Dual fluorescence sensor for trace oxygen and temperature with unmatched range and sensitivity

An optical dual sensor for oxygen and temperature is presented that is highly oxygen sensitive and covers a broad temperature range. Dual sensing is based on luminescence lifetime measurements. The novel sensor contains two luminescent compounds incorporated into polymer films. The temperature-sensitive dye (ruthenium tris-1,10-phenanthroline) has a highly temperature-dependent luminescence and is incorporated in poly(acrylonitrile) to avoid cross-sensitivity to oxygen. Fullerene C70 was used as the oxygen-sensitive probe owing to its strong thermally activated delayed fluorescence at elevated temperatures that is extremely oxygen sensitive. The cross-sensitivity of C70 to temperature is accounted for by means of the temperature sensor. C70 is incorporated into a highly oxygen-permeable polymer, either ethyl cellulose or organosilica. The two luminescent probes have different emission spectra and decay times, and their emissions can be discriminated using both parameters. Spatially resolved sensing is achieved by means of fluorescence lifetime imaging. The response times of the sensor to oxygen are short. The dual sensor exhibits a temperature operation range between at least 0 and 120 degrees C, and detection limits for oxygen in the ppbv range, operating for oxygen concentrations up to at least 50 ppmv. These ranges outperform all dual oxygen and temperature sensors reported so far. The dual sensor presented in this study is especially appropriate for measurements under extreme conditions such as high temperatures and ultralow oxygen levels. This dual sensor is a key step forward in a number of scientifically or commercially important applications including food packaging, for monitoring of hyperthermophilic microorganisms, in space technology, and safety and security applications in terms of detection of oxygen leaks.     

Fluorescence quenching of the europium tetracycline hydrogen peroxide complex by copper (II) and other metal ions

The europium-tetracycline complex [Eu(Tc)] is known to show only weak fluorescence with an emission maximum at 615 nm. On addition of hydrogen peroxide (HP), the strongly fluorescent [Eu(Tc)(HP)] complex is formed, which displays a 15-fold stronger luminescence intensity. This study describes the decrease in luminescence intensity of the [Eu(Tc)(HP)] complex in aqueous solution in the presence of Cu2+, Fe3+, Ag+, Al3+, Zn2+, Co2+, Ni2+, Mn2+, Ca2+, and Mg2+. Static and dynamic quenching can be induced by Cu2+, and these processes were quantified by means of their quenching constants. Stern-Volmer plots were also derived from lifetime imaging measurements accomplished by the rapid lifetime determination (RLD) technique based on microwell plate assays, and also by the time-correlated single photon counting (TCSPC) technique. According to those data, a time-resolved fluorescent method for copper determination can be proposed that is based on dynamic quenching of the [Eu(Tc)(HP)] complex by Cu2+ ions. The response to copper concentrations is linear up to 1.6 micromol L(-1), providing a detection limit of 0.2 micromol L(-1).     

Error analysis of the rapid lifetime determination method for double-exponential decays and new windowing schemes

The rapid lifetime determination method (RLD) is a mathematical technique for extremely rapid evaluations of lifetimes in exponential decays. It has been applied in luminescence microscopy and single-molecule lifetime evaluation. To date, the primary application has been in single-exponential evaluations. We present extensions of the method to double exponentials. Using Monte Carlo simulations, we assess the performance of both the double-exponential decay with known lifetimes and the double-exponential decay with unknown preexponential factors and lifetimes. Precision is evaluated as a function of the noise level (Poisson statistics), the ratios of the lifetimes, the ratios of their preexponential factors, and the fitting window. Optimum measurement conditions are determined. RLD is shown to work well over a wide range of practical experimental conditions. If the lifetimes are known, the preexponential factors can be determined with good precision even at low total counts (10(4)). With unknown preexponential factors and lifetimes, precisions decrease but are still acceptable. A new gating scheme (overlapped gating) is shown to offer improved precision for the case of a single-exponential decay. Theoretical predictions are tested against actual experimental data from a laser-based lifetime instrument.     

ESA-based in-fiber nanocavity for hydrogen-peroxide detection

A fiber-optic sensor sensitive to hydrogen peroxide has been designed based on the electrostatic layer-by-layer self-assembly method. Prussian blue has been deposited in a polymeric structure formed by Poly(allylamine hydrochloride) and poly(acrylic acid). The concentration that can be detected range between 10/sup -6/-10/sup -3/ M, and recovery of the sensor after immersion into a reductive agent was demonstrated. The response of the sensor is independent of the pH for values that range between 4-7.4. Some rules for estimation of the refractive index of the material deposited and the thickness of the bilayers are also presented.     

Optimized gating scheme for rapid lifetime determinations of single-exponential luminescence lifetimes

The rapid lifetime method (RLD) for determining excited-state lifetimes uses the ratio of the areas under two regions of the decay. To get good precision with the standard method, prior knowledge of the lifetime is essential to selecting the integration regions. As will be shown, the usual method of selecting integration regions is far from optimal. An optimal gating scheme that is more precise and much more forgiving in the selection of integration region than any of the prior methods will be shown. Monte Carlo simulations were performed to determine the optimal gating. Experimental data was used to confirm the capabilities of the optimized RLD. The speed of the optimal RLD is similar to the standard RLD but without the necessity of matching the integration interval to the lifetime for precise results.     

Synthesis,characterisation and catalytic activity of gold and silver nanoparticles in the bio-sensor application

Most of the hydrogen peroxide (H₂O₂) bio-sensors developed till date are based on enzymes and proteins causing them to have a limited lifetime. Moreover, complex procedures are followed for sensor fabrication. Therefore, an inorganic material-based sensor, with a simple design and longer shelf life is highly desirable. In this work, surfactant-metal (gold and silver) nanoparticles are prepared in aqueous solutions containing cetyltrimethylammonium bromide. The particle sizes of the metal nanoparticles obtained are characterised by UV–Vis, HRTEM, X-ray diffraction and FTIR; the average sizes of gold and silver nanoparticles are 8 and 10?±?0.2?nm, respectively. The nanoparticles are tested for H₂O₂ detection. The sensor is characterised and tested using samples from M to mM H₂O₂ range and a linear response is observed. Low-detection limits and high sensitivity are some of the advantages of this work. Same principle could be extended for the detection of other substrates as well.     

Cyanide detection using a substrate-regenerating, peroxidase-based biosensor

An enzyme-based, dual working electrode system is described for the sensing of cyanide. Horseradish peroxidase (HRP) is incorporated as the sensing element. A continuous monitoring of oxidative activity by the enzyme results through the generation and regeneration of substrates at the electrode surfaces. Thus, HRP is oxidized by hydrogen peroxide generated from dissolved oxygen, at the primary electrode, and then reduced through the secondary electrode by mediated electron transfer using ferrocene as a carrier. Ferrocene regeneration at this electrode is proportional to the intrinsic activity of HRP. The dynamics of the system are investigated by using a rotating ring-disk electrode. The enzyme is immobilized to provide better control over its catalytic activity and to increase the lifetime of the biosensor. Cyanide inhibition of current can be modeled by reversible binding kinetics. Detection of cyanide is possible in submicromolar (ppb) concentrations, with a half maximal response at 2 microM. The response time for detection of introduced cyanide is within 1 s. The sensor can be operated between 5 and 40 degrees C, and cyanide inhibition is unaffected by pH changes between 5 and 8. The sensor is reproducible for cyanide determination and is stable for over 6 months.     

Calibration approach for fluorescence lifetime determination for applications using time-gated detection and finite pulse width excitation

Time-gated techniques are useful for the rapid sampling of excited-state (fluorescence) emission decays in the time domain. Gated detectors coupled with bright, economical, nanosecond-pulsed light sources like flashlamps and nitrogen lasers are an attractive combination for bioanalytical and biomedical applications. Here we present a calibration approach for lifetime determination that is noniterative and that does not assume a negligible instrument response function (i.e., a negligible excitation pulse width) as does most current rapid lifetime determination approaches. Analogous to a transducer-based sensor, signals from fluorophores of known lifetime (0.5-12 ns) serve as calibration references. A fast avalanche photodiode and a GHz-bandwidth digital oscilloscope is used to detect transient emission from reference samples excited using a nitrogen laser. We find that the normalized time-integrated emission signal is proportional to the lifetime, which can be determined with good reproducibility (typically <100 ps) even for data with poor signal-to-noise ratios ( approximately 20). Results are in good agreement with simulations. Additionally, a new time-gating scheme for fluorescence lifetime imaging applications is proposed. In conclusion, a calibration-based approach is a valuable analysis tool for the rapid determination of lifetime in applications using time-gated detection and finite pulse width excitation.     

Luminescence Lifetime–Based In Vivo Detection with Responsive Rare Earth–Dye Nanocomposite

For years, luminescence lifetime imaging has served as a quantitative tool in indicating intracellular components and activities. However, very few studies involve the in vivo study of animals, especially in vivo stimuli‐responsive activities of animals, as both excitation and emission wavelengths should fall into the near‐infrared (NIR) optical transparent window (660–950 and 1000–1500 nm). Herein, this work reports a lifetime‐responsive nanocomposite with both excitation and emission in the NIR I window (800 nm) and lifetime in the microsecond region. The incorporation of Tm³⁺‐doped rare‐earth nanocrystals and NIR dye builds an efficient energy transfer pathway that enables a tunable luminescence lifetime range. The NaYF₄:Tm nanocrystal, which absorbs and emits photons at the same energy level, is found to be 33 times brighter than optimized core–shell upconversion nanocrystals, and proved to be an effective donor for NIR luminescence resonance energy transfer (LRET). The anti‐interference capability of luminescence lifetime signals is further confirmed by luminescence and lifetime imaging. In vivo studies also verify the lifetime response upon stimulation generated in an arthritis mouse model. This work introduces an intriguing tool for luminescence lifetime–based sensing in the microsecond region.     

Four-dimensional multiphoton microscopy with time-correlated single-photon counting

We report on the implementation of fluorescence-lifetime imaging in multiphoton excitation microscopy that uses PC-compatible modules for time-correlated single-photon counting. Four-dimensional data stacks are produced with each pixel featuring fluorescence-decay curves that consist of as many as 4096 bins. Fluorescence lifetime(s) and their amplitude(s) are extracted by statistical methods at each pixel or in arbitrarily defined regions of interest. When employing an avalanche photodiode the width of the temporal response function is 420 ps. Although this response confines the temporal resolution to values greater than several hundreds of picoseconds, the lifetime precision is determined by the signal-to-noise ratio and can be in the range of tens of picosconds. Lifetime changes are visualized in pulsed-laser-deposited fluorescent layers as well as in cyan fluorescent proteins that transfer energy to yellow fluorescent proteins in live mammalian cells.     

Application of some recently synthesized 9, 10-anthraquinone derivatives as new class of ionophores responsive to lead (II) ion

Lead-selective solvent polymeric membrane electrodes, based on some recently synthesized 9, 10-anthraquinone derivatives, are described. The electrode exhibits a good Nernstian response for Pb (II) ions over a wide concentration range of 1.0/spl times/10/sup -6/-1.0/spl times/10/sup -2/ M with a slope of 28.9 mV decade/sup -1/. The potential-pH profile of membrane based on 1-hydroxy-2-({2-[2-(2-hydroxyethoxy)ethoxy]ethoxy}methyl)anthra-9, 10-quinone (A/sub 3/) demonstrated a lack of H/sup +/ interference within a wide pH range (1.5-6.8). The detection limit is 6.7/spl times/10/sup -7/ M. The developed sensor has a very short response time (2.0 s), and it can be used as a working electrode in a flow injection system. The lifetime of the proposed sensor is 120 days (without any considerable divergence in potentials) with good reproducibility (SD=/spl plusmn/0.1 mV). The proposed sensor revealed good selectivity for Pb (II) over a wide variety of other metal ions. It can be used as an indicator electrode in the potentiometric titration of lead ions, with EDTA, oxalate, chromate, and hydroxide ions, and in direct determination of lead in a wastewater sample.     

Real-time detection of bacterial contamination in dynamic aqueous environments using optical sensors

Here we describe on-line, real-time detection of waterborne bacteria using an optical sensor based on a starburst dendrimer film containing a lipophilic fluorophore. The sensor is constructed via covalent coupling between amine-terminated polyamidoamine dendrimer and silanized glass through an amide bond. The reporter molecule is embedded in the dendrimer layer through host-guest interaction. Real-time automated detection and quantitation of the bacteria are realized by using a charge-coupled detector camera and customized imaging and analysis software. The sensor responds to bacteria introduced to an aqueous flow system within 1 min. The limit of detection is approximately 10(4)cells/mL. The operational lifetime is more than 64 h, and the storage lifetime of the sensor is at least 7 months.     

Disposable biosensor based on a hemoglobin colloidal gold-modified screen-printed electrode for determination of hydrogen peroxide

A disposable reagentless hydrogen peroxide biosensor based on the direct electrochemistry of hemoglobin immobilized on a colloidal gold-modified screen-printed carbon electrode (Hb-Au-SPCE) was proposed. The electrochemical behavior of immobilized Hb at a SPCE was studied for the first time. The electrode reaction of immobilized Hb showed a surface-controlled process with an electron transfer rate constant of (0.40 /spl plusmn/ 0.02) s/sup -1/ determined in the scan rate range from 25 to 200 mV s/sup -1/. The Hb-Au-SPCE exhibited an electrocatalytic activity toward the reduction of hydrogen peroxide with a K/sub M//sup app/ value of 1.8 mM, which was allowed to be used as a disposable sensor for determination of hydrogen peroxide with a linear range from 1.0 /spl times/ 10/sup -5/ M to 3.2 /spl times/ 10/sup -4/ M, a detection limit of 5.5 /spl times/ 10/sup -6/ M at 3/spl sigma/, a high sensitivity, fast response, and good selectivity, accuracy, and reproducibility. The disposable reagentless sensor was stable, low cost, and simple to use for detection of hydrogen peroxide in real samples.     

Improved electrodeposited iridium oxide pH sensor fabricated on etched titanium substrates

In the present paper, the preparation and characterization of an improved solid-state pH sensor are described. The sensor is based on anodically electrodeposited iridium oxide film, as a pH-sensing layer. Merits of the present sensor include (i) excellent adhesion of the pH sensitive layer to the substrate, (ii) excellent reproducibility of sensor fabrication, (iii) faster preparation procedure, and (iv) low cost of the titanium substrate. These advantages are realized by combining acid-etched titanium as the electrode substrate with an optimized electrodeposition solution consisting of IrCl4 as an iridium source, hydrogen peroxide, potassium oxalate, and potassium carbonate. Heating the electrodeposition solution to 90 degrees C reduced the time required for solution development from approximately 3 days to 10 min. The pH-sensing layer is protected with a layer of Nafion and a microporous polyester membrane. The improved sensor showed a super-Nernstian response (-73.7 +/- 1.2 mV/pH unit) in the pH range of 1.5-11.5. The present pH sensor, fabricated in a tubular form, is used as a detector in a flow injection analysis (FIA) system for pH measurements. Optimization of the FIA experimental parameters resulted in a linear dependence of peak heights on the pH of the injected samples in the pH range of 2-11.     

Temperature‐Dependent Exciton and Trap‐Related Photoluminescence of CdTe Quantum Dots Embedded in a NaCl Matrix: Implication in Thermometry

Temperature‐dependent optical studies of semiconductor quantum dots (QDs) are fundamentally important for a variety of sensing and imaging applications. The steady‐state and time‐resolved photoluminescence properties of CdTe QDs in the size range from 2.3 to 3.1 nm embedded into a protective matrix of NaCl are studied as a function of temperature from 80 to 360 K. The temperature coefficient is found to be strongly dependent on QD size, with the highest sensitivity obtained for the smallest size of QDs. The emission from solid‐state CdTe QD‐based powders is maintained with high color purity over a wide range of temperatures. Photoluminescence lifetime data suggest that temperature dependence of the intrinsic radiative lifetime in CdTe QDs is rather weak, and it is mostly the temperature‐dependent nonradiative decay of CdTe QDs which is responsible for the thermal quenching of photoluminescence intensity. By virtue of the temperature‐dependent photoluminescence behavior, high color purity, photostability, and high photoluminescence quantum yield (26%–37% in the solid state), CdTe QDs embedded in NaCl matrices are useful solid‐state probes for thermal imaging and sensing over a wide range of temperatures within a number of detection schemes and outstanding sensitivity, such as luminescence thermochromic imaging, ratiometric luminescence, and luminescence lifetime thermal sensing.     

Amperometric sensors for simultaneous superoxide and hydrogen peroxide detection

A two-channel sensor capable of almost instantaneous simultaneous detection of superoxide radical and hydrogen peroxide in the concentration range 10(-)(7)-10(-)(4) M is very important for understanding of a number of rapid kinetics processes. A glassy carbon working microelectrode covered by an electrodeposited polypyrrole/horseradish peroxidase (PPy/HRP) membrane was employed as a H(2)O(2) sensor. Another glassy carbon microelectrode covered by a composite membrane of an inside layer of PPy/HRP and an outside layer of superoxide dismutase was employed as a working electrode for superoxide detection. These two working electrodes with Pt counter and tungsten oxide (WO(3)) reference electrodes were contained in one 6 mm diameter Teflon cylinder. Simultaneous measurements were performed at a potential of -60 mV (vs WO(3) reference, pH 5.1). Additional sensor characterization was performed for pH 5.1-9.0. Superoxide sensor behavior as a function of membrane deposition conditions and coating time is reported. Sensors' mutual influence, selectivity, response times, linearity, stability, and sensitivity for hydrogen peroxide and superoxide are presented and discussed. A mathematical model of sensors' responses is proposed, with model calculation corresponding to experiment within 10%.     

In vivo imaging of hydrogen peroxide with chemiluminescent nanoparticles

The overproduction of hydrogen peroxide is implicated in the development of numerous diseases and there is currently great interest in developing contrast agents that can image hydrogen peroxide in vivo. In this report, we demonstrate that nanoparticles formulated from peroxalate esters and fluorescent dyes can image hydrogen peroxide in vivo with high specificity and sensitivity. The peroxalate nanoparticles image hydrogen peroxide by undergoing a three-component chemiluminescent reaction between hydrogen peroxide, peroxalate esters and fluorescent dyes. The peroxalate nanoparticles have several attractive properties for in vivo imaging, such as tunable wavelength emission (460-630 nm), nanomolar sensitivity for hydrogen peroxide and excellent specificity for hydrogen peroxide over other reactive oxygen species. The peroxalate nanoparticles were capable of imaging hydrogen peroxide in the peritoneal cavity of mice during a lipopolysaccharide-induced inflammatory response. We anticipate numerous applications of peroxalate nanoparticles for in vivo imaging of hydrogen peroxide, given their high specificity and sensitivity and deep-tissue-imaging capability.     

Effect of interfacial reaction rate on the morphogenesis of nanostructured coatings in a simulated electrodeposition process

Brownian dynamics simulations (BDSs) are performed to investigate the influence of interfacial electrochemical reaction rate on the evolution of coating morphology on circular fibres. The boundary condition for the fluid phase concentration, representing the balance between the rates of interfacial reaction and transport of ions by bulk diffusion, is incorporated into the BDS by using a reaction probability, P(s). Different modes of growth, ranging from diffusion limited ([Formula: see text]) to reaction controlled [Formula: see text], are studied. It is found that, consistent with experimental observations, two distinct morphological regimes exist, with a dense and uniform structure for [Formula: see text] (reaction limited deposition (RLD)) and an open and porous one as [Formula: see text] (diffusion limited deposition (DLD)). An analysis of the fractal dimension indicates that this morphological transition occurs at P(s)≈0.3. Long-time power-law scalings for the evolution of thickness [Formula: see text] and roughness (ξ) of the coating exist, i.e.?[Formula: see text] with 0.86≤α≤0.91 and 0.56≤β≤0.93 for 0.01≤P(s)≤1. These values are different from those reported for sequential, pseudo-time lattice simulations on planar surfaces, signifying the importance of multiparticle dynamics and surface curvature. The internal structure and porosity of the coating are characterized quantitatively by the radial density profile, pair correlation function, two-point probability function, void distribution function and pore area distribution. For RLD the radial density, ρ(n), remains nearly constant, while for DLD ρ(n) follows a power law, [Formula: see text]. The coating exhibits short ranged order in the RLD regime while a long range order is created by DLD. The void distribution function becomes broader with increasing P(s), indicating that in the RLD regime the coating consists of small and spherical pores, while in the DLD regime large and elongated pores are obtained. The pore area distribution shows narrower distributions in DLD for small pores, while the area of the largest pore increases by nearly three orders of magnitude as one moves from the RLD to the DLD regime. Such morphological diversity could be potentially exploited for applications such as percolation, catalysis and surface protection.     

Controlling the color space response of colorimetric luminescent oxygen sensors

The color space response of colorimetric luminescent oxygen sensors is described in terms of the Commission Internationale de l'Eclairage (CIE) x,y color coordinates. We show how the color change response to oxygen can be obtained, provided the quenching kinetics for all lumophores involved can be represented mathematically. The theory is illustrated by analysis of examples of theoretical sensors in which lumophores are quenched by Stern-Volmer kinetics to give red to green, green to red, and red to green to blue color responses as the partial pressure of oxygen is increased. The effects of lumophore emission lifetime and the permeability of the polymer matrix to oxygen are discussed in terms of the control of sensor response, which variations in these parameters offer.     

Evaluation of acridine in Nafion as a fluorescence-lifetime-based pH sensor

We report a novel fluorescence-lifetime-based pH sensing method that utilizes acridine incorporated into Nafion (AcNaf) as the fluorescent indicator. The AcNaf sensor is excited using a 380 nm light emitting diode (LED) and the fluorescence lifetimes are measured at 450 and 500 nm. The fluorescence behavior of acridine as a function of pH in aqueous phosphate buffers and incorporated into the Nafion membrane has been investigated. The results show that incorporating acridine into Nafion changes the apparent ground-state pKa from -5.45 to -9, while the apparent excited-state pKa* is only slightly changed (approximately 9.4 in 0.1 M phosphate buffer). The AcNaf film shows a good pH response with a change in average lifetime of approximately 19 ns (at an emission wavelength of 450 nm) over the pH 8 to 10 range. We also show that excited-state protonation does not occur in the AcNaf sensor film and that chloride quenching cannot occur because of the permselective nature of Nafion. We also discuss how the unique structure of Nafion affects the fluorescence behavior of acridine at various pH values and examine the impact of buffer concentration on apparent pKa and pH sensing ability.