Frequency locked micro disk resonator for real time and precise monitoring of refractive index.

We experimentally demonstrate a frequency modulation locked servo loop, locked to a resonance line of an on-chip microdisk resonator in a silicon nitride platform. By using this approach, we demonstrate real-time monitoring of refractive index variations with a precision approaching 10(-7) RIU, using a moderate Q factor of 10(4). The approach can be applied for intensity independent, dynamic and precise index of refraction monitoring for biosensing applications.

Pipeline Architecture and Parallel Computation-Based Real-Time Stereovision Tracking System for Surgical Navigation

This paper studies the development of a real-time stereovision system to track multiple infrared markers attached to a surgical instrument. Multiple stages of pipeline in field-programmable gate array (FPGA) are developed to recognize the targets in both left and right image planes and to give each target a unique label. The pipeline architecture includes a smoothing filter, an adaptive threshold module, a connected component labeling operation, and a centroid extraction process. A parallel distortion correction method is proposed and implemented in a dual-core DSP. A suitable kinematic model is established for the moving targets, and a novel set of parallel and interactive computation mechanisms is proposed to position and track the targets, which are carried out by a cross-computation method in a dual-core DSP. The proposed tracking system can track the 3-D coordinate, velocity, and acceleration of four infrared markers with a delay of 9.18 ms. Furthermore, it is capable of tracking a maximum of 110 infrared markers without frame dropping at a frame rate of 60 f/s. The accuracy of the proposed system can reach the scale of 0.37 mm RMS along the x- and y-directions and 0.45 mm RMS along the depth direction (the depth is from 0.8 to 0.45 m). The performance of the proposed system can meet the requirements of applications such as surgical navigation, which needs high real time and accuracy capability.

Pipeline Architecture and Parallel Computation-Based Real-Time Stereovision Tracking System for Surgical Navigation

This paper studies the development of a real-time stereovision system to track multiple infrared markers attached to a surgical instrument. Multiple stages of pipeline in field-programmable gate array (FPGA) are developed to recognize the targets in both left and right image planes and to give each target a unique label. The pipeline architecture includes a smoothing filter, an adaptive threshold module, a connected component labeling operation, and a centroid extraction process. A parallel distortion correction method is proposed and implemented in a dual-core DSP. A suitable kinematic model is established for the moving targets, and a novel set of parallel and interactive computation mechanisms is proposed to position and track the targets, which are carried out by a cross-computation method in a dual-core DSP. The proposed tracking system can track the 3-D coordinate, velocity, and acceleration of four infrared markers with a delay of 9.18 ms. Furthermore, it is capable of tracking a maximum of 110 infrared markers without frame dropping at a frame rate of 60 f/s. The accuracy of the proposed system can reach the scale of 0.37 mm RMS along the x- and y-directions and 0.45 mm RMS along the depth direction (the depth is from 0.8 to 0.45 m). The performance of the proposed system can meet the requirements of applications such as surgical navigation, which needs high real time and accuracy capability.

a real-time path planning approach without the computation of cspace obstacles

An important concept proposed in the early stage of robot path planning field is the shrinking of the robot to a point and meanwhile expanding of the obstacles in the workspace as a set of new obstacles. The resulting grown obstacles are called the Configuration Space (Cspace) obstacles. The find-path problem is then transformed into that of finding a collision free path for a point robot among the Cspace obstacles. However, the research experiences obtained so far have shown that the calculation of the Cspace obstacles is very hard work when the following situations occur: 1. both the robot and obstacles are not polygons and 2. the robot is allowed to rotate. This situation is even worse when the robot and obstacles are three dimensional (3D) objects with various shapes. Obviously a direct path planning approach without the calculation of the Cspace obstacles is strongly needed. This paper presents such a new real-time robot path planning approach which, to the best of our knowledge, is the first one in the robotic community. The fundamental ideas are the utilization of inequality and optimization technique. Simulation results have been presented to show its merits.

[Ru(bpy)2dppz]2+ electrochemiluminescence switch and its applications for DNA interaction study and label-free ATP aptasensor

[Ru(bpy)2dppz]2+ electrochemiluminescence (ECL) was studied, and it was used to investigate DNA interaction and develop a label-free ATP aptasensor for the first time. ECL of [Ru(bpy)2dppz]2+ is negligible in aqueous solution, and increases approximately 1000 times when [Ru(bpy)2dppz]2+ intercalates into the nucleic acid structure. The ECL switch behavior of [Ru(bpy)2dppz]2+ is ascribed to the intercalation that shields the phenazine nitrogens from the solvent and results in a luminescent excited state. The ECL switch by DNA was applied to investigate the interaction of [Ru(bpy)2dppz]2+ with herring sperm DNA. The calculated equilibrium constant (K) is 1.35 x 10(6) M(-1), and the calculated binding-site size (s) is 0.88 base pair, which is consistent with the reported values.

Label-Free Colorimetric Detection of Aqueous Mercury Ion (Hg2+) Using Hg2+-Modulated G-Quadruplex-Based DNAzymes

Mercury ion (Hg2+) is able to specifically bind to the thymine-thymine (T-T) base pair in a DNA duplex, thus providing a rationale for DNA-based selective detection of Hg2+ with various means. In this work, we for the first time utilize the Hg2+-mediated T-T base pair to modulate the proper folding of G-quadruplex DNAs and inhibit the DNAzyme activity, thereby pioneering a facile approach to sense Hg2+ with colorimetry. Two bimolecular DNA G-quadruplexes containing many T residues are adopted here, which function well in low- and high-salt conditions, respectively. These G-quadruplex DNAs are able to bind hemin to form the peroxidase-like DNAzymes in the folded state. Upon addition of Hg2+, the proper folding of G-quadruplex DNAs is inhibited due to the formation of T-Hg2+-T complex. Ibis is reflected by the notable change of the Soret band of hemin when investigated by using UV-vis absorption spectroscopy. As a result of Hg2+ inhibition, a sharp decrease in the catalytic activity toward the H2O2-mediated oxidation of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)diammonium salt (ABTS) is observed, accompanied by a change in solution color. Through this approach, aqueous Hg2+ can be detected at 50 nM (10 ppb) with colorimetry in a facile way, with high selectivity against other metal ions.

Real-time PCR microfluidic devices with concurrent electrochemical detection

Electrochemistry-based detection methods hold great potential towards development of hand-held nucleic-acid analyses instruments. In this work, we demonstrate the implementation of in situ electrochemical (EC) detection method in a microfluidic flow-through EC-qPCR (FTEC-qPCR) device, where both the amplification of the target nucleic-acid sequence and subsequent EC detection of the PCR amplicon are realized simultaneously at selected PCR cycles in the same device. The FTEC-qPCR device utilizes methylene blue (MB), an electroactive DNA intercalator, for electrochemical signal measurements in the presence of PCR reagent components. Our EC detection method is advantageous, when compared to other existing EC methods for PCR amplicon analysis, since FTEC-qPCR does not require probe-modified electrodes, or asymmetric PCR, or solid-phase PCR. Key technical issues related to surface passivation, electrochemical measurement, PCR inhibition by metal electrode, bubble-free PCR, were investigated. By controlling the concentration of MB and the exposure of PCR mixture to the bare metal electrode, we successfully demonstrated electrochemical measurement of MB in solution-phase, symmetric PCR by amplifying a fragment of lambda phage DNA.

Features of programmed cell death in intact Xenopus oocytes and early embryos revealed by near-infrared fluorescence and real-time monitoring.

Factors influencing apoptosis of vertebrate eggs and early embryos have been studied in cell-free systems and in intact embryos by analyzing individual apoptotic regulators or caspase activation in static samples. A novel method for monitoring caspase activity in living Xenopus oocytes and early embryos is described here. The approach, using microinjection of a near-infrared caspase substrate that emits fluorescence only after its proteolytic cleavage by active effector caspases, has enabled the elucidation of otherwise cryptic aspects of apoptotic regulation. In particular, we show that brief caspase activity (10 min) is sufficient to cause apoptotic death in this system. We illustrate a cytochrome c dose threshold in the oocyte, which is lowered by Smac, a protein that binds thereby neutralizing the inhibitor of apoptosis proteins. We show that meiotic oocytes develop resistance to cytochrome c, and that the eventual death of oocytes arrested in meiosis is caspase-independent. Finally, data acquired through imaging caspase activity in the Xenopus embryo suggest that apoptosis in very early development is not cell-autonomous. These studies both validate this assay as a useful tool for apoptosis research and reveal subtleties in the cell death program during early development. Moreover, this method offers a potentially valuable screening modality for identifying novel apoptotic regulators.

Real-time imaging of novel spatial and temporal responses to photodynamic stress.

Cells subjected to various forms of stress have been shown to induce bystander responses in nontargeted cells, thus extending the stress response to a larger population. However, the mechanism(s) of bystander responses remains to be clearly identified, particularly for photodynamic stress. Oxidative stress and cell viability were studied on the spatial and temporal levels after photodynamic targeting of a subpopulation of EMT6 murine mammary cancer cells in a multiwell plate by computerized time-lapse fluorescence microscopy. In the targeted population a dose-dependent loss of cell viability was observed in accordance with increased oxidative stress. This was accompanied by increased oxidative stress in bystander populations but on different time scales, reaching a maximum more rapidly in targeted cells. Treatment with extracellular catalase, or the NADPH oxidase inhibitor diphenyleneiodinium, decreased production of reactive oxygen species (ROS) in both populations. These effects are ascribed to photodynamic activation of NADPH-oxidase in the targeted cells, resulting in a rapid burst of ROS formation with hydrogen peroxide acting as the signaling molecule responsible for initiation of these photodynamic bystander responses. The consequences of increased oxidative stress in bystander cells should be considered in the overall framework of photodynamic stress.

A traffic differentiation add-on to the IEEE 802.15.4 protocol: implementation and experimental validation over a real-time operating system

The IEEE 802.15.4 is the most widespread used protocol for Wireless Sensor Networks (WSNs) and it is being used as a baseline for several higher layer protocols such as ZigBee, 6LoWPAN or WirelessHART. Its MAC (Medium Access Control) supports both contention-free (CFP, based on the reservation of guaranteed time-slots GTS) and contention based (CAP, ruled by CSMA/CA) access, when operating in beacon-enabled mode. Thus, it enables the differentiation between real-time and best-effort traffic. However, some WSN applications and higher layer protocols may strongly benefit from the possibility of supporting more traffic classes. This happens, for instance, for dense WSNs used in time-sensitive industrial applications. In this context, we propose to differentiate traffic classes within the CAP, enabling lower transmission delays and higher success probability to timecritical messages, such as for event detection, GTS reservation and network management. Building upon a previously proposed methodology (TRADIF), in this paper we outline its implementation and experimental validation over a real-time operating system. Importantly, TRADIF is fully backward compatible with the IEEE 802.15.4 standard, enabling to create different traffic classes just by tuning some MAC parameters.

Real-time communications over cluster-tree sensor networks with mobile sink behaviour

Modelling the fundamental performance limits of wireless sensor networks (WSNs) is of paramount importance to understand the behaviour of WSN under worst case conditions and to make the appropriate design choices. In that direction, this paper contributes with a methodology for modelling cluster tree WSNs with a mobile sink. We propose closed form recurrent expressions for computing the worst case end to end delays, buffering and bandwidth requirements across any source-destination path in the cluster tree assuming error free channel. We show how to apply our theoretical results to the specific case of IEEE 802.15.4/ZigBee WSNs. Finally, we demonstrate the validity and analyze the accuracy of our methodology through a comprehensive experimental study, therefore validating the theoretical results through experimentation.

Chitosan/AuNPs Modified Graphene Electrochemical Sensor for Label-Free Human Chorionic Gonadotropin Detection

A new immunosensor is presented for human chorionic gonadotropin (hCG), made by electrodepositing chitosan/gold-nanoparticles over graphene screen-printed electrode (SPE). The antibody was covalently bound to CS via its Fc-terminal. The assembly was controlled by electrochemical Impedance Spectroscopy (EIS) and followed by Fourier Transformed Infrared (FTIR). The hCG-immunosensor displayed linear response against the logarithm-hCG concentration for 0.1–25 ng/mL with limit of detection of 0.016 ng/mL. High selectivity was observed in blank urine and successful detection of hCG was also achieved in spiked samples of real urine from pregnant woman. The immunosensor showed good detection capability, simplicity of fabrication, low-cost, high sensitivity and selectivity.

Label-free human chorionic gonadotropin detection at picogram levels using oriented antibodies bound to graphene screen-printed electrodes

Human chorionic gonadotropin (hCG) is a key diagnostic marker of pregnancy and an important biomarker for cancers in the prostate, ovaries and bladder and therefore of great importance in diagnosis. For this purpose, a new immunosensor of screen-printed electrodes (SPEs) is presented here. The device was fabricated by introducing a polyaniline (PANI) conductive layer, via in situ electropolymerization of aniline, onto a screen-printed graphene support. The PANI-coated graphene acts as the working electrode of a three terminal electrochemical sensor. The working electrode is functionalised with anti-hCG, by means of a simple process that enabled oriented antibody binding to the PANI layer. The antibody was attached to PANI following activation of the –COOH group at the Fc terminal. Functionalisation of the electrode was analysed and optimized using Electrochemical Impedance Spectroscopy (EIS). Chemical modification of the surface was characterised using Fourier transform infrared, and Raman spectroscopy with confocal microscopy. The graphene–SPE–PANI devices displayed linear responses to hCG in EIS assays from 0.001 to 50 ng mL−1 in real urine, with a detection limit of 0.286 pg mL−1. High selectivity was observed with respect to the presence of the constituent components of urine (urea, creatinine, magnesium chloride, calcium chloride, sodium dihydrogen phosphate, ammonium chloride, potassium sulphate and sodium chloride) at their normal levels, with a negligible sensor response to these chemicals. Successful detection of hCG was also achieved in spiked samples of real urine from a pregnant woman. The immunosensor developed is a promising tool for point-of-care detection of hCG, due to its excellent detection capability, simplicity of fabrication, low-cost, high sensitivity and selectivity.

Permutation entropy based real-time chatter detection using audio signal in turning process

Machine tool chatter is an unfavorable phenomenon during metal cutting, which results in heavy vibration of cutting tool. With increase in depth of cut, the cutting regime changes from chatter-free cutting to one with chatter. In this paper, we propose the use of permutation entropy (PE), a conceptually simple and computationally fast measurement to detect the onset of chatter from the time series using sound signal recorded with a unidirectional microphone. PE can efficiently distinguish the regular and complex nature of any signal and extract information about the dynamics of the process by indicating sudden change in its value. Under situations where the data sets are huge and there is no time for preprocessing and fine-tuning, PE can effectively detect dynamical changes of the system. This makes PE an ideal choice for online detection of chatter, which is not possible with other conventional nonlinear methods. In the present study, the variation of PE under two cutting conditions is analyzed. Abrupt variation in the value of PE with increase in depth of cut indicates the onset of chatter vibrations. The results are verified using frequency spectra of the signals and the nonlinear measure, normalized coarse-grained information rate (NCIR).

EFFECTIVE NETWORKS FOR REAL-TIME DISTRIBUTED PROCESSING

This paper applies the concepts and methods of complex networks to the development of models and simulations of master-slave distributed real-time systems by introducing an upper bound in the allowable delivery time of the packets with computation results. Two representative interconnection models are taken into account: Uniformly random and scale free (Barabasi-Albert), including the presence of background traffic of packets. The obtained results include the identification of the uniformly random interconnectivity scheme as being largely more efficient than the scale-free counterpart. Also, increased latency tolerance of the application provides no help under congestion.