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At least 55 records · Page 3

Fluorescent compositions

An article that includes a fluorescent composition having at least one of a fluorescent sensor compound and organic reporter molecules encapsulated in a microsphere structure. When encapsulated, the fluorescent sensor compound and the organic reporter molecules are distributed in a liquid organic matrix. When non-encapsulated, the remaining one of the fluorescent sensor compound and the organic reporter molecules reside in the matrix. In response to a force applied to the composition sufficient to break at least a portion of the microsphere structure, the fluorescent sensor compound and the organic reporter molecules are transformed into a non-reversible fluorescent state exhibiting a quantum yield greater than 0.2. The fluorescent state is objectively visually verifiable without physically contacting the composition.

Smartt, Heidi A.↗

Fluorescence Enhancement in the Solid State by Isolating Perylene Fluorophores in Metal–Organic Frameworks

Polycyclic aromatic hydrocarbons such as perylene, pyrene, and their derivatives are highly emissive fluorophores in solution. Furthermore, the practical applications of these materials in the field of molecular electronic and light emitting devices are often hindered by self-quenching effects due to the formation of non-fluorescent aggregates in concentrated solutions or in the solid state. Herein, we demonstrate that aggregation caused quenching of perylenes can be minimalized by molecular incorporation into metalorganic frameworks (MOFs). This study utilized a stable Zr 6 cluster-based MOF, UiO-67, as a matrix. Linear linkers containing the photo-responsive moieties were designed and incorporated into the parent UiO-67 scaffold through the partially replacement of the non-fluorescent linkers of a similar length, forming mixed-linker MOFs. Here, the average distance between perylene moieties was tuned by changing the linker ratios, thus controlling the fluorescence intensity, emission wavelength, and quantum yield. Molecular modeling was further adopted to correlate the number of isolated perylene linkers within the framework to the ratio between the two linkers, therefore rationalizing the change in the observed fluorescent properties. Taking advantage of the tunable fluorescence, inherent porosity, and high chemical stability of this MOF platform, it was applied as a fluorescent sensor for oxygen detection in the gas phase, a model reaction, showing fast response and good recyclability.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Imaging of multiple fluorescent proteins in canopies enables synthetic biology in plants

Reverse genetics approaches have revolutionized plant biology and agriculture. Phenomics has the prospect of bridging plant phenotypes with genes, including transgenes, to transform agricultural fields. Genetically encoded fluorescent proteins (FPs) have revolutionized plant biology paradigms in gene expression, protein trafficking and plant physiology. While the first instance of plant canopy imaging of green fluorescent protein (GFP) was performed over 25 years ago, modern phenomics has largely ignored fluorescence as a transgene expression device despite the burgeoning FP colour palette available to plant biologists. Here, we show a new platform for stand-off imaging of plant canopies expressing a wide variety of FP genes. The platform-the fluorescence-inducing laser projector (FILP)-uses an ultra-low-noise camera to image a scene illuminated by compact diode lasers of various colours, coupled with emission filters to resolve individual FPs, to phenotype transgenic plants expressing FP genes. Each of the 20 FPs screened in plants were imaged at >3 m using FILP in a laboratory-based laser range. We also show that pairs of co-expressed fluorescence proteins can be imaged in canopies. The FILP system enabled a rapid synthetic promoter screen: starting from 2000 synthetic promoters transfected into protoplasts to FILP-imaged agroinfiltrated Nicotiana benthamiana plants in a matter of weeks, which was useful to characterize a water stress-inducible synthetic promoter. FILP canopy imaging was also accomplished for stably transformed GFP potato and in a split-GFP assay, which illustrates the flexibility of the instrument for analysing fluorescence signals in plant canopies.

abiotic stress↗

Advances in multi-dimensional cellulose-based fluorescent carbon dot composites

Recently, fluorescent materials composed of carbon dots (CDs) have attracted increasing attention in diverse fields, including the environment, food, biology, and medicine. Cellulose has emerged as a promising class of materials for carriers due to its low cost, fascinating biodegradability, and various morphologies, especially in available different dimensions. By taking advantage of the respective superiority of fluorescent materials and cellulose carriers, the performance of fluorescence composite materials can be optimized and their application may be extended. Here, in this review, the study emphasizes the synthesis and applications of cellulose-based fluorescent materials with different dimensions. Moreover, some effective strategies and potential challenges for further development of cellulose-based fluorescent materials are discussed. A deeper understanding will provide a guide for the materials with ideal fluorescent performances.

42 ENGINEERING↗

Fluorescence Signatures of Rare Earth Metals during Precipitation in Various Conditions

Fluorescence spectroscopy is a widely used sensor methodology that analyzes light emitted from a compound or element as it decays from an excited state. This technique is very sensitive and selective, which is ideal to characterize analytes at lower limits of detection. Key example targets of significant industry and research interest include rare earth elements (REEs) such as dysprosium (Dy) and europium (Eu). These are widely used in advanced technologies including semiconductors, electric vehicle motors, lasers, and permanent magnets. Identifying new sources and responsible reutilization of REEs is essential, and new approaches to extract and recycle REEs could be notably enhanced through the integration of on-line sensors. The sensors can support faster process design, informed scale-up, and cost-effective deployment. This study covers the initial exploration of applying fluorescence-based on-line monitoring to REEs within a precipitation process. This study demonstrates the successful scale-up of a fluorescence -based sensing approach, from stationary cuvettes and small-volume microfluidic devices to continuous flow systems operating at the bench scale (10-25mL). This work also provides initial insight into the challenges of signal’s effects and utility within a turbid environment. Using a modular design for monitoring flowing solutions in a flow tube, fluorescence can be characterized for a variety of analytical targets. In this study, detection performance parameters between the cuvette and flow tube system were compared. Additionally, the response of Dy during precipitation by sodium bicarbonate in the two measurement designs was explored. This letter represents a starting point to bridge the gap between traditional fluorescence sensor measurements in a cuvette to future developments that explore the ability to integrate fluorescence sensors into extraction and separation processes at industrially relevant scales.

fluorescence↗

Fluorescence Enhancement of Lignin-Based Carbon Quantum Dots by Concentration-Dependent and Electron-Donating Substituent Synergy and Their Cell Imaging Applications

Black liquor is an important pollutant in the pulp industry, but it also has the potential for high-value utilization. In this study, lignin extracted from black liquor was hydrothermally prepared into lignin-based carbon quantum dots (L-CQDs) using a one-pot method. Physicochemical characterization suggested that the L-CQDs exhibited a lamellar core–shell multilayered graphene structure surrounded by oxygen-containing functional groups. Here, the fluorescence intensity of the L-CQDs was strengthened depending on their own concentration dependence and the doping of external groups. The fluorescence intensity of L-CQDs varied between 89.09 and 183.66 under different concentrations, and the most intense fluorescence (183.66) was obtained at 0.1 mg mL –1 . At hydroxyl and amino adsorption capacities of 11.08 and 0.98 mmol g –1 , the hydroxylated RL-CQDs-5 and aminated NL-CQDs-3 exhibited the highest fluorescence intensities at 689.22 and 605.39, respectively. Moreover, when pristine L-CQDs were sequentially aminated and hydroxylated, the NRL-CQDs’ fluorescence intensity reached 1224.92. Cell imaging experiments proved that cells cultivated with NRL-CQDs have brighter fluorescence compared with L-CQDs. The results will render L-CQDs more suitable for practical applications.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Excitation spectral microscopy for highly multiplexed fluorescence imaging and quantitative biosensing

The multiplexing capability of fluorescence microscopy is severely limited by the broad fluorescence spectral width. Spectral imaging offers potential solutions, yet typical approaches to disperse the local emission spectra notably impede the attainable throughput. Here we show that using a single, fixed fluorescence emission detection band, through frame-synchronized fast scanning of the excitation wavelength from a white lamp via an acousto-optic tunable filter, up to six subcellular targets, labeled by common fluorophores of substantial spectral overlap, can be simultaneously imaged in live cells with low (~1%) crosstalks and high temporal resolutions (down to ~10 ms). The demonstrated capability to quantify the abundances of different fluorophores in the same sample through unmixing the excitation spectra next enables us to devise novel, quantitative imaging schemes for both bi-state and Förster resonance energy transfer fluorescent biosensors in live cells. We thus achieve high sensitivities and spatiotemporal resolutions in quantifying the mitochondrial matrix pH and intracellular macromolecular crowding, and further demonstrate, for the first time, the multiplexing of absolute pH imaging with three additional target organelles/proteins to elucidate the complex, Parkin-mediated mitophagy pathway. Together, excitation spectral microscopy provides exceptional opportunities for highly multiplexed fluorescence imaging. The prospect of acquiring fast spectral images without the need for fluorescence dispersion or care for the spectral response of the detector offers tremendous potential.

59 BASIC BIOLOGICAL SCIENCES↗

Speckle contrast of interfering fluorescence X-rays

With the development of X-ray free-electron lasers (XFELs), producing pulses of femtosecond durations comparable with the coherence times of X-ray fluorescence, it has become possible to observe intensity–intensity correlations due to the interference of emission from independent atoms. This has been used to compare durations of X-ray pulses and to measure the size of a focused X-ray beam, for example. Here it is shown that it is also possible to observe the interference of fluorescence photons through the measurement of the speckle contrast of angle-resolved fluorescence patterns. Speckle contrast is often used as a measure of the degree of coherence of the incident beam or the fluctuations of the illuminated sample as determined from X-ray diffraction patterns formed by elastic scattering, rather than from fluorescence patterns as addressed here. Commonly used approaches to estimate speckle contrast were found to suffer when applied to XFEL-generated fluorescence patterns due to low photon counts and a significant variation of the excitation pulse energy from shot to shot. A new method to reliably estimate speckle contrast under such conditions, using a weighting scheme, is introduced. The method is demonstrated by comparing the speckle contrast of fluorescence observed with pulses of 3 fs to 15 fs duration.

73 NUCLEAR PHYSICS AND RADIATION PHYSICS↗

Nanomechanical and Fluorescence Characterizations of Weathered PV Module Encapsulation

Nanoindentation and fluorescence spectroscopy provide spatially resolved mechanical and chemical characterization for degradation of poly (ethylene-co-vinyl acetate) (EVA) encapsulants aged under accelerated weathering and in field deployed photovoltaic (PV) modules. For the accelerated weathering tests, two different EVA formulations in the glass/EVA/glass-laminated coupons were exposed to ultraviolet (UV) light for 180 days at three different temperatures (40, 60, and 80 °C) and UV intensities (0.4, 2.2, and 2.9 W/m2/nm at 340 nm). The fluorescence spectra and moduli were uniform through the cross section of both EVA formulations at the lowest temperature and UV intensity; however, under the highest temperature and UV intensity, the UV irradiated sides showed lower moduli and higher fluorescence compared to the opposite sides. For EVA obtained from PV modules deployed 27 years in Sacramento, CA, USA, changes in fluorescence intensity across the thickness were relatively greater than changes in modulus. On the other hand, for EVA obtained from the same type of module stored in the shed for 27 years, no fluorescence was observed, and a uniform modulus was seen through the EVA layer, but with substantially higher values than those obtained from the field-deployed modules. The different extent of changes between laboratory and field exposures suggests the importance of considering effects of temperature and UV intensity for designing a reliable accelerated testing. This study uniquely demonstrates the capability of microscale characterization using nanoindentation and fluorescence microscopy to monitor mechanical and chemical degradation of EVA encapsulants in PV modules.

14 SOLAR ENERGY↗

Optimization of fluorescence and surface adsorption of citric acid/ethanolamine carbon nanoparticles for subsurface tracers

The tunable fluorescence and biocompatibility of fluorescent carbon nanoparticles (FCNs) makes them appealing tracers in geothermal, environmental, and clinical applications. Here, we address FCNs synthesized from reaction and pyrolysis of citric acid and ethanolamine. We examine the fluorescence and the adsorption of these FCNs as critical parameters that influence their use in applications. FCN fluorescence was observed to develop during pyrolysis and measured at pyrolysis temperatures ranging from 190 to 250 °C. Transmission electron microscopy shows that the FCN average diameter of approximately 13 nm is unaffected by pyrolysis temperature. However, pyrolysis temperature strongly affects fluorescence, which peaks at 210 °C. Here, the surface hydrophobicity of the FCNs increases with pyrolysis temperature as measured by retention on reverse phase chromatography. Together, these results suggest that particle surface functionalities provide both fluorescence and hydrophilicity, and that these surface functionalities are destroyed with excessive pyrolysis. To examine their surface adsorption, quartz crystal microbalance measurements demonstrate that pyrolysis temperature increases FCN adsorption on mineral surfaces. Surfaces with greater hydrophobicity show greater FCN adsorption. Adsorption is unaffected by the presence of mono- or divalent ions in solution. This suggests adsorption by hydrophobic interaction for the Fe 2 O 3 , SiO 2 , and AlSiO surfaces studied.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

A fluorescent-protein spin qubit

Quantum bits (qubits) are two-level quantum systems that support initialization, readout and coherent control1. Optically addressable spin qubits form the foundation of an emerging generation of nanoscale sensors. The engineering of these qubits has mainly focused on solid-state systems. However, fluorescent proteins, rather than exogenous fluorescent probes, have become the gold standard for in vivo microscopy because of their genetic encodability. Although fluorescent proteins possess a metastable triplet state, they have not been investigated as qubits. Here we realize an optically addressable spin qubit in enhanced yellow fluorescent protein. A near-infrared laser pulse enables triggered readout of the triplet state with up to 20% spin contrast. Using coherent microwave control of the enhanced-yellow-fluorescent-protein spin at liquid-nitrogen temperatures, we measure a (16 ± 2) μs coherence time under Carr–Purcell–Meiboom–Gill decoupling. We express the qubit in mammalian cells, maintaining contrast and coherent control despite the complex intracellular environment. Finally, we demonstrate optically detected magnetic resonance in bacterial cells at room temperature with contrast up to 8%. Our results introduce fluorescent proteins as a powerful qubit platform that paves the way for applications in the life sciences, such as nanoscale field sensing and spin-based imaging modalities.

Feder, Jacob S. [Univ. of Chicago, IL (United Stat↗

Detection and quantification of γ-H2AX using a dissociation enhanced lanthanide fluorescence immunoassay

Phosphorylation of the histone protein H2AX to form γ-H2AX foci directly represents DNA double-strand break formation. Traditional γ-H2AX detection involves counting individual foci within individual nuclei. The novelty of this work is the application of a time-resolved fluorescence assay using dissociation-enhanced lanthanide fluorescence immunoassay for quantitative measurements of γ-H2AX. For comparison, standard fluorescence detection was employed and analyzed either by bulk fluorescent measurements or by direct foci counting using BioTek Spot Count algorithm and Gen 5 software. Etoposide induced DNA damage in A549 carcinoma cells was compared across all test platforms. Time resolved fluorescence detection of europium as a chelated complex enabled quantitative measurement of γ-H2AX foci with nanomolar resolution. Comparative bulk fluorescent signals achieved only micromolar sensitivity. Lanthanide based immunodetection of γ-H2AX offers superior detection and a user-friendly workflow. These approaches have the potential to improve screening of compounds that either enhance DNA damage or protect against its deleterious effects.

59 BASIC BIOLOGICAL SCIENCES↗

Mixed metal zero-mode guides (ZMWs) for tunable fluorescence enhancement

Zero-mode waveguides (ZMWs) are capable of modifying fluorescence emission through interactions with surface plasmon modes leading to either plasmon-enhanced fluorescence or quenching. Enhancement requires spectral overlap of the plasmon modes with the absorption or emission of the fluorophore. Thus, enhancement is limited to fluorophores in resonance with metals (e.g. Al, Au, Ag) used for ZMWs. The ability to tune interactions to match a wider range of fluorophores across the visible spectra would significantly extend the utility of ZMWs. We fabricated ZMWs composed of aluminum and gold individually and also in mixtures of three different ratios, (Al : Au; 75 : 25, 50 : 50, 25 : 75). We characterized the effect of mixed-metal ZMWs on single-molecule emission for a range fluorophores across the visible spectrum. Mixed metal ZMWs exhibited a shift in the spectral range where they exhibited the maximum fluorescence enhancement allowing us to match the emission of fluorophores that were nonresonant with single metal ZMWs. We also compared the effect of mixed-metal ZMWs on the photophysical properties of fluorescent molecules due to metal–molecule interactions. We quantified changes in fluorescence lifetimes and photostability that were dependent on the ratio of Au and Al. Tuning the enhancement properties of ZMWs by changing the ratio of Au and Al allowed us to match the fluorescence of fluorophores that emit in different regions of the visible spectrum.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

CRAGE-mediated insertion of fluorescent chromosomal markers for accurate and scalable measurement of co-culture dynamics in Escherichia coli

Monitoring population dynamics in co-culture is necessary in engineering microbial consortia involved in distributed metabolic processes or biosensing applications. However, it remains difficult to measure strain-specific growth dynamics in high-throughput formats. This is especially vexing in plate-based functional screens leveraging whole-cell biosensors to detect specific metabolic signals. Here, we develop an experimental high-throughput co-culture system to measure and model the relationship between fluorescence and cell abundance, combining chassis-independent recombinase-assisted genome engineering (CRAGE) and whole-cell biosensing with a PemrR-green fluorescent protein (GFP) monoaromatic reporter used in plate-based functional screening. CRAGE was used to construct Escherichia coli EPI300 strains constitutively expressing red fluorescent protein (RFP) and the relationship between RFP expression and optical density (OD600) was determined throughout the EPI300 growth cycle. A linear equation describing the increase of normalized RFP fluorescence during deceleration phase was derived and used to predict biosensor strain dynamics in co-culture. Measured and predicted values were compared using flow cytometric detection methods. Induction of the biosensor lead to increased GFP fluorescence normalized to biosensor cell abundance, as expected, but a significant decrease in relative abundance of the biosensor strain in co-culture and a decrease in bulk GFP fluorescence. Taken together, these results highlight sensitivity of population dynamics to variations in metabolic activity in co-culture and the potential effect of these dynamics on the performance of functional screens in plate-based formats. The engineered strains and model used to evaluate these dynamics provide a framework for optimizing growth of synthetic co-cultures used in screening, testing and pathway engineering applications.

59 BASIC BIOLOGICAL SCIENCES↗

Localization of fluorescent targets in deep tissue with expanded beam illumination for studies of cancer and the brain

Imaging fluorescence through millimeters or centimeters of tissue has important in vivo applications, such as guiding surgery and studying the brain. Often, the important information is the location of one of more optical reporters, rather than the specifics of the local geometry, motivating the need for a localization method that provides this information. We present an optimization approach based on a diffusion model for the fast localization of fluorescent inhomogeneities in deep tissue with expanded beam illumination that simplifies the experiment and the reconstruction. We show that the position of a fluorescent inhomogeneity can be estimated while assuming homogeneous tissue parameters and without having to model the excitation profile, reducing the computational burden and improving the utility of the method. We perform two experiments as a demonstration. First, a tumor in a mouse is localized using a near infrared folate-targeted fluorescent agent (OTL38). This result shows that localization can quickly provide tumor depth information, which could reduce damage to healthy tissue during fluorescence-guided surgery. Second, another near infrared fluorescent agent (ATTO647N) is injected into the brain of a rat, and localized through the intact skull and surface tissue. This result will enable studies of protein aggregation and neuron signaling.

60 APPLIED LIFE SCIENCES↗

Waveform-dependent air fluorescence from neutral and ionic nitrogen molecules

Laser-induced air fluorescence in the ultraviolet regime is primarily attributed to transitions between the C and B states in excited neutral N 2 molecules and between the B and X states in N$^+_2$ ions. However, the mechanism underlying the former remains contentious, as direct population to the C state by light fields is forbidden by electron spin constraints. In this work, we investigate the mechanism of air fluorescence from excited neutral N 2 molecules by carrier-envelope phase–stabilized sub–4 femtosecond pulses. Our results show that fluorescence from N$^+_2$ ions reaches a maximum with cosine-like pulses, while fluorescence from excited neutral N 2 molecules peaks with sine-like pulses. In addition, by scanning the chirp of the driving pulse, we find that ionic fluorescence is maximized with chirp-free pulses, whereas neutral fluorescence favors negatively chirped pulses. These observations, supported by classical trajectory Monte Carlo simulations, support the mechanism of intersystem crossing from excited spin-singlet states, which are populated via recollision-induced strong-field excitation.

Science & Technology - Other Topics↗

Biomimetic tissue phantoms for neurosurgical near-infrared fluorescence imaging

Neurosurgical fluorescence imaging is a well-established clinical approach with a growing range of indications for use. However, this technology lacks effective phantom-based tools for development, performance testing, and clinician training. Our primary aim was to develop and evaluate 3D-printed phantoms capable of optically and morphologically simulating neurovasculature under fluorescence angiography. Volumetric digital maps of the circle of Willis with basilar and posterior communicator artery aneurysms, along with surrounding cerebral tissue, were generated. Phantoms were fabricated with a stereolithography printer using custom photopolymer composites, then visualized under white light and near-infrared fluorescence imaging. Feature sizes of printed components were found to be within 13% of digital models. Phantoms exhibited realistic optical properties and convincingly recapitulated fluorescence angiography scenes. Methods identified in this study can facilitate the development of realistic phantoms as powerful new tools for fluorescence imaging.

3D modeling↗

Fluorescent Particle Vertical Profiles (Counts and Fractions) at SAIL

This dataset contains vertically resolved fluorescent aerosol number concentrations and fluorescent fractions of total aerosol number concentrations measured with the ARM Tethered Balloon System during the Surface Atmosphere Integrated Field Laboratory campaign near Gothic, Colorado. Measurements were collected during April and June 2023 as part of the ARM–EMSL FICUS project “Characterization of the Aerobiome during the ARM Surface-Atmosphere Integrated Laboratory Campaign,” which was designed to characterize the vertical distribution of bioaerosol-relevant particles in the Colorado River Basin. The dataset includes profiles of fluorescent particle concentrations measured with a Fluorescent Portable Optical Particle Sizer (FPOPS) in the approximately 0.5–3 µm diameter range. For fluorescent fractions of total aerosol number concentrations, we compare FPOPS measurements to POPS on the same TBS flight.

Fluorescent particle concentration↗