High Fluorescence Resonance Energy Transfer Research Articles

Engineering highly efficient NIR-II FRET platform for Background-Free homogeneous detection of SARS-CoV-2 neutralizing antibodies in whole blood

Förster or fluorescence resonance energy transfer (FRET) enables to probe biomolecular interactions, thus playing a vital role in bioassays. However, conventional FRET platforms suffer from limited sensitivity due to the low FRET efficiency and poor anti-interference of existing FRET pairs. Here we report a NIR-II (1000–1700 nm) FRET platform with extremely high FRET efficiency and exceptional anti-interference capability. This NIR-II FRET platform is established based on a pair of lanthanides downshifting nanoparticles (DSNPs) by employing Nd3+ doped DSNPs as an energy donor and Yb3+ doped DSNPs as an energy acceptor. The maximum FRET efficiency of this well-engineered NIR-II FRET platform reaches up to 92.2%, which is much higher than most commonly used ones. Owing to the all-NIR advantage (λex = 808 nm, λem = 1064 nm), this highly efficient NIR-II FRET platform exhibits extraordinary anti-interference in whole blood, and thus enabling background-free homogeneous detection of SARS-CoV-2 neutralizing antibodies in clinical whole blood sample with high sensitivity (limit of detection = 0.5 μg/mL) and specificity. This work opens up new opportunities for realizing highly sensitive detection of various biomarkers in biological samples with severe background interference.

DNA tetrahedron-based split aptamer probes for reliable imaging of ATP in living cells

Accurate detection and imaging of adenosine triphosphate (ATP) expression levels in living cells is of great value for understanding cell metabolism, physiological activities, and pathologic mechanisms. Here, we developed a DNA tetrahedron-based split aptamer probe (TD probe) for ratiometric fluorescence imaging of ATP in living cells. The TD probe is constructed by hybridizing two split ATP aptamer probes (Apt-a and Apt-b) to a DNA tetrahedron assembled by four DNA oligonucleotides (T1, T2, T3, and T4). In the presence of ATP, the TD probe will alter its structure from the open to closed state, thus bringing the separated donor and acceptor fluorophores into close proximity for high fluorescence resonance energy transfer (FRET) signals. The TD probe exhibits low cytotoxicity, efficient cell internalization and good biological stability. Moreover, based on the FRET “off” to “on” signal output mode, the TD probe can effectively avoid false-positive signals from complex biological matrices, which is significant for long-term reliable imaging in living cells. In addition, by changing the split aptamers attached to DNA tetrahedron, the proposed strategy may be extended for detecting various intracellular targets. Collectively, this strategy provides a valuable sensing platform for biomarkers analysis in living cells, thus having great potential for early clinical diagnosis and therapeutic evaluation.

Photoswitchable Thermally Activated Delayed Fluorescence Nanoparticles for “Double‐Check” Confocal and Time‐Resolved Luminescence Bioimaging

AbstractThermally activated delayed fluorescence (TADF) materials have attracted increasing attention due to their great potential in time‐resolved luminescence imaging (TRLI) in bioimaging. However, the triplet state (T1) of delay fluorescence is quenched readily by oxygen and the autofluorescence of biological systems remains inevitable interference in luminescence imaging, which often reduces signal‐to‐noise ratio during biodetection. To address this issue, the authors design a two‐component photoswitchable TADF polymeric nanoparticle (PDFPNs) by covalently incorporating a well‐designed TADF molecule (PDC‐DA) and a photochromic spiropyran derivative (SPMA). The as‐prepared PDFPNs with a unique luminescent “double‐check” strategy exhibit dual‐color fluorescence and variable fluorescence lifetime upon fluorescence resonance energy transfer (FRET) process through external lights irradiation. The obtained PDFPNs display the advantages of negligible oxygen‐sensitivity, high FRET efficiency, rapid photoresponsiveness, outstanding photoreversibility, and prominent long‐term fluorescence stability. The established PDFPNs show promising application in the high‐contrast dual‐color confocal and time‐resolved luminescence bioimaging in oxygenic living cells.

A ratiometric fluorescent nanoprobe for signal amplification monitoring of intracellular telomerase activity.

Telomerase is considered a valuable diagnostic and prognostic cancer biomarker. Accurate and reliable detection of telomerase activity is of great value in clinical diagnosis, screening of inhibitors, and therapeutics. Here, we developed a novel amplified fluorescence resonance energy transfer (FRET) nanoprobe for highly sensitive and reliable monitoring of intracellular telomerase activity. The nanoprobe (QDSA@DNA) was composed of a streptavidin-modified quantum dot (QDSA) which was functionalized with a telomerase primer sequence (TP) and Cy5-tagged signal switching sequence (SS) through biotin-streptavidin interaction. When the nanoprobe was assembled, the Cy5 was in close proximity to the QDSA, resulting in high FRET efficiency from the QDSA to Cy5. In the presence of telomerase, the TP could be extended to produce telomeric repeat units, which was complementary to the loop of SS. Thus, the SS could hybridize with elongated sequences to form a rigid double-stranded structure, which forced the Cy5 away from the surface of the QDSA, causing low FRET efficiency. Furthermore, due to the production of multiple repeat units by telomerase, multiple hairpin structures could be opened, yielding significant fluorescence ratio (FQDsa/FCy5) enhancement for sensing of telomerase activity. In this way, the combination of a FRET and target-assisted strategy in a nanoprobe improved the detection accuracy and amplified the detection signal, respectively. The QDSA@DNA nanoprobe also showed high selectivity, excellent nuclease stability, and good biocompatibility. More importantly, this nanoprobe was found to be an excellent platform for efficient monitoring of intracellular telomerase activity, providing a potential platform in tumor diagnosis and screening of telomerase-related inhibitors.

Photon upconversion of all-inorganic CsPbX3 quantum dots based on fluorescence resonance energy transfer in hetero-structured perovskite/upconversion nanocomposites

Photo upconversion that converts low-energy light into high-energy photons holds great promise for boosting the energy conversion efficiencies particularly for photovoltaic devices based on all-inorganic CsPbX3 (X = Cl, Br, and I) perovskite quantum dots. However, the realization of nonlinear upconversion luminescence (UCL) from these all-inorganic CsPbX3 perovskites has been met with very limited success until now. Herein, we propose a rational perovskite/upconversion hetero-structured design strategy to achieve excitonic UCL of all-inorganic CsPbX3 perovskites based on fluorescence resonance energy transfer (FRET), where all-inorganic CsPbX3 perovskites and upconverting CaF2:Ln (Ln = Yb3+/Er3+, Yb3+/Ho3+, and Yb3+/Tm3+) nanocrystals are closely integrated with each other as a whole by using a modified two-step epitaxial growth approach, forming the unique upconverting CsPbX3/CaF2:Ln hetero-structured nanocomposites (HSNCs). Benefitting from the extremely high FRET efficiency of ∼99.7%, excitonic UCL from CsPbX3 perovskites can be realized upon low-power density 980-nm diode laser irradiation. More importantly, all the HSNCs are found to outperform their pure CsPbX3 counterparts in terms of air-stability, thermal-stability, and excitonic downconversion luminescence intensity, clearly demonstrating the overwhelming advantages of upconverting HSNCs we prepared over the pure CsPbX3 perovskites for high-performance optoelectronic and photovoltaic applications.

Albumin-based fluorescence resonance energy transfer nanoprobes for multileveled tumor tissue imaging and dye release imaging

Tumor tissue imaging and drug release imaging are both crucial for tumor imaging and image-guided drug delivery. It is urgent to develop a multileveled tumor imaging platform to realize the multiple imaging applications. In this work, we synthesized an albumin-based fluorescence resonance energy transfer (FRET) probe Cy5/7@HSA NPs containing two near-infrared cyanine dyes (CyBI5 and CyBI7) with high FRET efficiency (97 %). Excellent brightness and efficient FRET inside Cy5/7@HSA NPs enabled high-sensitive cell imaging and tumor imaging. This unique nanoprobe imaged 4T1 tumor-bearing mice with high sensitivity (TBR = 5.2) at 24 h post-injection and the dyes penetrated the tumor interior around 4 h post-injection. The release of dyes from nanoprobes was also tracked. This result shows the strong potential of this albumin-based FRET nanoprobe as multileveled tumor imaging platform for in vivo tumor imaging, drug delivery and image-guided surgery.

A sensitive biomolecules detection device with catalytic hairpin assembly and cationic conjugated polymer-assisted dual signal amplification strategy

A simple, sensitive, selective, and enzyme-free homogeneous fluorescent biosensing device for DNA and protein detection is fabricated based on catalytic hairpin assembly (CHA), cationic conjugated polymer (CCP), and graphene oxide (GO). In this biosensing device, CCP together with CHA, provides dual signal amplification, and GO suppresses the background when the target is absent. Thus, this CHA/CCP/GO-based biosensor shows improved sensitivity compared with conventional CHA-based biosensors. In the biosensor, two 6-carboxyfluorescein (FAM)-labeled hairpin DNA probes (H1 and H2) are designed, and in the initial state, they could absorb on the surface of GO, leading the system to produce a low background fluorescence signal. When the target DNA appears, it continually catalyzes the formation of H1–H2 double-stranded DNA (dsDNA) complex by CHA reaction, which could be regarded as the first-step amplification. At the same time, the H1–H2 dsDNA complex departures from the surface of GO and interacts with CCP through electrostatic interaction. Then, CCP provides the second-step amplification due to its high fluorescence resonance energy transfer (FRET) efficiency from CCP to FAM. The limit of detection (LOD) and the limit of quantification (LOQ) for the target DNA could reach 32 pM and 1 nM, respectively. The linear range was from 0.1 to 40 nM, and relative standard deviation (RSD) for the points on the calibration curve ranged from 2.8% to 13.9%. This strategy could also be applied to protein detection potentially by integrating the aptamer of the target protein into the hairpin DNA. As proof of concept, thrombin was detected, and the LOD and LOQ was 11 pM and 33 pM, respectively. The linear range was from 3 to 54 nM, and RSD ranged from 3.3% to 10.4%. It showed good selectivity for thrombin compared to equal concentrations of interferences. It was also applied to quantify the thrombin (5, 10, 20 nM) in 1% spiked human serum, which showed satisfying recovery in the range of 94.7 ± 5.3 to 103.7 ± 4.9%.

Simplified Instrument Calibration for Wide-Field Fluorescence Resonance Energy Transfer (FRET) Measured by the Sensitized Emission Method.

Fӧrster (or fluorescence) resonance energy transfer (FRET) is a quantifiable energy transfer in which a donor fluorophore nonradiatively transfers its excitation energy to an acceptor fluorophore. A change in FRET efficiency indicates a change of proximity and environment of these fluorophores, which enables the study of intermolecular interactions. Measurement of FRET efficiency using the sensitized emission method requires a donor-acceptor calibrated system. One of these calibration factors named the G factor, which depends on instrument parameters related to the donor and acceptor measurement channels and on the fluorophores quantum efficiencies, can be determined in several different ways and allows for conversion of the raw donor and acceptor emission signals to FRET efficiency. However, the calculated value of the G factor from experimental data can fluctuate significantly depending on the chosen experimental method and the size of the sample. In this technical note, we extend the results of Gates et al. (Cytometry Part A 95A (2018) 201-213) by refining the calibration method used for calibration of FRET from image pixel data. Instead of using the pixel histograms of two constructs with high and low FRET efficiency to determine the G factor, we use pixel histogram data from one construct of known efficiency. We validate this method by determining the G factor with the same constructs developed and used by Gates et al. and comparing the results from the two approaches. While the two approaches are equivalent theoretically, we demonstrate that the use of a single construct with known efficiency provides a more precise experimental measurement of the G factor that can be attained by collecting a smaller number of images. © 2020 International Society for Advancement of Cytometry.

Ultrasound and Near-Infrared Light Dual-Triggered Upconversion Zeolite-Based Nanocomposite for Hyperthermia-Enhanced Multimodal Melanoma Therapy via a Precise Apoptotic Mechanism.

High reactive oxygen species (ROS) generation efficiency and induction of targeted cell apoptosis are recognized as key objectives to achieve a highly efficient strategy for cancer therapy with minimum side effects of inflammatory reactions. However, it is still a challenge to realize higher therapeutic efficiency with a cell apoptosis model. Herein, we present strong upconversion luminescent biosafe cores derived from Linde Type A (LTA) zeolites and modification with targeted/therapeutic drugs for multimodal therapy, in which sonodynamic therapy (SDT) combined with photodynamic therapy (PDT) increases therapeutic efficiency especially in deep sites of tumor via producing cytoplasmic ROS and mitochondrial superoxide and photothermal therapy (PTT) enhances PDT effects via higher fluorescence resonance energy transfer (FRET) efficacy attributed to an increased temperature. Furthermore, the transcriptomic analysis reveals that cellular internalization of the nanosystem can lead to tumor ablation via cell apoptosis. We expect that the multimodal therapy based on LTA zeolite drug nanocarriers could be applied in the cancer therapeutics in the near future.

Quaternary Supramolecular Nanoparticles as a Photoerasable Luminescent Ink and Photocontrolled Cell‐Imaging Agent

AbstractPhotoswitchable luminescent supramolecular assemblies based on cyclodextrins have attracted considerable attention owing to their potential applications as smart materials, but most of the assemblies reported to date emit green or blue light with low contrast and high interference. In this study, novel photoluminescent red‐luminescent quaternary supramolecular nanoparticles (2) are constructed from a dithienylethene derivative (1), a β‐cyclodextrin‐functionalized ruthenium complex (Ru‐HOP‐CD), Pluronic F‐127, and cetrimonium bromide. Compared with the binary assembly 1@Ru‐HOP‐CD, the quaternary nanoparticles exhibit high fluorescence resonance energy transfer efficiency, with Ru‐HOP‐CD acting as the donor and 1 as the acceptor. Owing to the reversible photoswitched interconversion of the two forms of the dithienylethene component, the fluorescence of the nanoparticles could be switched on/off by irradiation with UV or visible light, both in solution and in the solid state. As a result, the nanoparticles could be used as a photoerasable red‐luminescent ink and as a photocontrolled cell‐imaging agent. These functional nanoparticles can be expected to be useful in the fields of information security and biology.

N-Glycans Mediate the Ebola Virus-GP1 Shielding of Ligands to Immune Receptors and Immune Evasion.

The Ebola Virus (EBOV) glycoprotein (GP) sterically shields cell-membrane ligands to immune receptors such as human leukocyte antigen class-1 (HLA-I) and MHC class I polypeptide-related sequence A (MICA), thus mediating immunity evasion. It was suggested that the abundant N-glycosylation of the EBOV-GP is involved in this steric shielding. We aimed to characterize (i) the GP N-glycosylation sites contributing to the shielding, and (ii) the effect of mutating these sites on immune subversion by the EBOV-GP. The two highly glycosylated domains of GP are the mucin-like domain (MLD) and the glycan cap domain (GCD) with three and six N-glycosylation sites, respectively. We mutated the N-glycosylation sites either in MLD or in GCD or in both domains. We showed that the glycosylation sites in both the MLD and GCD domains contribute to the steric shielding. This was shown for the steric shielding of either HLA-I or MICA. We then employed the fluorescence resonance energy transfer (FRET) method to measure the effect of N-glycosylation site removal on the distance in the cell membrane between the EBOV-GP and HLA-I (HLA.A*0201 allele). We recorded high FRET values for the interaction of CFP-fused HLA.A*0201 and YFP-fused EBOV-GP, demonstrating the very close distance (<10 nm) between these two proteins on the cell membrane of GP-expressing cells. The co-localization of HLA-I and Ebola GP was unaffected by the disruption of steric shielding, as the removal of N-glycosylation sites on Ebola GP revealed similar FRET values with HLA-I. However, these mutations directed to N-glycosylation sites had restored immune cell function otherwise impaired due to steric shielding over immune cell ligands by WT Ebola GP. Overall, we showed that the GP-mediated steric shielding aimed to impair immune function is facilitated by the N-glycans protruding from its MLD and GCD domains, but these N-glycans are not controlling the close distance between GP and its shielded proteins.

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.

Enhanced Energy Transfer in a Donor-Acceptor Photosensitizer Triggers Efficient Photodynamic Therapy.

Photosensitizers (PSs) play a vital role in photodynamic therapy (PDT) for combating bacterial resistance and treating tumor. In this study, we report new donor-acceptor porphyrin PSs with a cationic conjugated oligomer (OPV) as a donor unit and porphyrin (TPP) as an acceptor unit by covalent linkage and achieved a fluorescence resonance energy transfer efficiency of 99% owing to their strong spectral overlap and short distance. The 1O2 yield of porphyrin derivatives is 121% (rose bengal as the standard reference) by virtue of OPVs' excellent light-harvesting ability and high fluorescence resonance energy transfer efficiency, greatly exceeding those of oligomer and porphyrin derivatives reported in the literature. Additionally, the cationic donors significantly improved the water solubility, decreased the aggregation of porphyrin, and promoted the adherence of the PSs to cell membranes through electrostatic interactions. As a result, the D-A porphyrin PSs exhibit dramatic PDT treatment efficiency. The half-inhibitory concentration is as low as 33 and 88 nM for methicillin-resistant Staphylococcus aureus and Escherichia coli, respectively. Therefore, this study provides a new strategy to construct PSs with high 1O2 yield and an excellent treatment effect at a low dose of PSs, which is promising for application in PDT used to treat cancer and microbial infections.

Nanolantern-Based DNA Probe and Signal Amplifier for Tumor-Related Biomarker Detection in Living Cells.

The introduction of nanotechnology can overcome some inherent drawbacks of traditional DNA probes, thus promoting their applications in living cells. Herein, a three-dimensional DNA nanostructure, a DNA nanolantern, was prepared via simple nucleotide hybridization of four short-stranded oligonucleotides and successfully applied to the construction of a novel DNA probe and signal amplifier. Compared to most reported DNA nanostructures, a DNA nanolantern shows the distinct advantages of low cost, easy design and preparation, more and arbitrary adjusted probe numbers, and high fluorescence resonance energy transfer (FRET) signal readout. Compared to traditional DNA probes, the constructed nanolantern-based one has improved cell internalization efficiency, enhanced biostability, accelerated reaction kinetics, excellent biocompatibility, and greatly reduced false-positive output and was demonstrated to work well for probing the expression level of tumor-related mRNA and microRNA in living cells. The DNA nanolantern can also be easily integrated with some reported signal amplification strategies, e.g., isothermal hybridization chain reaction (HCR), and the obtained signal amplifier combines the advantages of the DNA nanolantern and the HCR, enabling sensitive imaging detection of ultralow abundance targets in living cells. This work demonstrated that this simple DNA nanostructure can not only improve the performance of traditional DNA probes but can also be easily integrated with reported DNA-based strategy and technology, thus showing a broad application prospect.

Cell-Surface-Anchored Ratiometric DNA Nanoswitch for Extracellular ATP Imaging.

The precise detection of extracellular ATP, although a challenging task, is of great significance for understanding the related cell processes. Herein, we developed a ratiometric DNA nanoswitch by employing a DNA tweezer and split aptamer. The nanoswitch is composed of three specially designed ssDNA strands, namely, the central strands O1, O2, and O3. This nanoswitch can be anchored on the cell membrane by cholesterol labeled at the 3' end of O3. Initially, the DNA tweezer adopts an open state, separating the dual fluorophores and giving rise to a low FRET (fluorescence resonance energy transfer) ratio. The presence of ATP induces the binding of the two split aptamers to alter the structure of the nanoswitch from the open state to the closed state, bringing the donor and the acceptor closer together and generating high FRET efficiency. The results demonstrated that the ratiometric DNA nanoswitch can be applied for quantitative analysis and real-time monitoring of the changes in extracellular ATP. We believe that the cell surface-anchored DNA nanoswitch has promising prospects for use as a powerful tool for the understanding of different ATP-related physiological activities.

Development of Novel Intramolecular FRET-Based ABC Transporter Biosensors to Identify New Substrates and Modulators.

Multidrug resistance protein 1 (MRP1) can efflux a wide variety of molecules including toxic chemicals, drugs, and their derivatives out of cells. Substrates of MRP1 include anti-cancer agents, antibiotics, anti-virals, anti-human immunodeficiency virus (HIV), and many other drugs. To identify novel substrates and modulators of MRP1 by exploiting intramolecular fluorescence resonance energy transfer (FRET), we genetically engineered six different two-color MRP1 proteins by changing green fluorescent protein (GFP) insertion sites, while keeping the red fluorescent protein (RFP) at the C-terminal of MRP1. Four of six recombinant proteins showed normal expression, localization, and transport activity. We quantified intramolecular FRET using ensemble fluorescence spectroscopy in response to binding of known substrate or ATP alone, substrate/ATP, and trapping of the transporter in closed conformation by vanadate. Recombinant MRP1 proteins GR-881, GR-888, and GR-905 exhibited reproducible and higher FRET changes under all tested conditions and are very promising for use as MRP1 biosensors. Furthermore, we used GR-881 to screen 40 novel anti-cancer drugs and identified 10 hits that potentially directly interact with MRP1 and could be substrates or modulators. Profiling of drug libraries for interaction with MRP1 can provide very useful information to improve the efficacy and reduce the toxicity of various therapies.

From a Molecular Toolbox to a Toolbox for Photoswitchable Fluorescent Polymeric Nanoparticles

AbstractSwitchable organic fluorescent materials are attracting much interest in many fields including flexible display, information storage, anti‐counterfeit, and bioimaging, due to their prominent properties and relatively low cost. Herein, a class of novel photoswitchable fluorescent polymeric nanoparticles (PFPNs) toolbox that shows switchable full‐colored emission including white light is described. This nanoparticles toolbox consists of a series of reversibly photoswitchable and non‐PFPNs with red, green, and blue fluorescence, and is built up from three primary fluorescent dyes and a photochromic diarylethene molecule, through a facile one‐pot miniemulsion method. The as‐prepared PFPNs display the merits of high fluorescence resonance energy transfer efficiency and fluorescence quantum yield, rapid responsiveness, prominent photoreversibility, and brilliant long‐term fluorescence stability (≈6 weeks). They enable switchable emission between white light and any visible color on‐demand in both solution and film states. Their potential in complex fluorescent encryption and photoswitchable white light‐emitting diode is described and its great potential in anti‐counterfeiting technology, data encryption, and the next generation of optoelectronic materials is foreseen.

Two-Photon Excited FRET Dyads for Lysosome-Targeted Imaging and Photodynamic Therapy.

Two-photon excitable fluorescent dyes with integrated functions of targeted imaging and photodynamic therapy (PDT) are highly desired for the development of cancer theranostic agents. Herein, fluorescence resonance energy transfer (FRET) dyads, AceDAN-H2Por-Lyso (1a) and AceDAN-ZnPor-Lyso (1b), were developed for two-photon excited (TPE) lysosome-targeted fluorescence imaging and PDT of cancer cells. Under one-photon or two-photon excitation, the AceDAN donor can effectively transfer the excited state energy to the porphyrin acceptor via high efficient FRET, leading to the generation of deep-red fluorescence and singlet oxygen for cell imaging and PDT, respectively. 1a and 1b exhibit high photocytotoxicity and low dark cytotoxicity, in addition to strong lysosomal targeting capability in living cells. By taking the advantages of the two-photon absorption properties of the AceDAN donor and the properly distributed S1 and T1 states of the porphyrin acceptor, the AceDAN-porphyrin dyads 1a and 1b have been successfully applied to TPE-fluorescence imaging for tracking the significant morphology changes of cancer cells under two-photon laser irradiation.

A Ratiometric Fluorescent Bioprobe Based on Carbon Dots and Acridone Derivate for Signal Amplification Detection Exosomal microRNA.

Recently, sensitive and selective detection of exosomal microRNAs (miRNAs) has been garnering significant attention, because it is related to many complex diseases, including cancer. Herein, we report a ratiometric fluorescent bioprobe based on DNA-labeled carbon dots (DNA-CDs) and 5,7-dinitro-2-sulfo-acridone (DSA) coupling with the target-catalyzing signal amplification for the detection of exosomal miRNA-21. There was high fluorescence resonance energy transfer (FRET) efficiency between carbon dots (CDs) and DSA when the bioprobe was assembled. However, in the presence of the target, with disassembling of the fluorescent bioprobe, the fluorescence intensities of CDs and DSA were changed simultaneously. Because of the ratio of dual fluorescence intensities, this ratiometric fluorescent bioprobe was able to cancel out environmental fluctuations by calculating emission intensity ratio at two different wavelengths, being robust and stable enough for detection of exosomal miRNA-21. In addition, we displayed that a single miRNA-21 can catalyze the disassembly of multiple CDs with DSA theoretically, yielding significant change in the fluorescence ratio for the detection of miRNA-21. With this signal amplification strategy, the limit of detection was as low as 3.0 fM. Furthermore, because of the introduction of lock nucleic acid to mediate the strand displacement reaction, the selectivity of this strategy was improved remarkably, even against single base mismatch sequence. More importantly, our strategy could monitor the dynamic change of exosomal miRNA-21, which maybe becomes a potential tool to distinguish cancer exosomes and nontumorigenic exosomes. In a short, this ratiometric fluorescence bioprobe possessed high stability, sensitivity and selectivity coupling with ease of operation and cost efficiency, leading to great potential for wide application.

Synthesis, photophysics of a novel green light emitting 1,3,4-oxadiazole and its application in FRET with ZnSe/ZnS QDs donor

The design and synthesis of a novel molecule 2-(4-((E)-2-(5-((E)-4-(5-(4-tert-butylphenyl)-1,3,4-oxadiazol-2-yl)styryl)thiophen-2-yl)vinyl)-5-(4-tert-butylphenyl)1,3,4-oxadiazole (2TVPO) based on Donor-π-Acceptor strategy through the Wittig reaction as well as characterization by 1H, 13C NMR spectroscopy, high resolution mass spectrometry and FT-IR spectroscopy form the central theme of the present work. Thermal (TGA and DSC), photophysical and electrochemical properties have been studied in detail for this novel probe. In addition, fluorescence resonance energy transfer (FRET) employing steady state and time resolved spectroscopy methods has been studied using core-shell ZnSe/ZnS QDs (7 and 8nm) and 2TVPO fluorescent probe, respectively, as the energy donor and acceptor. Results of these studies reveal that (i) FRET pairs show a strong dipole-dipole interaction tendency to transfer energy from donor to acceptor; (ii) they possess high FRET efficiency and (iii) strong dependence of the acceptor emission on the overlap integral, quantum dot size and donor-acceptor distance. Further, the type of quenching is analyzed by Stern–Volmer plots.