Real-time detection of multiple moving objects in complex image sequences

In this article, a real-time multistage method for detecting multiple objects moving in real scenes is presented. At the first level, a rough focus-of-attention mechanism is used to individuate areas of the input image that show remarkable differences with a real-time updated background image. Binary statistical morphology (BMS) operators are applied to individuate image pixels, which can be associated with real objects moving into the scene. High stability to noise is obtained by tuning the smoothing effects of the BSM operators according to the noise level present in the original image sequence. Then, at the second level, a composition of BSM is applied to eliminate isolated points and to favor dense agglomerate of changed pixels, i.e., blobs. The last level attempts to describe changes in terms of motion of blobs by allowing blobs to merge, split, appear, and vanish. A blob-matching procedure is used for tracking blobs over consecutive frames. Experimental results on real scenes, which demonstrate the advantages of the proposed method with respect to existing change detection methods, are given. © 1999 John Wiley & Sons, Inc. Int J Imaging Syst Technol 10, 305–317, 1999     

一种实时的复杂户外场景下目标检测算法

根据实际系统中高帧频摄像机背景图像运动不大而目标有一定运动的特点,首先分离奇偶帧图像,隔帧相减并用自适应阈值技术二值化图像来检测运动目标,经过形态学处理得到许多连通的区域,然后利用目标面积最大等特征识别目标并用外接矩形定位目标,得到目标运动的真实轨迹。仿真结果表明：该算法能简单有效地检测机动目标且易于硬件并行实现。     

Possible mercury speciation in urine samples using potentiometric methods

The strong dependence of heavy metal toxicity on metal physical–chemical structure has directed the interest in the qualitative and quantitative measurement of metal species in different environments. Mercury is ubiquitous in the environment, and occurs in three oxidation states in several physical and chemical forms. Among these, Hg(II) can severely damage the kidneys and the gastrointestinal tract, and is generally a serious hazard to human health. Potentiometric sensors based on ion-selective electrodes provide a simple, highly selective, precise, and economical method for online Hg(II) concentration monitoring in a variety of environments. Due to their selectivity, neutral carrier-based potentiometric sensors are routinely used for measuring ions directly in complex biological and environmental samples, and therein, several Hg(II)-selective electrodes have been studied in literature. This work presents a Hg(II) ion-selective electrode that uses a 1,4,8,11-tetrathiacyclotetradecane derivative entrapped inside a membrane for the measurement of Hg(II) in urine samples. Alumina modified dimethyl sulfoxide (AMDMSO) was used to separate Hg(II) from matrices, a mechanism already suggested in literature explaining the unique uptake of Hg(II) ions by binding as neutral at pH values < 3.     

New polyaniline-based potentiometric biosensor for pesticides detection

A cholinesterase potentiometric sensor based on a glassy carbon electrode modified with processible polyaniline has been developed and explored for the detection of organophosphorus and carbamic pesticides. Acetyl- and butyryl-cholinesterase from various sources were immobilized on the surface of a polyaniline modified electrode by cross-linking with glutaraldehyde. Polyaniline modification made it possible to obtain a reversible pH response of about 86-mV/pH unit and to improve the analytical and operational characteristics of substrate and inhibitor detection. The sensitivity of pesticide detection depends on both the source of the enzyme and its activity and on pesticide hydrophobicity. The detection limits of the pesticides investigated (Trichlorfon, 1.5/spl middot/10/sup -7/ mol/l, Coumaphos, 5/spl middot/10/sup -9/ mol/l, Methiocarb, 8/spl middot/10/sup -7/ mol/l, Aldicarb, 2/spl middot/10/sup -7/ mol/l) were found lower than those obtained with other similar cholinesterase sensors. The possibility for the detection of the trace amounts of pesticides in river waters and grape juice with cholinesterase sensors developed has been established on model samples.     

Improved detection limits and sensitivities of potentiometric titrations

Zero-current ion fluxes through polymer membranes of ion-selective electrodes (ISEs) may lead to biased endpoints of potentiometric titrations. The bias is eliminated, and the sensitivity of the end-point detection is improved through reducing transmembrane ion fluxes by an appropriate choice of the inner solution. Surprisingly, ISE membranes that have a significant primary ion flux toward the inner solution show much larger sensitivities than expected by titration theory; however, depending on the experimental conditions, their application may bias the endpoint. With the optimal systems, endpoint detection is now possible with total sample concentrations below 10(-6) M, as demonstrated by the titration of EDTA with a Pb2+ solution.     

Mass amplifying probe for sensitive fluorescence anisotropy detection of small molecules in complex biological samples

Fluorescence anisotropy (FA) is a reliable and excellent choice for fluorescence sensing. One of the key factors influencing the FA value for any molecule is the molar mass of the molecule being measured. As a result, the FA method with functional nucleic acid aptamers has been limited to macromolecules such as proteins and is generally not applicable for the analysis of small molecules because their molecular masses are relatively too small to produce observable FA value changes. We report here a molecular mass amplifying strategy to construct anisotropy aptamer probes for small molecules. The probe is designed in such a way that only when a target molecule binds to the probe does it activate its binding ability to an anisotropy amplifier (a high molecular mass molecule such as protein), thus significantly increasing the molecular mass and FA value of the probe/target complex. Specifically, a mass amplifying probe (MAP) consists of a targeting aptamer domain against a target molecule and molecular mass amplifying aptamer domain for the amplifier protein. The probe is initially rendered inactive by a small blocking strand partially complementary to both target aptamer and amplifier protein aptamer so that the mass amplifying aptamer domain would not bind to the amplifier protein unless the probe has been activated by the target. In this way, we prepared two probes that constitute a target (ATP and cocaine respectively) aptamer, a thrombin (as the mass amplifier) aptamer, and a fluorophore. Both probes worked well against their corresponding small molecule targets, and the detection limits for ATP and cocaine were 0.5 μM and 0.8 μM, respectively. More importantly, because FA is less affected by environmental interferences, ATP in cell media and cocaine in urine were directly detected without any tedious sample pretreatment. Our results established that our molecular mass amplifying strategy can be used to design aptamer probes for rapid, sensitive, and selective detection of small molecules by means of FA in complex biological samples.     

Metal nanoparticle-based electrochemical stripping potentiometric detection of DNA hybridization

A new nanoparticle-based electrical detection of DNA hybridization, based on electrochemical stripping detection of the colloidal gold tag, is described. In this protocol, the hybridization of a target oligonucleotide to magnetic bead-linked oligonucleotide probes is followed by binding of the streptavidin-coated metal nanoparticles to the captured DNA, dissolution of the nanometer-sized gold tag, and potentiometric stripping measurements of the dissolved metal tag at single-use thick-film carbon electrodes. An advanced magnetic processing technique is used to isolate the DNA duplex and to provide low-volume mixing. The influence of relevant experimental variables, including the amounts of the gold nanoparticles and the magnetic beads, the duration of the hybridization and gold dissolution steps, and the parameters of the potentiometric stripping operation upon the hybridization signal, is examined and optimized. Transmission electron microscopy micrographs indicate that the hybridization event leads to the bridging of the gold nanoparticles to the magnetic beads. Further signal amplification, and lowering of the detection limits to the nanomolar and picomolar domains, are achieved by precipitating gold or silver, respectively, onto the colloidal gold label. The new electrochemical stripping metallogenomagnetic protocol couples the inherent signal amplification of stripping metal analysis with discrimination against nonhybridized DNA, the use of microliter sample volumes, and disposable transducers and, hence, offers great promise for decentralized genetic testing.     

Triangle-programmed coulometric nanotitrations completed by continuous flow with potentiometric detection

Coulometric nanotitrations were realized in a microchannel system using a continuous-flow titration technique with a triangle current-time profile. Redox and acid-base titrations were carried out on Fe(II) and nitric acid samples, respectively, with the same nanotitrator device. A linear relation between the concentration and the coulometric current transferred to the solution was found. The advantages of this universally applicable nanotitrator are fast response, low sample volume, high sensitivity, and high reproducibility as well as the convenience of handling an automated analyzer of the flow-through type.     

Trace detection and discrimination of explosives using electrochemical potentiometric gas sensors

In this article, selective and sensitive detection of trace amounts of pentaerythritol tetranitrate (PETN), 2,4,6-trinitrotoluene (TNT) and cyclotrimethylenetrinitramine (RDX) is demonstrated. The screening system is based on a sampling/concentrator front end and electrochemical potentiometric gas sensors as the detector. Preferential hydrocarbon and nitrogen oxide(s) mixed potential sensors based on lanthanum strontium chromite and Pt electrodes with yttria stabilized zirconia (YSZ) solid electrolyte were used to capture the signature of the explosives. Quantitative measurements based on hydrocarbon and nitrogen oxide sensor responses indicated that the detector sensitivity scaled proportionally with the mass of the explosives (1-3 μg). Moreover, the results showed that PETN, TNT, and RDX samples could be discriminated from each other by calculating the ratio of nitrogen oxides to hydrocarbon integrated area under the peak. Further, the use of front-end technology to collect and concentrate the high explosive (HE) vapors make intrinsically low vapor pressure of the HE less of an obstacle for detection while ensuring higher sensitivity levels. In addition, the ability to use multiple sensors each tuned to basic chemical structures (e.g., nitro, amino, peroxide, and hydrocarbon groups) in HE materials will permit the construction of low-cost detector systems for screening a wide spectrum of explosives with lower false positives than present-day technologies.     

Modified carbon paste electrode for potentiometric determination of diquat dibromide pesticide in water and urine samples

A carbon paste electrode for diquat dibromide (Dq.2Br) was prepared and fully characterized in terms of composition, usable pH range, response time and temperature. The electrode was applied to the potentiometric determination of diquat ions in water and urine samples with average recoveries of 97.5-104.0% and relative standard deviations of 0.30-4.73%. The electrode is based on the ion pair, namely, diquat-phosphotungstate dissolved in 2-nitrophenyloctyl ether (2-NPOE) as pasting liquid with 1.0% Na-TPB as an additive. The modified electrode showed a near-Nernstian slope of 30.8 mV over the concentration range of 3.8 × 10^− 6to 1.0 × 10^− 3 M with the limit of detection 9.0 × 10^− 7 over the pH range 4.5-9.5. The electrode exhibits good selectivity for Dq cations with respect to a large number of inorganic cations, organic cations, sugars and amino acids. The proposed potentiometric method offers the advantages of simplicity, accuracy, automation feasibility and applicability to turbid and colored sample solutions.     

Real-time action detection and temporal segmentation in continuous video

Temporal segmentation of actions has been under intensive focus in the field of computer vision for a prolonged period. The present study proposed a template-based framework to resolve the issues concerning timeliness and real-time performance in the temporal segmentation in a continuous video. A complete action can be detected, based on the previous frames, and the action can be segmented immediately without waiting for the follow-up frames. Herein, characteristic frames are selected by a martingale-based method, followed by the formation of the corresponding motion history through backtracking along the characteristic frames, and the final segmentation is determined according to the recognition model trained by the extreme learning machine. In the experiment on the IXMAS database, the average rate of the detection of action reached 91%, and the accuracy in the frame level reached 83.5%. In the experiment on the 3D skeleton data based on Kinect, the detection rate reached 94%.     

Real-time rolling circle amplification for protein detection

Real-time nucleic acid amplification methods can be extremely useful for the identification and quantitation of nucleic acid analytes, but are more difficult to adapt to protein or other analytes. To facilitate the development of real-time rolling circle amplification (RCA) for protein targets, we have developed a novel type of conformation-switching aptamer that can be circularized upon interaction with its protein target, the platelet-derived growth factor (PDGF). Using the structure-switching aptamer, real-time RCA can be used to specifically quantitate PDGF down to the low-nanomolar range (limit of detection, 0.4 nM), even against a background of cellular lysate. The aptamer can also be adapted to RCA on surfaces, although quantitation proved to be more difficult. One of the great advantages of the method described herein is that it can be immediately adapted to almost any aptamer and does not require two or more affinity reagents as do sandwich or proximity assays.     

Turbidimetric detection method in flow-assisted separation of dispersed samples

Characterization of dispersed samples is an outstanding trend in analytical science. Among flow-assisted separation techniques for dispersed samples, size exclusion chromatography, hydrodynamic chromatography, and field-flow fractionation are the most widely applied. With dispersed analytes separated by these techniques, the UV/vis spectrophotometric detectors work as turbidimeters. To directly convert the analytical signal for quantitative analysis, the extinction properties of the dispersed analyte must be known. A new method is proposed to experimentally obtain-by single-run, flow-assisted separation with UV/vis diode-array detectors-the mass-size (or number-size) distribution function of the analytes when a retention-to-size relationship is either theoretically or empirically available for the chosen separation technique. This approach needs neither standards nor reliance on a method to predict the optical properties of the analytes. Theory and original algorithms are presented. Algorithms are then tested to optimize the numerical routines. Accuracy and robustness of the method are evaluated by simulation, and limitations for the application to experimental data are described. Finally, first application to field-flow fractionation shows validity of the method when applied to a few real cases.     

Limit of detection determination for censored samples

The problem of setting the limit of detection is considered for censored samples and heterogeneous errors. After formal definitions of the critical level and of the method detection limit, we obtain simplified maximum likelihood-type estimators. The estimation problem of the truncation parameter and its uncertainty are reviewed. Upper confidence bounds for the limit of detection are derived and some simulation results are given.     

Fringe detection in noisy complex interferograms

A new algorithm to estimate the two-dimensional local frequencies of phase interferometric data is described. With a complex sine-wave model, demonstration is given that a conventional multiple-signal classification (MUSIC) algorithm can be used in spite of multiplicative noise perturbations. A faster algorithm dedicated to the processing of interferograms is developed and a measure of confidence in the estimate is proposed. We studied numerical performances using synthetic fringes. As a result of the frequency estimation, knowledge of the fringe local width and orientation can be applied to restore noisy phase data. Results of a complex phase filter are presented for real interferograms obtained from synthetic aperture radar images.     

Ultrasensitive reporter protein detection in genetically engineered bacteria

We demonstrate the use of laser-induced fluorescence confocal spectroscopy to measure analyte-stimulated enhanced green fluorescent protein (egfp) synthesis by genetically modified Escherichia coli bioreporter cells. Induction is measured in cell lysates and, since the spectroscopic focal volume is approximately the size of one bioreporter cell, also in individual live bacteria. This is, to our knowledge, the first ever proof-of-concept work utilizing instrumentation with single-molecule detection capability to monitor bioreporter response. Although we use arsenic inducible bioreporters here, the method is extensible to gfp/egfp bioreporters that are responsive to other substances.     

Antimicrobial peptides for detection of bacteria in biosensor assays

Bacteria, plants, and higher and lower animals have evolved an innate immune system as a first line of defense against microbial invasion. Some of these organisms produce antimicrobial peptides (AMPs) as a part of this chemical immune system. AMPs exert their antimicrobial activity by binding to components of the microbe's surface and disrupting the membrane. The overall goal of this study was to apply the AMP magainin I as a recognition element for Escherichia coli O157:H7 and Salmonella typhimurium detection on an array-based biosensor. We immobilized magainin I on silanized glass slides using biotin-avidin chemistry, as well as through direct covalent attachment. Cy5-labeled, heat-killed cells were used to demonstrate that the immobilized magainin I can bind Salmonella with detection limits similar to analogous antibody-based assays. Detection limits for E. coli were higher than in analogous antibody-based assays, but it is expected that other AMPs may possess higher affinities for this target. The results showed that both specific and nonspecific binding strongly depend on the method used for peptide immobilization. Direct attachment of magainin to the substrate surface not only decreased nonspecific cell binding but also resulted in improved detection limits for both Salmonella and E. coli.     

Real-time detection and elimination of nonorthogonality error in interference fringe processing

In the measurement system of interference fringe, the nonorthogonality error is a main error source that influences the precision and accuracy of the measurement system. The detection and elimination of the error has been an important target. A novel method that only uses the cross-zero detection and the counting is proposed to detect and eliminate the nonorthogonality error in real time. This method can be simply realized by means of the digital logic device, because it does not invoke trigonometric functions and inverse trigonometric functions. And it can be widely used in the bidirectional subdivision systems of a Moiré fringe and other optical instruments.