Fluorescence Resonance Energy Transfer System Research Articles

Detection of Al3+ and Fe3+ Ions with Phthalocyanine‐Merocyanine 540 Dye‐Based Fluorescence Resonance Energy Transfer

In this study, a sensor system was designed to detect metal ions based on the fluorescence resonance energy transfer (FRET) process between free‐phthalocyanine (PC) and merocyanine 540 (MC540). Following the determination of parameters and appropriate conditions for energy transfer, the sensor system was tested with different metal ions. The conclusion was that the system showed high selectivity to iron and aluminum ions. In this FRET system, limit of detection (LOD) values for metal ions were calculated as 437 nM for Al3+ and 32.1 nM for Fe3+. The system was able to detect iron and aluminum in very low concentrations. It also showed good selectivity for the two metal ions.

A Fluorescence Resonance Energy Transfer Biosensor Based on Graphene Quantum Dots and Protoporphyrin IX for the Detection of Melamine.

Graphene quantum dots (GQDs), which have high photostability, anti-photobleaching and scintillation, good biocompatibility and low toxicity, are important member of the fluorescent material family, and have attracted extensive research interest. In this paper, a fluorescence resonance energy transfer (FRET) biosensor based on protoporphyrin IX (PpIX) and GQDs was developed for melamine detection. PpIX was bound to the surface of GQDs to produce self-assembled nanosensors, and a FRET process occurred between GQDs and PpIX. However, due to the combination of melamine and PpIX, the FRET process was shut down in the presence of melamine. The FRET system could quickly and accurately detect melamine with a detection range of 1.0 × 10-8 to 2.0 × 10-6mol/L based on the fluorescence intensity ratio of PpIX and GQDs, and the detection limit was 3.6 × 10-9mol/L. This method obtained satisfactory results when it was employed to the determination of melamine in milk samples.

Fluorescence resonance energy transfer between carbon quantum dots and silver nanoparticles: Application to mercuric ion sensing.

Fluorescence resonance energy transfer (FRET) process as a practical and competitive sensing strategy was utilized between carbon quantum dots (C-dots) and silver nanoparticles (Ag NPs) for the determination of mercuric ions. The novel synthesized C-dots with the quantum yield of 84% acted as the donor and Ag NPs operated as the acceptor in the FRET process leading to the fluorescence quenching of the C-dots. In the presence of Hg(II) ions, the FRET-quenched fluorescence emission of the C-dots-Ag NPs system was recovered owing to oxidation of Ag NPs by Hg(II) ions, so that the turn-on fluorescence intensity was directly proportional to the Hg(II) ion concentration. Accordingly, combination of the FRET system with the redox reaction was firstly utilized to construct an innovative turn-off/on fluorescent sensor for the quantification of Hg(II) ion. The calibration plot was linear in the concentration range 0.5-500.0nmolL-1 with a determination coefficient (R2) of 0.9965. The limit of detection and limit of quantification were 0.10 and 0.35nmolL-1, respectively, according to the IUPAC definition. The method was applied for the determination of Hg(II) ion in lake water, wastewater and tea samples, and the proper relative recoveries (98.0-104.0%) were obtained for the spiked samples. The method has high potential to diagnose trace values of mercuric ions in real samples with high sensitivity and repeatability.

Etching-controlled suppression of fluorescence resonance energy transfer between nitrogen-doped carbon dots and Ag nanoprisms for glucose assay and diabetes diagnosis.

Numerous methods have been developed for glucose detection, only few cases can be really applied in clinical diagnosis. Herein, we report a new approach to achieve the detection of glucose in clinical samples and distinguishing the diabetic patients with healthy ones. Specifically, a fluorescence resonance energy transfer (FRET) system is established first, where nitrogen-doped carbon dots (N-CDs) and Ag nanoprisms (AgNPRs) with good spectral overlap act as energy donor and acceptor, respectively. Then, the FRET can be inhibited through oxidative etching of the energy acceptor in the presence of glucose and glucose oxidase, where hydrogen peroxide is generated to transform AgNPRs into Ag+ ions. Based on the turn-on fluorescent signal versus glucose concentration, a new method for quantitative detection of glucose is developed. This etching-induced analytical method is simple, reliable, robust and cost-effective, which is promising to assist the doctors to clinically diagnose diabetes and other diseases related to metabolic disorders.

"Plug and Play" logic gate construction based on chemically triggered fluorescence switching of gold nanoparticles conjugated with Cy3-tagged aptamer.

Gold nanoparticles (AuNPs) conjugated with Cy3-tagged aptamer which can specifically recognize chloramphenicol (CAP) (referred to as AuNPs-AptCAP) are described. CAP can trigger the configuration change of CAP binding aptamer, and thus switching the fluorescence of AuNPs-AptCAP through changing the efficiency of the fluorescence resonance energy transfer (FRET) system with Cy3 as donors and AuNPs as recipients. AuNPs-AptCAP exhibits a linear range of CAP concentrations from 26.0 to 277μgL-1 with a limit of detection of 8.1μgL-1 when Cy3 was excited at 530nm and emission was measured at 570nm. More importantly, AuNPs-AptCAP can be utilized as signal transducers for the build-up of a series of logic gates including YES, PASS 0, INH, NOT, PASS 1, and NAND. Utilizing the principle of ametal ion-mediated fluorescence switch together with astrong metal ion chelator, the fluorescence of AuNPs-AptCAP could be modulated by adding metal ions and EDTA sequentially. Therefore, a "Plug and Play" logic system based on AuNPs-AptCAP has been realized by simply adding other components to create new logic functions. This work highlights theadvantages of simple synthesis and facile fluorescence switching properties, which will provide useful knowledge for the establishment of molecular logic systems. Graphical abstract.

A ratiometric and far-red fluorescence "off-on" sensor for sequential determination of copper(II) and L-histidine based on FRET system between N-acetyl-L-cysteine-capped AuNCs and N,S,P co-doped carbon dots.

Afar-red fluorescence "off-on" sensing strategy is describedfor sequential ratiometric determination of Cu2+ and L-histidine (L-His) based on fluorescence resonance energy transfer (FRET) system. N,S,P co-doped carbon dots (N,S,P-CDs) and N-acetyl-L-cysteine functionalized gold nanoclusters (NAC-AuNCs) are used in the FRET system, which serve as energy donor and acceptor, respectively. After adding NAC-AuNCs into the solution of N,S,P-CDs, the fluorescence of N,S,P-CDs is effectively quenched, while the far-red fluorescence of NAC-AuNCs appears. Cu2+ can decrease fluorescence of NAC-AuNCs, and then L-His can effectively recover the fluorescence of NAC-AuNCs. The possible reason is that the stronger affinity between Cu2+ and L-His can pull Cu2+ away from the surface of NAC-AuNCs. Through it all, the emission intensity of N,S,P-CDs remains nearly constant, so the ratio of fluorescence intensities at 485 and 625nm exhibits a linear correlation to the Cu2+ and L-His concentration, respectively. The sensing platform shows good selectivity towards Cu2+ and L-His with a linear range of 0.65-26.58μM and 3.13-56.25μM and determination limits of 0.50μM and 0.374μM, respectively. The proposed method has been successfully used for Cu2+ and L-His determination in real samples with satisfying results. Graphical abstract.

Phototuning Energy Transfer in Self‐Organized Luminescent Helical Superstructures for Photonic Applications

AbstractStimuli‐triggered changes in the emission color of functional materials are in demand for a variety of potential applications. Herein, a strategy to reversibly phototune fluorescence color in cholesteric liquid crystal (CLC) with self‐organized luminescent helical superstructure via fluorescence resonance energy transfer (FRET) is developed. This dynamic FRET system is constructed by doping with 1,2‐dithienyldicyanoethene‐based chiral fluorescent photoswitch (switch 7) as an acceptor and a conventional coumarin dye C6 as a donor into a nematic liquid crystal (LC) host. The efficient energy transfer from C6 to switch 7 in the LC media, which is evidenced by both of emission color shift and enhanced circularly polarized luminescence, can be modulated on the basis of reversible trans–cis photoisomerization of switch 7 upon visible/UV light irradiations, leading to the emission color change from orange to green. Noteworthily, the FRET efficiency can be significantly improved when CLCs are confined in the polymer microtubes. Preliminary demonstrations of the LC film and LC microtubes array for information display and encryption with ease of electrical operation between planar, homeotropic, and focal conic states or light manipulation are described.

Human Neutrophil Elastase Activated Fluorescent Probe for Pulmonary Diseases Based on Fluorescence Resonance Energy Transfer Using CdSe/ZnS Quantum Dots.

There is an increasing demand for effective noninvasive diagnosis against common pulmonary diseases, which are rising sharply due to the serious air pollution. Human neutrophil elastase (HNE), a typical protease highly involved in pulmonary inflammatory diseases and lung cancer, is a potential predictor for disease progression. Currently, few of the HNE-targeting probes are applicable in vivo due to the limitation in sensitivity and biocompatibility. Herein, we reported the achievement of in vitro detection and in vivo imaging of HNE by incorporating the HNE-specific peptide substrate, quantum dots (QDs), and organic dyes into the fluorescence resonance energy transfer (FRET) system. The refined nanoprobe, termed QDP, could specifically measure the HNE with excellent sensitivity of 7.15 pM in aqueous solution and successfully image the endogenous and exogenous HNE in living cells. In addition, this nanoprobe enabled HNE imaging in mouse models of lung cancer and acute lung injury, and the HNE activity at high temporal and spatial resolution was continuously monitored. Most importantly, QDP successfully discriminated the serums of patients with lung diseases from those of the healthy controls based on the HNE activity determination. Overall, this study demonstrates the advantages of a FRET-system-based nanoprobe in imaging performance and provides an applicable tool for in vivo HNE detection and pulmonary disease diagnosis.

An upconversion nanoparticle-based photostable FRET system for long-chain DNA sequence detection

The fluorescence resonance energy transfer (FRET)-based diagnosis method has been widely used in fast and accurate diagnosis. However, the traditional FRET-based diagnosis method is unable to detect long-chain DNA sequences, due to the limitation of the distance between the donor and acceptor, while the long-chain DNA sequence enables higher selectivity and is quite important for confirming many major diseases. Therefore, it is urgently needed to develop an efficient FRET system for long-chain DNA detection. Herein a ‘head-to-tail’ structure was developed using NaYF4:Yb,Er nanoparticles as the energy donor and gold nanoparticles (AuNPs) as the acceptor to detect long-chain oligonucleotides sequences (i.e., HIV DNA, 52 bp). We modified NaYF4:Yb,Er nanoparticles with carboxylic acid groups by using poly(acrylic acid) to enhance its hydrophilic and then covalently attached 5 ‘end of capture oligonucleotides strand to the surface of the particles. In the presence of target HIV DNA, gold nanoparticles with reported DNA were brought close to NaYF4:Yb,Er nanoparticles upon ‘head-to-tail’ sandwich hybridization with target HIV DNA, resulting in an efficient FRET. Moreover, benefited from both photostable nanoparticles of UCNPs and AuNPs, the photobleaching issue has also been settled down. This developed method possesses high selectivity, high sensitivity, and reached a nanomolar limitation level. To our knowledge, it is the first time to report a ‘head-to-tail’ structure FRET system for detecting long-chain DNA sequences.

Application of maleimide modified graphene quantum dots and porphyrin fluorescence resonance energy transfer in the design of ‘‘turn-on’’ fluorescence sensors for biothiols

Two novel fluorescent probes were designed to detect the biothiol in foods using the highly efficient Michael addition reaction between maleimide-derived probes and the biothiol. First, maleimide functionalized GQDs (M-GQDs) were synthesized and used for biothiol identification according to the Michael addition principle. The biothiol can be detected in the range of 5 × 10−9 to 4 × 10−7 mol/L and the detection limit was 1.69 × 10−9 mol/L. Then, a fluorescence resonance energy transfer (FRET) system between M-GQDs and tetrakis (4-aminophenyl) porphyrin (TAPP) for biothiol detection was developed. However, the process of FRET was switched off in the presence of biothiols due to the switch of M-GQDs fluorescence emission to the“ON” mode following the Michael addition mechanism. The system could quickly and accurately detect the biothiol with a detection range of 6.7 × 10−10 to 2 × 10−7 mol/L and a detection limit of 2.34 × 10−10 mol/L. Compared to the single detection system, the FRET system had a wider detection range and lower detection limit, and the related biomolecules did not interfere with the quantitative identification of the biothiol. The proposed method was successfully applied for the determination of the biothiol in foods and human blood samples.

A highly selective “Turn On” fluorescent probe based on FRET mechanism for hydrogen sulfide detection in living cells

A novel class of fluorescent “turn on” probe based on the fluorescence resonance energy transfer (FRET) approach was designed, and successfully synthesized for the detection of hydrogen sulfide (H2S) both in vitro and in vivo. This new H2S responsive fluorescent probe was developed based on the basis of the dabsyl and fluorescein FRET system. In the presence of H2S, a fluorescence enhancement was markedly observed mainly due to the inhibition of the FRET process. This probe could quantitatively measure the level of H2S with the detection limit of 0.02 μM. Cell imaging results demonstrated the potential applicability of a dabsylfluorescein probe for H2S detection in living cells.

Self-Assembled FRET Nanoprobe with Metal-Organic Framework As a Scaffold for Ratiometric Detection of Hypochlorous Acid.

Metal-organic framework (MOF) has been extensively explored in a number of fields due to its diverse properties. In this work, we demonstrated the potential of MOF in the establishment of a self-assembled fluorescence resonance energy transfer (FRET) system for developing ratiometric fluorescent nanoprobe. For this purpose, zeolitic imidazolate framework-8 (ZIF-8) was selected as a MOF model to entrap carbon dot (CD) and curcumin (CCM) during its self-assembly, which produces CD/CCM@ZIF-8. Benefiting from the confinement effect of ZIF-8, the loaded CD and CCM can be brought in close proximity for energy transfer to occur. Under optimal conditions, a high FRET efficiency of 68.7% can be obtained. Importantly, compared with traditional FRET systems, the fabrication process of CD/CCM@ZIF-8 is much more simple and straightforward, which does not involve the elaborate design and complicated synthesis of molecular linkers. However, in the presence of hypochlorous acid (HClO), the FRET process from CD to CCM will be disrupted, rendering CD/CCM@ZIF-8 to display a ratiometric response to HClO. This finding led to a method for ratiometric fluorescent detection of HClO with a detection limit of 67 nM and excellent selectivity over other reactive oxygen species. We believe that this study can give a new insight into the rational design and application of FRET-based nanoprobes.

CdSe quantum dots-sensitized FRET system for ciprofloxacin detection

The prepared CdSe quantum dots (QDs) were applied to selectively and sensitively assay ciprofloxacin as a fluorescence sensor based on fluorescence resonance energy transfer (FRET) mechanism. The efficiency of FRET between CdSe QDs and ciprofloxacin can be modulated by the degree of spectral overlap between the excitation peak of CdSe QDs and the emission peak of ciprofloxacin. Within the range of 0 to 120 μmol·L−1, the good linearity was obtained (R = 0.99561) and the detection limit reached to 0.6 μmol·L−1. This method is simple and rapid and can be used in environmental water and milk samples.

A highly sensitive fluorescent immunosensor for sensitive detection of nuclear matrix protein 22 as biomarker for early stage diagnosis of bladder cancer.

A novel strategy is reported for highly sensitive, rapid, and selective detection of nuclear matrix protein NMP22 using two-color quantum dots based on fluorescence resonance energy transfer (FRET). Quantum dots (QDs) are highly advantageous for biological imaging and analysis, particularly when combined with (FRET) properties of semiconductor quantum dot (QDs) are ideal for biological analysis to improve sensitivity and accuracy. In this FRET system narrowly dispersed green emitting quantum dot CdTe core is used as a donor and labelled by monoclonal (mAb) antibody, while orange emitting quantum dot CdTe/CdS core shell is used as an accepter and labelled by polyclonal (pAb) antibody. The quantum dots are labelled by antibodies using EDC/NHS as crosslinking agent. Bovine serum albumin (BSA) solution was added to block nonspecific binding sites. The fluorescence intensity of QDs accepter decreased linearly with the increasing concentrations of NMP22 from 2–22 pg mL−1 due to FRET system and fluoroimmunoassay reaction. This method has good regression coefficient (R2 = 0.998) and detection limit was 0.05 pg mL−1. The proposed FRET-based immunosensor provides a quick, simple and sensitive immunoassay tool for protein detection, and can be considered as a promising approach for clinical applications. The proposed FRET-based immunosensor provides a quick, simple and sensitive immunoassay tool for protein detection, and can be considered as a promising approach for clinical applications.

Reactive Quantum Dot-Based FRET Systems for Target-Catalyzed Detection of RNA.

Oligonucleotide-templated reactions (OTRs) between two reactive hybridization probes allow for the detection of a DNA or RNA of interest by exploiting the target molecule as a catalyst of chemical reactions. The product of such a reaction commonly exhibits distinct fluorescence properties and can be detected by the means of fluorescence spectroscopy. The vast majority of OTR systems utilize organic dyes as fluorescent reporters. However, the use of brighter emitters, such as semiconductor quantum dots (QDs), has potential to improve the sensitivity of detection by providing brighter signals and permitting the use of probes at very low concentrations. Here we report an RNA-templated reaction between two fluorescently labeled peptide nucleic acid (PNA)-based probes, which proceeds on the surface of a QD. The QD-bound PNA probe bears a cysteine functionality, while the other PNA is functionalized with an organic dye as a thioester. OTR between these probes proceeds through a transfer of the organic dye to the QD and can be conveniently monitored via fluorescence resonance energy transfer (FRET) from the QD to the Cy5. The reaction was performed in a conventional fluorescence microplate reader and permits the detection of RNA in the picomolar range.

Real-Time Fluorescence-Based Approaches to Disentangle Mechanisms of a Protein's RNA Chaperone Activity.

RNA-binding proteins with an RNA chaperone activity exert either one or both of the following catalytic activities: (1) RNA annealing, i.e., the protein supports intra- as well as intermolecular RNA-RNA interactions and (2) strand displacement, i.e., the protein mediates the exchange of individual strands of a preexisting RNA duplex. To discriminate and further characterize these activities, it requires defined assay systems. These are based on entirely or partially complementary RNA oligonucleotides that are labeled with fluorescent and/or quencher dyes. The non-catalyzed and the protein-supported associations of the RNA molecules are followed by a real-time fluorescence resonance energy transfer (FRET) system. By site-specific modification of the RNAs or the protein, the substrate- and protein-specific parameters of the RNA chaperone activity can be explored and identified.In this chapter, we present strategies on the design of labeled RNA molecules to be used to characterize the activities of an RNA-binding protein and explain how to monitor progress curves of RNA annealing and strand displacement reactions in single cuvette or well-plate scales. We provide sets of equations and models to determine and analyze different types of reactions, e.g., by calculation of first- and second-order rate constants. Likewise, we demonstrate how to exploit these simple experimental setups to elucidate elementary principles of the reaction mechanisms performed by the protein of interest by applying basic kinetic applications, such as ARRHENIUS and linear free energy relationship analyses. These approaches will be explained by providing example plots and graphs from experiments investigating the RNA chaperone activities of the RNA-binding proteins NF90-NF45 and AUF1 p45.

Multiplexed detection of micro-RNAs based on microfluidic multi-color fluorescence droplets.

In this work, simple, rapid, and low-cost multiplexed detection of tumor-related micro-RNAs (miRNAs) was achieved based on multi-color fluorescence on a microfluidic droplet chip, which simplified the complexity of light path to a half. A four-T-junction structure was fabricated to form uniform nano-volume droplet arrays with customized contents. Multi-color quantum dots (QDs) used as the fluorescence labels were encapsulated into droplets to develop the multi-path fluorescence detection module. We designed an integrated multiplex fluorescence resonance energy transfer system assisted by multiple QDs (four colors) and one quencher to detect four tumor-related miRNAs (miRNA-20a, miRNA-21, miRNA-155, and miRNA-221). The qualitative analysis of miRNAs was realized by the color identification of QDs, while the quantitative detection of miRNAs was achieved based on the linear relationship between the quenching efficiency of QDs and the concentration of miRNAs. The practicability of the multiplex detection device was further confirmed by detecting four tumor-related miRNAs in real human serum samples. The detection limits of four miRNAs ranged from 35 to 39pmol/L was achieved without any target amplification. And the linear range was from 0.1nmol/L to 1μmol/L using 10nL detection volume (one droplet) under the detection speed of 320 droplets per minute. The multiple detection system for miRNAs is simple, fast, and low-cost and will be a powerful platform for clinical diagnostic analysis. Graphical abstract.

A smart bioresponsive nanosystem with dual-modal imaging for drug visual loading and targeted delivery

Abstract Most of present studies are focusing on improving cancer therapy effect by synthesizing new carriers or drugs, while little attention is paid to develop new drug loading patterns and improve the efficiency. Herein, a novel and smart nanosystem that not only realized targeted delivery and responsive degradation but also achieved facilely visual drug loading and release, was reported for the first time. To build this system, highly fluorescent carbon quantum dots (CQDs) were synthesized and served as an effective indicator and carrier to achieve visual doxorubicin (DOX) loading by the strong fluorescence resonance energy transfer (FRET) interactions between them. Biodegradable nano-MnO2 was then synthesized by biomineralization with folic acid modified bovine serum albumin, which was employed as the other fluorescence quencher for CQDs and DOX and established the other FRET system to visually monitor drug loading. The double-FRET nanosystem could be specifically ingested by cancer cells and responsively degraded by intracellular H+/H2O2 or glutathione. Meanwhile, this triggered drug release behavior was visually detected after gradual dissociation of the two FRET systems by monitoring the fluorescence and magnetic resonance changes. More importantly, both in vitro and in vivo experiments demonstrated that the novel nanosystem exhibited much better cancer therapy effects than pure DOX, and we also reported its first experimental example to treat cancer migration. This work established a new pattern to improve and monitor drug loading, delivery and release, and demonstrated its good performance in cancer therapy and anti-migration.

Broad Spectrum Tunable Photoluminescent Material Based on Cascade Fluorescence Resonance Energy Transfer between Three Fluorophores Encapsulated within the Self-Assembled Surfactant Systems.

A broad spectrum tunable photoluminescent material with dual encryption based on a two-step fluorescence resonance energy transfer (FRET) between pyrene (Py), coumarin 480 (Cou480), and rhodamine 6G (R6G) in micelles of SDS and bmimDS is presented. The phenomenon is achievable due to the encapsulation of the fluorophores within these micelles. The transfer of energy as FRET between the pair Py and Cou480 showed ON at 336 nm and OFF at 402 nm in contrast to the FRET observed between the pair Cou480 and R6G that showed ON at 402 nm and OFF at 336 nm. However, the transfer of energy as FRET occurs from Py to R6G in the presence of Cou480 when excited at 336 nm, thereby making it a chain of three fluorophores with Cou480 acting as a relay fluorophore receiving energy from Py and transferring it to R6G. The different FRET scenarios between the three fluorophores in micelles provide a window for the generation of a matrix of colors, which occupies a significant 2D area in the chromaticity diagram, having potential applications in security printing. The different fluorophoric ratios generate different colors based on their individual photonic emissions and the FRET processes taking place between them. Writing tests were carried out using varied ratios of the fluorophores in the micellar systems producing different colored outputs under the UV light with insignificant visibility under the white light. We envision that this as-discovered three fluorophoric FRET system could form the basis for the future development of multi-FRET light-harvesting devices and anti-counterfeiting security inks based on much simpler non-covalent interaction aided encapsulation of the fluorophores within the self-assembled soft systems.

Fabricating a fluorescence resonance energy transfer system with AIE molecular for sensitive detection of Cu(II) ions.

The aggregation-induced emission (AIE) luminogens has exhibited strong potential in fabricating the fluorescence resonance energy transfer (FRET) system. In this paper one efficient FRET system was fabricated in aqueous solution based on an AIE molecular (T) and Nile Red (NiR) dyes: T acts as the energy donor and NiR acts as the energy acceptor with a ratio of 250:1. The energy-transfer efficiency from the donor to acceptor is 82.52%, and the antenna effect is 24.9. Base on this data, a very low detection limit for Cu2+ was calculated to be 35.5 pM. This method displays penitential application on fluorescence probe for small ions or molecular detection by light-harvesting system based on a simple AIE donor under physiological conditions.