Fluorescent Semiconducting Polymer Dots Research Articles

Ratiometric fluorescent semiconducting polymer dots for temperature sensing.

Semiconducting polymer dots (Pdots) have received much attention due to their unique characteristics, including high water solubility, good light stability, excellent biocompatibility, and low cost. Herein, we report a ratiometric nanoprobe based on Pdots-Eu for temperature sensing in vitro. The Pdots-Eu thermometer was composed of a blue temperature-insensitive semiconducting polymer, poly(9-vinylcarbazole) (PVK), a red temperature-sensitive complex tris(dibenzoylmethane)mono(5-amino-1,10-phenanthroline)europium (III) (Eu complex), and an amphiphilic polymer polystyrene graft ethylene oxide functionalized with carboxyl groups (PS-PEG-COOH). The Pdots-Eu thermometer showed two peaks at 368 nm from PVK and 611 nm from the Eu complex. The red/blue fluorescence intensity ratio of Pdots-Eu decreased with an increase in temperature, which could be used for the ratiometric monitoring of temperature change. The results showed that the red/blue fluorescence intensity ratio demonstrated a good linear relationship to the change of temperature from 25 °C to 55 °C. Impressively, the ratiometric probe featured good accuracy and high sensitivity for temperature detection in vitro, which could be used for monitoring temperature change in cells.

Development of Stereo NIR-II Fluorescence Imaging System for 3D Tumor Vasculature in Small Animals.

Near-infrared-II (NIR-II, 1000–1700 nm) fluorescence imaging boasts high spatial resolution and deep tissue penetration due to low light scattering, reduced photon absorption, and low tissue autofluorescence. NIR-II biological imaging is applied mainly in the noninvasive visualization of blood vessels and tumors in deep tissue. In the study, a stereo NIR-II fluorescence imaging system was developed for acquiring three-dimension (3D) images on tumor vasculature in real-time, on top of the development of fluorescent semiconducting polymer dots (IR-TPE Pdots) with ultra-bright NIR-II fluorescence (1000–1400 nm) and high stability to perform long-term fluorescence imaging. The NIR-II imaging system only consists of one InGaAs camera and a moving stage to simulate left-eye view and right-eye view for the construction of 3D in-depth blood vessel images. The system was validated with blood vessel phantom of tumor-bearing mice and was applied successfully in obtaining 3D blood vessel images with 0.6 mm- and 5 mm-depth resolution and 0.15 mm spatial resolution. The NIR-II stereo vision provides precise 3D information on the tumor microenvironment and blood vessel path.

Fluorescent Polymer Dot-Based Multicolor Stimulated Emission Depletion Nanoscopy with a Single Laser Beam Pair for Cellular Tracking.

Stimulated emission depletion (STED) nanoscopy provides subdiffraction resolution while preserving the benefits of fluorescence confocal microscopy in live-cell imaging. However, there are several challenges for multicolor STED nanoscopy, including sophisticated microscopy architectures, fast photobleaching, and cross talk of fluorescent probes. Here, we introduce two types of nanoscale fluorescent semiconducting polymer dots (Pdots) with different emission wavelengths: CNPPV (poly[2-methoxy-5-(2-ethylhexyloxy)-1,4-(1-cyanovinylene-1,4-phenylene)]) Pdots and PDFDP (poly[{9,9-dihexyl-2,7-bis(1-cyanovinylene)fluorene}-alt-co-{2,5-bis (N,N'-diphenylamino)-1,4-phenylene}]) Pdots, for dual-color STED bioimaging and cellular tracking. Besides bright fluorescence, strong photostability, and easy bioconjugation, these Pdots have large Stokes shifts, which make it possible to share both excitation and depletion beams, thus requiring only a single pair of laser beams for the dual-color STED imaging. Long-term tracking of cellular organelles by the Pdots has been achieved in living cells, and the dynamic interaction of endosomes derived from clathrin-mediated and caveolae-mediated endocytic pathways has been monitored for the first time to propose their interaction models. These results demonstrate the promise of Pdots as excellent probes for live-cell multicolor STED nanoscopy.

Dextran Functionalization of Semiconducting Polymer Dots and Conjugation with Tetrameric Antibody Complexes for Bioanalysis and Imaging.

Brightly fluorescent semiconducting polymer dots (Pdots) are emerging as very useful probes for bioanalysis and imaging. Unfortunately, Pdot materials often suffer from limitations such as poor colloidal and physical stability, nonspecific adsorption, and relatively few reported surface chemistries and bioconjugate chemistries. To help address these limitations, we have developed dextran-functionalized Pdots (Dex-Pdots). This functionalization improves particle stability over a range of pH and at high ionic strength, hinders surface-induced unfolding, and enables the preparation of immunoconjugates via tetrameric antibody complexes (TAC). The utility of TAC-conjugated Dex-Pdots is demonstrated through a proof-of-concept fluorescence-linked immunosorbent assay (FLISA) for human erythropoietin (EPO), and through immunolabeling of human epidermal growth factor receptor 2 (HER2)-positive SK-BR3 breast cancer cells. The conjugates exhibited less nonspecific binding and greater specific binding than Pdots without dextran functionalization. Overall, dextran functionalization is a highly promising surface chemistry for biological applications of Pdots.

Sensitive fluorometric determination of glutathione using fluorescent polymer dots and the dopamine-melanin nanosystem.

A bioinspired fluorometric method has been developed for the detection of glutathione (GSH) in biological fluids. It is based on the use of near-infrared fluorescent semiconducting polymer dots (P-dots) and of the dopamine (DA)-melanin nanosystem. The P-dots were prepared from poly(styrene-co-maleic anhydride), the semiconducting polymer poly[(9,9'-dioctyl-2,7-divinylenefluorenylene)-alt-2-methoxy-5-(2-ethyl-hexyloxy)-1,4-phenylene] and the fluorescent dye tetraphenylporphyrin. They have excitation/emission maxima at 458/656nm, and this enables measurement to be performed with low autofluorescence and scattering background. DA can self-polymerize on the surface of the P-dots to yield a poly-DA coating. This coating, at weak alkaline pH values, causes the quenching of the fluorescence of the P-dots. However, the polymerization of DA is inhibited by GSH. Hence, quenching of fluorescence is prevented. This effect was used to design a fluorometric assay for GSH that has good selectivity and sensitivity. Under optimal conditions, the method has a linear response in the 0.2 to 20μM GSH concentration range and a 60nM detection limit. It was successfully applied to the determination of GSH in HepG2 cells and in spiked human serum. Graphical abstract Schematic representation of using aNIR fluorescent P-dots and dopamine (DA)-melanin nanohybridas a probe for glutathione (GSH) detection. The P-dots were prepared from poly(styrene-co-maleic anhydride) (PSMA), the semiconducting polymer poly[(9,9'-dioctyl-2,7-divinylenefluorenylene)-alt-2-methoxy-5-(2-ethyl-hexyloxy)-1,4-phenylene] (PEPV) and the fluorescent dye tetraphenylporphyrin (TPP).The GSH can inhibit the dopamine self-polymerization and prevented the formation of PDA and fluorescence quenching of P-dots.

Nile red-doped fluorescent semiconducting polymer dots as a highly sensitive hydrophobicity probe: protein conformational changes detection and plasma membrane imaging

One-pot synthesis of a photostable, hydrophobic and green fluorescent probe remains a challenge to develop luminescent biomaterials as well as biomedia technology. Semiconducting polymer dots (PDs) hold a great promise as fluorescence nanoprobes due to their photostability, biocompatibility and high quantum yield. Here, the synthesis and characterization of Nile red-doped PFBT PDs as a highly sensitive hydrophobicity probe has been reported. Nile red, a phenoxazone dye, fluoresces intensely in organic solvents and hydrophobic lipids and in varying color; however its fluorescence is totally quenched in water. First, Nile red-doped poly[(9,9-dioctylfluorenyl-2,7-diyl)-alt-co-(1,4-benzo-(2,1′,3)-thiadiazole)] (PFBT) PDs were synthesized through a nano-precipitation technique. Energy transfer from PFBT to Nile red happens through FRET mechanism. The PDs exhibited a red fluorescence with a peak centered at 620 nm (photo-excited at 460 nm). In aqueous media, the fluorescence strongly quenched, but by increasing the hydrophobicity the fluorescence of PDs significantly enhances as the polarity of the medium decreases. This can happen by adding organic solvents or nonpolar species to the medium. The Nile red@PFBT PDs have remarkable specification over Nile red dye. Compared to Nile red, this probe is soluble in water, more photostable, and more brightness due to light-harvesting ability of PDs and less wavelength shift in different medium polarity. We used this probe to investigate polarity of different organic solvents, to study surface hydrophobicity changes of BSA and also Ca2+ determination due to conformational changes of calmodulin. Also we used this probe for imaging of lipid bilayers and cellular membrane.

Dual Colorimetric and Fluorescent Imaging of Latent Fingerprints on Both Porous and Nonporous Surfaces with Near-Infrared Fluorescent Semiconducting Polymer Dots.

Semiconducting polymer dots (Pdots) have recently been proven as a novel type of ultrabright fluorescent probes that can be extensively used in analytical detection. Here, we developed a dual visual sensor based on Pdots for fingerprint imaging. We first designed and synthesized two types of near-infrared (NIR) fluorescent polymers and then embedded ninhydrin into the Pdot matrix. The resulting Pdot assays showed the colorimetric and fluorescent dual-readout abilities to detect latent fingerprints on both porous and nonporous surfaces. The developed fingerprints clearly revealed first-, second-, and third-level details with high contrast, high selectivity, and low background interference. We also grafted the chemical groups on the nanoparticle surface to investigate the mechanisms involved in the fingerprint development processes. We further utilized this assay in note paper and checks for latent fingerprint imaging. We believe that this dual-readout method based on Pdots will create a new avenue for research in fingerprint detection and anticounterfeiting technology.

Near-infrared fluorescent semiconducting polymer dots with high brightness and pronounced effect of positioning alkyl chains on the comonomers.

In recent years, semiconducting polymer dots (Pdots) have emerged as a novel class of extraordinarily bright fluorescent probes with burgeoning applications in bioimaging and sensing. While the desire for near-infrared (NIR)-emitting agents for in vivo biological applications increases drastically, the direct synthesis of semiconducting polymers that can form Pdots with ultrahigh fluorescence brightness is extremely lacking due to the severe aggregation-caused quenching of the NIR chromophores in Pdots. Here we describe the synthesis of dithienylbenzoselenadiazole (DBS)-based NIR-fluorescing Pdots with ultrahigh brightness and excellent photostability. More importantly, the fluorescence quantum yields of these Pdots could be effectively increased by the introduction of long alkyl chains into the thiophene rings of DBS to significantly inhibit the aggregation-caused emission quenching. Additionally, these new series of DBS-based Pdots can be excited by a commonly used 488 nm laser and show a fluorescence quantum yield as high as 36% with a Stokes shift larger than 200 nm. Single-particle analysis indicates that the per-particle brightness of the Pdots is at least 2 times higher than that of the commercial quantum dot (Qdot705) under identical laser excitation and acquisition conditions. We also functionalized the Pdots with carboxylic acid groups and then linked biomolecules to Pdot surfaces to demonstrate their capability for specific cellular labeling without any noticeable nonspecific binding. Our results suggest that these DBS-based NIR-fluorescing Pdots will be very practical in various biological imaging and analytical applications.

Yellow Fluorescent Semiconducting Polymer Dots with High Brightness, Small Size, and Narrow Emission for Biological Applications.

Cross-linked polymer dots with intense and narrow yellow emission were designed using boron-dipyrromethene (BODIPY) polymer as the acceptor and poly[9,9-dioctylfluorenyl-2,7-diyl-co-1,4-benzo-{2,1′-3}-thiadiazole] (PFBT) polymer as the donor. The emission fwhm’s of the polymer dots (Pdots) were 37 nm. CL-BODIPY 565 Pdots were about 5 times brighter than commercial quantum dots (Qdots) 565 under identical experimental conditions. Specific cellular targeting indicated that the small, bright, and narrow emissive CL-BODIPY 565 Pdots are promising probes for biological applications.

Polydiacetylene-enclosed near-infrared fluorescent semiconducting polymer dots for bioimaging and sensing.

Semiconducting polymer dots (P-dots) recently have emerged as a new type of ultrabright fluorescent probe with promising applications in biological imaging and detection. With the increasing desire for near-infrared (NIR) fluorescing probes for in vivo biological measurements, the currently available NIR-emitting P-dots are very limited and the leaching of the encapsulated dyes/polymers has usually been a concern. To address this challenge, we first embedded the NIR dyes into the matrix of poly[(9,9-dioctylfluorene)-co-2,1,3-benzothiadiazole-co-4,7-di(thiophen-2-yl)-2,1,3-benzothiadiazole] (PF-BT-DBT) polymer and then enclosed the doped P-dots with polydiacetylenes (PDAs) to avoid potential leakage of the entrapped NIR dyes from the P-dot matrix. These PDA-enclosed NIR-emitting P-dots not only emitted much stronger NIR fluorescence than conventional organic molecules but also exhibited enhanced photostability over CdTe quantum dots, free NIR dyes, and gold nanoclusters. We next conjugated biomolecules onto the surface of the resulting P-dots and demonstrated their capability for specific cellular labeling without any noticeable nonspecific binding. To employ this new class of material as a facile sensing platform, an easy-to-prepare test paper, obtained by soaking the paper into the PDA-enclosed NIR-emitting P-dot solution, was used to sense external stimuli such as ions, temperature, or pH, depending on the surface functionalization of PDAs. We believe these PDA-coated NIR-fluorescing P-dots will be very useful in a variety of bioimaging and analytical applications.

ChemInform Abstract: Highly Fluorescent Semiconducting Polymer Dots for Biology and Medicine.

AbstractReview: 176 refs.

Highly Fluorescent Semiconducting Polymer Dots for Biology and Medicine

In recent years, semiconducting polymer nanoparticles have attracted considerable attention because of their outstanding characteristics as fluorescent probes. These nanoparticles, which primarily consist of π-conjugated polymers and are called polymer dots (Pdots) when they exhibit small particle size and high brightness, have demonstrated utility in a wide range of applications such as fluorescence imaging and biosensing. In this review, we summarize recent findings of the photophysical properties of Pdots which speak to the merits of these entities as fluorescent labels. This review also highlights the surface functionalization and biomolecular conjugation of Pdots, and their applications in cellular labeling, in vivo imaging, single-particle tracking, biosensing, and drug delivery. We discuss the relationship between the physical properties and performance, and evaluate the merits and limitations of the Pdot probes for certain imaging tasks and fluorescence assays. We also tackle the current challenges of Pdots and share our perspective on the future directions of the field.

Multicolor Fluorescent Semiconducting Polymer Dots with Narrow Emissions and High Brightness

Fluorescent semiconducting polymer dots (Pdots) have attracted great interest because of their superior characteristics as fluorescent probes, such as high fluorescence brightness, fast radiative rates, and excellent photostability. However, currently available Pdots generally exhibit broad emission spectra, which significantly limit their usefulness in many biological applications involving multiplex detections. Here, we describe the design and development of multicolor narrow emissive Pdots based on different boron dipyrromethene (BODIPY) units. BODIPY-containing semiconducting polymers emitting at multiple wavelengths were synthesized and used as precursors for preparing the Pdots, where intraparticle energy transfer led to highly bright, narrow emissions. The emission full width at half-maximum of the resulting Pdots varies from 40 to 55 nm, which is 1.5-2 times narrower than those of conventional semiconducting polymer dots. BODIPY 520 Pdots were about an order of magnitude brighter than commercial Qdot 525 under identical laser excitation conditions. Fluorescence imaging and flow cytometry experiments indicate that the narrow emissions from these bright Pdots are promising for multiplexed biological detections.