Method for simultaneous luminescence sensing of two species using optical probes of different decay time, and its application to an enzymatic reaction at varying temperature

Chemical sensing, imaging and microscopy based on the use of fluorescent probes has so far been limited almost exclusively to the detection of a single parameter at a time. We present a scheme that can overcome this limitation by enabling optical sensing of two parameter simultaneously and even at identical excitation and emission wavelengths of two probes provided (a) their decay times are different enough to enable two time windows to be recorded, and (b) the emission of the shorter-lived probe decays to below the detectable limit while that of the other still can be measured. We refer to this new scheme as the dual lifetime determination (DLD) method and show that it can be widely varied by appropriate choice of probes and experimental settings. DLD is demonstrated to work by sensing oxygen and temperature independently from each other by making use of two probes, one for oxygen (a platinum porphyrin dissolved in polystyrene), and one for temperature [a europium complex dissolved in poly(vinyl methylketone)]. DLD was applied to monitor the consumption of oxygen in the glucose oxidase-catalyzed oxidation of glucose at varying temperatures. The scheme is expected to have further applications in cellular assays and biophysical imaging. Figure Principle behind the dual lifetime determination (DLD) method     

Dual sensing of oxygen and temperature using quantum dots and a ruthenium complex

A scheme for the simultaneous determination of oxygen and temperature using quantum dots and a ruthenium complex is demonstrated. The luminescent complex [Ru(II)-tris(4,7-diphenyl-1,10-phenanthroline)]²⁺ is immobilized in a non-hydrolytic sol-gel matrix and used as the oxygen sensor. The temperature information is provided by the luminescent emission of core-shell CdSe-ZnS semiconductor nanocrystals immobilized in the same material. Measurements of oxygen and temperature could be performed with associated errors of ±2% of oxygen concentration and ±1 °C, respectively. In addition, it is shown that while the dye luminescence intensity is quenched both by oxygen and temperature, the nanocrystals luminescent emission responds only to temperature. Results presented demonstrate that the combined luminescence response allows the simultaneous assessment of both parameters using a single optical fiber system. In particular, it was shown that a 10% error in the measured oxygen concentration, induced by a change in the sample temperature, could be compensated using the nanocrystals temperature information and a correction function.     

An optical sensor for mercury ion based on the fluorescence quenching of tetra(p-dimethylaminophenyl)porphyrin

An optical sensor for mercury ion (Hg²⁺), based on quenching the fluorescence of the sensing reagent porphyrin immobilized in plasticized poly(vinyl chloride) (PVC) membrane, has been developed. The responses to mercury ion were compared for the sensors modified with three porphyrin compounds including 5,10,15,20-tetraphenylporphyrin (TPP), tetra(p-dimethylaminophenyl)porphyrin (TDMAPP) and tetra(N-phenylpyrazole) porphyrin (TPPP). Among them, TDMAPP showed the most remarkable response to Hg²⁺. The drastic decrease of the TDMAPP fluorescence intensity was attributed to the formation of a complex between TDMAPP and Hg²⁺, which has been utilized as the fabrication basis of a Hg²⁺-sensitive fluorescence sensor. The analytical performance characteristics of the TDMAPP modified sensor was investigated. The response mechanism, especially involving the response difference of three porphyrin compounds, was discussed in detail. The sensor can be applied to the quantification of Hg²⁺ with a linear range covering from 4.0 × 10⁻⁸ mol L⁻¹ to 4.0 × 10⁻⁶ mol L⁻¹. The limit of detection was 8.0 × 10⁻⁹ mol L⁻¹. The sensor exhibited excellent reproducibility, reversibility and selectivity. Also, the TDMAPP-based sensor was successfully used for the determination of Hg²⁺ in environmental water samples.     

Optimising oxygen-sensitivity of optical sensor using pyrene carboxylic acid by myristic acid co-chemisorption onto anodic oxidized aluminium plate

Optical oxygen-sensitivity using pyrene carboxylic acid with long alkyl chain (1-pyrenedecanoic acid and 1-pyrenedodecanoic acid) and myristic acid co-chemisorption layer was controlled by varying the molar ratio of myristic acid to pyrene carboxylic acid. The ratio I₀/I₁₀₀, where I₀ and I₁₀₀ represent the detected fluorescence intensities from a substrate exposed to 100% argon and 100% oxygen, respectively, is used as an indicator of the sensitivity of the sensing film. At a composition ratio of 1 pyrene carboxylic acid to 10 myristic acids, the I₀/I₁₀₀ attained its maximum value and then the ratio decreased with increase in the molar ratio of myristic acid to pyrene carboxylic acid. The Stern-Volmer constant (K_SV) also attained its maximum value at a composition ratio of one pyrene carboxylic acid to ten myristic acids and then the ratio decreased with increase in the molar ratio of myristic acid to pyrene carboxylic acid. The oxygen-sensitivity of optical sensor using pyrene carboxylic acid is optimized by myristic acid co-chemisorption.     

A selective optical chemical sensor for 2,6-dinitrophenol based on fluorescence quenching of a novel functional polymer

A bifurcated optical fiber based chemical sensor for continuous monitoring of 2,6-dinitrophenol (2,6-DNP) has been proposed based on the reversible chemical reaction between a novel functional poly(vinyl chloride) (PVC) as the sensing material and the analytes. The functional PVC (FPVC), containing a fluorescent curcumin moiety, was synthesized by the nucleophilic substitution of a fraction of the chlorine atoms bound to the PVC backbone by curcumin. When plasticized in a membrane of 5 μm thickness, FPVC extracts 2,6-DNP from aqueous solution into the bulk membrane phase and reacts with the analyte to form a complex with low fluorescence efficiency through hydrogen bonding. Formation of the complex gave a significant fluorescence quenching which is suitable for signalling the occurrence of the host-guest interaction. At pH 3.50, the sensor exhibits a dynamic detection range from 2.5 × 10⁻⁶ to 7.0 × 10⁻³ mol L⁻¹ with a limit of detection of 1.0 × 10⁻⁶ mol L⁻¹. As 2,6-DNP can provide an optimal space geometry matches to the formation of hydrogen bonds, the sensor shows excellent selectivity for 2,6-DNP over other nitrophenols. The forward and reverse response time (t₉₅) of the sensor both was within 1 min. The repeatability, reproducibility, and lifetime of the sensor were also satisfied. The sensor was applied to determine 2,6-DNP in water samples successfully.     

Novel oxygen sensitive complexes for optical oxygen sensing

Novel optical sensing films for oxygen based on highly luminescent iridium (III) and ruthenium (II) complexes have been developed. These demonstrate excellent long-term photostability (several months) when incorporated into polystyrene membranes. The influence of different plasticizers on the specific luminescence quantum yield, the Stern-Volmer constant, the reversibility and the response time were evaluated. Additionally the sensing films can be sterilized by chemical cleaning and gamma-ray irradiation.     

Optical oxygen sensor based on fluorescence change of pyrene-1-butyric acid chemisorption film on an anodic oxidation aluminium plate

An optical oxygen-sensing material based on the fluorescence intensity changes of pyrene-1-butyric acid (PBA) chemisorption film has been developed and characterised. The fluorescence intensity of PBA film decreased with increase of oxygen concentration. The I₀/I₁₀₀ value of PBA film is estimated to be 6.14±0.15 and large Stern-Volmer constant (K_SV=0.028±0.13 Torr⁻¹) is obtained. After irradiation for 24 h with 150 W tungsten lamp, little changes of oxygen-sensing properties were observed. These results indicate that PBA film is highly oxygen-sensitive and photostability device. The response times of the PBA chemisorption film were 10.0 s for switching from argon to oxygen, and 53.0 s for switching from oxygen to argon. Moreover, the optical sensor based on the PBA chemisorption film was applied to the measurement of oxygen concentration in aqueous solution.     

Photocured thiol-ene based optical fluorescence sensor for determination of gold(III)

This study describes the preparation and the characterization of a new thiol-ene based polymeric fluorescence sensor by photo initiated polymerization of trimethylolpropane tris(3-mercaptopropionate), 2-hydroxyethylacrylate, and 2,4,6-triallyloxy-1,3,5-triazine which are used as monomers and also a photo initiator (2,2-dimethoxy-2-phenylacetophenone) for its usage as optical sensor for gold ions. The thiol-ene based polymeric membrane sensor was characterized by using attenuated total reflectance-fourier transform infrared spectroscopy (ATR-FTIR) and scanning electron microscopy (SEM). The response characteristics of the sensors including dynamic range, pH effect, response time, and the effect of foreign ions were investigated. Fluorescence spectra showed that the excitation/emission maxima of the membrane were at 379/425 nm, respectively     

茜素红S-硼酸体系对单糖的双光谱识别

利用硼酸与茜素红S和糖中的邻二羟基可逆结合的特点,以硼酸为中介运用竞争结合作用机理构建单糖分析法.在pH7.4的KH2PO4-NaOH缓冲溶液中,茜素红S作为指示剂与硼酸结合生成ARS-BA配合物,其结合常数为5.09×102L/mol.糖与指示剂ARS竞争结合硼酸使指示剂游离出来,产生明显的颜色变化,据此建立糖的识别方法.考察了D-葡萄糖、D-山梨醇、D-半乳糖、D-甘露糖、D-果糖、D-阿拉伯糖和L-阿拉伯糖对上述ARS-BA体系光谱的影响.结果显示:该体系对D-山梨醇和D-果糖有较好的光谱响应,其光谱变化灵敏度依D-山梨醇＞D-果糖＞D-阿拉伯糖～D-半乳糖＞D-葡萄糖＞D-甘露糖＞L-阿拉伯糖之序.     

Sol-gel-based optical sensor for the detection of aqueous amines

We present an optical sensor for the detection of aqueous amines obtained by incorporating chromoionophore XV (ETH^T 4001) into sol-gel thin films. Acid- and base-catalyzed sol-gel processes were studied to prepare stable ormosil layers using various amounts of organically modified sol-gel precursor such as methyltriethoxysilane (MTriEOS). The sensor layers were coated with a protective layer of microporous white polytetrafluoroethylene (PTFE) in order to prevent interference from ions and ambient light. The measurements were carried out in a flow-through cell in the reflection mode. Acid-catalyzed ormosil layers (pH 1) based on the copolymerization of tetraethoxysilane (TEOS) and MTriEOS did not show any change in signal upon exposure to aqueous amine solutions, while base-catalyzed sensor layers (pH 3 and 13) showed significant changes in signal. The response time (t ₁₀₀) for the base-catalyzed sensor layer L3 (pH 13) upon exposure to different solutions containing 0–608 mmol L⁻¹ aqueous propylamine was 20–30 s, the regeneration time was 70 s and the detection limit was 0.1 mmol L⁻¹. The sensor response was reproducible and reversible. The porous ormosil layers permit dry sensor storage conditions.     

An optical pH sensor with extended detection range based on fluoresceinamine covalently bound to sol–gel support

An optical pH sensor was developed based on the fluorophor, fluoresceinamine isomer II (FA), covalently immobilized in a sol–gel matrix. This sol–gel matrix was created by the copolymerization of two precursors, methyltriethoxysilane (MTES) and 3-glycidoxypropyltrimethoxysilane (GPTMS), in an ethanolic solution. Fluoresceinamine was covalently bound to the glycidoxypropyl chain group of GPTMS, thereby preventing it from leaching. Moreover, the immobilization of the fluoresceinamine also extended its linear detection range. The sensor showed good repeatability, a short response time of less than 8 s, high long-term stability and no temperature effect in the biologically relevant range. In the pH range of 4–10, the sensor was very sensitive and its linear range was found to be between pH 6 and 9 (R² = 0.995).     

Design of an optical sensor for indirect determination of isoniazid

The characterization of an optical sensor membrane is described for indirect determination of isoniazid. The sensing membrane was consisted of immobilized 3-(2-pyridyl)-5,6-diphenyl-1,2,4-triazine (PDT) on a triacetylcellulose membrane. The procedure is based on the reaction of Fe(III) with isoniazid in the presence of PDT. Fe(III) is reduced by isoniazid to Fe(II) which forms a complex with PDT. The complex shows an absorption maximum at 558nm. By measuring the absorbance of the complex at this wavelength, isoniazid can be determined in the range of 0.62-6.15mugmL(-1). This method was applied to the determination of isoniazid in pharmaceutical formulation and enabled the determination of isoniazid in microgram quantities.     

A selective optical sensor for picric acid assay based on photopolymerization of 3-(N-methacryloyl) amino-9-ethylcarbazole

A novel optical sensor based on covalent immobilization for picric acid assay has been described. To improve the stability of the sensor, a terminal double bond was attached to the fluorescent compound, 3-amino-9-ethylcarbazole (AEC), via methacryloyl chloride. The resultant compound, 3-(N-methacryloyl) amino-9-ethylcarbazole (MAEC) was copolymerized with 2-hydroxypropyl methacrylate on surface-modified quartz glass plates by UV irradiation. The resulting optical sensor (optode membrane) was used to determine picric acid based on fluorescence quenching. It shows a linear response toward picric acid in the concentration range of 9.33 × 10⁻⁸ to 9.33 × 10⁻⁵ mol l⁻¹, with rapid response, high stability and good selectivity to picric acid.     

试纸-光反射传感仪快速测定痕量甲醛的研究

在KOH溶液中，利用甲醛与4-氨基-3-联氨-5-巯基-1、2、4-三氮杂茂(AHMT)的缩合，并经KIO4氧化后的显色反应原理制备出检测甲醛的试纸条，并将该检测试纸与研制光反射传感仪联用，实现了对海产品及其浸泡液中痕量甲醛的现场、快速、定量检测。该方法的检出限为0．4μg/mL，对鱿鱼、海参以及水样中甲醛进行测定，其RSD均小于5％，与国家标准方法进行了对照，结果一致。该法用于甲醛的检测可实现对痕量甲醛进行现场快速定量检测，值得推广应用。     

Poly(norbornene)s as matrix materials for platinum tetrakis(pentafluorophenyl)porphyrin based optical oxygen sensors

A set of poly(norbornenes) was prepared using ring opening metathesis polymerization (ROMP) and used as matrix material for the preparation of optical oxygen sensor layers based on platinum tetrakis(pentafluorophenyl)porphyrin (PtTFPP) as the sensitive dye. Different polymers were prepared and investigated in order to retrieve information on the influence of the anchor group and the side chain attached to the polymer backbone on their performance as matrix material for the dye. Bulky side groups increased the oxygen permeability through ROM polymer layers, especially when the bulky group was directly attached via an anchor group to the polymer backbone without any aliphatic spacer in between. Sensor layers made of poly(endo,exo[2.2.1]bicyclo-5-heptene-2,3-dicarboxyclic acid di-tert-butylester) and PtTFPP exhibited the highest τ₀/τ ratio and responded strongly to small amounts of oxygen.     

Dual electrochemical microsensor for simultaneous measurements of nitric oxide and oxygen: Fabrication and characterization

A planar-type amperometric dual microsensor was developed for the simultaneous measurement of the nitric oxide (NO) and oxygen (O₂) concentrations. The sensor (overall diameter = 500 μm) consisted of a dual working electrode (WE) containing two platinized platinum microdisks (25 μm diameter, WE1, WE2, distance between two disks > 330 μm) and a Ag/AgCl wire reference electrode covered with an expanded poly(tetrafluoroethylene) gas-permeable membrane. The differentiation and concurrent measurements of NO and O₂ were obtained successfully using two sensing WEs with different applied potentials (+0.75 V for WE1 and −0.4 V for WE2). Cross-talk between WE1 and WE2 was eliminated with an optimized internal solution composition. Linear dynamic range, selectivity, sensitivity, detection limit (<5 nM for NO; <500 nM for O₂), and stability (>50 h) were evaluated.     

Bifunctional sensor of nitric oxide and oxygen based on hematite nanotubes embedded in chitosan matrix

A novel hybrid bifunctional sensing platform for simultaneous determination of NO and O₂ has been developed, whereby hematite nanotubes are immobilized into the chitosan matrix onto a gold electrode (labeled as HeNTs-Chi/Au). The HeNTs distributed in porous-structured chitosan matrix not only offer abundant active sites for bifunctional sensing of NO and O₂, but also facilitate oxidation of NO and reduction of O₂ dramatically. Straight calibration curves are achieved in analyte concentration ranges of 5.0 × 10⁻⁸ to 1.25 × 10⁻⁶ mol L⁻¹ for NO and 2.5 × 10⁻⁷ to 6.0 × 10⁻⁶ mol L⁻¹ for O₂. Also, the detection limits are low of 8.0 × 10⁻⁹ mol L⁻¹ for NO and 5.0 × 10⁻⁸ mol L⁻¹ for O₂. Such an efficient bifunctional sensor for NO and O₂ offers great potential in quantitation of NO levels in biological and medical systems, since NO level is highly regulated by various reactive oxygen species.     

A new optical platform for biosensing based on fluorescence anisotropy

A novel fluorescence-based optical platform for the interrogation of an optical biochip was designed and developed. The optical biochip was made of poly(methyl methacrylate) (PMMA) formed by two pieces of PMMA appropriately shaped in order to obtain four microchannels that are 500-μm wide and 400-μm high. The lower part includes the microchannels and the inlet and outlet for the fluidics, while the sensing biolayer was immobilized on the upper part. The optical signal comprised the fluorescence emitted by the biolayer, which was anisotropically coupled to the PMMA cover and suitably guided by the PMMA chip. The potentiality of the optical chip as a biosensor was investigated by means of a direct IgG/anti-IgG interaction carried out inside the flow channels. The mouse-IgG was covalently immobilized on the internal wall of the PMMA cover, and the Cy5-labelled anti-mouse IgG was used for the specific interaction. Several chemical treatments of the PMMA surface were investigated, poly(L-lactic acid), Eudragit L100 and NaOH, in order to obtain the most effective distribution of carboxylic groups useful for the covalent immobilisation of the mouse-IgG. The treatment with Eudragit L100 was found to be the most successful. Limits of detection and quantification of 0.05 μg mL⁻¹ and 0.2 μg mL⁻¹, respectively, were obtained with the configuration described.     

Design and characterization of a Li-selective optical sensor

An optical ion-sensing system based on a PVC membrane containing a Li⁺ ionophore in combination with a H⁺ selective chromoionophore (ETH 5294) is presented. It exhibits an absorbance response to Li⁺ concentration in pH buffered solutions. The dynamic range and Li⁺/Na⁺ selectivity of this new optode system are discussed.     

An optical chemical sensor for mercury ion based on 2-mercaptopyrimidine in PVC membrane

An optical chemical sensor based on 2-mercaptopyrimidine (2-MP) in plasticized poly(vinyl chloride) (PVC) membrane incorporating (N,N-diethyl-5-(octadecanoylimino)-5H benzo[a]phenoxazine-9-amine (ETH 5294) and sodium tetraphenyl borate (NaTPB) for batch and flow-through determination of mercury ion is described. The response of the sensor is based on selective complexation of Hg²⁺ with 2-MP in the membrane phase, resulting in an ion exchange process between H⁺ in the membrane and Hg²⁺ in the sample solution. The influences of several experimental parameters, such as membrane composition, pH, and type and concentration of the regenerating reagent, were investigated. The sensor has a response range of 2.0 × 10⁻⁹ to 2.0 × 10⁻⁵ mol L⁻¹ Hg²⁺ with a detection limit of 4.0 × 10⁻¹⁰ mol L⁻¹ and a response time of ≤45 s at optimum pH of 6.5 with high measurement repeatability and sensor-to-sensor reproducibility. It shows high selectivity for Hg²⁺ over several transition metal ions, including Ag⁺, Cd²⁺, Co²⁺, Cr³⁺, Cu²⁺, Fe³⁺, Mn²⁺, Ni²⁺, and common alkali and alkaline earth ions such as Na⁺, K⁺, Mg²⁺, Ca²⁺, and Pb²⁺. The sensor membrane can be easily regenerated with dilute acid solutions. The sensor has been used for the determination of mercury ion concentration in water samples.