Standoff analysis of laser-produced plasmas using laser-induced fluorescence
Not Available
Harilal, S. S. (ORCID:0000000322667976)↗
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Results for “FLUORESCENCE”
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Optimization-Based Approach for Joint X-Ray Fluorescence and Transmission Tomographic Inversion
Fluorescence tomographic reconstruction, based on the detection of photons coming from fluorescent emission, can be used for revealing the internal elemental composition of a sample. On the other hand, conventional X-ray transmission tomography can be used for reconstructing the spatial distribution of the absorption coefficient inside a sample. In this work, we integrate both X-ray fluorescence and X-ray transmission data modalities and formulate a nonlinear optimization-based approach for reconstruction of the elemental composition of a given object. This model provides a simultaneous reconstruction of both the quantitative spatial distribution of all elements and the absorption effect in the sample. Mathematically speaking, we show that compared with the single-modality inversion (i.e., the X-ray transmission or fluorescence alone), the joint inversion provides a better-posed problem, which implies a better recovery. Therefore, the challenges in X-ray fluorescence tomography arising mainly from the effects of self-absorption in the sample are partially mitigated. The use of this technique is demonstrated on the reconstruction of several synthetic samples.
46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗
A new fluorescence-based method to monitor the pH in the thylakoid lumen using GFP variants
The ΔpH-dependent/Tat pathway is unique for using only the proton motive force for driving proteins transport across the thylakoid membrane in chloroplasts. 9-aminoacridine fluorescence quenching is widely used to monitor the ΔpH developed across the thylakoid membrane in the light. However, this method suffers from limited sensitivity to low ΔpH values and to spurious fluorescence signals due to membrane binding. In order to develop a more sensitive method for monitoring the real pH of the thylakoid lumen without these problems we transformed Arabidopsis thaliana with a ratiometric pH-sensitive GFP variant (termed pHluorin) targeted to the lumen by the prOE17 transit peptide. Positive transgenic plants displayed localization of pHluorin in the chloroplast by confocal microscopy, and fractionation experiments revealed that it is in the lumen. The pHluorin signal was the strongest in very young plants and diminished as the plants matured. The pHluorin released from the lumen displayed the expected fluorescence intensity changes in response to pH titration. The fluorescence signal in isolated chloroplasts responded to illumination in a manner consistent with light-dependent lumen acidification. Future experiments will exploit the use of this new pH-indicating probe of the thylakoid lumen to examine the influence of the thylakoid ΔpH on ATP synthesis and protein transport.
60 APPLIED LIFE SCIENCES↗
Size-dependent fluorescence of bioaerosols: Mathematical model using fluorescing and absorbing molecules in bacteria
This paper uses a mathematical model of fluorescent biological particles composed of bacteria and/or proteins (mostly as in Hill et al., 2013 [23]) to investigate the size-dependence of the total fluorescence emitted in all directions. The model applies to particles which have negligible reabsorption of fluorescence within the particle. The specific particles modeled here are composed of ovalbumin and of a generic Bacillus. The particles need not be spherical, and in some cases need not be homogeneous. However, the results calculated in this paper are for spherical homogeneous particles. Light absorbing and fluorescing molecules included in the model are amino acids, nucleic acids, and several coenzymes. Here the excitation wavelength is 266 nm. The emission range, 300 to 370 nm, encompasses the fluorescence of tryptophan. The fluorescence cross section (C F ) is calculated and compared with one set of published measured values. We investigate power law (Ad y ) approximations to C F , where d is diameter, and A and y are parameters adjusted to fit the data, and examine how y varies with d and composition, including the fraction as water. The particle's fluorescence efficiency (Q F =C F /geometric-cross-section) can be written for homogeneous particles as Q abs R F , where Q abs is the absorption efficiency, and R F , the fraction of the absorbed light emitted as fluorescence, is independent of size and shape. When Q F is plotted vs. m i d or mi(m r -1)d, where m=m r +im i is the complex refractive index, the plots for different fractions of water in the particle tend to overlap.
54 ENVIRONMENTAL SCIENCES↗
Probing the predissociated levels of the S 1 state of acetylene via H-atom fluorescence and photofragment fluorescence action spectroscopy
Here, we report two new experimental schemes to obtain rotationally resolved high-resolution spectra of predissociated S 1 acetylene levels in the 47 000-47 300 cm -1 energy region (~1200 cm -1 above the predissociation threshold). The two new detection schemes are compared to several other detection schemes (employed at similar laser power, molecular beam temperature, and number of signal averages) that have been used in our laboratory to study predissociated S 1 acetylene levels, both in terms of the signal-to-noise ratio (S/N) of the resultant spectra and experimental simplicity. In the first method, H-atoms from the predissociated S 1 acetylene levels are probed by two-photon laser-induced fluorescence (LIF). The H-atoms are pumped to the 3d level by the two-photon resonance transition at 205.14 nm. The resulting 3d–2p fluorescence (654.5 nm) is collected by a photomultiplier. The S/N of the H-atom fluorescence action spectrum is consistently better by ~3× than that of the more widely used H-atom resonance-enhanced multiphoton ionization (REMPI) detection. Laser alignment is also considerably easier in H-atom fluorescence detection than H-atom REMPI detection due to the larger number-density of molecules that can be used in fluorescence vs. REMPI detection schemes. In the second method, fluorescence from electronically excited C 2 and C 2 H photofragments of S 1 acetylene is detected. In contrast to the H-atom detection schemes, the detected C 2 and C 2 H photofragments are produced by the same UV laser as is used for the ÖX acetylene excitation. As a result, laser alignment is greatly simplified for the photofragment fluorescence detection scheme, compared to both H-atom detection schemes. Using the photofragment fluorescence detection method, we are able to obtain action spectra of predissociated S 1 acetylene levels with S/N ~2× better than the HCCH REMPI detection and ~10× better than H-atom and HCCH LIF detection schemes.
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
Site‐selective fluorescence and spectroscopic properties of Yb‐doped lanthanum titanate glasses
Abstract Ytterbium‐doped lanthanum titanate glasses were prepared by levitation melting for the detailed characterization of the spectroscopic properties in the rare‐earth titanate glass host. Low‐temperature fluorescence spectroscopy reveals distinct site‐selectivity in both static and lifetime fluorescence measurements suggesting an absence of clustering as well as significant variation of local ytterbium environments. Typical site‐selectivity behavior of a shrinking Stark manifold with lower excitation energy is observed. At 77 K, both the mean emission frequency and the fluorescence lifetime initially increase as the excitation energy decreases from about 11100 to 10750 and then slightly decrease at lower excitation energy. Temperature‐dependent lifetime measurements between 77 and 420 K show a decreasing lifetime with increasing temperature and are well described by a two‐level thermal activation model. The temperature‐dependent fluorescence spectroscopy coupled with a room temperature white light absorption measurement allow the determination of the Stark energy levels of in lanthanum titanate glass as well as the calculation of the laser cross‐sections.
Materials Science↗
FSVPy: A python-based package for fluorescent streak velocimetry (FSV)
Predictive constitutive equations that connect easy-to-measure transport properties (e.g., viscosity and conductivity) with system performance variables (e.g., power consumption and efficiency) are needed to design advanced thermal and electrical systems. In this work, we explore the use of fluorescent particle-streak analysis to directly measure the local velocity field of a pressure-driven flow, introducing a new Python package (FSVPy) to perform the analysis. Fluorescent streak velocimetry combines high-speed imaging with highly fluorescent particles to produce images that contain fluorescent streaks, whose length and intensity can be related to the local flow velocity. By capturing images throughout the sample volume, the three-dimensional velocity field can be quantified and reconstructed. We demonstrate this technique by characterizing the channel flow profiles of several non-Newtonian fluids: micellar Cetylpyridinium Chloride solution, Carbopol 940, and Polyethylene Glycol. We then explore more complex flows, where significant acceleration is created due to microscale features encountered within the flow. We demonstrate the ability of FSVPy to process streaks of various shapes and use the variable intensity along the streak to extract position-specific velocity measurements from individual images. Thus, we demonstrate that FSVPy is a flexible tool that can be used to extract local velocimetry measurements from a wide variety of fluids and flow conditions.
Mechanics↗
Optimizing detector geometry for trace element mapping by X-ray fluorescence
Trace metals play critical roles in a variety of systems, ranging from cells to photovoltaics. X-Ray Fluorescence (XRF) microscopy using X-ray excitation provides one of the highest sensitivities available for imaging the distribution of trace metals at sub-100 nm resolution. With the growing availability and increasing performance of synchrotron light source based instruments and X-ray nanofocusing optics, and with improvements in energy-dispersive XRF detectors, what are the factors that limit trace element detectability? To address this question, we describe an analytical model for the total signal incident on XRF detectors with various geometries, including the spectral response of energy dispersive detectors. This model agrees well with experimentally recorded X-ray fluorescence spectra, and involves much shorter calculation times than with Monte Carlo simulations. With such a model, one can estimate the signal when a trace element is illuminated with an X-ray beam, and when just the surrounding non-fluorescent material is illuminated. From this signal difference, a contrast parameter can be calculated and this can in turn be used to calculate the signal-to-noise ratio (S/N) for detecting a certain elemental concentration. We apply this model to the detection of trace amounts of zinc in biological materials, and to the detection of small quantities of arsenic in semiconductors. We conclude that increased detector collection solid angle is (nearly) always advantageous even when considering the scattered signal. However, given the choice between a smaller detector at 90° to the beam versus a larger detector at 180° (in a backscatter-like geometry), the 90° detector is better for trace element detection in thick samples, while the larger detector in 180° geometry is better suited to trace element detection in thin samples.
46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗
Simultaneous cryo X-ray ptychographic and fluorescence microscopy of green algae
Trace metals play important roles in normal and in disease-causing biological functions. X-ray fluorescence microscopy reveals trace elements with no dependence on binding affinities (unlike with visible light fluorophores) and with improved sensitivity relative to electron probes. However, X-ray fluorescence is not very sensitive for showing the light elements that comprise the majority of cellular material. Here we show that X-ray ptychography can be combined with fluorescence to image both cellular structure and trace element distribution in frozen-hydrated cells at cryogenic temperatures, with high structural and chemical fidelity. Ptychographic reconstruction algorithms deliver phase and absorption contrast images at a resolution beyond that of the illuminating lens or beam size. Using 5.2-keV X-rays, we have obtained sub-30-nm resolution structural images and similar to 90-nm-resolution fluorescence images of several elements in frozen-hydrated green algae. This combined approach offers a way to study the role of trace elements in their structural context.
Deng, Junjing↗
Use of a Novel Rover-mounted Fluorescence Imager and Fluorescent Probes to Detect Biological Material in the Atacama Desert in Daylight
We have developed an imaging system, the Fluorescence Imager (FI), for detecting fluorescence signals from sparse microorganisms and biofilms during autonomous rover exploration. The fluorescence signals arise both from naturally occurring chromophores, such as chlorophyll of cyanobacteria and lichens, and from fluorescent probes applied to soil and rocks. Daylight imaging has been accomplished by a novel use of a high-powered flashlamp synchronized to a CCD camera. The fluorescent probes are cell permanent stains that have extremely low intrinsic fluorescence (quantum yields less than 0.01) and a large fluorescence enhancement (quantum yields greater than 0.4) when bound to the target. Each probe specifically targets either carbohydrates, proteins, nucleic acids or membrane lipids, the four classes of macromolecules found in terrestrial life. The intent of the probes is to interrogate the environment for surface and endolithic life forms.
Weinstein, S.↗
Development and Calibration of Electron Density Measurements in Argon Plasma Using Laser Collision-Induced Fluorescence.
Abstract not provided.
Barnat, Edward V.↗
Dye Encapsulation in Fluorescent Core–Shell Silica Nanoparticles as Probed by Fluorescence Correlation Spectroscopy
Synthetic advances in the formation of ultrasmall (<10 nm) fluorescent poly(ethylene glycol)-coated (PEGylated) core–shell silica nanoparticles (SNPs), enabling improved particle size and surface chemical property control, have led to successful clinical translation of SNPs as diagnostic probes in oncology. Despite the success of such probes, details of the dye incorporation and resulting silica architecture are still poorly understood. Here, we employ afterpulse-corrected fluorescence correlation spectroscopy (FCS) to monitor fast fluorescence fluctuations (lag times <10 –5 s) of the negatively charged cyanine dye Cy5 as a probe to study such details for dye encapsulation in 5 nm silica cores of PEGylated core–shell SNPs (C dots). Upon deposition of additional silica shells over the silica core, we find that the amplitude of photoinduced cis–trans isomerization decreases, suggesting that the Cy5 dyes are located near or on the surface of the original SNP cores. In combination with time-correlated fluorescence decay measurements, we deduce radiative and nonradiative rates of the Cy5 dye in these particles. Finally, results demonstrate that FCS is a well-suited tool to investigate aspects of the photophysics of fluorescent nanoparticles, and that conformational changes of cyanine dyes like Cy5 are excellent indicators for the local dye environment within ultrasmall SNPs.
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
Testing Fluorescence Lifetime Standards using Two-Photon Excitation and Time-Domain Instrumentation: Fluorescein, Quinine Sulfate and Green Fluorescent Protein
It is essential for everyone working with experimental science to be certain that their instruments produce reliable results, and for fluorescence lifetime experiments, information about fluorescence lifetime standards is crucial. A large part of the literature on lifetime standards dates back to the 1970s and 1980s, and the use of newer and faster measuring devices may deem these results unreliable. We have tested the three commonly used fluorophores fluorescein, quinine sulfate and green fluorescent protein for their suitability to serve as lifetime standards, especially to be used with two-photon excitation measurements in the time-domain. We measured absorption and emission spectra for the fluorophores to determine optimal wavelengths to use for excitation and detector settings. Fluorescence lifetimes were measured for different concentrations, ranging from 10{sup − 3} − 10{sup − 5} M, as well as for various solvents. Fluorescein was soluble in both ethanol, methanol and sulfuric acid, while quinine sulfate was only soluble in sulfuric acid. Green fluorescent protein was prepared in a commercial Tris-HCl, EDTA solution, and all three fluorophores produced stable lifetime results with low uncertainties. No siginificant variation with concentration was measured for any of the fluorophores, and all showed single-exponential decays. All lifetime measurements were carried out using two-photon excitation and lifetime data was obtained in the time-domain using time-correlated single-photon counting.
74 ATOMIC AND MOLECULAR PHYSICS↗
A Strongly Fluorescing Anaerobic Reporter and Protein-Tagging System for Clostridium Organisms Based on the Fluorescence-Activating and Absorption-Shifting Tag Protein (FAST)
Visualizing protein localization and characterizing gene expression activity in live Clostridium cells is limited for lack of a real-time, highly fluorescent, oxygen-independent reporter system. Enzymatic reporter systems have been used successfully for many years with Clostridium spp.; however, these assays do not allow for real-time analysis of gene expression activity with flow cytometry or for visualizing protein localization through fusion proteins. Commonly used fluorescent reporter proteins require oxygen for chromophore maturation and cannot be used for most strictly anaerobic Clostridium organisms. Here we show that the fluorescence-activating and absorption-shifting tag protein (FAST), when associated with the fluorogenic ligand 4-hydroxy-3-methylbenzylidene-rhodanine (HMBR; now commercially available) and other commercially available ligands, is highly fluorescent in Clostridium acetobutylicum under anaerobic conditions. Using flow cytometry and a fluorescence microplate reader, we demonstrated FAST as a reporter system by employing the promoters of the C. acetobutylicum thiolase (thl), acetoacetate decarboxylase (adc), and phosphotransbutyrylase (ptb) metabolic genes, as well as a mutant P thl and modified ribosome binding site (RBS) versions of P adc and P ptb . Flow cytometry-based sorting was efficient and fast in sorting FAST-expressing cells, and positively and negatively sorted cells could be effectively recultured. FAST was also used to tag and examine protein localization of the predicted cell division FtsZ partner protein, ZapA, to visualize the divisome localization in live C. acetobutylicum cells. Our findings suggest that FAST can be used to further investigate Clostridium divisomes and more broadly the localization and expression levels of other proteins in Clostridium organisms, thus enabling cell biology studies with these organisms.
59 BASIC BIOLOGICAL SCIENCES↗
Two-dimensional fluorescence spectroscopy of uranium isotopes in femtosecond laser ablation plumes
Here, we demonstrate measurement of uranium isotopes in femtosecond laser ablation plumes using two-dimensional fluorescence spectroscopy (2DFS). The high-resolution, tunable CW-laser spectroscopy technique clearly distinguishes atomic absorption from 235 U and 238 U in natural and highly enriched uranium metal samples. We present analysis of spectral resolution and analytical performance of 2DFS as a function of ambient pressure. Simultaneous measurement using time-resolved absorption spectroscopy provides information on temporal dynamics of the laser ablation plume and saturation behavior of fluorescence signals. The rapid, non-contact measurement is promising for in-field, standoff measurements of uranium enrichment for nuclear safety and security.
11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗
Thermally Activated Delayed Fluorescence Sensitization for Highly Efficient Blue Fluorescent Emitters
Hyperfluorescence is emerging as a powerful strategy to develop optoelectronic devices with high-color purity and enhanced stability. It requires appropriate integration of a sensitizer displaying efficient thermally activated delayed fluorescence (TADF) and an emitter displaying strong, narrow-band fluorescence. Here, through a joint computational and experimental approach, we provide an unprecedented, end-to-end systems level description of the electronic and optical processes that take place in a hyperfluorescent emissive layer composed of a TADF sensitizer, bis(2,5-di(9H-carbazol-9-yl)phenyl)-1,3,4-oxadiazole (4CzDPO), and a fluorescent emitter, 2,5,8,11-tetra-tert-butylperylene (TBPe). We combine the photophysical properties measurement of the emissive layer with the computational determination of the electronic properties, film morphology, and excitation transfer phenomena. The Förster resonance energy transfer rates from 4CzDPO to TBPe are on the order of 1011 s–1, considerably higher than the radiative and non-radiative recombination rates for 4CzDPO. These features ensure nearly complete energy transfer to TBPe, leading to a five-fold increase in the photoluminescence quantum yields in the 4CzDPO:TBPe system in comparison to neat films of 4CzDPO. Furthermore, our approach highlights the factors that can provide efficient energy transfer from TADF molecules to fluorescent emitters, suppress energy transfer among TADF molecules, and avoid the need for a host material within the emissive layer.
36 MATERIALS SCIENCE↗
Investigation of Sampling-Probe Distorted Temperature Fields with X-Ray Fluorescence Spectroscopy
Flame-sampling experiments, especially in conjunction with laminar low-pressure premixed flames, are routinely used in combustion chemistry studies to unravel the identities and quantities of key intermediates and their pathways. In many instances, however, an unambiguous interpretation of the experimental and modeling results is hampered by the uncertainties about the probe-induced, perturbed temperature profile. To overcome this limitation, two-dimensional perturbations of the temperature field caused by sampling probes with different geometries have been investigated using synchrotron-based X-ray fluorescence spectroscopy. In these experiments, which were performed at the 7-BM beamline of the Advanced Photon Source (APS) at the Argonne National Laboratory, a continuous beam of hard X-rays at 15 keV was used to excite krypton atoms that were added in a concentration of 5 vol.-% to the unburnt gas mixture and the resulting krypton fluorescence at 12.65 keV was subsequently collected. The highly spatially resolved signal was converted into the local flame temperature to obtain temperature fields at various burner-probe separations as functions of the distance to the burner surface and the radial distance from the centerline. Multiple measurements were performed with different probe geometries and because of the observed impact on the temperature profiles the results clearly revealed the need to specify the sampling probe design to enable quantitative and meaningful comparisons of modeling results with flame-sampled mole fraction data. (C) 2018 The Combustion Institute. Published by Elsevier Inc. All rights reserved.
Hansen, N.↗