Fluorescent AIE dots encapsulated organically modified silica (ORMOSIL) nanoparticles for two-photon cellular imaging

2,3-Bis(4-(phenyl(4-(1,2,2-triphenylvinyl)phenyl)amino)phenyl) fumaronitrile (TPE-TPA-FN or TTF), which possesses aggregation-induced emission (AIE) characteristic, is doped in organically modified silica (ORMOSIL) nanoparticles. By increasing the weight ratio of TTF to the precursor of silica nanoparticles (the quantities of the precursors were kept the same), the fluorescence intensity of nanoparticles increased correspondingly, due to the formation of larger AIE dots in the cores of ORMOSIL nanoparticles. The fluorescent and biocompatible nanoprobes were then utilized for in vitro imaging of HeLa cells. Two-photon fluorescence microscopy clearly illustrated that the nanoparticles have the capacity of nucleus permeability, as well as cytoplasm staining towards tumor cells. Our experimental results may offer a promising method for fast and bright fluorescence imaging, as well as bio-molecule/drug delivery to cell nucleus.     

Light-Induced Self-Escape of Spherical Nucleic Acid from Endo/Lysosome for Efficient Non-Cationic Gene Delivery

Developing non-cationic gene carriers and achieving efficient endo/lysosome escape of functional nucleic acids in cytosol are two major challenges faced by the field of gene delivery. Herein, we demonstrate the concept of self-escape spherical nucleic acid (SNA) to achieve light controlled non-cationic gene delivery with sufficient endo/lysosome escape capacity. In this system, Bcl-2 antisense oligonucleotides (OSAs) were conjugated onto the surface of aggregation-induced emission (AIE) photosensitizer (PS) nanoparticles to form core–shell SNA. Once the SNAs were taken up by tumor cells, and upon light irradiation, the accumulative ¹O₂ produced by the AIE PSs ruptured the lysosome structure to promote OSA escape. Prominent in vitro and in vivo results revealed that the AIE-based core–shell SNA could downregulate the anti-apoptosis protein (Bcl-2) and induce tumor cell apoptosis without any transfection reagent.     

Biocompatible and noncytotoxic nucleoside-based AIEgens sensor for lighting-up nucleic acids

Aggregation-induced emission luminogens (AIEgens) have been used in biomacromolecules detection. Herein, TPE-dC and TPE-dU acted as the nucleoside-based AIEgens sensors in the first case, which can be used to detect ctDNA and rRNA in vitro and light up the nucleus in vivo depending on the intermolecular binding affinity. This AIE process enables the quantitative analysis or visualization of nucleic acids in solution or gels state, respectively. Furthermore, confocal laser scanning microscopy (CLSM) images of L929 cells stained with TPE-dC or TPE-dU clearly shows that nucleoside-based AIEgens bio-probes can pass the cell membranes to reach the cell nucleus, without cytotoxicity at the imaging condition (incubation time > 12 h, and 10 μmol/L of concentration). Since the nucleus is rich in DNA/RNA, fluorescence turn-on mode has a great potential in nucleus imaging and clinical diagnosis.     

Conjugated polymer nanoparticles with aggregation induced emission characteristics for intracellular Fe3+ sensing

In this article, a novel zwitterionic conjugated polyelectrolyte containing tetraphenylethene unit was synthesized via Pd‐catalyzed Sonogashira reaction. The resulting polymer (P2), which exhibited typical aggregation‐induced emission (AIE) properties, was weakly fluorescent in dilute DMSO solution and showed bright fluorescence emissions when aggregated in DMSO/water mixtures or fabricated into conjugated polymer nanoparticles (CPNs). The nanoparticles from P2 could be prepared by reprecipitation method with an average diameter around 23 nm. Notably, the cell‐staining efficiencies of lipid‐P2 nanoparticles could be enhanced with lipid encapsulation and these nanoparticles were endocytosed via caveolae‐mediated and clathrin‐mediated endocytosis pathways. Furthermore, the lipid‐P2 nanoparticles with low cytotoxicity, high photostability and efficient cell staining ability could be employed for in vitro detection of Fe³⁺ ions in A549 cells. © 2016 Wiley Periodicals, Inc. J. Polym. Sci., Part A: Polym. Chem. 2016 , 54, 1686–1693     

Determination of nucleic acids based on the fluorescence quenching of Hoechst 33258 at pH 4.5

It was found the strong fluorescence emitted by the bis-benzimidazole derivative Hoechst 33258 at 490 nm could be efficiently quenched in pH 4.5 buffer when nucleic acids were added. Analysis of fluorescence intensity showed that the procedure was a static quenching dominated one, which was also demonstrated by the electron absorption spectra and lifetime of the excited state. The binding constant and numbers of binding sites were obtained from the Scatchard plot. The decreased fluorescence intensity was in proportion to the concentration of nucleic acids in the range 40-1800 ng ml⁻¹ for dsDNA and 26-1700 ng ml⁻¹ for ssDNA. The limits of detection were 12 and 8 ng ml⁻¹, respectively. The sensitivity of the method was about 3.4 times higher for dsDNA detection and 5.4 times higher for ssDNA detection compared with the widely used fluorescence enhancement method using the same dye. Application results to synthetic samples showed simplicity, rapidity and satisfactory reproducibility of the presented method. Measurement of real samples extracted from leaves of Crassula argentea and E. coli genome also gave satisfactory results, which were in good agreement with those obtained using spectrophotometric method.     

Tuning the singlet-triplet energy gap: a unique approach to efficient photosensitizers with aggregation-induced emission (AIE) characteristics

The efficiency of the intersystem crossing process can be improved by reducing the energy gap between the singlet and triplet excited states (ΔE _ST), which offers the opportunity to improve the yield of the triplet excited state. Herein, we demonstrate that modulation of the excited states is also an effective strategy to regulate the singlet oxygen generation of photosensitizers. Based on our previous studies that photosensitizers with aggregation-induced emission characteristics (AIE) showed enhanced fluorescence and efficient singlet oxygen production in the aggregated state, a series of AIE fluorogens such as TPDC, TPPDC and PPDC were synthesized, which showed ΔE _ST values of 0.48, 0.35 and 0.27 eV, respectively. A detailed study revealed that PPDC exhibited the highest singlet oxygen efficiency (0.89) as nanoaggregates, while TPDC exhibited the lowest efficiency (0.28), inversely correlated with their ΔE _ST values. Due to their similar optical properties, TPDC and PPDC were further encapsulated into nanoparticles (NPs). Subsequent surface modification with cell penetrating peptide (TAT) yielded TAT–TPDC NPs and TAT–PPDC NPs. As a result of the stronger singlet oxygen generation, TAT–PPDC NPs showed enhanced cancer cell ablation as compared to TAT–TPDC NPs. Fine-tuning of the singlet-triplet energy gap is thus proven to be an effective new strategy to generate efficient photosensitizers for photodynamic therapy.     

Self-assembled fluorescent tripeptide nanoparticles for bioimaging and drug delivery applications

Peptide self-assembled nanomaterials have attracted more and more attention due to their wide applications such as drug delivery, cell imaging, and real-time drug monitoring. However, the application of the peptide is still limited by its inherent optical properties. Here we proposed and prepared a series of fluorescent tripeptide nanoparticles (TPNPs) through π-π stacking and zinc coordination. The experimental results show that the nanoparticles (TPNPs1) formed by the self-assembly of the tripeptide tryptophan-tryptophan-tryptophan have the highest fluorescence intensity, uniform and appropriate size, and low cytotoxicity. Furthermore, there was fluorescence resonance between TPNPs1 and doxorubicin, which has been successfully applied for real-time cell imaging and drug release monitoring.     

Efficient enhancement of fluorescence emission via TPE functionalized cationic pillar[5]arene-based host–guest recognition-mediated supramolecular self-assembly

A tetraphenylethene (TPE) functionalized cationic pillar[5]arene (CWP5-TPE) was successfully synthesized, and the intramolecular rotation of the TPE motif was restricted via cationic pillar[5]arene-based host–guest recognition-mediated supramolecular self-assembly in water, resulting in the efficient enhancement of fluorescence emission based on the aggregation induced emission (AIE) mechanism. CWP5-TPE self-assembled into nanoribbons while the host–guest inclusion complex formed into supramolecular amphiphile nanoparticles in water.     

A self-assembled tetrapeptide that acts as a “turn-on” fluorescent sensor for Hg2+ ion

The tetrapeptide (Bz-$\mathrm{\Delta }$Phe(p-NPh₂)-l-DOPA(protected)-l-Phe-l-Phe-OMe was designed to incorporate seven phenyl rings so that it’s conformation, self-assembly and application in Hg²⁺ ions sensing could be studied. Peptide molecules adopted an overlapping β-turn of type III/III conformation in crystals. The peptide showed a highly selective turn-on response towards mercuric ion over other metal ions with a 10-fold enhancement in fluorescence intensity. This intensity change coupled with the selectivity of the peptide towards mercury allowed us to demonstrate simple colorimetric dip sensing of Hg²⁺ ions. The technique provides a highly selective and effective way to detect Hg²⁺ ions. The peptide also self-assembled into nanospheres with diameter ranges from 100 to 500?nm. Mercuric ion coordination enabled these peptide nanospheres to aggregate into well-defined nanoparticles. The enhanced fluorescence upon Hg²⁺ addition demonstrates that peptide scaffolds can be exploited in the development of different selective sensors.     

Bioactivated in vivo assembly(BIVA) peptide-tetraphenylethylene(TPE) probe with controllable assembled nanostructure for cell imaging

The emergence of fluorescent light-up molecular probe, which can specifically turn on their fluorescent in the presence of stimulation factors, has open up a new opportunity to advance biosensing and bioimaging. In this work, we designed and synthesized a peptide-AIE conjugate probe for cell imaging with controlled in situ assembled nanostructures. The modular designed probe is consisted of a selfassembled peptide-tetraphenylethene(TPE) motif, a fibroblast activation protein alpha(FAP-α)responsive motif, a hydrophilic motif and a targeting motif. The probe exhibits typically turn-on fluorescence property specifically triggered by FAP-α, which is a significant overexpressed membrane protein on pancreatic tumor cells. Interestingly, the peptide modified the TPE dramatically impacts the assembled nanostructure, which can be modulated by peptide sequences. As a result, the peptide FF(PhePhe) modification of TPE as the self-assembled motif provides a suitable balance of the probe with lightup property and nanofiber assembled structure in situ. Finally, our probe could effectively detect the FAP-α on tumor cells with high specificity. Meantime, the nanofibers in situ assembled on the surface of CAFs enhanced the probe accumulation and prolonged the retention for cell imaging. We envision that this study may inspire new insights into the design of nanostructure controlled AIE light-up bio-probe.     

Biocompatible organic dots with aggregation-induced emission for in vitro and in vivo fluorescence imaging

Fluorescent probes play a key role in modern biomedical research. As compared to inorganic quantum dots (QDs) composed with heavy metal elements, organic dye-based fluorescent nanoparticles have higher biocompatibility and are richer in variety. However, traditional organic fluorophores tend to quench fluorescence upon aggregation, which is known as aggregation-caused quenching (ACQ) effect that hinders the fabrication of highly emissive fluorescent nanoparticles. In this work, we demonstrate the synthesis of organic fluorescent dots with aggregation-induced emission (AIE) in far-red/near-infrared (FA/NIR) region. A conventional ACQ-characteristic fluorescent dye, 3,4:9,10-tetracarboxylic perylene bisimide (PBI), is converted into an AIE fluorogen through attaching two tetraphenylethylene (TPE) moieties. The fluorescent dots with surface folic acid groups are fabricated from PBI derivative (DTPEPBI), showing specific targeting effect to folate receptor-overexpressed cancer cells. In vivo studies also suggest that the folic acid-functionalized AIE dots preferentially accumulate in the tumor site through enhanced permeability and retention (EPR) effect and folate receptor-mediated active targeting effect. The low cyto-toxicity, good FR/NIR contrast and excellent targeting ability in in vitro/in vivo imaging indicate that the AIE dots have great potentials in advanced bioimaging applications.     

One-step preparation of branched PEG functionalized AIE-active luminescent polymeric nanoprobes

The synthesis of amphiphilic aggregation-induced emission(AIE) dyes based organic nanoparticles has recently attracted increasing attention in the biomedical fields. These AIE dyes based nanoparticles could effectively overcome the aggregation caused quenching effect of conventional organic dyes, making them promising candidates for fabrication of ultrabright organic luminescent nanomaterials. In this work, AIE-active luminescent polymeric nanoparticles(4-NH_2-PEG-TPE-E LPNs) were facilely fabricated through Michael addition reaction between tetraphenylethene acrylate(TPE-E) and 4-arm-poly(ethylene glycol)-amine(4-NH_2-PEG) in rather mild ambient. The 4-NH_2-PEG can not only endow these AIE-active LPNs good water dispersibility, but also provide functional groups for further conjugation reaction. The size, morphology and luminescent properties of 4-NH_2-PEG-TPE-E LPNs were characterized by a series of techniques in detail. Results suggested that these AIE-active LPNs showed spherical morphology with diameter about 100–200 nm. The obtained 4-NH_2-PEG-TPE-E LPNs display high water dispersibility and strong fluorescence intensity because of their self assembly and AIE properties of TPE-E.Biological evaluation results demonstrated that 4-NH_2-PEG-TPE-E LPNs showed negative toxicity toward cancer cells and good fluorescent imaging performance. All of these features make 4-NH_2-PEG-TPE-E LPNs promising candidates for biological imaging and therapeutic applications.     

Mapping Live Cell Viscosity with an Aggregation‐Induced Emission Fluorogen by Means of Two‐Photon Fluorescence Lifetime Imaging

Intracellular viscosity is a crucial parameter that indicates the functioning of cells. In this work, we demonstrate the utility of TPE‐Cy, a cell‐permeable dye with aggregation‐induced emission (AIE) property, in mapping the viscosity inside live cells. Owing to the AIE characteristics, both the fluorescence intensity and lifetime of this dye are increased along with an increase in viscosity. Fluorescence lifetime imaging of live cells stained with TPE‐Cy reveals that the lifetime in lipid droplets is much shorter than that from the general cytoplasmic region. The loose packing of the lipids in a lipid droplet results in low viscosity and thus shorter lifetime of TPE‐Cy in this region. It demonstrates that the AIE dye could provide good resolution in intracellular viscosity sensing. This is also the first work in which AIE molecules are applied in fluorescence lifetime imaging and intracellular viscosity sensing.     

Highlights on the Road towards Highly Emitting Solid‐State Luminophores: Two Classes of Thiazole‐Based Organoboron Fluorophores with the AIEE/AIE Effect

Developing a novel, small‐sized molecular building block that may be capable of emitting light in the solid state is a challenging task and has rarely been reported in the literature. BF₂‐containing dyes seem to be promising candidates towards this aim. Two series of new N^NBF₂ complexes showing aggregation‐induced emission (AIE) and aggregation‐induced emission enhancement (AIEE) were designed and synthesized by means of a new protocol, which improved on the traditional method by employing microwave irradiation. The optical and photophysical properties of the BF₂ complexes were investigated in depth. The synthesized complexes showed fluorescence in both solution and the solid state and, in a mixture of tetrahydrofuran/water, may aggregate into fluorescent nanoparticles. The experimental investigation was supported by quantum mechanical calculations. Their availability, stability, large Stokes shifts, and aggregation capabilities, along with their solid‐state emission capability, render this new class of BF₂ complexes promising AIEE/AIE fluorophores for further applications in the fields of fluorescence imaging and materials science.     

Novel fluorescent colloids as a DNA fluorescence probe

Fluorescent perylene colloids in the 80–90 nm size range have been prepared by the reprecipitation method. These nanoparticles were modified by cetyltrimethylammonium bromide (CTAB) which inhibited their growth. The nanoparticles also readily interacted with DNA. The fluorescence emission was measured at _ex/_em=400/565 nm. The fluorescence decrease of colloid–CTAB in aqueous solution was measured in the presence of nucleic acids. Under the optimum conditions, the ratio of fluorescence intensity in the absence and presence of nucleic acids was proportional to the concentration of nucleic acids over the range 0.02–5.1 µg mL^–1 for FS (fish sperm) DNA or CT (calf thymus) DNA. The detection limits were 0.01 µg mL^–1 for FS DNA and 0.012 µg mL^–1 for CT DNA, respectively. Based on this approach, a new quantitative method for DNA assay is presented in this paper.     

Peptide nucleic acid stabilized perovskite nanoparticles for nucleic acid sensing

Nanostructural hybrid organic-inorganic metal halide perovskites offer a wide range of potential applications including photovoltaics, solar cells, and light emitting diodes. Up to now the surface stabilizing ligands were used solely to obtain the optimal properties of nanoparticles in terms of dimensionality and stability, however their possible additional functionality was rarely considered. In the present work, hybrid lead bromide perovskite nanoparticles (PNP) were prepared using a unique approach where a peptide nucleic acid is used as a surface ligand. Methylammonium lead bromide perovskite colloidal nanoparticles stabilized by thymine-based peptide nucleic acid monomer (PNA-M) and relevant trimer (PNA-T) were prepared exhibiting the size below 10 nm. Perovskite structure and crystallinity were verified by X-ray powder diffraction spectroscopy and high resolution transmission electron microscopy. PNP-PNA-M and PNP-PNA-T colloidal dispersions in chloroform and toluene possessed green-blue fluorescence, while Fourier-transform infrared spectroscopy (FT-IR) and quantum chemical calculations showed that the PNA coordinates to the PNP surface through the primary amine group. Additionally, the sensing ability of the PNA ligand for adenine nucleic acid was demonstrated by photoluminescence quenching via charge transfer. Furthermore, PNP thin films were effectively produced by the centrifugal casting. We envision that combining the unique, tailored structure of peptide nucleic acids and the prospective optical features of lead halide perovskite nanoparticles could expand the field of applications of such hybrids exploiting analogous ligand chemistry.     

Design and synthesis of an AIE-active fluorogen with red emission and its biological application

A novel triphenylamine-base derivative L containing pyridine and terpyridine was designed and synthesized. Compound L exhibited distinct aggregation-induced emission (AIE) behavior in water–ethanol and also displayed a threefold increase in the intensity of luminescence at 608 nm. Furthermore, confocal microscopy imaging demonstrated that compound L displays low toxicity and brights red fluorescence in mitochondria in living HepG2 cells. Inherent from the mitochondrial-targeting ability of pyridine moiety and the AIE characteristic of triphenylamine group, compound L could be employed as a fluorescent probe in the near-infrared region for living cell imaging.     

Peptide nucleic acid molecular beacons for the detection of PCR amplicons in droplet-based microfluidic devices

The use of droplet-based microfluidics and peptide nucleic acid molecular beacons for the detection of polymerase chain reaction (PCR)-amplified DNA sequences within nanoliter-sized droplets is described in this work. The nanomolar–attomolar detection capabilities of the method were preliminarily tested by targeting two different single-stranded DNA sequences from the genetically modified Roundup Ready soybean and the Olea europaea genomes and detecting the fluorescence generated by peptide nucleic acid molecular beacons with fluorescence microscopy. Furthermore, the detection of 10 nM solutions of PCR amplicon of DNA extracted from leaves of O. europaea L. encapsulated in nanoliter-sized droplets was performed to demonstrate that peptide nucleic acid molecular beacons can discriminate O. europaea L. cultivar species carrying different single-nucleotide polymorphisms.

Highly enantioselective recognition of a wide range of carboxylic acids based on enantioselectively aggregation-induced emission

A new chiral fluorescence receptor, designed to exploit aggregation-induced emission (AIE), exhibited not only an exceptionally high enantioselectivity but was also applicable to a wide range of chiral carboxylic acids. The fluorescence intensity ratio of two enantiomers ranged from 10 to 1.6 × 10(4) and could be used to determine the purity of the enantiomers.     

TPE based aggregation induced emission fluorescent sensors for viscosity of liquid and mechanical properties of hydrogel

Two amphiphilic TPE E/Z isomers with aggregation induced emission(AIE) property have been synthesized and characterized. The logarithmic fluorescent intensity of the two molecules was in positive relationship with logarithmic viscosity of liquid. To note, the Z-TPE isomer exhibited more sensitivity in the viscosity of liquid sensing in comparison with the corresponding E-TPE counterpart(around 1.80 folds).Furthermore, two molecules could be used as fluorescent sensors for mechanical properties(v...