Hyperbranched Polymer Dots with Strong Absorption and High Fluorescence Quantum Yield for In Vivo NIR-II Imaging.

In order to improve the fluorescence quantum yield (QY) of NIR-II-emitting nanoparticles, D-A-D fluorophores are typically linked to intramolecular rotatable units to reduce aggregation-induced quenching. However, incorporating such units often leads to a twisted molecular backbone, which affects the coupling within the D-A-D unit and, as a result, lowers the absorption. Here, we overcome this limitation by cross-linking the NIR-II fluorophores to form a 2D polymer network, which simultaneously achieves a high QY by well-controlled fluorophore separation and strong absorption by restricting intramolecular distortion. Using the strategy, we developed polymer dots with the highest NIR-II single-particle brightness among reported D-A-D-based nanoparticles and applied them for imaging of hindlimb vasculatures and tumors as well as fluorescence-guided tumor resection. The high brightness of the polymer dots offered exceptional image quality and excellent surgical results, showing a promising performance for these applications.

High-Precision Mapping of Membrane Proteins on Synaptic Vesicles using Spectrally Encoded Super-Resolution Imaging.

The spatial resolution of single-molecule localization microscopy is limited by the photon number of a single switching event because of the difficulty of correlating switching events dispersed in time. Here we overcome this limitation by developing a new class of photoswitching semiconducting polymer dots (Pdots) with structured and highly dispersed single-particle spectra. We imaged the Pdots at the first and the second vibronic emission peaks and used the ratio of peak intensities as a spectral coding. By correlating switching events using the spectral coding and performing 4-9 frame binning, we achieved a 2-3 fold experimental resolution improvement versus conventional superresolution imaging. We applied this method to count and map SV2 and proton ATPase proteins on synaptic vesicles (SVs). The results reveal that these proteins are trafficked and organized with high precision, showing unprecedented level of detail about the composition and structure of SVs.

Multimode Time-Resolved Superresolution Microscopy Revealing Chain Packing and Anisotropic Single Carrier Transport in Conjugated Polymer Nanowires.

Here, we developed a novel, multimode superresolution method to perform full-scale structural mapping and measure the energy landscape for single carrier transport along conjugated polymer nanowires. Through quenching of the local emission, the motion of a single photogenerated hole was tracked using blinking-assisted localization microscopy. Then, utilizing binding and unbinding dynamics of quenchers onto the nanowires, local emission spectra were collected sequentially and assembled to create a superresolution map of emission sites throughout the structure. The hole polaron trajectories were overlaid with the superresolution maps to correlate structures with charge transport properties. Using this method, we compared the efficiency of inter- and intrachain hole transport inside the nanowires and for the first time directly measured the depth of carrier traps originated from torsional disorder and chemical defects.

Improving the Accuracy of Pdot-Based Continuous Glucose Monitoring by Using External Ratiometric Calibration.

Continuous glucose monitoring (CGM) allows type I and II diabetes patients to track changes in their glucose levels, allowing detection of impending hypoglycemia or hyperglycemia. Polymer dots (Pdots) are candidates for use in implanted CGM systems due to their exceptional brightness, photostability, sensitivity, and biocompatibility. However, Pdot glucose transducers are oxygen-dependent, and changes in tissue oxygen levels affect their measurement accuracy. Here, we describe an external ratiometric calibration method that corrects for changes in tissue oxygen levels to improve measurement accuracy. This method uses the ratio of oxygen concentrations inside and outside the Pdot glucose transducer as an indicator of glucose concentration to correct for signal deviations caused by tissue oxygen fluctuations. A second oxygen-sensitive Pdot that is not conjugated with glucose oxidase is used to measure the oxygen concentration outside the Pdot glucose transducer. We describe the theoretical basis for this approach and demonstrate its effectiveness experimentally in a subcutaneous mouse implant model. This external ratiometric system achieves higher accuracy glucose measurements than previous Pdot-based CGM systems and comparable accuracy to current commercial CGM systems, demonstrating the utility of the external ratiometric calibration strategy.

High-Throughput Counting and Superresolution Mapping of Tetraspanins on Exosomes Using a Single-Molecule Sensitive Flow Technique and Transistor-like Semiconducting Polymer Dots.

A method for high-throughput counting and superresolution mapping of surface proteins on exosomes is described. The method combines a single-molecule sensitive flow technique and an adaptive superresolution imaging method. Exosomes stained with membrane dye and dye-conjugated antibodies were analyzed using a microfluidic platform at a flow rate of 100 exosome s-1 to determine size and protein copy number. Superresolution mapping was performed with exosomes labeled with novel transistor-like, semiconducting polymer dots (Pdots), which exhibit spontaneous blinking with <5 nm localization error and a broad range of optical-adjustable duty cycles. Based on the copy numbers extracted from the flow analysis, the switch-on frequency of the Pdots were finely adjusted so that structures of hundreds of exosomes were obtained within five minutes. The high throughput and high sensitivity of this method offer clear advantages for characterization of exosomes and similar biological vesicles.

Reversible Ratiometric NADH Sensing Using Semiconducting Polymer Dots.

Reduced nicotinamide adenine dinucleotide (NADH) is a key coenzyme in living cells due to its role as an electron carrier in redox reactions, and its concentration is an important indicator of cell metabolic state. Abnormal NADH levels are associated with age-related metabolic diseases and neurodegenerative disorders, creating a demand for a simple, rapid analytical method for point-of-care NADH sensing. Here we develop a series of NADH-sensitive semiconducting polymer dots (Pdots) as nanoprobes for NADH measurement, and test their performance in vitro and in vivo. NADH sensing is based on electron transfer from semiconducting polymer chains in the Pdot to NADH upon UV excitation, quenching Pdot fluorescence emission. In polyfluorene-based Pdots, this mechanism resulted in an on-off NADH sensor; in DPA-CNPPV Pdots, UV excitation resulted in NADH-sensitive emission at two wavelengths, enabling ratiometric detection. Ratiometric NADH detection using DPA-CNPPV Pdots exhibits high sensitivity (3.1 µM limit of detection), excellent selectivity versus other analytes, reversibility, and a fast response (less than 5 s). We demonstrate applications of the ratiometric NADH-sensing Pdots including smartphone-based NADH imaging for point-of-care use.

Monitoring Metabolites Using an NAD(P)H-sensitive Polymer Dot and a Metabolite-Specific Enzyme.

We introduce an NAD(P)H-sensitive polymer dot (Pdot) biosensor for point-of-care monitoring of metabolites. The Pdot is combined with a metabolite-specific NAD(P)H-dependent enzyme that catalyzes the oxidation of the metabolite, generating NAD(P)H. Upon UV illumination, the NAD(P)H quenches the fluorescence emission of Pdot at 627 nm via electron transfer, and also fluoresces at 458 nm, resulting in a shift from red to blue emission at higher NAD(P)H concentrations. Metabolite concentration is quantified ratiometrically-based on the ratio of blue-to-red channel emission intensities, with a digital camera-with high sensitivity and specificity. We demonstrate phenylalanine biosensing in human plasma for a phenylketonuria screening test, quantifying several other disease-related metabolites (lactate, glucose, glutamate, and ß-hydroxybutyrate), and a paper-based assay with smartphore imaging for point-of-care use.

Dual-Mode Superresolution Imaging Using Charge Transfer Dynamics in Semiconducting Polymer Dots.

In a conjugated polymer-based single-particle heterojunction, stochastic fluctuations of the photogenerated hole population lead to spontaneous fluorescence switching. We found that 405 nm irradiation can induce charge recombination and activate the single-particle emission. Based on these phenomena, we developed a novel class of semiconducting polymer dots that can operate in two superresolution imaging modes. The spontaneous switching mode offers efficient imaging of large areas, with <10 nm localization precision, while the photoactivation/deactivation mode offers slower imaging, with further improved localization precision (ca. 1 nm), showing advantages in resolving small structures that require high spatial resolution. Superresolution imaging of microtubules and clathrin-coated pits was demonstrated, under both modes. The excellent localization precision and versatile imaging options provided by these nanoparticles offer clear advantages for imaging of various biological systems.

Light-Harvesting and Amplified Energy Transfer in Conjugated Polymer Nanoparticles.

Conjugated polymer nanoparticles are a class of nanoparticles with many useful and interesting properties, including very high fluorescence brightness, excellent photostability, and sensing capabilities. They also exhibit interesting and potentially useful phenomena, such as highly efficient energy transfer, anomalous single particle blinking, and twinkling phenomena associated with polaron motion. As little as one dye molecule per nanoparticle can efficiently quench the fluorescence of hundreds of polymer chromophore units. Similarly, loss of a single electron can result in quenching of hundreds of chromophores. These phenomena and properties are dictated by the nature of interactions between chromophores in this dense, nanoscale multichromophoric system, and are characterized as amplified energy transfer or multiple energy transfer. In this review, we summarize the key aspects of conjugated polymer nanoparticles optical properties and phenomena, and discuss the current understanding of exciton dynamics in these and related systems. In particular, our current understanding and theoretical models for amplified or multiple energy transfer based on exciton theory and Förster resonance energy transfer are explored.

A BODIPY-Based Donor/Donor-Acceptor System: Towards Highly Efficient Long-Wavelength-Excitable Near-IR Polymer Dots with Narrow and Strong Absorption Features.

Bright long-wavelength-excitable semiconducting polymer dots (LWE-Pdots) are highly desirable for in vivo imaging and multiplexed in vitro bioassays. LWE-Pdots have been obtained by incorporating a near-infrared (NIR) emitter into the backbone of a polymer host to develop a binary donor-acceptor (D-A) system. However, they usually suffer from severe concentration quenching and a trade-off between fluorescence quantum yield (Φf ) and absorption cross-section (σ). Herein, we describe a ternary component (D1 /D2 -A) strategy to achieve ultrabright, green laser-excitable Pdots with narrow-band NIR emission by introducing a BODIPY-based assistant polymer donor as D1 . The D1 /D2 -A Pdots possess improved Φf and σ compared to corresponding binary D2 -A Pdots. Their Φf is as high as 40.2 %, one of the most efficient NIR Pdots reported. The D1 /D2 -A Pdots show ultrahigh single-particle brightness, 83-fold brighter than Qdot 705 when excited by a 532 nm laser. When injected into mice, higher contrast in vivo tumor imaging was achieved using the ternary Pdots versus the binary D-A Pdots.

Single-Molecule Flow Platform for the Quantification of Biomolecules Attached to Single Nanoparticles.

We describe here a flow platform for quantifying the number of biomolecules on individual fluorescent nanoparticles. The platform combines line-confocal fluorescence detection with near nanoscale channels (1-2 µm in width and height) to achieve high single-molecule detection sensitivity and throughput. The number of biomolecules present on each nanoparticle was determined by deconvolving the fluorescence intensity distribution of single-nanoparticle-biomolecule complexes with the intensity distribution of single biomolecules. We demonstrate this approach by quantifying the number of streptavidins on individual semiconducting polymer dots (Pdots); streptavidin was rendered fluorescent using biotin-Alexa647. This flow platform has high-throughput (hundreds to thousands of nanoparticles detected per second) and requires minute amounts of sample (∼5 µL at a dilute concentration of 10 pM). This measurement method is an additional tool for characterizing synthetic or biological nanoparticles.

Hyper strength, high sensitivity integrated wearable signal sensor based on non-covalent interaction of an ionic liquid and bacterial cellulose for human behavior monitoring.

Ion-sensing hydrogels exhibit electrical conductivity, softness, and mechanical and sensory properties akin to human tissue, rendering them an ideal material for mimicking human skin. In the realm of fabricating sensors for detecting human physiological activities, they present an ideal alternative to traditional rigid metal conductors. Nevertheless, achieving ionic hydrogels with outstanding tensile properties, toughness, ionic conductivity, and transport stability poses a significant challenge. This paper describes a simple method of forming a basic network by free radical polymerization of acrylamide, and then bacterial cellulose (BC) and 1-ethyl-3-methylimidazolium chloride ([EMIM]Cl) were introduced into the basic network. The polyhydrogen bonds and electrostatic interactions in the system gave the hydrogel notable tensile properties (3271 ± 37%), toughness (7.39 ± 0.13 MJ m-3), and high ultimate tensile stress (385.1 ± 7.2 kPa). In addition, the combination of BC and [EMIM]Cl collaboratively enhanced the mechanical properties and electrical conductivity. Ion sensing hydrogels have a wide operating strain range (≈1000%) and high sensitivity (gage factor (GF) = 11.85), and are therefore considered promising candidates for next-generation gel-based strain sensor platforms.

Preparation and optimization of dummy molecularly imprinted polymer-based solid-phase extraction system for selective enrichment of p-toluene sulfonate esters genotoxic impurities.

Sulfonate esters, one class of genotoxic impurities (GTIs), have gained significant attention in recent years due to their potential to cause genetic mutations and cancer. In the current study, we employed the dummy template molecular imprinting technology with a dummy template molecule replacing the target molecule to establish a pretreatment method for samples containing p-toluene sulfonate esters. Through computer simulation and ultraviolet-visible spectroscopy analysis, the optimal functional monomer acrylamide and polymerization solvent chloroform were selected. Subsequently, a dummy template molecularly imprinted polymer (DMIP) was prepared by the precipitation polymerization method, and the polymer was characterized in morphology, particle size, and composition. The results of the adsorption and enrichment study demonstrated that the DMIP has high adsorption capability (Q = 7.88 mg/g) and favorable imprinting effects (IF = 1.37); Further, it could simultaneously adsorb three p-toluene sulfonate esters. The optimal adsorption conditions were obtained by conditional optimization of solid-phase extraction (SPE). A pH 7 solution was selected as the loading condition, the methanol/1 % phosphoric acid solution (20:80, v/v) was selected as the washing solution, and acetonitrile containing 10 % acetic acid in 6 mL was selected as the elution solvent. Finally, we determined methyl p-toluene sulfonate alkyl esters, ethyl p-toluene sulfonate alkyl esters, and isopropyl p-toluene sulfonate alkyl esters in tosufloxacin toluene sulfonate and capecitabine at the 10 ppm level (relative to 1 mg/mL active pharmaceutical ingredient (API) samples) by using DMIP-based SPE coupled with HPLC. This approach facilitated the selective enrichment of p-toluene sulfonate esters GTIs from complex API samples.

Super-stretching and high-performance ionic thermoelectric hydrogels based on carboxylated bacterial cellulose coordination for self-powered sensors.

Self-powered sensors that do not require external power sources are crucial for next-generation wearable electronics. As environment-friendly ionic thermoelectric hydrogels can continuously convert the low-grade heat of human skin into electricity, they can be used in intelligent human-computer interaction applications. However, their low thermoelectric output power, cycling stability, and sensitivity limit their practical applications. This paper reports a 2,2,6,6-tetramethylpiperidine-1-oxyl radical (TEMPO)-oxidized carboxylated bacterial cellulose (TOBC) coordination double-network ionic thermoelectric hydrogel with lithium bis(trifluoromethane) sulfonimide (LiTFSI) as an ion provider for thermodiffusion, as LiTFSI exhibits excellent thermoelectric properties with a maximum power output of up to 538 nW at a temperature difference of 20 K. The interactions between the ions and the hydrogel matrix promote the selective transport of conducting ionic ions, producing a high Seebeck coefficient of 11.53 mV K-1. Hydrogen bonding within the polyacrylamide (PAAm) network and interactions within the borate ester bond within the TOBC confer excellent mechanical properties to the hydrogel such that the stress value at a tensile deformation of 3100 % is reaches 0.85 MPa. The combination of the high ionic thermovoltage and excellent mechanical properties ionic thermoelectric hydrogels provides an effective solution for the design and application of self-powered sensors based on hydrogels.

Ultrabright Pdots with a Large Absorbance Cross Section and High Quantum Yield.

Semiconducting polymer dots (Pdots) are increasingly used in biomedical applications due to their extreme single-particle brightness, which results from their large absorption cross section (σ). However, the quantum yield (Φ) of Pdots is typically below 40% due to aggregation-induced self-quenching. One approach to reducing self-quenching is to use FRET between the donor (D) and acceptor (A) groups within a Pdot; however, Φ values of FRET-based Pdots remain low. Here, we demonstrate an approach to achieve ultrabright FRET-based Pdots with simultaneously high σ and Φ. The importance of self-quenching was revealed in a non-FRET Pdot: adding 30 mol % of a nonabsorbing polyphenyl to a poly(9,9-dioctylfluorene) (PFO) Pdot increased Φ from 13.4 to 71.2%, yielding an ultrabright blue-emitting Pdot. We optimized the brightness of FRET-based Pdots by exploring different D/A combinations and ratios with PFO and poly[(9,9-dioctylfluorenyl-2,7-diyl)-co-(1,4-phenylene)] (PFP) as donor polymers and poly[(9,9-dioctyl-2,7-divinylenefluorenylene)-alt-co-(1,4-phenylene)] (PFPV) and poly[(9,9-dioctylfluorenyl-2,7-diyl)-alt-co-(1,4-benzo-{2,1',3}-thiadiazole)] (PFBT) as acceptor polymers, with a fixed concentration of poly(styrene-co-maleic anhydride) as surfactant polymer. Ultrabright blue-emitting Pdots possessing high Φ (73.1%) and σ (σR = σabs/σall, 97.5%) were achieved using PFP/PFPV Pdots at a low acceptor content (A/[D + A], 2.5 mol %). PFP/PFPV Pdots were 1.8 times as bright as PFO/PFPV Pdots due to greater coverage of acceptor absorbance by donor emission─a factor often overlooked in D/A pair selection. Ultrabright green-emitting PFO Pdots (Φ = 76.0%, σR = 92.5%) were obtained by selecting an acceptor (PFBT) with greater spectral overlap with PFO. Ultrabright red-emitting Pdots (Φ = 64.2%, σR = 91.0%) were achieved by blending PFO, PFBT, and PFTBT to create a cascade FRET Pdot at a D:A1:A2 molar ratio of 61:5:1. These blue, green, and red Pdots are among the brightest Pdots reported. This approach of using a small, optimized amount of FRET acceptor polymer with a large donor-acceptor spectral overlap can be generalized to produce ultrabright Pdots with emissions that span the visible spectrum.

Bacterial cellulose-based dual chemical reaction coupled hydrogel thermocells for efficient heat harvesting.

Reasonable and efficient utilization of low-grade thermal energy in nature is the choice for sustainable energy development. We demonstrate a bacterial cellulose (BC) hydrogel thermocell (TEC) based on BC electrolyte combined with carbon fiber paper and copper composite electrode sheets. The large specific surface area of carbon fiber paper provides a large number of active sites for thermoelectric ions, which drives the redox reaction inside the electrolyte and stimulates the chemical reaction between the electrolyte and the electrode. The combination of the two chemical reactions significantly improves the thermoelectric performance of the thermocell. The thermopower of the BC-TEC reaches 5.9 mV·K-1 at a temperature difference of 50 K. The TEC consisting of 6-units in series produces an open-circuit voltage of about 2 V and a peak power of 535 µW. The TEC shows new potential and prospects in ambient thermoelectric energy conversion by rationally designing the power generation principle.

Degradable collagen/sodium alginate/polyvinyl butyral high barrier coating with water/oil-resistant in a facile and effective approach.

Degradable bio-based materials have been widely considered as functional coatings, however, it is a great challenge to fabricate biodegradable coatings with high barrier, water- and oil- resistance. In this work, such coatings were fabricated by using collagen fibers (CF), sodium alginate (SA), and polyvinyl butyral (PVB). CF and SA were mixed evenly and coated on Ca2+ pretreated filter paper. It was mainly due to the electrostatic adsorption between collagen fibers and sodium alginate, and the crosslinking between the adsorption products and Ca2+. By coating PVB solution, the barrier performance was further improved. Notably, the composite exhibited excellent water vapor resistance (48 g/m2·24 h), water resistance (31 g/m2), oil resistance (kit rating: 12/12) and good mechanical properties. This degradable, environmentally friendly, and simple composite paper method has excellent barrier properties, mechanical properties and fluorine-free properties, and will have many applications in the food and packaging fields.

Strong Bacterial Cellulose-Based Films with Natural Laminar Alignment for Highly Sensitive Humidity Sensors.

Humidity sensors have been widely used for humidity monitoring in industry and agriculture fields. However, the rigid structure, nondegradability, and large dimension of traditional humidity sensors significantly restrict their applications in wearable fields. In this study, a flexible, strong, and eco-friendly bacterial cellulose-based humidity sensor (BPS) was fabricated using a two-step method, involving solvent evaporation-induced self-assembly and electrolyte permeation. Rapid evaporation of organic solvent induces the formation of nanopores of the bacterial cellulose (BC) surface and promotes structural densification. Furthermore, the successful embedding of potassium hydroxide into the sophisticated network of BC effectively enhanced the sensing performance of BPS. The BPS exhibits an excellent humidity sensing response of more than 103 within the relative humidity ranging from 36.4 to 93% and strong (66.4 MPa) and high flexibility properties owing to the ultrafine fiber network and abundant hydrophilic functional groups of BC. Besides being strong and thin, BPS is also highly flexible, biodegradable, and humidity-sensitive, making it a potential candidate in wearable electronics, human health monitoring, and noncontact switching.

In vitro gastrointestinal digestibility of corn oil-in-water Pickering emulsions stabilized by three types of nanocellulose.

The effect of three nanocellulose (various in crystalline allomorph and morphology) on lipid in vitro gastrointestinal digestibility was investigated. Corn oil-in-water emulsions were prepared by CNCs-I, CNCs-II and CNFs respectively. The variations of droplets diameter D[4,3], zeta potential, and microstructure were measured during gastrointestinal digestion (mouth, stomach and small intestine), and the free fatty acid (FFA) released in the small intestine phase were examined. The FFA-released test results indicated that both crystalline allomorph and morphology of nanocellulose affected the degree of lipid digestion, especially the morphology. FFA released amount was ranked in the order of CNCs-I (56.60%), CNCs-II (48.67%) and CNFs (28.21%). This is mainly due to the difference in the self-assembly behavior of nanocellulose at the interface. Our findings provide an innovative solution that using nanocellulose as food-grade particle stabilizer to modulate the digestion of Pickering emulsified lipids, which would benefit the development of given functional foods.

Pickering emulsions stabilized by spherical cellulose nanocrystals.

Nano cellulose has attracted more attentions as promising stabilizer candidate for Pickering emulsions due to its renewable and biocompatible properties. In this work, spherical cellulose nanocrystals (S-CNCs) (diameter 30-60 nm) were prepared by mixed acid hydrolysis of mercerized microcrystalline cellulose under the treatment of ultrasonic. We characterized the Pickering emulsions stabilized by various S-CNCs concentration in the aqueous phase and visualized their distribution in emulsions system. The diameter of emulsion droplets has no increase after 7 days storage. The emulsions present ultra-low viscosity even at a high S-CNCs concentration in the aqueous phase (5 g/L). Furthermore, noting that Pickering emulsions droplets stabilized by S-CNCs exhibited high stability even pH, ionic strengths, and temperatures changed in wide range. Thus, the S-CNCs acted as a new stabilizer for Pickering emulsion open opportunities for the development of biomedicine, cosmetics, and food applications.