critical oscillators in hearing

    NASA Astrophysics Data System (ADS)

    Julicher, Frank

    2004-03-01

    The ear posesses exquisit abilities to detect sounds over a wide range of frequencies with an extraordinary dynamic range. From the faintest sounds that we hear to the loudest noises, the sound pressure varies over twelve orders of magnitude. It has been first proposed by Thomas Gold in 1948 that passive resonating elements alone cannot explain these abilities of the ear. The observation, that the ears of seemingly all vertebrates exhibit spontaneous sound emissions, so called oto-acoustic emissions, adds evidence to the idea that hearing is based on active mechanisms for signal amplification and detection which for different species are based on the same principles. It has been suggested that the active properties and the observed nonlinearities in the ear's response can be explained by assuming that the cochlea contains dynamical systems operating in the vicinity of the critical point of an oscillating instability. A sound stimulus excites deformations of the basilar membrane which travel along the choclea towards the apex. The combination of this travelling wave with properties of critical oscillators leads to a scenario where sound excites nonlinear waves in the cochlea. The large dynamic range of hearing then results from compressive power law responses of these nonlinear excitations.

  20. Modeling and optimizing of low-repetition-rate high-energy pulse amplification in high-concentration erbium-doped fiber amplifiers

    NASA Astrophysics Data System (ADS)

    Li, Jianfeng; Dai, Zhiyong; Ou, Zhonghua; Zhang, Lixun; Liu, Yongzhi; Liu, Yong

    2009-09-01

    Starting from the modeling of isolated ions and ion-pairs, a closed form rate and power evolution equations for pulse amplification in high-concentration erbium-doped fiber amplifiers (EDFAs) are constructed. According to the equations, the effects of ion-pairs on the performance of a high-concentration EDFA in steady state including upper-state population, ASE powers without input signal are analyzed numerically. Furthermore, the effects of ion-pairs on the dynamic characteristics of low-repetition-rate pulse amplification in the EDFA including the storied energy, output pulse energy and evolution of pulse waveform distortion are systematically studied by using the finite-difference method. The results show that the presence of the ion-pairs deteriorates amplifier performance, such as the upper-state population, ASE power, storied energy, output pulse energy, and saturated gain, etc. For the high-concentration EDFA, the optimum fiber length should be modified to achieve a better performance. The relations between the evolution of pulse waveform distortion or output pulse energy and the input pulse peak power are also discussed. The results can provide important guide for the design and optimization of the low-repetition-rate pulse amplification in high-concentration EDFAs.

  1. Ultrasensitive electrochemical detection of nucleic acid by coupling an autonomous cascade target replication and enzyme/gold nanoparticle-based post-amplification.

    PubMed

    Liu, Shufeng; Wei, Wenji; Wang, Yanqun; Fang, Li; Wang, Li; Li, Feng

    2016-06-15

    Owing to the intrinsic importance of nucleic acid as bio-targets, the development of isothermal and ultrasensitive electrochemical DNA biosensor is very essential for biological studies and medical diagnostics. Herein, the autonomous cascade DNA replication strategy was effectively married with the enzyme/gold nanoparticle-based post-amplification strategy to promote the detection performance toward target DNA. A hairpin DNA probe (HP) is designed that consists of an overhang at 3'-end as the recognition unit for target DNA, a recognition site for nicking endonuclease, and an alkane spacer to terminate polymerization reaction. The autonomous DNA replication-scission-displacement reaction operated by the nicking endonuclease/KF polymerase induced the autocatalytic opening of HP, which was then specifically bound by the enzyme/gold nanoparticles for further dual-signal amplification toward target-related sensing events. A low detection limit of 0.065 fM with an excellent selectivity toward target DNA could be achieved. The proposed biosensor could be also easily regenerated for target detection. The developed biosensor creates an opportunity for the effective coupling of the target replication with post-amplification strategies and thus opens a promising avenue for the detection of nucleic acid with low abundance in bioanalysis and clinical biomedicine.

  2. Persistent nicotine treatment potentiates amplification of the dihydrofolate reductase gene in rat lung epithelial cells as a consequence of Ras activation.

    PubMed

    Guo, Jinjin; Chu, Michelle; Abbeyquaye, Tetteh; Chen, Chang-Yan

    2005-08-26

    Although nicotine has been suggested to promote lung carcinogenesis, the mechanism of its action in this process remains unknown. The present investigation demonstrates that the treatment of rat lung epithelial cells with nicotine for various periods differentially mobilizes multiple intracellular pathways. Protein kinase C and phosphoinositide 3-OH-kinase are transiently activated after the treatment. Also, Ras and its downstream effector ERK1/2 are activated after long term exposure to nicotine. The activation of Ras by nicotine treatment is responsible for the subsequent perturbation of the methotrexate (MTX)-mediated G1 cell cycle restriction as well as an increase in production of reactive oxygen species. When p53 expression is suppressed by introducing E6, persistent exposure to nicotine enables dihydrofolate reductase gene amplification in the presence of methotrexate (MTX) and the formation of the MTX-resistant colonies. Altering the activity of phosphoinositide 3-OH-kinase has no effect on dihydrofolate reductase amplification. However, the suppression of protein kinase C dramatically affects the colony formation in soft agar. Thus, our data suggest that persistent exposure to nicotine perturbs the G1 checkpoint and causes DNA damage through the increase of the production of reactive oxygen species. However, a third element rendered by loss of p53 is required for the initiation of the process of gene amplification. Under p53-deficient conditions, the establishment of a full oncogenic transformation, in response to long term nicotine exposure, is achieved through the cooperation of multiple signaling pathways. PMID:15983034

  3. 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.

  4. 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.

  5. 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. PMID:26563112

  6. 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.

  7. Enhanced Amplification and Fan-Out Operation in an All-Magnetic Transistor

    NASA Astrophysics Data System (ADS)

    Barman, Saswati; Saha, Susmita; Mondal, Sucheta; Kumar, Dheeraj; Barman, Anjan

    2016-09-01

    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.

  8. 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

  9. Enhanced Amplification and Fan-Out Operation in an All-Magnetic Transistor.

    PubMed

    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

  10. Point-of-care multiplexed assays of nucleic acids using microcapillary-based loop-mediated isothermal amplification.

    PubMed

    Zhang, Yi; Zhang, Lu; Sun, Jiashu; Liu, Yulei; Ma, Xingjie; Cui, Shangjin; Ma, Liying; Xi, Jianzhong Jeff; Jiang, Xingyu

    2014-07-15

    This report demonstrates a straightforward, robust, multiplexed and point-of-care microcapillary-based loop-mediated isothermal amplification (cLAMP) for assaying nucleic acids. This assay integrates capillaries (glass or plastic) to introduce and house sample/reagents, segments of water droplets to prevent contamination, pocket warmers to provide heat, and a hand-held flashlight for a visual readout of the fluorescent signal. The cLAMP system allows the simultaneous detection of two RNA targets of human immunodeficiency virus (HIV) from multiple plasma samples, and achieves a high sensitivity of two copies of standard plasmid. As few nucleic acid detection methods can be wholly independent of external power supply and equipment, our cLAMP holds great promise for point-of-care applications in resource-poor settings. PMID:24937125

  11. 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.

  12. A dual-wavelength amplifier that enables the simultaneous chirped-pulse amplification of femtosecond 1562 nm pulses and continuous wave 1064 nm radiation for applications in difference frequency generation

    NASA Astrophysics Data System (ADS)

    Krzempek, Karol; Sobon, Grzegorz; Sotor, Jaroslaw; Abramski, Krzysztof

    2016-10-01

    We present the first demonstration of a chirped-pulse amplification system that enables the simultaneous amplification of femtosecond pulses from the erbium band and continuous wave radiation centered at 1064 nm. The setup generated 470 fs laser pulses, centered at 1562 nm, with an average output power of 590 mW. By exploiting the unique dual-wavelength design an additional gain of 18.08 dB was achieved for the auxiliary 1064 nm seed signal required in the frequency conversion part of the setup. The usability of the constructed source was verified by setting up a difference frequency generation experiment in which the unique characteristics of the mode-locked laser pulses were transferred into the mid-infrared wavelength region.

  13. FGFR-1 amplification in metastatic lymph-nodal and haematogenous lobular breast carcinoma

    PubMed Central

    2012-01-01

    Background Lobular breast carcinoma usually shows poor responsiveness to chemotherapies and often lacks targeted therapies. Since FGFR1 expression has been shown to play pivotal roles in primary breast cancer tumorigenesis, we sought to analyze the status of FGFR1 gene in a metastatic setting of lobular breast carcinoma, since promising FGFR1 inhibitors has been recently developed. Methods Fifteen tissue metastases from lobular breast carcinomas with matched primary infiltrative lobular breast carcinoma were recruited. Eleven cases showed loco-regional lymph-nodal and four haematogenous metastases. FGFR-1 gene (8p12) amplification was evaluated by chromogenic in situ hybridization (CISH) analysis. Her-2/neu and topoisomerase-IIα gene status was assessed. E-cadherin and Hercept Test were also performed. We distinguished amplification (>6 or cluster of signals) versus gains (3–6 signals) of the locus specific FGFR-1 gene. Results Three (20%) primary lobular breast carcinomas showed >6 or cluster of FGFR1 signals (amplification), six cases (40%) had a mean of three (range 3–6) chromogenic signals (gains) whereas in 6 (40%) was not observed any abnormality. Three of 15 metastasis (20%) were amplified, 2/15 (13,4%) did not. The ten remaining cases (66,6%) showed three chromogenic signals. The three cases with FGFR-1 amplification matched with those primary breast carcinomas showing FGFR-1 amplification. The six cases showing FGFR-1 gains in the primary tumour again showed FGFR-1 gains in the metastases. Four cases showed gains of FGFR-1 gene signals in the metastases and not in the primary tumours. Her-2/neu gene amplification was not observed in all cases but one (6%) case. Topoisomerase-IIα was not amplified in all cases. Conclusions 1) a subset of metastatic lobular breast carcinoma harbors FGFR-1 gene amplification or gains of chromogenic signals; 2) a minor heterogeneity has been observed after matching primary and metastatic carcinomas; 3) in the era of

  14. Optical parametric amplification via non-Hermitian phase matching.

    PubMed

    El-Ganainy, R; Dadap, J I; Osgood, R M

    2015-11-01

    We introduce the notion of dissipative optical parametric amplifiers (DOPA) and demonstrate that, even in the absence of the Hermitian phase-matching condition in these structures, the signal beam can be amplified when the idler mode suffers optical attenuation. We discuss the optical implementation of this concept in waveguide platforms, and we propose different methods to control the optical loss of these configurations only at the wavelength of the idler component. Surprisingly, this spectrally selective dissipation process allows the signal beam to draw more energy from the pump and, as a result, attains net amplification. Similar results also apply if the losses are introduced only to the signal component. This intriguing feature can open new avenues for building long wavelength light sources and parametric amplifiers by using semiconductor planar structures, where Hermitian phase-matching requirements can be difficult to satisfy without adding stringent geometric constraints or relatively complex fabrication steps. PMID:26512525

  15. Raman Amplification and Tunable Pulse Delays in Silicon Waveguides

    SciTech Connect

    Rukhlenko, Ivan D.; Premaratne, Malin; Garanovich, Ivan L.; Sukhorukov, Andrey A.; Agrawal, Govind P.

    2010-10-07

    The nonlinear process of stimulated Raman scattering is important for silicon photonics as it enables optical amplification and lasing. However, generally employed numerical approaches provide very little insight into the contribution of different silicon Raman amplifier (SRA) parameters. In this paper, we solve the coupled pump-signal equations analytically and derive an exact formula for the envelope of a signal pulse when picosecond optical pulses are amplified inside a SRA pumped by a continuous-wave laser beam. Our solution is valid for an arbitrary pulse shape and fully accounts for the Raman gain-dispersion effects, including temporal broadening and group-velocity reduction. Our results are useful for optimizing the performance of SRAs and for engineering controllable signal delays.

  16. 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. PMID:22449695

  17. Microfluidic beads-based immunosensor for sensitive detection of cancer biomarker proteins using multienzyme-nanoparticle amplification and quantum dots labels.

    PubMed

    Zhang, He; Liu, Lian; Fu, Xin; Zhu, Zhenjun

    2013-04-15

    This study reports the development of a microfluidic beads-based immunosensor for sensitive detection of cancer biomarker α-fetoprotein (AFP) that uses multienzyme-nanoparticle amplification and quantum dots labels. This method utilizes microbeads functionalized with the capture antibodies (Ab₁) and modified electron rich proteins as sensing platform that was fabricated within a microfluidic channel, and uses gold nanoparticles (AuNPs) functionalized with the horseradish peroxidase (HRP) and the detection antibodies (Ab₂) as label. Greatly enhanced sensitivity for the cancer biomarker is based on a dual signal amplification strategy: first, the large surface area of Au nanoparticle carrier allows several binding events of HRP on each nanosphere. Enhanced sensitivity was achieved by introducing the multi-HRP-antibody functionalized AuNPs onto the surface of microbeads through "sandwich" immunoreactions and subsequently multiple biotin moieties could be deposited onto the surface of beads resulted from the oxidation of biotin-tyramine by hydrogen peroxide. Streptavidin-labeled quantum dots were then allowed to bind to the deposited biotin moieties and displayed the signal. Secondly, enhanced mass transport capability inherent from microfluidics leads to higher capture efficiency of targets because continuous flow within micro-channel delivers fresh analyte solution to the reaction site which maintains a high concentration gradient differential to enhance mass transport. Based on the dual signal amplification strategy, the developed microfluidic bead-based immunosensor could discriminate as low as 0.2 pg mL⁻¹ AFP in 10 μL of undiluted calf serum (0.2 fg/chip), and showed a 500-fold increase in detection limit compared to the off-chip test and 50-fold increase in detection limit compared to microfluidic beads-based immunoassay using single label HRP-Ab₂. The immunosensor showed acceptable repeatability and reproducibility. This microfluidic beads

  18. 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.

  19. 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. PMID:26274635

  20. 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). PMID:27393827

  1. Ambient Noise Observables from USArray: Site Amplification, Scattering and Attenuation

    NASA Astrophysics Data System (ADS)

    Bowden, D. C.; Tsai, V. C.; Lin, F. C.

    2015-12-01

    Previous work has demonstrated the usefulness of tracking wavefronts from ambient noise cross correlations across small, dense arrays (e.g., the Long Beach array). We now apply a similar method to USArray data to explore the information available on a continental scale. The approach measures the travel times and amplitudes of a wave propagating across the array and through numerical computations of spatial gradients corrects for geometric spreading, focusing and defocusing; any remaining signal is attributed to local site amplification effects, intrinsic attenuation, or the presence of sources/scatterers within the array. Because the USArray was active in different regions at different times and therefore subject to a variable background noise field, we are careful to divide the US into smaller regions and ensure that noise correlation pairs can be stacked over an identical time period before measuring a given spatial gradient. USArray observations of distant earthquakes have been shown to recover site amplification terms and attenuation through helmholtz tomography, but the use of ambient noise allows for such measurements at shorter periods and with greater flexibility. Site amplification maps produced with ambient noise show promising variations correlated with local geology, tectonics and velocity models, for periods at least as low as 12 seconds. Additionally, source differences between the incoming and outgoing wavefronts of ambient noise (negative and positive lags) may provide the ability to image noise sources or local scatterers within the continental US. These types of observations, especially site amplification and attenuation, may be used to supplement or improve traditional velocity-based tomographic methods, as they provide different constraints on temperature and density. They may also be useful for seismic hazard studies as they provide direct observations of site effects independent of complex inversions and forward model simulations.

  2. Profiling in situ microbial community structure with an amplification microarray.

    PubMed

    Chandler, Darrell P; Knickerbocker, Christopher; Bryant, Lexi; Golova, Julia; Wiles, Cory; Williams, Kenneth H; Peacock, Aaron D; Long, Philip E

    2013-02-01

    The objectives of this study were to unify amplification, labeling, and microarray hybridization chemistries within a single, closed microfluidic chamber (an amplification microarray) and verify technology performance on a series of groundwater samples from an in situ field experiment designed to compare U(VI) mobility under conditions of various alkalinities (as HCO(3)(-)) during stimulated microbial activity accompanying acetate amendment. Analytical limits of detection were between 2 and 200 cell equivalents of purified DNA. Amplification microarray signatures were well correlated with 16S rRNA-targeted quantitative PCR results and hybridization microarray signatures. The succession of the microbial community was evident with and consistent between the two microarray platforms. Amplification microarray analysis of acetate-treated groundwater showed elevated levels of iron-reducing bacteria (Flexibacter, Geobacter, Rhodoferax, and Shewanella) relative to the average background profile, as expected. Identical molecular signatures were evident in the transect treated with acetate plus NaHCO(3), but at much lower signal intensities and with a much more rapid decline (to nondetection). Azoarcus, Thaurea, and Methylobacterium were responsive in the acetate-only transect but not in the presence of bicarbonate. Observed differences in microbial community composition or response to bicarbonate amendment likely had an effect on measured rates of U reduction, with higher rates probable in the part of the field experiment that was amended with bicarbonate. The simplification in microarray-based work flow is a significant technological advance toward entirely closed-amplicon microarray-based tests and is generally extensible to any number of environmental monitoring applications.

  3. Profiling In Situ Microbial Community Structure with an Amplification Microarray

    PubMed Central

    Knickerbocker, Christopher; Bryant, Lexi; Golova, Julia; Wiles, Cory; Williams, Kenneth H.; Peacock, Aaron D.; Long, Philip E.

    2013-01-01

    The objectives of this study were to unify amplification, labeling, and microarray hybridization chemistries within a single, closed microfluidic chamber (an amplification microarray) and verify technology performance on a series of groundwater samples from an in situ field experiment designed to compare U(VI) mobility under conditions of various alkalinities (as HCO3−) during stimulated microbial activity accompanying acetate amendment. Analytical limits of detection were between 2 and 200 cell equivalents of purified DNA. Amplification microarray signatures were well correlated with 16S rRNA-targeted quantitative PCR results and hybridization microarray signatures. The succession of the microbial community was evident with and consistent between the two microarray platforms. Amplification microarray analysis of acetate-treated groundwater showed elevated levels of iron-reducing bacteria (Flexibacter, Geobacter, Rhodoferax, and Shewanella) relative to the average background profile, as expected. Identical molecular signatures were evident in the transect treated with acetate plus NaHCO3, but at much lower signal intensities and with a much more rapid decline (to nondetection). Azoarcus, Thaurea, and Methylobacterium were responsive in the acetate-only transect but not in the presence of bicarbonate. Observed differences in microbial community composition or response to bicarbonate amendment likely had an effect on measured rates of U reduction, with higher rates probable in the part of the field experiment that was amended with bicarbonate. The simplification in microarray-based work flow is a significant technological advance toward entirely closed-amplicon microarray-based tests and is generally extensible to any number of environmental monitoring applications. PMID:23160129

  4. Linking Arctic amplification and local feedbacks

    NASA Astrophysics Data System (ADS)

    Balcerak, Ernie

    2011-11-01

    Climate simulations show that as the Earth warms, the Arctic warms more than the average global warming. However, models differ on how much more the Arctic warms, and although scientists have proposed a variety of mechanisms to explain the Arctic warming amplification, there is no consensus on the main reasons for it. To shed light on this issue, Hwang et al. investigated the relationship between Arctic amplification and poleward energy transport and local Arctic feedbacks, such as changes in cloud cover or ice loss, across a group of models. The researchers noted that differences in atmospheric energy transport did not explain the ranges of polar amplification; rather, models with more amplification showed less energy transport into high latitudes. The authors found that decreasing energy transport is due to a coupled relationship between Arctic amplification and energy transport: Arctic amplification reduces the equator-to-pole temperature gradient, which strongly decreases energy transport. They suggest that this coupled relationship should be taken into account in studies of Arctic amplification. (Geophysical Research Letters, doi:10.1029/2011GL048546, 2011)

  5. 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.

  6. Molecular beacon probes combined with amplification by NASBA enable homogeneous, real-time detection of RNA.

    PubMed

    Leone, G; van Schijndel, H; van Gemen, B; Kramer, F R; Schoen, C D

    1998-05-01

    Molecular beacon probes can be employed in a NASBA amplicon detection system to generate a specific fluorescent signal concomitantly with amplification. A molecular beacon, designed to hybridize within the target sequence, was introduced into NASBA reactions that amplify the genomic RNA of potato leafroll virus (PLRV). During amplification, the probe anneals to the antisense RNA amplicon generated by NASBA, producing a specific fluorescent signal that can be monitored in real-time. The assay is rapid, sensitive and specific. As RNA amplification and detection can be carried out in unopened vessels, it minimizes the risk of carry-over contaminations. Robustness has been verified on real-world samples. This homogeneous assay, called AmpliDet RNA, is a significant improvement over current detection methods for NASBA amplicons and is suitable for one-tube applications ranging from high-throughput diagnostics to in vivo studies of biological activities.

  7. 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

  8. 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.

  9. 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

    2014-08-01

    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.

  10. 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.

  11. Balanced amplification: a new mechanism of selective amplification of neural activity patterns.

    PubMed

    Murphy, Brendan K; Miller, Kenneth D

    2009-02-26

    In cerebral cortex, ongoing activity absent a stimulus can resemble stimulus-driven activity in size and structure. In particular, spontaneous activity in cat primary visual cortex (V1) has structure significantly correlated with evoked responses to oriented stimuli. This suggests that, from unstructured input, cortical circuits selectively amplify specific activity patterns. Current understanding of selective amplification involves elongation of a neural assembly's lifetime by mutual excitation among its neurons. We introduce a new mechanism for selective amplification without elongation of lifetime: "balanced amplification." Strong balanced amplification arises when feedback inhibition stabilizes strong recurrent excitation, a pattern likely to be typical of cortex. Thus, balanced amplification should ubiquitously contribute to cortical activity. Balanced amplification depends on the fact that individual neurons project only excitatory or only inhibitory synapses. This leads to a hidden feedforward connectivity between activity patterns. We show in a detailed biophysical model that this can explain the cat V1 observations.

  12. 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...

  13. 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.

  14. Molecular Zipper: a fluorescent probe for real-time isothermal DNA amplification.

    PubMed

    Yi, Jizu; Zhang, Wandi; Zhang, David Y

    2006-01-01

    Rolling-circle amplification (RCA) and ramification amplification (RAM, also known as hyperbranched RCA) are isothermal nucleic acid amplification technologies that have gained a great application in in situ signal amplification, DNA and protein microarray assays, single nucleotide polymorphism detection, as well as clinical diagnosis. Real-time detection of RCA or RAM products has been a challenge because of most real-time detection systems, including Taqman and Molecular Beacon, are designed for thermal cycling-based DNA amplification technology. In the present study, we describe a novel fluorescent probe construct, termed molecular zipper, which is specially designed for quantifying target DNA by real-time monitoring RAM reactions. Our results showed that the molecular zipper has very low background fluorescence due to the strong interaction between two strands. Once it is incorporated into the RAM products its double strand region is opened by displacement, therefore, its fluorophore releases a fluorescent signal. Applying the molecular zipper in RAM assay, we were able to detect as few as 10 molecules within 90 min reaction. A linear relationship was observed between initial input of targets and threshold time (R2 = 0.985). These results indicate that molecular zipper can be applied to real-time monitoring and qualification of RAM reaction, implying an amenable method for automatic RAM-based diagnostic assays.

  15. Molecular Zipper: a fluorescent probe for real-time isothermal DNA amplification

    PubMed Central

    Yi, Jizu; Zhang, Wandi; Zhang, David Y.

    2006-01-01

    Rolling-circle amplification (RCA) and ramification amplification (RAM, also known as hyperbranched RCA) are isothermal nucleic acid amplification technologies that have gained a great application in in situ signal amplification, DNA and protein microarray assays, single nucleotide polymorphism detection, as well as clinical diagnosis. Real-time detection of RCA or RAM products has been a challenge because of most real-time detection systems, including Taqman and Molecular Beacon, are designed for thermal cycling-based DNA amplification technology. In the present study, we describe a novel fluorescent probe construct, termed molecular zipper, which is specially designed for quantifying target DNA by real-time monitoring RAM reactions. Our results showed that the molecular zipper has very low background fluorescence due to the strong interaction between two strands. Once it is incorporated into the RAM products its double strand region is opened by displacement, therefore, its fluorophore releases a fluorescent signal. Applying the molecular zipper in RAM assay, we were able to detect as few as 10 molecules within 90 min reaction. A linear relationship was observed between initial input of targets and threshold time (R2 = 0.985). These results indicate that molecular zipper can be applied to real-time monitoring and qualification of RAM reaction, implying an amenable method for automatic RAM-based diagnostic assays. PMID:16822854

  16. Multiple displacement amplification for preimplantation genetic diagnosis of fragile X syndrome.

    PubMed

    Lee, H-S; Kim, M J; Lim, C K; Cho, J W; Song, I O; Kang, I S

    2011-01-01

    Preimplantation genetic diagnosis (PGD) has become an assisted reproductive technique for couples that have genetic risks. Despite the many advantages provided by PGD, there are several problems, including amplification failure, allele drop-out and amplification inefficiency. We evaluated multiple displacement amplification (MDA) for PGD of the fragile X syndrome. Whole genome amplification was performed using MDA. MDA products were subjected to fluorescent PCR of fragile X mental retardation-1 (FMR1) CGG repeats, amelogenin and two polymorphic markers. In the pre-clinical tests, the amplification rates of the FMR1 CGG repeat, DXS1215 and FRAXAC1 were 84.2, 87.5 and 75.0%, respectively, while the allele dropout rates were 31.3, 57.1 and 50.0%, respectively. In two PGD treatment cycles, 20 embryos among 30 embryos were successfully diagnosed as 10 normal embryos, four mutated embryos and six heterozygous carriers. Three healthy embryos were transferred to the uterus; however, no clinical pregnancy was achieved. Our data indicate that MDA and fluorescent PCR with four loci can be successfully applied to PGD for fragile X syndrome. Advanced methods for amplification of minuscule amounts of DNA could improve the sensitivity and reliability of PGD for complicated single gene disorders. PMID:22095609

  17. Multiple displacement amplification for preimplantation genetic diagnosis of fragile X syndrome.

    PubMed

    Lee, H-S; Kim, M J; Lim, C K; Cho, J W; Song, I O; Kang, I S

    2011-11-17

    Preimplantation genetic diagnosis (PGD) has become an assisted reproductive technique for couples that have genetic risks. Despite the many advantages provided by PGD, there are several problems, including amplification failure, allele drop-out and amplification inefficiency. We evaluated multiple displacement amplification (MDA) for PGD of the fragile X syndrome. Whole genome amplification was performed using MDA. MDA products were subjected to fluorescent PCR of fragile X mental retardation-1 (FMR1) CGG repeats, amelogenin and two polymorphic markers. In the pre-clinical tests, the amplification rates of the FMR1 CGG repeat, DXS1215 and FRAXAC1 were 84.2, 87.5 and 75.0%, respectively, while the allele dropout rates were 31.3, 57.1 and 50.0%, respectively. In two PGD treatment cycles, 20 embryos among 30 embryos were successfully diagnosed as 10 normal embryos, four mutated embryos and six heterozygous carriers. Three healthy embryos were transferred to the uterus; however, no clinical pregnancy was achieved. Our data indicate that MDA and fluorescent PCR with four loci can be successfully applied to PGD for fragile X syndrome. Advanced methods for amplification of minuscule amounts of DNA could improve the sensitivity and reliability of PGD for complicated single gene disorders.

  18. 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.

  19. Detection of Chlamydia trachomatis by isothermal ramification amplification method: a feasibility study.

    PubMed

    Zhang, Wandi; Cohenford, Menashi; Lentrichia, Brian; Isenberg, Henry D; Simson, Elkin; Li, Hengjin; Yi, Jizu; Zhang, David Y

    2002-01-01

    Chlamydia trachomatis is the leading cause of sexually transmitted disease in the United States. Effective screening for this agent can facilitate prompt treatment and prevent its sequelae. The recent introduction of liquid-based cytology has made possible the simultaneous screening of cervical intraepithelial lesions and detection of C. trachomatis in a single collection vial. In this study we determined whether cytological fluid could support DNA-based amplification for the detection of C. trachomatis. Three methods were compared, including ramification amplification (RAM), real-time PCR with molecular beacon, and Abbott's ligase chain reaction (LCx). RAM is a novel, recently introduced, isothermal DNA amplification technique that utilizes a circular probe for target detection and achieves exponential amplification through the mechanism of primer extension, strand displacement, and ramification. Our results show that RAM can detect as few as 10 C. trachomatis elementary bodies in less than 2 h, comparable to results with real-time PCR. Thirty clinical specimens collected in PreservCyt solution were tested by LCx, real-time PCR, and RAM. Among 30 specimens, 15 were positive by PCR and LCx and 14 were positive by RAM. One specimen missed by RAM had an inadequate amount of residual cellular material. Our results show that nucleic acid amplification methods can serve to detect C. trachomatis and presumably other sexually transmitted agents in cytological fluid and that the RAM assay can be an alternative to PCR and LCx because of its simplicity and isothermal amplification.

  20. 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.