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At least 793 records · Page 44

The origin of bimodal luminescence of β-SiAlON:Eu{sup 2+} phosphors as revealed by fluorescence microscopy and cathodoluminescence analysis

Graphical abstract: - Highlights: • Bimodal emission is originated from β-SiAlON grains with z ≥ 2. • Coexistence of two kinds of emission centers in the β-SiAlON phase is definite. • Fluorescence microscopy shows influence of the z value on emission of β-SiAlON. - Abstract: Eu{sup 2+}-doped SiAlON phosphors with the composition of Eu{sub x}Si{sub 6−z}Al{sub z}O{sub z}N{sub 8−z} (0.5 ≤ z ≤ 3) at a fixed x = 0.01 were synthesized by the gas pressure sintering method. Dependence of luminescence properties on the phase compositions in β-SiAlON:Eu{sup 2+} phosphors has been examined via fluorescence microscope and scanning electron microscope equipped with a cathodoluminescence spectrometer and an energy dispersive spectrometer. Bimodal emission (green and violet) from β-SiAlON phase is observed in the samples with z ≥ 2, indicating co-existence of two different kinds of coordination for Eu{sup 2+} ions in the host lattice.

36 MATERIALS SCIENCE↗

Room temperature biosynthesis of greatly stable fluorescent ZnO quantum dots for the selective detection of Cr{sup 3+} ions

Graphical abstract: Biosynthesized fluorescent ZnO quantum dots are extremely stable and are able to sense up to 2 nM of Cr3+ ions in aqueous solution. - Highlights: • ZnO quantum dotes (QDs) are rapidly synthesized from the bio waste at room temperature. • The capping and reducing agent is from natural source hence biodegradable. • The water soluble fluorescent ZnO quantum dots are highly stable. • The QDs are able to sense 2 nM of Cr{sup 3+} ions in aqueous solution. - Abstract: Synthesis of extremely stable and water soluble quantum dots (QDs) would be smart if the method is rapid and eco-friendly. Here in, we have employed a simple strategy for the preparation of well dispersed, luminescent ZnO quantum dots by using pomegranate peel extract at room temperature. This method excludes the use of strong commercial stabilizing agents, different pH conditions, higher temperature and inert atmosphere during the synthesis. The microscopic study has revealed the ZnO QDs to be of a narrow size with an average size distribution of about 4 nm. The water soluble QDs exhibited a strong emission peak at 490 nm and were stable for twelve months. The emission from ZnO QDs are selectively quenched upon addition of Cr{sup 3+} ions. The interaction between the Cr{sup 3+} and the biomolecules on the exterior of ZnO QDs, enables the system to detect up to 2 nM Cr{sup 3+} ions in aqueous solution.

36 MATERIALS SCIENCE↗

Synthesis of dual-emitting (Gd,Eu)2O3-PEI@CD composite and its potential as ratiometric fluorescent sensor for curcumin

Highlights: • Dual-emitting (Gd,Eu){sub 2}O{sub 3}-PEI@CD composite were synthesized. • Under UV irradiation, the composite exhibits both blue CD emission and red Eu{sup 3+} emission. • a novel self-referencing fluorescent sensor for curcumin was designed. • The limit of detection is 0.0615 μg/ml at the signal to noise ratio of 3. - Abstract: Carbon dots (CD)-coated (Gd,Eu){sub 2}O{sub 3}-PEI nanospheres were successfully prepared via a facile electrostatic absorption method. Under UV irradiation, the composite simultaneously exhibits both blue CD emission band and red Eu{sup 3+} emission peaks. It is found that blue CD emission could be quenched by curcumin, while red Eu{sup 3+} emission is hardly influenced. Thus, a novel self-referencing fluorescent sensor for curcumin with high sensitivity and superior signal discriminability was designed. The ratio of two emissions (F{sub 445}/F{sub 611}) shows good linear relationship with curcumin concentration (R{sup 2} = 0.9973). The limit of detection is determined to be 0.0615 μg/ml at the signal to noise ratio of 3. These attractive merits enable this composite in ratiometric sensing for medicines in the future.

77 NANOSCIENCE AND NANOTECHNOLOGY↗

Unpacking the drivers of diurnal dynamics of sun-induced chlorophyll fluorescence (SIF): Canopy structure, plant physiology, instrument configuration and retrieval methods

Sun-induced chlorophyll fluorescence (SIF) from spaceborne sensors is a promising tool for global carbon cycle monitoring, but its application is constrained by insufficient understanding of the drivers underlying diurnal SIF dynamics. SIF measurements from ground-based towers can reveal diurnal SIF dynamics across biomes and environmental conditions; however, meaningful interpretation of diurnal variations requires disentangling impacts from canopy structure, plant physiology, instrument configuration and retrieval methods, which often interact with and confound each other. This study aims to unpack these drivers using 1) concurrent ground and airborne canopy-scale and leaf-scale measurements at a corn field, 2) a mechanistic SIF model that explicitly considers the dynamics of photochemistry (via the fraction of open photosystem II reaction centers, qL) and photoprotection (via nonphotochemical quenching, NPQ) as well as their interactive dependence on the sub-canopy light environment, and 3) cross-comparison of SIF instrument configurations and retrieval methods. We found that crop row orientations and sun angles can introduce a distinctive midday dip in SIF in absence of stress, due to a midday drop of absorbed photosynthetically active radiation (APAR) when crop rows are north-south oriented. Canopy structure caused distinctive responses in both qL and NPQ at different positions within the vertical canopy that collectively influenced fluorescence quantum yield (Φ F ) at the leaf scale. Once integrated at the canopy scale, diurnal dynamics of both APAR and canopy escape probability (ε) are critical for accurately shaping diurnal SIF variations. While leaf-level qL and NPQ exhibited strong diurnal dynamics, their influence was attenuated at the canopy scale due to opposing effects on SIF at different canopy layers. Furthermore, different system configurations (i.e., bi-hemispherical vs. hemispherical-conical) and retrieval methods can bias the SIF magnitude and distort its diurnal shapes, therefore confounding the interpretation of inherent strength and dynamics of SIF emission. Our findings demonstrate the importance of crop row structures, interactive variations in canopy structure and plant physiology, instrument configuration, and retrieval method in shaping the measured dynamics of diurnal SIF. This study highlights the necessity to account for these factors to accurately interpret satellite SIF, and informs future synthesis work with different SIF instrumentation and retrieval methods across sites.

59 BASIC BIOLOGICAL SCIENCES↗

Enzymatically hydrolyzed fluorescence-based chemical probe enables in situ mapping of chitinase activity in the rhizosphere

Rhizosphere microbiomes and root exudates play a pivotal biochemical role in helping to catalyze chitin catabolism. Chitin is a recalcitrant and ubiquitous soil biopolymer, estimated to be the second most abundant organic soil biopolymer on Earth. Despite its abundance, role as a source of C and N in soil, and importance to ecosystem function, the biochemical mechanisms controlling chitin fate in the rhizosphere are elusive and poorly understood. To enable spatial mapping of chitinase activity in the rhizosphere, we designed and synthesized an enzymatically activated fluorogenic substrate, chitotriose-TokyoGreen (chitotriose-TG), by incorporating a fluorescein derivative (TG) onto the trimeric unit of chitin. This non-fluorescent substrate is selectively hydrolyzed by chitinase to release TG and yield a high fluorescence signal, which can be used to spatially image and measure chitinase activity in the rhizosphere. To demonstrate the application of this technique, we grew switchgrass (Panicum virgatum) in rhizoboxes amended with a horizontal layer of chitin. We extracted mobile proteins from the rhizobox using a nitrocellulose membrane blotting technique which offer non-destructive enzyme extraction while preserving the 2D spatial position of the enzymes. We then subjected these membranes to the synthesized chitotriose-TG stain to spatially visualize the distribution of chitinase activity within the rhizosphere. Furthermore, we observed increased chitinase activity near plant roots and higher activity within the soil zone enriched in chitin, showing an adaptive response of chitinase activity with spatial focusing in areas of higher chitin abundance. Thus, the enzyme extraction and visualization strategy we describe here can help enlighten efforts to better understand spatial controls on chitin breakdown in rhizosphere, further elucidating the role of chitin as a C and N source in these systems.

59 BASIC BIOLOGICAL SCIENCES↗

Exploring chemistry with single-molecule and -particle fluorescence microscopy

The evolution of single-molecule and -particle fluorescence microscopy imaging techniques for the investigation of chemical reactions yielded achievements at all fronts, from synthetic organic chemistry to materials science. The removal of ensemble averaging effects uncovers unique, otherwise hidden reaction intermediates, reactivity, and kinetics. Thus, new insights can be obtained even for well-explored chemical reactions and processes. The aim of this article is to serve as a guide and a source of inspiration for scientists who wish to develop single-molecule and -particle fluorescence microscopy to answer questions in chemistry. Here, we describe the current status of technical accomplishments in the field and point out technical and conceptual obstacles for overcoming in the near future, with focus on improvement of bench-scale synthetic reactions.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Live cell imaging of interactions between replicase and capsid protein of Brome mosaic virus using Bimolecular Fluorescence Complementation: Implications for replication and genome packaging

In Brome mosaic virus, it was hypothesized that a physical interaction between viral replicase and capsid protein (CP) is obligatory to confer genome packaging specificity. Here we tested this hypothesis by employing Bimolecular Fluorescent Complementation (BiFC) as a tool for evaluating protein–protein interactions in living cells. The efficacy of BiFC was validated by a known interaction between replicase protein 1a (p1a) and protein 2a (p2a) at the endoplasmic reticulum (ER) site of viral replication. Additionally, co-expression in planta of a bona fide pair of interacting protein partners of p1a and p2a had resulted in the assembly of a functional replicase. Subsequent BiFC assays in conjunction with mCherry labeled ER as a fluorescent cellular marker revealed that CP physically interacts with p2a, but not p1a, and this CP:p2a interaction occurs at the cytoplasmic phase of the ER. The significance of the CP:p2a interaction in BMV replication and genome packaging is discussed. - Highlights: • YFP fusion proteins of BMV p1a and p2a are biologically active. • Self-interaction was observed for p1a, p2a and CP. • CP interacts with p2a but not p1a. • Majority of reconstituted YFP resulting from bona fide fusion protein partners localized on ER.

60 APPLIED LIFE SCIENCES↗

Development of a reverse genetics system to generate a recombinant Ebola virus Makona expressing a green fluorescent protein

Previous studies have demonstrated the potential application of reverse genetics technology in studying a broad range of aspects of viral biology, including gene regulation, protein function, cell entry, and pathogenesis. Here, we describe a highly efficient reverse genetics system used to generate recombinant Ebola virus (EBOV) based on a recent isolate from a human patient infected during the 2014–2015 outbreak in Western Africa. We also rescued a recombinant EBOV expressing a fluorescent reporter protein from a cleaved VP40 protein fusion. Using this virus and an inexpensive method to quantitate the expression of the foreign gene, we demonstrate its potential usefulness as a tool for screening antiviral compounds and measuring neutralizing antibodies. - Highlights: • Recombinant Ebola virus (EBOV) derived from Makona variant was rescued. • New protocol for viral rescue allows 100% efficiency. • Modified EBOV expresses a green fluorescent protein from a VP40-fused protein. • Modified EBOV was tested as tool to screen antiviral compounds and measure neutralizing antibodies.

60 APPLIED LIFE SCIENCES↗

Determining heavy metals in spent compact fluorescent lamps (CFLs) and their waste management challenges: Some strategies for improving current conditions

Highlights: • Heavy metals in spent compact fluorescent lamps (CFLs) determined. • Current waste management condition of CFLs in Iran assessed. • Currently, waste of CFLs is disposed by municipal waste stream in waste landfills. • We propose extended producer responsibility (EPR) for CFLs waste management. - Abstract: From environmental viewpoint, the most important advantage of compact fluorescent lamps (CFLs) is reduction of green house gas emissions. But their significant disadvantage is disposal of spent lamps because of containing a few milligrams of toxic metals, especially mercury and lead. For a successful implementation of any waste management plan, availability of sufficient and accurate information on quantities and compositions of the generated waste and current management conditions is a fundamental prerequisite. In this study, CFLs were selected among 20 different brands in Iran. Content of heavy metals including mercury, lead, nickel, arsenic and chromium was determined by inductive coupled plasma (ICP). Two cities, Tehran and Tabriz, were selected for assessing the current waste management condition of CFLs. The study found that waste generation amount of CFLs in the country was about 159.80, 183.82 and 153.75 million per year in 2010, 2011 and 2012, respectively. Waste generation rate of CFLs in Iran was determined to be 2.05 per person in 2012. The average amount of mercury, lead, nickel, arsenic and chromium was 0.417, 2.33, 0.064, 0.056 and 0.012 mg per lamp, respectively. Currently, waste of CFLs is disposed by municipal waste stream in waste landfills. For improving the current conditions, we propose by considering the successful experience of extended producer responsibility (EPR) in other electronic waste management. The EPR program with advanced recycling fee (ARF) is implemented for collecting and then recycling CFLs. For encouraging consumers to take the spent CFLs back at the end of the products’ useful life, a proportion of ARF (for example, 50%) can be refunded. On the other hand, the government and Environmental Protection Agency should support and encourage recycling companies of CFLs both technically and financially in the first place.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Multicolor Three-Dimensional Tracking for Single-Molecule Fluorescence Resonance Energy Transfer Measurements

Single-molecule fluorescence resonance energy transfer (smFRET) remains a widely utilized and powerful tool for quantifying heterogeneous interactions and conformational dynamics of biomolecules. However, traditional smFRET experiments either are limited to short observation times (typically less than 1 ms) in the case of “burst” confocal measurements or require surface immobilization which usually has a temporal resolution limited by the camera framing rate. We developed a smFRET 3D tracking microscope that is capable of observing single particles for extended periods of time with high temporal resolution. The confocal tracking microscope utilizes closed-loop feedback to follow the particle in solution by recentering it within two overlapping tetrahedral detection elements, corresponding to donor and acceptor channels. We demonstrated the microscope’s multicolor tracking capability via random walk simulations and experimental tracking of 200 nm fluorescent beads in water with a range of apparent smFRET efficiency values, 0.45-0.69. We also demonstrated the microscope’s capability to track and quantify double-stranded DNA undergoing intramolecular smFRET in a viscous glycerol solution. In future experiments, the smFRET 3D tracking system will be used to study protein conformational dynamics while diffusing in solution and native biological environments with high temporal resolution.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Nanofocused Scanning X-ray Fluorescence Microscopy Revealing an Effect of Heterozygous Hemoglobin S and C on Biochemical Activities in Plasmodium falciparum -Infected Erythrocytes

While there is ample evidence suggesting that carriers of heterozygous hemoglobin S and C are protected from life-threatening malaria, little is known about the underlying biochemical mechanisms at the single cell level. In this work, using nanofocused scanning X-ray fluorescence microscopy, we quantify the spatial distribution of individual elements in subcellular compartments, including Fe, S, P, Zn, and Cu, in Plasmodium falciparum -infected ( P. falciparum -infected) erythrocytes carrying the wild type or variant hemoglobins. Our data indicate that heterozygous hemoglobin S and C significantly modulate biochemical reactions in parasitized erythrocytes, such as aberrant hemozoin mineralization and a delay in hemoglobin degradation. The label-free scanning X-ray fluorescence imaging has great potential to quantify the spatial distribution of elements in subcellular compartments of P. falciparum -infected erythrocytes and unravel the biochemical mechanisms underpinning disease and protective traits.

59 BASIC BIOLOGICAL SCIENCES↗

Advanced Analysis of X-ray Fluorescence Measurements of the Interfacial Density of Eu Ions at Liquid–Liquid Interfaces for Solvent Back-Extraction

X-ray fluorescence near total reflection (XFNTR) is the primary technique used to measure the element-specific interfacial density of ions at liquid−liquid interfaces. Fluorescence from ions in the bulk liquids can complicate the determination of the interfacial density; consequently, measurements have been previously limited to samples without ions in the upper phase and with concentrations on the order of 10 μM or less in the lower phase. We modify the analysis of XFNTR data to account for ions in both bulk phases, then demonstrate its use in the context of rare-earth separations processes. In a model of ion stripping (i.e., back-extraction), dodecane solutions of di(2-ethylhexyl)- phosphoric acid (HDEHP) loaded with Eu(III) at the 1 mM level are placed in contact with either pure water or aqueous solutions of nitric or citric acid at pH 3. XFNTR measurements of equilibrated, quiescent samples reveal that citric acid solutions produce a disproportionately large depletion of ions from the interface compared to that from the bulk organic solution, whereas stripping by pure water or nitric acid solutions is negligible. As a result, this advance in the methodology of XFNTR may have broad applicability to the investigation of metal ions in chemical and biological processes at liquid−liquid interfaces.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

Small Molecule Sorting: A Fluorescence Study of Microemulsions

The application of microemulsions to a wide range of industries relies on their ability to solubilize small molecules with vastly different structures. Herein, we use multiple fluorescence techniques to probe ionic (rhodamine 6g, r6g), polar (coumarin 153, c153), nonpolar (diphenylanthracene, DPA), and amphiphilic (laurdan) small molecules in a nonionic, bicontinuous microemulsion of varying hydration. All fluorophores investigated were found to associate with the surfactant region despite their different structures and properties. The hydration of the surfactant layer was found to increase linearly with water addition, but while this initially increases the fluidity of the surfactant layer, fluorescence anisotropy of c153 and r6g indicates a stiffening of the surfactant at water content >60%. This stiffening of the surfactant layer at higher water content correlates with a morphological change in the microemulsion from a bicontinuous structure to droplets. In contrast, the nonpolar DPA shows a change in partitioning as hydration changes, increasing its association with the oil domain. Altogether, these studies elucidate not only the capability of these microemulsions to host a range of small molecules in the surfactant layer with tunable position but also the ability to probe the driving force of bulk structural changes in these heterogeneous fluids.

25 ENERGY STORAGE↗

Solvent-Induced Swelling Behaviors of Microphase-Separated Polystyrene- block -Poly(ethylene oxide) Thin Films Investigated Using In Situ Spectroscopic Ellipsometry and Single-Molecule Fluorescence Microscopy

Block copolymers have attracted considerable interest in the fields of nanoscience and nanotechnology, because these polymers afford well-defined nanostructures via self-assembly. An in-depth understanding of solvent effects on the physicochemical properties of these microdomains is crucial for their preparation and utilization. Herein, we employed in situ spectroscopic ellipsometry and single-molecule fluorescence techniques to gain detailed insights into microdomain properties in polystyrene-block-poly(ethylene oxide) (PS-b-PEO) films exposed to ethanol- and water-saturated N 2 . We observed a quick increase and a subsequent gradual decrease in the ellipsometric thickness of PS-b-PEO films upon exposure to ethanol-saturated N 2 . This observation was unexpected because ethanol-saturated N 2 induced negligible thickness change for PS and PEO homopolymer films. The similarity in maximum thickness gain observed under ethanol- and water-saturated N 2 implied the swelling of PEO microdomains. Ethanol vapor permeation through the PEO microdomains was supported by the red-shift of the ensemble and single-molecule fluorescence emission of Nile red in PS-b-PEO films. Single-molecule tracking data showed the initial enhancement and subsequent reduction of the diffusion of hydrophilic sulforhodamine B molecules in PS-b-PEO films upon exposure to ethanol-saturated N 2 , consistent with the spectroscopic ellipsometry results. Furthermore, the higher ethanol susceptibility of the PEO microdomains was attributable to their amorphous nature, as shown by FTIR data.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Single-Molecule Fluorescence Investigations of Solute Transport Dynamics in Nanostructured Membrane Separation Materials

Many materials used for membrane separations comprise of nanoscale structures such as pores and domains. Such nanostructures often control the solute permeability and selectivity of separation membranes. Thus, for future development of highly efficient separation membranes, it is important to understand the structural and chemical properties of these nanostructures, and also their influences on solute transport dynamics. For the last two decades, single-molecule fluorescence techniques have been used to measure the detailed dynamics of solute molecules diffusing in various nanostructured materials, giving valuable insights into molecular transport mechanisms influenced by nanoscale material heterogeneity. This Perspective will discuss recent single-molecule fluorescence studies on solute diffusion in materials relevant to membrane separations, including dense polymer films and nanoporous materials. These studies have revealed the formation and properties of nanostructures, and unique transport dynamics of solute molecules manipulated by their confinement and partitioning to the nanostructures, which play key roles in membrane separations. Furthermore, this Perspective will also point out scientific challenges toward thorough understanding of molecular-level mechanisms in membrane separations.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Fluorescence-Enabled Self-Reporting for Redox Flow Batteries

Monitoring battery health is challenging. Self-reporting enables the rapid health assessment of redox flow batteries (RFBs) and provides insight into degradation mechanisms of electrochemically active molecules (redoxmers). In this study, we introduce fluorescence as an orthogonal property to probe this chemistry in situ. An established anolyte redoxmer, 2,1,3-benzothiadiazole, is rendered an efficient blue-green fluorophore through minimalistic derivatization. One of the derivatives is electrochemically reversible with a long lifetime and cycling stability in the charged state. Spectroscopic measurements on this new redoxmer reveal strong effects of the electrolyte cation on the fluorescence that are useful for probing the solvent microenvironment. Using this probe, we demonstrate proof-of-concept in situ crossover sensing and characterize the effects of electrolyte composition on this crossover. In this way, real-time tracing of redoxmers in a flow cell is demonstrated, paving the way to include still more self-reporting functions into the redoxmers.

25 ENERGY STORAGE↗

Localizing Exciton Recombination Sites in Conformationally Distinct Single Conjugated Polymers by Super-resolution Fluorescence Imaging

To thoroughly elucidate how molecular conformation and photophysical properties of conjugated polymers (CPs) are related requires simultaneous probing of both. Here, previous efforts used fluorescence imaging with one nanometer accuracy (FIONA) to image CPs, which allowed simultaneous estimation of molecular conformation and probing of fluorescence intensity decay. We show that calculating the molecular radius of gyration for putative folded and unfolded poly(2-methoxy-5-(2'-ethylhexyloxy)1,4-phenylenevinylene) (MEH-PPV) molecules using FIONA underestimates molecular extension by averaging over emitters during localization. In contrast, employing algorithms based on single molecule high resolution imaging with photobleaching (SHRImP), including an approach we term all-frames SHRImP, allows localization of individual emitters. SHRImP processing corroborates that compact MEH-PPV molecules have distinct photophysical properties from extended ones. Estimated radii of gyration for isolated 168 kDa MEH-PPV molecules immobilized in polystyrene and exhibiting either stepwise or continuous intensity decays are found to be 12.6 and 25.3 nm, respectively, while the distance between exciton recombination sites is estimated to be ~10 nm independent of molecular conformation.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Developing Fluorescence-Based Sensors to Support Rare Earth Element Separation

Rare earth elements (REEs) are essential to most renewable energy technologies. Unfortunately, as we transition to sustainable energy production, the demand for REEs is rapidly growing well beyond current rates of production. As a result, novel means of efficient, scalable, and easily adaptable methods for processing primary and recycle feedstocks are needed. Development and integration of sensors for highly selective in-line monitoring can support more efficient design and testing of such novel separation processes, as well as more cost-effective deployment of those separation flowsheets. Work here will explore the application of fluorescence spectroscopy, a highly sensitive and selective technique, to quantify multiple lanthanides in complex mixtures including known interferents or quenching agents. Results include identification of the optimal excitation wavelength and the limit of detection of various rare earth elements as well as the performance of data-science-based quantification approaches in streams where “unknowns” are present. Overall, the data science tools in conjunction with optical sensor data were able to quantify analytes in the presence of other lanthanides which can be anticipated in the actual industrial stream. Here we include characterization of lanthanides in a microfluidic device similar to those used in new process development. This study demonstrates the capability of utilizing fluorescence spectroscopy to quantify analytes in a complicated solution matrix, suggesting this is a successful approach for in-line monitoring to optimize the separation efficiency in an industrial stream.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗