FRET Processes Research Articles

A multi-wavelength cross-reactive fluorescent sensor ensemble for fingerprinting flavonoids in serum and urine

Flavonoids are a kind of natural polyphenols which are closely related to human health, and the identification of flavonoids with similar structures is an important but difficult issue. We herein easily constructed a powerful fluorescent sensor ensemble by using surfactant cetyltrimethylammoniumbromide (CTAB) encapsulating two commercially available fluorescent probes (F1 and F2) with multi-wavelength emission. Fluorescence measurements illustrate the present sensor ensemble exhibits turn-off responses to flavones and flavonols but ratiometric responses to isoflavones, owing to different FRET processes. The heat map and linear discriminant analysis (LDA) results show that this single sensor can effectively distinguish 6 flavonoids belong to three subgroups by collecting the fluorescence variation at four typical wavelengths. Moreover, it can be applied to identify different flavonoids even in biofluids like serum and urine, providing potential practical application.

Chiroptically active quantum nanonails.

In recent years, extensive research efforts have been dedicated to the investigation of CdSe/CdS-based quantum-confined nanostructures, driven by their distinctive properties. The morphologies of these nanostructures have been shown to directly affect their properties, an area which has proven to be an important field of study. Herein, we report a new morphology of CdSe/CdS core-shell heterostructures in the form of a 'nanonail' - a modified nanorod-like morphology, in which a distinctive triangular head can be observed at one end of the structure. In-depth studies of this morphology reveal a material with tuneable rod length and width, as well as exceptional photoluminescent properties. Following this, we have demonstrated the ability to induce chiroptical activity via ligand exchange, revealing the important role of the specific morphology, shell thickness and chiral ligand concentration in the effect of ligand induced chirality. In addition, the cellular uptake and cytotoxicity of obtained chiral nanostructures were evaluated on human lung-derived A549 cancer cells, revealing a significant enantioselectivity in biological activity. Finally, analysis on monolayers of the material demonstrate the complete absence of FRET processes. Overall, this CdSe/CdS heterostructure is another tuneable morphology of a very important nanomaterial, one which shows great advantages and a range of potential applications.

Exploiting the photophysical features of DMAN template in ITQ-51 zeotype in the search for FRET energy transfer.

The combination between photoactive molecules and inorganic structures is of great interest for the development of advanced materials in the field of optics. Particularly, zeotypes with extra-large pore size are attractive because they allow the encapsulation of bulky dyes. The microporous aluminophoshate Mg-ITQ-51 (IFO-type structure) represents an ideal candidate because of the synergic combination of two crucial features: the IFO framework itself, which is composed of non-interconnected one-dimensional extra-large elliptical channels with a diameter up to 11 Å able to host bulky guest species, and the particular organic structure-directing agent used for the synthesis (1,8-bis(dimethylamino)naphthalene, DMAN), which efficiently fills the IFO pores, and is itself a photoactive molecule with interesting fluorescence properties in the blue range of the visible spectrum, thus providing a densely-incorporated donor species for FRET processes. Besides, occlusion of DMAN dye in the framework triggers a notable improvement of its fluorescence properties by confinement effect. To extend the action of the material and to mimic processes such as photosynthesis in which FRET is essential, two robust laser dyes with bulky size, rhodamine 123 and Nile Blue, have been encapsulated for the first time in a zeolitic framework, together with DMAN, in a straightforward one-pot synthesis. Thus, photoactive systems with emission in the entire visible range have been achieved due to a partial FRET between organic chromophores protected in a rigid aluminophosphate matrix.

A more flexible long-chain bis(salamo)-like fluorescent probe based on FRET and ICT effects for the identification of aluminum ion in the environment and plants

A more flexible long-chain bis(salamo)-like fluorescent probe DS was designed and synthesized. The unique coupling systems of FRET and ICT effects were designed for DS, which was demonstrated to be capable of detecting aluminum ion through the FRET effect, changing the fluorescence of the solution from green to bright blue (55 nm blue-shift) with good selectivity and fast response, and detecting Al3+ over a wide pH range. The recognition mechanism was determined based on spectral characterization, in the sense that the coordination of Al3+ with the probe DS molecule turned on the intramolecular FRET and ICT processes. ESP analysis and DFT calculations validated the binding sites and fluorescence recognition mechanism. Finally, good results have been achieved in practical applications. The test paper experiment confirmed that DS can be prepared into portable Al3+ test paper, the fluorescence imaging experiment of mung bean sprouts showed that DS can be used for the fluorescence imaging of Al3+ in plants, and the environmental water samples test proved that various ions in the actual water samples hardly interfere with the identification of Al3+, and it is expected to be applied in the environmental testing and other fields.

Target-independent hybridization chain reaction-fluorescence resonance energy transfer for sensitive assay of ctDNA based on Cas12a

Circulating tumor DNA (ctDNA) is a noninvasive biomarker which offer valuable information for cancer diagnosis and prognosis. In this study, a target-independent fluorescent signal system, Hybridization chain reaction-Fluorescence resonance energy transfer (HCR-FRET) system, is designed and optimized. Combined with CRISPR/Cas12a system, a fluorescent biosensing protocol was developed for sensing assay of T790 M. When the target is absent, the initiator remains intact, opens the fuel hairpins and triggers the following HCR-FRET. At presence of the target, the Cas12a/crRNA binary complex specifically recognizes the target, and the Cas12a trans-cleavage activity is activated. As a result, the initiator is cleaved and subsequent HCR responses and FRET processes are attenuated. This method showed detection range from 1 pM to 400 pM with a detection limit of 316 fM. The target independent property of the HCR-FRET system endows this protocol a promising potential to transplant to the assay of other DNA target in parallel.

Dual-ratiometric fluorescence probe for viscosity and hypochlorite based on AIEgen with mitochondria-targeting ability.

Viscous microenvironments are ubiquitous in living organisms. An oxidative stress attack could cause the fluctuation of lymph or blood viscosity, which is closely related to many common diseases. However, the dual-function detection of reactive oxygen species and viscosity is still ineffective due to design defects. Herein, by rational linking a propeller-shaped triphenylamine rotator and a rigid planar anthracene, we report the first dual-ratiometric-response probe for ClO- and viscosity detection. The probe itself shows weak fluorescence background in low-viscous solution or at non-aggregate state, which can largely switch on its blue (414nm) and yellow fluorescence (600nm) respectively in response to ClO- and viscosity. Their sensing mechanisms were elucidated through the interruption of FRET and TICT processes for ClO- and viscosity detection, respectively. In addition to its mitochondria targeting ability, TPP-AN was employed for mitochondrial viscosity and ClO- imaging in HeLa cells, providing a potentially useful probe for clinical assays.

Tandem rigidification and π-extension as a key tool for the development of a narrow linewidth yellow hyperfluorescent OLED system.

Hyperfluorescence (HF), a relatively new phenomenon utilizing the transfer of excitons between two luminophores, requires careful pairwise tuning of molecular energy levels and is proposed to be the crucial step towards the development of new, highly effective OLED systems. To date, barely few HF yellow emitters with desired narrowband emission but moderate external quantum efficiency (EQE < 20%) have been reported. This is because a systematic strategy embracing both Förster resonance energy transfer (FRET) and triplet to singlet (TTS) transition as complementary mechanisms for effective exciton transfer has not yet been proposed. Herein, we present a rational approach, which allows, through subtle structural modification, a pair of compounds built from the same donor and acceptor subunits, but with varied communication between these ambipolar fragments, to be obtained. The TADF-active dopant is based on a naphthalimide scaffold linked to the nitrogen of a carbazole moiety, which through the introduction of an additional bond leads not only to π-cloud enlargement, but also rigidifies and inhibits the rotation of the donor. This structural change prevents TADF, and guides bandgaps and excited state energies to simultaneously pursue FRET and TTS processes. New OLED devices utilizing the presented emitters show excellent external quantum efficiency (up to 27%) and a narrow full width at half maximum (40 nm), which is a consequence of very good alignment of energy levels. The presented design principles prove that only a minor structural modification is needed to obtain commercially applicable dyes for HF OLED devices.

Fluorene based fluorescent and colorimetric sensors for ultrasensitive detection of nitroaromatics in aqueous medium

New butterfly-shaped π-extended sensors 4 and 5 were synthesized through the Knoevenagel condensation and the Suzuki-Miyaura coupling reaction. Sensors were employed for sensitive detection of nitroaromatics. Among nitroaromatic compounds (NACs), sensors 4 and 5 exhibited ultrasensitive fluorescence quenching response to picric acid (PA) with quenching efficiency >45 and >260 folds and limit of detection (LOD) 22.0 and 0.23 ppt (parts per trillion), respectively. This ultra-sensitivity was contributed by FRET and IFE processes. Selective interaction of sensors with PA was evaluated by fluorescence, UV–Vis., 1H NMR titration, dynamic light scattering (DLS), and density functional theory (DFT) calculations. These sensors were successfully employed for detection of picric acid in real water samples. Interestingly, sensors 4 and 5 showed selective colorimetric recognition of picric acid. Moreover, sensors’ coated test strips were fabricated for an immediate naked eye detection of PA that demonstrated rapidity, simplicity, and convenience for on-site detection of PA. Robust sensitivity for PA illustrated that these sensors can be employed in the development of new detection tools for PA analysis in real samples.

Mechanochemical Preparation and Self-Assembly of Protein:Dye Hybrids for White Luminescence.

Protein nanofibrils (PNFs) functionalized with multiple dyes are prepared by a combination of mechanochemistry and liquid-phase self-assembly. The three employed dyes are Fluorescent Brightener 378 (F378), 2-butyl-6-(butylamino)-1H-benzo[de]isoquinoline-1,3(2H)-dione (Fluorol 555), and Nile red (NR). F378 acts as the donor with Fluorol 555 as the acceptor. F555 in turn acts as the donor and NR as the acceptor. This enables a FRET cascade that enables conversion of UV light to white light. The efficiency of FRET can be influenced by the details of the self-assembly process. If proteins milled with different dyes are mixed prior to self-assembly, nanofibrils are formed containing all three dyes, thus favoring FRET processes. By tuning the ratio of the three luminescent dyes, PNF dispersions are obtained that display bright white light emission. Moreover, the PNF dispersions can be converted into white luminescent films and gels where the PNFs may help to organize dye molecules. Additionally, the PNF materials can be employed as coatings on commercial LEDs, enabling emission of white light.

High-Efficiency FRET Processes in BODIPY-Functionalized Quantum Dot Architectures.

Efficient FRET systems are developed combining colloidal CdSe quantum dots (QDs) donors and BODIPY acceptors. To promote effective energy transfer in FRET architectures, the distance between the organic fluorophore and the QDs needs to be optimized by a careful system engineering. In this context, BODIPY dyes bearing amino-terminated functionalities are used in virtue of the high affinity of amine groups in coordinating the QD surface. A preliminary QD surface treatment with a short amine ligand is performed to favor the interaction with the organic fluorophores in solution. The successful coordination of the dye to the QD surface, accomplishing a short donor-acceptor distance, provides effective energy transfer already in solution, with efficiency of 76 %. The efficiency further increases in the solid state where the QDs and the dye are deposited as single coordinated units from solution, with a distance between the fluorophores down to 2.2 nm, demonstrating the effectiveness of the coupling strategy.

Mapping heterogeneous polarity in multicompartment nanoparticles

Understanding polarity gradients inside nanomaterials is essential to capture their potential as nanoreactors, catalysts or in drug delivery applications. We propose here a method to obtain detailed, quantitative information on heterogeneous polarity in multicompartment nanostructures. The method is based on a 2-steps procedure, (i) deconvolution of complex emission spectra of two solvatochromic probes followed by (ii) spectrally resolved analysis of FRET between the same solvatochromic dyes. While the first step yields a list of polarities probed in the nanomaterial suspension, the second step correlates the polarities in space. Colocalization of polarities falling within few nanometer radius is obtained via FRET, a process called here nanopolarity mapping. Here, Prodan and Nile Red are tested to map the polarity of a water-dispersable, multicompartment nanostructure, named PluS nanoparticle (NPs). PluS NPs are uniform core-shell nanoparticles with silica cores (diameter ~10 nm) and Pluronic F127 shell (thickness ~7 nm). The probes report on a wide range of nanopolarities among which the dyes efficiently exchange energy via FRET, demonstrating the coexistence of a rich variety of environments within nanometer distance. Their use as a FRET couple highlights the proximity of strongly hydrophobic sites and hydrated layers, and quantitatively accounts for the emission component related to external water, which remains unaffected by FRET processes. This method is general and applicable to map nanopolarity in a large variety of nanomaterials.

Synthesis and photophysical properties of novel 1,8-naphthalimide light-harvesting antennae based on benzyl aryl ether architecture

Two novel benzyl aryl ether light-harvesting antennae accomplishing high effective energy transfer, core and peripherally decorated with fluorescent 1,8-naphthalimide units were successfully synthesized. The peripheral blue-emitting benzyl aryl ether intermediate wedge showed considerable probe potential for determination of water content in organic solvents probably as a result of a TICT process. The core of one of the two light-harvesting antennae was configured on the “fluorophore-spacer-receptor” format. Due to the simultaneous operation of FRET and PET processes that system showed excellent pH sensing characteristics and high potential as a probe for monitoring the pH variations in environmental and biological samples.

A multi-responsive diarylethene-rhodamine 6G derivative for sequential detection of Cr3+ and CO32−

A new unsymmetrical diarylethene derivative (1o) with rhodamine 6G as a functional group has been designed and synthesized. It displayed good physicochemical properties induced by lights and chemical stimuli. 1o could sensitively detect towards Cr3+ with a 1:1 stoichiometry, and exhibit an obviously fluorescence (from dark to light cyan) and color (from colorless to pink) changes during the recognition process. The limit of detection was determined to be 27 nM and 8.5 nM via UV/vis and fluorescence methods, respectively. More importantly, the resulting complex 1o-Cr3+ (1o') could be served as a potential fluorescent probe to selectively and sensitively recognize toward CO32−, the limit of detection was determined to be 0.88 μM and 0.26 μM via UV/vis and fluorescence methods, respectively. Moreover, the quenching of fluorescence intensity can reach 95% due to the perfect FRET processes between the excited open-ring rhodamine 6G moiety and the closed-ring diarylethene unit.

Doubly Caged Linker for AND-Type Fluorogenic Construction of Protein/Antibody Bioconjugates and In Situ Quantification.

In situ quantification of the conjugation efficiency of azide-terminated synthetic polymers/imaging probes and thiol-functionalized antibodies/proteins/peptides was enabled by a doubly caged profluorescent and heterodifunctional core molecule C1 as a self-sorting bridging unit. Orthogonal dual "click" coupling of C1 with azide- and thiol-functionalized precursors led to highly fluorescent bioconjugates, whereas single-click products remained essentially nonfluorescent. Integration with FRET processes was also possible. For the construction of antibody-probe conjugates from an anti-carcinoembryonic antigen and a quinone-caged profluorescent naphthalimide derivative, the dual "click" coupling process with C1 was monitored on the basis of the emission turn-on of C1, whereas prominent changes in FRET ratios occurred for antibody-imaging-probe conjugates when specifically triggered by quinone oxidoreductase (NQO1), which is overexpressed in various types of cancer cells.

Doubly Caged Linker for AND‐Type Fluorogenic Construction of Protein/Antibody Bioconjugates and In Situ Quantification

AbstractIn situ quantification of the conjugation efficiency of azide‐terminated synthetic polymers/imaging probes and thiol‐functionalized antibodies/proteins/peptides was enabled by a doubly caged profluorescent and heterodifunctional core molecule C1 as a self‐sorting bridging unit. Orthogonal dual “click” coupling of C1 with azide‐ and thiol‐functionalized precursors led to highly fluorescent bioconjugates, whereas single‐click products remained essentially nonfluorescent. Integration with FRET processes was also possible. For the construction of antibody–probe conjugates from an anti‐carcinoembryonic antigen and a quinone‐caged profluorescent naphthalimide derivative, the dual “click” coupling process with C1 was monitored on the basis of the emission turn‐on of C1, whereas prominent changes in FRET ratios occurred for antibody–imaging‐probe conjugates when specifically triggered by quinone oxidoreductase (NQO1), which is overexpressed in various types of cancer cells.

A new PET and FRET-based molecular logic circuit mimicking the three-state logic gate

A new dyad composed of a naphthalimide unit and a spiropyran unit with a piperazine unit as the bridge has been synthesized to mimick the Three State Logic (TSL) gate and the luminescence was recorded as the output. Inputs including Protonation, deprotonation, UV irradiation and Fe3+ coordination were employed to modulate the emission because of their distinctive outputs. Transient fluorescence and theoretical calculation were performed to further investigate the PET and FRET processes.

A selective fluorescent and colorimetric probe for cyanide based on dual-site controlled intramolecular charge transfer–photoinduced electron transfer–Fluorescence resonance energy transfer

A novel dual-site controlled intramolecular charge transfer–photoinduced electron transfer–Fluorescence resonance energy transfer (ICT–PET–FRET) probe 6b and its reference ICT–FRET probe 6a were designed and synthesized. The probes show high selective fluorescent and colorimetric sensing for cyanide ions. The NH and CH groups connected by the π-extension framework in probe 6b were simultaneously employed as the two sensing sites and controlled the fluorescence of naphthalimide unites by PET, ICT followed by the corresponding FRET processes between coumarin and naphthalimide fluorophores.

Dual Site-Controlled and Lysosome-Targeted Intramolecular Charge Transfer-Photoinduced Electron Transfer-Fluorescence Resonance Energy Transfer Fluorescent Probe for Monitoring pH Changes in Living Cells.

Acidic pH is a critical physiological factor for controlling the activities and functions of lysosome. Herein, we report a novel dual site-controlled and lysosome-targeted intramolecular charge transfer-photoinduced electron transfer-Fluorescence resonance energy transfer (ICT-PET-FRET) fluorescent probe (CN-pH), which was essentially the combination of a turn-on pH probe (CN-1) and a turn-off pH probe (CN-2) by a nonconjugated linker. Coumarin and naphthalimide fluorophores were selected as donor and acceptor to construct the FRET platform. Hydroxyl group and morpholine were simultaneously employed as the two pH sensing sites and controlled the fluorescence of coumarin and naphthalimide units by ICT and PET, respectively. The sensing mechanism of CN-pH to pH was essentially an integration of ICT, PET, and FRET processes. Meanwhile, the morpholine also can serve as a lysosome-targeted group. By combining the two data analysis approaches of the ratios of the two emission intensities (R) and the reverse ratio R' (R' = 1/R), the fluorescent ratio of CN-pH can show proportional relationship to pH values in a very broad range from pH 4.0 to 8.0 with high sensitivity. The probe has been successfully applied for the fluorescence imaging of the lysosomal pH values, as well as ratiometrically visualizing chloroquine-stimulated changes of intracellular pH in living cells. These features demonstrate that the probe can afford practical application in biological systems.

The influence of fluorescent silica nanoparticle surface chemistry on the energy transfer processes with lipid bilayers

A study of 3D and 2D intracellular FRET processes at the interface between surface tailored silica nanoparticles and lipid bilayers.

A ratiometric fluorescence sensor for Fe(3+) based on FRET and PET processes.

A rhodamine/coumarin-based ratiometric fluorescent Fe(3+) sensor has been designed and synthesized. The sensor exhibits a good response to Fe(3+) ions with high sensitivity, selectivity and a large shift in the emission spectra (>100 nm), which shows Fe(3+) -induced FRET OFF-ON and PET ON-OFF behavior.