Fluorescent Sensor Molecules Research Articles

Fluorescence-Based Portable Assays for Detection of Biological and Chemical Analytes.

Fluorescence-based detection techniques are part of an ever-expanding field and are widely used in biomedical and environmental research as a biosensing tool. These techniques have high sensitivity, selectivity, and a short response time, making them a valuable tool for developing bio-chemical assays. The endpoint of these assays is defined by changes in fluorescence signal, in terms of its intensity, lifetime, and/or shift in spectrum, which is monitored using readout devices such as microscopes, fluorometers, and cytometers. However, these devices are often bulky, expensive, and require supervision to operate, which makes them inaccessible in resource-limited settings. To address these issues, significant effort has been directed towards integrating fluorescence-based assays into miniature platforms based on papers, hydrogels, and microfluidic devices, and to couple these assays with portable readout devices like smartphones and wearable optical sensors, thereby enabling point-of-care detection of bio-chemical analytes. This review highlights some of the recently developed portable fluorescence-based assays by discussing the design of fluorescent sensor molecules, their sensing strategy, and the fabrication of point-of-care devices.

Expedite Fluorescent Sensor Prototype for Hydrogen Peroxide Detection with Long-Life Test Substrates.

We report a practical fluorescent sensor device for the trace amount detection of hydrogen peroxide vapor. In this paper, we have significantly improved the performance of fluorescence analysis for the detection of peroxides by solving the problems of packaging and storage of active materials and transferring the chemical experiment phenomenon to the actual project output. The fluorescent sensor molecule, test substrates, mixing methods, and the way to improve the life time are carefully studied. Combined with the design of circuit and programming, a field-test prototype was designed for peroxide explosives and its performance and algorithm were screened and optimized. In the detection of traces of H2O2 generated by ultraviolet separation or leaked as inherent impurities, the high-efficiency and rapid detection of peroxide-based explosives is achieved. The detection limit of H2O2 is expected to reach 2 ppb, and the response time can reach <0.5 s.

Silk: A bio-derived coating for optical fiber sensing applications

Optical fiber chemical sensing is generally achieved through attachment of sensor molecules to the fiber, a process that involves chemicals that are not biologically compatible or are limited to thin monolayer coatings. To address these limitations and enable in-vivo biosensing, we report here, for the first time, silk fibroin coating of optical fibers for encapsulating fluorescent sensor molecules. Silica exposed core fiber (ECF) samples were coated with a thin layer of silk – a naturally derived biopolymer composed entirely of proteins and amino acids. The silk was doped with the fluorophore 5,6-carboxynapthofluorescein (CNF), which allows optical measurement of pH by a robust ratiometric fluorescence method. The fluorescent signal from the doped-silk layer is coupled into the core of the ECF, enabling remote measurement of pH along the length of the fiber. We have demonstrated real time in vivo pH sensing measurements in a mouse model of hypoxia. Our results showed a continuous drop in the subcutaneous pH in the mouse lumbar area as hypoxia developed. The work explores, for the first time, the potential of a natural silk protein coating to perform fiber sensing inside the body.

Mechanical Activation of Terpyridine Metal Complexes in Polymers

The mechanical addressability of specific chemical bonds holds a high potential for the improvement of polymeric materials. While in many cases, mechanical forces applied to polymers lead to bond scissoring and materials failure, including mechanophores into the polymer structure can lead to stimuli-responsive materials reacting to an applied force in a predefined manner. In this contribution, the mechanical addressability of bis-terpyridine metal complexes embedded into a polymer structure is investigated. The activation of the transition metal complexes in the metallopolymer is monitored by adding a fluorescent sensor molecule to the metallopolymer solution during ultrasonication. Upon sonication, the activation of the complexes leads to fluorescence-quenching of the sensor. The dependency of the metal ion and the type of polymer as well as their molar mass is investigated in detail, showing that this concept could possibly be used in further application of stimuli-responsive or self-healing materials.

Optically active crown ether-based fluorescent sensor molecules: A mini-review.

This mini-review focuses on fluorescent optically active crown ethers (polymeric derivatives are not included) reported in the literature (according to our knowledge), of which enantiomeric recognition ability, and in some cases, also inorganic cation complexation properties, were investigated by the sensitive and versatile fluorescence spectroscopy. These crown ether-based chemosensors contain various fluorophore signaling units such as binaphthyl, anthracene, pyrene, tryptophan, benzimidazole, terpyridine, acridine, phenazine, acridone, BODIPY, and another conjugated aromatic one.

Synthesis and potential use of 1,8-naphthalimide type 1O2 sensor molecules

New double (fluorescent and spin) sensor molecules containing 4-amino substituted 1,8-naphthalimide as a fluorophore and a sterically hindered amine (pre-nitroxide) or pyrroline nitroxide as a quencher and radical capturing moiety were synthesized. All sensors were substituted with a diethylaminoethyl side-chain to increase the water solubility. Steady state fluorescence properties of these compounds and their responses to ROS in vitro are reported with perspectives of plant physiology use in vivo.

Ternary complex formation and competition quench fluorescence of ZnAF family zinc sensors

Our current understanding of the intracellular thermodynamics and kinetics of Zn(ii) ions is largely based on the application of fluorescent sensor molecules, used to study and visualize the concentration, distribution and transport of Zn(ii) ions in real time. Such agents are designed for high selectivity for zinc in respect to other biological metal ions. However, the issue of their sensitivity to physiological levels of low molecular weight Zn(ii) ligands (LMWLs) has not been addressed. We followed the effects of eight such compounds on the fluorescence of ZnAF-1 and ZnAF-2F, two representatives of the ZnAF family of fluorescein-based zinc sensors containing the N,N-bis(2-pyridylmethyl)ethylenediamine chelating unit. Fluorescence titrations of equimolar Zn(ii)-ZnAF-1 and Zn(ii)-ZnAF-2F solutions with acetate, phosphate, citrate, glycine, glutamic acid, histidine, ATP and GSH demonstrated strong fluorescence quenching. These results are interpreted in terms of an interplay of the formation of the [ZnAF-Zn(ii)-LMWL] ternary complexes and the competition for Zn(ii) between ZnAF and LMWLs. UV-vis spectroscopic titrations revealed the existence of supramolecular interactions between the fluorescein moiety of ZnAF-1 and ATP and His, which, however, did not contribute to fluorescence quenching. Therefore, the obtained results show that the ZnAF sensors, other currently used zinc sensors containing the N,N-bis(2-pyridylmethyl)ethylenediamine unit, and, in general, all sensors that do not saturate the Zn(ii) coordination sphere may co-report cellular metabolites and Zn(ii) ions, leading to misrepresentations of the concentrations and fluxes of biological zinc.

Mercury detection in a microfluidic device by using a molecular sensor soluble in organoaqueous solvent

A series of fluorescent sensor molecules based on a phosphane sulfide derivative that is soluble in an organoaqueous solvent were designed and synthesized. The structure of the fluorophore has been optimized in order to have the best compromise in terms of solubility and photophysical properties. The obtained properties are in full agreement with quantum chemical calculations. A fluorescent molecular sensor containing one polyoxoethylene group has been synthesized and an efficient quenching upon mercury complexation has been observed. Finally, this sensing molecule has been introduced in a microfluidic chip in which fluorescence detection has been integrated. An efficient fluorescence response was observed upon mercury addition.

Fluorescence turn-on of easily prepared fluorescein derivatives by zinc cation in water and living cells

By combining a sulfonated β-naphtol as receptor, a hydrophilic fluorescence probe with extreme sensitivity (7 ppb) and selectivity for zinc ion has been designed and synthesized in a simple and efficient way. Based on the inhibition of internal charge transfer (ICT) process, This “turn-on” type fluorescent sensor molecule features high fluorescence increase (13 fold) at 508 nm as well as a red-shift from 371 to 430 nm over most competitive heavy and transition metal (HTM) ions in water (pH 7.4). Furthermore, cell imaging experiments show that this fluorescence probe might be used for monitoring Zn 2+ within biological samples. The role of this sensor molecule in living cells was further confirmed with the treatment of N, N, N′, N′- Tetrakis( 2-pyridylmethyl) ethylenediamine. And the impact of water-solubility and liposolubility of such fluorescence probe on biological application has been deeply discussed.

Adsorption and molecular dynamics of spin probes on hydrophobizated silica gel microparticles: EPR spin-label study

The molecular dynamics parameters and hydrophobicity of the surface of Silasorb silica gel particles covalently modified with alkyl CnH2n + 1 groups differing in length (n = 2, 8, 18) were studied by electron paramagnetic resonance using spin probes of different structures, sizes, and electrical charges. It has been shown that the hydrocarbon shells of the silica gel microparticles in liquid suspensions provide high rotational and conformational mobility and a very high hydrophobicity of the environment for the incorporated molecules. Some regularities in the dependences of the rotational mobility, orientation ordering, and environmental hydrophobicity have on the hydrocarbon shell thickness have been established for spin probes of different structures: a radical containing an aromatic residue which models a fluorescent sensor molecule, lipid spin probes sensitive to the microviscosity and orientation ordering, and positively charged probes sensitive to electrostatic interactions. It has been found that the rotational mobility of the lipid spin probes sharply decreases upon removing the solvent and recovers in the presence of water or water-ethanol vapors. The high values of rotational mobility and environmental hydrophobicity for the spin probes located in the hydrocarbon shell of the Silasorb particles, which are dispersed in suspensions or in the solvent vapors, allow one to conclude that these microparticles can be perspective matrices for molecular optical chemosensors for analytes in solutions or the gas phase.

Design and Synthesis of a Highly Sensitive Off–On Fluorescent Chemosensor for Zinc Ions Utilizing Internal Charge Transfer

Fluorescence imaging is a powerful tool for the visualization of biological molecules in living cells, tissue slices, and whole bodies, and is important for elucidating biological phenomena. Furthermore, zinc (Zn(2+)) is the second most abundant heavy metal ion in the human body after iron, and detection of chelatable Zn(2+) in biological studies has attracted much attention. Herein, we present a novel, highly sensitive off-on fluorescent chemosensor for Zn(2+) by using the internal charge transfer (ICT) mechanism. The rationale of our approach to highly sensitive sensor molecules is as follows. If fluorescence can be completely quenched in the absence of Zn(2+), chemosensors would offer a better signal-to-noise ratio. However, it is difficult to quench the fluorescence completely before Zn(2+) binding, and most sensor molecules still show very weak fluorescence in the absence of Zn(2+). But even though the sensor shows a weak fluorescence in the absence of Zn(2+), this fluorescence can be further suppressed by selecting an excitation wavelength that is barely absorbed by the Zn(2+)-free sensor molecule. Focusing on careful control of ICT within the 4-amino-1,8-naphthalimide dye platform, we designed and synthesized a new chemosensor (1) that shows a pronounced fluorescence enhancement with a blueshift in the absorption spectrum upon addition of Zn(2+). The usefulness of 1 for monitoring Zn(2+) changes was confirmed in living HeLa cells. There have been several reports on 4-amino-1,8-naphthalimide-based fluorescent sensor molecules. However, 1 is the first Zn(2+)-sensitive off-on fluorescent sensor molecule that employs the ICT mechanism; most off-on sensor molecules for Zn(2+) employ the photoinduced electron transfer (PeT) mechanism.

Highly selective and sensitive Hg2+ fluorescent sensors based on a phosphane sulfide derivative

A series of fluorescent sensor molecules based on a phosphane sulfide derivative were designed and synthesized. The effect of the distance between the complex entities as well as the number and the length of fluorophores was investigated using both steady-state and time-resolved fluorescence methods. The complexation behavior of these sensor molecules, which have a distinct affinity for Hg(2+), is reported. The coordination of Hg(2+) induces a photoinduced electron transfer from fluorophore to the complexed mercury and results in a significant decrease of the fluorescence. Theses sensors exhibit very low detection limits in CH(3)CN/H(2)O (80/20 v:v) and excellent sensitivity to Hg(2+) over other potentially interfering cations such as Na(+), K(+), Mg(2+), Ca(2+), Cu(2+), Ag(+), Zn(2+), Cd(2+) and Pb(2+).

Fluorescent Iminodiacetamide Derivatives as Potential Ionophores for Optical Zinc Ion-selective Sensors

Fluorescent sensor molecules were synthesized by conjugation of iminodiacetamide derivatives with fluorescent moieties of different structures and their UV-visible and fluorescent properties were characterized in acetonitrile solvent. The fluorescent measurements revealed that the N-(2-naphthyl) and N-phenyl derivatives exhibit a distinct zinc ion-selectivity over alkali and alkaline earth metal ions, while N-(anthrylmethyl) and N-(3-methoxyphenyl) derivatives do not possess any ion-selectivities. In contrast to the fluorescent measurements, all ligands show Zn(2+) selectivity over Ca(2+) and Mg(2+) ions in plasticized PVC membranes using potentiometric signal transduction. This observation found for N-(anthrylmethyl) and N-(3-methoxyphenyl) derivatives can be ascribed to the more hindered interaction between the signalling group of the ionophore and the central metal ion in PVC membranes than in acetonitrile solution upon complexation. From the fluorescent measurements it can also be concluded that the ligands with metal ions form complexes mainly with 2:1 stoichiometry (L(2)M). On complex formation a considerable decrease in the fluorescent intensity was observed for all ligands except the N-(anthrylmethyl) derivative, where a 25 - 30 fold fluorescence enhancement was found, which is explained by the photoinduced electron transfer (PET) mechanism. All ionophores exhibited serious hydrogen ion interference, therefore complexation-induced spectral changes were measured in aprotic acetonitrile solution.

A new series of 2,5-bis(4-methylphenyl)-1,3,4-oxadiazole derivatives: their synthesis and fluorescence properties for anion sensors

A new series of 2,5-bis(4-methylphenyl)-1,3,4-oxadiazole derivatives containing various substituted groups on the ortho-position to oxadiazole ring was synthesized and their fluorescent sensor properties were investigated. The fluorescent sensor molecules showed UV absorption shift as well as fluorescence emission shift upon exposure to fluoride anion in DMF solution, which was considerably dependent on the substituent attached on the phenyl group. The new sensory compound, 1d can be used as a fluoride anion sensor in terms of naked-eye detection and fluorescent sensing with high selectivity.

Novel fluorescence assay for tracking molecular and cellular allergen–protein interactions

T cells recognizing nickel (Ni) are key mediators in human Ni allergy, which represents the most common form of human contact hypersensitivity. In contrast to well-characterized Ni-specific human T cell clones, molecular knowledge about the extra- and intracellular route(s) of antigen/allergen presentation and processing of Ni-specific epitopes is still fragmentary. Here, we demonstrate a new metal-specific fluorescent technique to detect and quantify metal ions, like Ni 2+, while they are associated with isolated metalloproteins. Moreover, utilizing the fluorescent metal sensor molecule Newport Green® (NPG) a novel method has been developed, which permits the metal-specific detection of Ni 2+ binding to surface or intracellular structures of individual human antigen presenting cells by flow cytometry. We expect such metal-specific fluorescent analyses to contribute to a better basic understanding of molecular and cellular immune processes involved in Ni-specific T cell epitope generation and the pathogenesis of human nickel allergy.

A Series of Polyamide Receptor Based PET Fluorescent Sensor Molecules: Positively Cooperative Hg2+ Ion Binding with High Sensitivity

[structure: see text] A series of PET fluorescent sensor molecules were designed and synthesized based on BODIPY fluorophore and polyamide receptors. Comparison of the photophysical properties of these sensor molecules, equipped with di-, tri-, and tetraamide receptor, provided a deep insight into the polyamide-Hg2+ interactions, and an unusual positively cooperative tetraamide-Hg2+ complexation was disclosed. In addition, sensor S3 displayed several favorable sensing properties.

Visualization of biological compounds using fluorescent sensor molecules

Visualization of biological compounds using fluorescent sensor molecules

On the Development of Sensor Molecules that Display FeIII-amplified Fluorescence

Incorporation of a tailor-made size-restricted dithia-aza-oxa macrocycle, 1-oxa-4,10-dithia-7-aza-cyclododecane, via a phenyl linker into two fluorescent sensor molecules with electronically decoupled, rigidly fixed, and sterically preoriented architectures, a 1,3,5-triaryl-Delta2-pyrazoline and a meso-substituted boron-dipyrromethene (BDP), yields amplified fluorescence in the red-visible spectral range upon binding of Fe(III) ions. The response to Fe(III) and potentially interfering metal ions is studied in highly polar aprotic and protic solvents for both probes as well as in neat and buffered aqueous solution for one of the sensor molecules, the BDP derivative. In organic solvents, the fluorescence of both indicators is quenched by an intramolecular charge or electron transfer in the excited state and coordination of Fe(III) leads to a revival of their fluorescence without pronounced spectral shifts. Most remarkably, the unbound BDP derivative shows dual emission in water and can be employed for the selective ratiometric signaling of Fe(III) in buffered aqueous solutions.

Recent advances in the design of small molecule-based FRET sensors for cell biology

Fluorescence resonance energy transfer (FRET) has been used extensively as the designing principle for fluorescent sensor molecules. One of the most significant advantages of designing sensor molecules with FRET modulation is that it can enable ratiometric measurement in living cells, which reduces artifacts from microscopic imaging system. We describe the design strategy for the development of small-molecule FRET sensor in terms of avoiding close contact of donor fluorophore and acceptor fluorophore in aqueous solution. Furthermore, we introduce a strategy to design FRET sensors with a modulating overlap integral of donor and acceptor.

Luminescence and Structural Comparisons of Strong-Acid Sensor Molecules. 2

Anthraquinone-containing cyclic polyether hosts form 1:1 complexes with hydronium ion, producing large enhancements in luminescence via inversion of npi* and pipi* exited states. We have characterized the binding of hydronium ion within these host molecules and have synthesized a large variety of analogous hosts that contain different structural and electronic features that allow better understanding of what controls binding and luminescence capacity in this class of fluorescent sensor molecules. X-ray crystallography of an anthraquinone host that contains terminal amine functional groups rather than terminal ether groups is investigated, and complete proton transfer to carbonyl groups is observed in concentrated sulfuric acid media that also produces a previously unobserved luminescence.