Fast widefield techniques for fluorescence and phase endomicroscopy

NASA Astrophysics Data System (ADS)

Ford, Tim N.

Endomicroscopy is a recent development in biomedical optics which gives researchers and physicians microscope-resolution views of intact tissue to complement macroscopic visualization during endoscopy screening. This thesis presents HiLo endomicroscopy and oblique back-illumination endomicroscopy, fast wide-field imaging techniques with fluorescence and phase contrast, respectively. Fluorescence imaging in thick tissue is often hampered by strong out-of-focus background signal. Laser scanning confocal endomicroscopy has been developed for optically-sectioned imaging free from background, but reliance on mechanical scanning fundamentally limits the frame rate and represents significant complexity and expense. HiLo is a fast, simple, widefield fluorescence imaging technique which rejects out-of-focus background signal without the need for scanning. It works by acquiring two images of the sample under uniform and structured illumination and synthesizing an optically sectioned result with real-time image processing. Oblique back-illumination microscopy (OBM) is a label-free technique which allows, for the first time, phase gradient imaging of sub-surface morphology in thick scattering tissue with a reflection geometry. OBM works by back-illuminating the sample with the oblique diffuse reflectance from light delivered via off-axis optical fibers. The use of two diametrically opposed illumination fibers allows simultaneous and independent measurement of phase gradients and absorption contrast. Video-rate single-exposure operation using wavelength multiplexing is demonstrated.

Probing the Mechanisms of an Air Amplifier using a LTQ-FT-ICR-MS and Fluorescence Spectroscopy

PubMed Central

Dixon, R. Brent; Muddiman, David C.; Hawkridge, Adam M.; Fedorov, A. G.

2008-01-01

We report the first quantitative assessment of electrosprayed droplet/ion focusing enabled by the use of a voltage-assisted air amplifier between an electrospray ionization emitter and a hybrid linear ion trap Fourier transform ion cyclotron resonance mass spectrometer (ESI-LTQ-FT-ICR-MS). A solution of fluorescent dye was electrosprayed with a stainless steel mesh screen placed in front of the MS inlet capillary acting as a gas-permeable imaging plate for fluorescence spectroscopy. Without use of the air amplifier no detectable FT-ICR signal was observed, as well as no detectable fluorescence on the screen upon imaging using a fluorescence scanner. When the air amplifier was turned ON while electrospraying the fluorescent dye, FT-ICR mass spectra with high signal to noise ratio were obtained with an average ion injection time of 21 milliseconds for an AGC target value of 5 × 105. Imaging of the screen using a fluorescence scanner produced a distinct spot of cross-sectional area ~33.5 mm2 in front of the MS inlet capillary. These experimental results provide direct evidence of aerodynamic focusing of electrosprayed droplets/ions enabled by an air amplifier, resulting in improved electrospray droplet/ion capture efficiency and reduced ion injection time. A second set of experiments was carried out to explore whether the air amplifier assists in desolvation. By electrospraying a mix of quaternary amines, ratios of increasingly hydrophobic molecules were obtained. Observation of the solvophobic effect associated with electrospray ionization resulted in a higher abundance of the hydrophobic molecule. This bias was eliminated when the air amplifier was turned ON and a response indicative of the respective component concentrations of the molecules in the bulk solution was observed. PMID:17855111

Uranium speciation in biofilms studied by laser fluorescence techniques.

PubMed

Arnold, Thuro; Grossmann, Kay; Baumann, Nils

2010-03-01

Biofilms may immobilize toxic heavy metals in the environment and thereby influence their migration behaviour. The mechanisms of these processes are currently not understood, because the complexity of such biofilms creates many discrete geochemical microenvironments which may differ from the surrounding bulk solution in their bacterial diversity, their prevailing geochemical properties, e.g. pH and dissolved oxygen concentration, the presence of organic molecules, e.g. metabolites, and many more, all of which may affect metal speciation. To obtain such information, which is necessary for performance assessment studies or the development of new cost-effective strategies for cleaning waste waters, it is very important to develop new non-invasive methods applicable to study the interactions of metals within biofilm systems. Laser fluorescence techniques have some superior features, above all very high sensitivity for fluorescent heavy metals. An approach combining confocal laser scanning microscopy and laser-induced fluorescence spectroscopy for study of the interactions of biofilms with uranium is presented. It was found that coupling these techniques furnishes a promising tool for in-situ non-invasive study of fluorescent heavy metals within biofilm systems. Information on uranium speciation and uranium redox states can be obtained.

Status of the radio technique for cosmic-ray induced air showers

NASA Astrophysics Data System (ADS)

Schröder, Frank G.

2016-10-01

Radio measurements yield calorimetric information on the electromagnetic shower component around the clock. However, until recently it was not clear whether radio measurements can compete in accuracy with established night-time techniques like air-Cherenkov or air-fluorescence detection. Due to recent progress in the radio technique as well as in the understanding of the emission mechanisms, the performance of current radio experiments has significantly improved. Above 100 PeV, digital, state-of-the-art antenna arrays achieve a reconstruction accuracy for the energy similar to that of other techniques, and can provide an independent measurement of the absolute energy scale. Furthermore, radio measurements are sensitive to the mass composition of the primary particles: First, the position of the shower maximum can be reconstructed from the radio signal. Second, in combination with muon detectors the measurement of the electromagnetic component provides complementary information on the primary mass. Since the radio footprint is huge for inclined showers, and the radio signal does not suffer absorption in the atmosphere, future radio arrays either focus on inclined showers at the highest energy, or on ultra-high precision measurements with extremely dense arrays. This proceeding reviews the current status of radio experiments and simulations as well as future plans.

Fluorescence correlation spectroscopy: novel variations of an established technique.

PubMed

Haustein, Elke; Schwille, Petra

2007-01-01

Fluorescence correlation spectroscopy (FCS) is one of the major biophysical techniques used for unraveling molecular interactions in vitro and in vivo. It allows minimally invasive study of dynamic processes in biological specimens with extremely high temporal and spatial resolution. By recording and correlating the fluorescence fluctuations of single labeled molecules through the exciting laser beam, FCS gives information on molecular mobility and photophysical and photochemical reactions. By using dual-color fluorescence cross-correlation, highly specific binding studies can be performed. These have been extended to four reaction partners accessible by multicolor applications. Alternative detection schemes shift accessible time frames to slower processes (e.g., scanning FCS) or higher concentrations (e.g., TIR-FCS). Despite its long tradition, FCS is by no means dated. Rather, it has proven to be a highly versatile technique that can easily be adapted to solve specific biological questions, and it continues to find exciting applications in biology and medicine.

Fluorescence and Raman Spectroscopy of Doped Nanodiamonds

NASA Astrophysics Data System (ADS)

Kudryavtsev, O. S.; Khomich, A. A.; Sedov, V. S.; Ekimov, E. A.; Vlasov, I. I.

2018-05-01

Raman and fluorescence spectroscopic techniques were used to study doped nanodiamonds synthesized at high pressure and high temperature (HPHT technique) and by chemical vapor deposition from the gas phase (CVD technique). For the CVD diamonds, a hundred-fold increase in fluorescence intensity of the silicon-vacancy centers normalized to the volume of the probe material was observed with an increase in synthesized diamond particle diameter from 150 to 300 nm. Graphitization temperature upon heating in the air significantly lower than for detonation nanodiamonds was found for the boron-doped HPHT nanodiamonds.

Neutron, fluorescence, and optical imaging: An in situ combination of complementary techniques

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wagner, D.; Egelhaaf, S. U.; Hermes, H. E.

2015-09-15

An apparatus which enables the simultaneous combination of three complementary imaging techniques, optical imaging, fluorescence imaging, and neutron radiography, is presented. While each individual technique can provide information on certain aspects of the sample and their time evolution, a combination of the three techniques in one setup provides a more complete and consistent data set. The setup can be used in transmission and reflection modes and thus with optically transparent as well as opaque samples. Its capabilities are illustrated with two examples. A polymer hydrogel represents a transparent sample and the diffusion of fluorescent particles into and through this polymermore » matrix is followed. In reflection mode, the absorption of solvent by a nile red-functionalized mesoporous silica powder and the corresponding change in fluorescent signal are studied.« less

In Vivo Mitochondrial Oxygen Tension Measured by a Delayed Fluorescence Lifetime Technique

PubMed Central

Mik, Egbert G.; Johannes, Tanja; Zuurbier, Coert J.; Heinen, Andre; Houben-Weerts, Judith H. P. M.; Balestra, Gianmarco M.; Stap, Jan; Beek, Johan F.; Ince, Can

2008-01-01

Mitochondrial oxygen tension (mitoPO2) is a key parameter for cellular function, which is considered to be affected under various pathophysiological circumstances. Although many techniques for assessing in vivo oxygenation are available, no technique for measuring mitoPO2 in vivo exists. Here we report in vivo measurement of mitoPO2 and the recovery of mitoPO2 histograms in rat liver by a novel optical technique under normal and pathological circumstances. The technique is based on oxygen-dependent quenching of the delayed fluorescence lifetime of protoporphyrin IX. Application of 5-aminolevulinic acid enhanced mitochondrial protoporphyrin IX levels and induced oxygen-dependent delayed fluorescence in various tissues, without affecting mitochondrial respiration. Using fluorescence microscopy, we demonstrate in isolated hepatocytes that the signal is of mitochondrial origin. The delayed fluorescence lifetime was calibrated in isolated hepatocytes and isolated perfused livers. Ultimately, the technique was applied to measure mitoPO2 in rat liver in vivo. The results demonstrate mitoPO2 values of ∼30–40 mmHg. mitoPO2 was highly sensitive to small changes in inspired oxygen concentration around atmospheric oxygen level. Ischemia-reperfusion interventions showed altered mitoPO2 distribution, which flattened overall compared to baseline conditions. The reported technology is scalable from microscopic to macroscopic applications, and its reliance on an endogenous compound greatly enhances its potential field of applications. PMID:18641065

Fluorescence hyperspectral imaging technique for foreign substance detection on fresh-cut lettuce.

PubMed

Mo, Changyeun; Kim, Giyoung; Kim, Moon S; Lim, Jongguk; Cho, Hyunjeong; Barnaby, Jinyoung Yang; Cho, Byoung-Kwan

2017-09-01

Non-destructive methods based on fluorescence hyperspectral imaging (HSI) techniques were developed to detect worms on fresh-cut lettuce. The optimal wavebands for detecting the worms were investigated using the one-way ANOVA and correlation analyses. The worm detection imaging algorithms, RSI-I (492-626)/492 , provided a prediction accuracy of 99.0%. The fluorescence HSI techniques indicated that the spectral images with a pixel size of 1 × 1 mm had the best classification accuracy for worms. The overall results demonstrate that fluorescence HSI techniques have the potential to detect worms on fresh-cut lettuce. In the future, we will focus on developing a multi-spectral imaging system to detect foreign substances such as worms, slugs and earthworms on fresh-cut lettuce. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

Alternate gram staining technique using a fluorescent lectin.

PubMed Central

Sizemore, R K; Caldwell, J J; Kendrick, A S

1990-01-01

Fluorescence-labeled wheat germ agglutinin binds specifically to N-acetylglucosamine in the outer peptidoglycan layer of gram-positive bacteria. The peptidoglycan layer of gram-negative bacteria is covered by a membrane and is not labeled by the lectin. By exploiting this phenomenon, an alternative Gram staining technique has been developed. Images PMID:1697149

Laser-induced fluorescence from N2(+) ions generated by a corona discharge in ambient air.

PubMed

Konthasinghe, Kumarasiri; Fitzmorris, Kristin; Peiris, Manoj; Hopkins, Adam J; Petrak, Benjamin; Killinger, Dennis K; Muller, Andreas

2015-09-01

In this work, we present the measurement of laser-induced fluorescence from N2(+) ions via the B(2)Σu(+)-X(2)Σg(+) band system in the near-ultraviolet. The ions were generated continuously by a plasma glow discharge in low pressure N2 and by a corona discharge in ambient air. The fluorescence decay time was found to rapidly decrease with increasing pressure leading to an extrapolated decay rate of ≍10(10) s(-1) at atmospheric pressure. In spite of this quenching, we were able to observe laser induced fluorescence in ambient air by means of a time-gated spectral measurement. In the process of comparing the emission signal with that of N2 spontaneous Raman scattering, ion concentrations in ambient air of order 10(8-)10(10) cm(-3) were determined. With moderate increases in laser power and collection efficiency, ion concentrations of less than 10(6) cm(-3) may be measurable, potentially enabling applications in atmospheric standoff detection of ionizing radiation from hazardous radioactive sources.

The small contribution of molecular Bremsstrahlung radiation to the air-fluorescence yield of cosmic ray shower particles

NASA Astrophysics Data System (ADS)

Al Samarai, Imen; Deligny, Olivier; Rosado, Jaime

2016-10-01

A small contribution of molecular Bremsstrahlung radiation to the air-fluorescence yield in the UV range is estimated based on an approach previously developed in the framework of the radio-detection of showers in the gigahertz frequency range. First, this approach is shown to provide an estimate of the main contribution of the fluorescence yield due to the de-excitation of the C 3Πu electronic level of nitrogen molecules to the B 3Πg one amounting to Y[ 337 ] =(6.05 ± 1.50) MeV-1 at 800 hPa pressure and 293 K temperature conditions, which compares well to previous dedicated works and to experimental results. Then, under the same pressure and temperature conditions, the fluorescence yield induced by molecular Bremsstrahlung radiation is found to be Y[330-400]MBR = 0.10 MeV-1 in the wavelength range of interest for the air-fluorescence detectors used to detect extensive air showers induced in the atmosphere by ultra-high energy cosmic rays. This means that out of ≃175 photons with wavelength between 330 and 400 nm detected by fluorescence detectors, one of them has been produced by molecular Bremsstrahlung radiation. Although small, this contribution is not negligible in regards to the total budget of systematic uncertainties when considering the absolute energy scale of fluorescence detectors.

Fluorescence probe techniques to monitor protein adsorption-induced conformation changes on biodegradable polymers.

PubMed

Benesch, Johan; Hungerford, Graham; Suhling, Klaus; Tregidgo, Carolyn; Mano, João F; Reis, Rui L

2007-08-15

The study of protein adsorption and any associated conformational changes on interaction with biomaterials is of great importance in the area of implants and tissue constructs. This study aimed to evaluate some fluorescent techniques to probe protein conformation on a selection of biodegradable polymers currently under investigation for biomedical applications. Because of the fluorescence emanating from the polymers, the use of monitoring intrinsic protein fluorescence was precluded. A highly solvatochromic fluorescent dye, Nile red, and a well-known protein label, fluorescein isothiocyanate, were employed to study the adsorption of serum albumin to polycaprolactone and to some extent also to two starch-containing polymer blends (SPCL and SEVA-C). A variety of fluorescence techniques, steady state, time resolved, and imaging were employed. Nile red was found to leach from the protein, while fluorescein isothiocyanate proved useful in elucidating a conformational change in the protein and the observation of protein aggregates adsorbed to the polymer surface. These effects were seen by making use of the phenomenon of energy migration between the fluorescent tags to monitor interprobe distance and the use of fluorescence lifetime imaging to ascertain the surface packing of the protein on polymer.

Fluorescence light microscopy of pulmonary surfactant at the air-water interface of an air bubble of adjustable size.

PubMed Central

Knebel, D; Sieber, M; Reichelt, R; Galla, H-J; Amrein, M

2002-01-01

The structural dynamics of pulmonary surfactant was studied by epifluorescence light microscopy at the air-water interface of a bubble as a model close to nature for an alveolus. Small unilamellar vesicles of dipalmitoylphosphatidylcholine, dipalmitoylphosphatidylglycerol, a small amount of a fluorescent dipalmitoylphosphatidylcholine-analog, and surfactant-associated protein C were injected into the buffer solution. They aggregated to large clusters in the presence of Ca(2+) and adsorbed from these units to the interface. This gave rise to an interfacial film that eventually became fully condensed with dark, polygonal domains in a fluorescent matrix. When now the bubble size was increased or decreased, respectively, the film expanded or contracted. Upon expansion of the bubble, the dark areas became larger to the debit of the bright matrix and reversed upon contraction. We were able to observe single domains during the whole process. The film remained condensed, even when the interface was increased to twice its original size. From comparison with scanning force microscopy directly at the air-water interface, the fluorescent areas proved to be lipid bilayers associated with the (dark) monolayer. In the lung, such multilayer phase acts as a reservoir that guarantees a full molecular coverage of the alveolar interface during the breathing cycle and provides mechanical stability to the film. PMID:12080141

Real-time quantitative fluorescence imaging using a single snapshot optical properties technique for neurosurgical guidance

NASA Astrophysics Data System (ADS)

Valdes, Pablo A.; Angelo, Joseph; Gioux, Sylvain

2015-03-01

Fluorescence imaging has shown promise as an adjunct to improve the extent of resection in neurosurgery and oncologic surgery. Nevertheless, current fluorescence imaging techniques do not account for the heterogeneous attenuation effects of tissue optical properties. In this work, we present a novel imaging system that performs real time quantitative fluorescence imaging using Single Snapshot Optical Properties (SSOP) imaging. We developed the technique and performed initial phantom studies to validate the quantitative capabilities of the system for intraoperative feasibility. Overall, this work introduces a novel real-time quantitative fluorescence imaging method capable of being used intraoperatively for neurosurgical guidance.

Ambient measurements of biological aerosol particles near Killarney, Ireland: a comparison between real-time fluorescence and microscopy techniques

NASA Astrophysics Data System (ADS)

Healy, D. A.; Huffman, J. A.; O'Connor, D. J.; Pöhlker, C.; Pöschl, U.; Sodeau, J. R.

2014-08-01

Primary biological aerosol particles (PBAPs) can contribute significantly to the coarse particle burden in many environments. PBAPs can thus influence climate and precipitation systems as cloud nuclei and can spread disease to humans, animals, and plants. Measurement data and techniques for PBAPs in natural environments at high time- and size resolution are, however, sparse, and so large uncertainties remain in the role that biological particles play in the Earth system. In this study two commercial real-time fluorescence particle sensors and a Sporewatch single-stage particle impactor were operated continuously from 2 August to 2 September 2010 at a rural sampling location in Killarney National Park in southwestern Ireland. A cascade impactor was operated periodically to collect size-resolved particles during exemplary periods. Here we report the first ambient comparison of a waveband integrated bioaerosol sensor (WIBS-4) with a ultraviolet aerodynamic particle sizer (UV-APS) and also compare these real-time fluorescence techniques with results of fluorescence and optical microscopy of impacted samples. Both real-time instruments showed qualitatively similar behavior, with increased fluorescent bioparticle concentrations at night, when relative humidity was highest and temperature was lowest. The fluorescent particle number from the FL3 channel of the WIBS-4 and from the UV-APS were strongly correlated and dominated by a 3 μm mode in the particle size distribution. The WIBS FL2 channel exhibited particle modes at approx. 1 and 3 μm, and each was correlated with the concentration of fungal spores commonly observed in air samples collected at the site (ascospores, basidiospores, Ganoderma spp.). The WIBS FL1 channel exhibited variable multimodal distributions turning into a broad featureless single mode after averaging, and exhibited poor correlation with fungal spore concentrations, which may be due to the detection of bacterial and non-biological fluorescent

Standoff detection: classification of biological aerosols using laser induced fluorescence (LIF) technique

NASA Astrophysics Data System (ADS)

Hausmann, Anita; Duschek, Frank; Fischbach, Thomas; Pargmann, Carsten; Aleksejev, Valeri; Poryvkina, Larisa; Sobolev, Innokenti; Babichenko, Sergey; Handke, Jürgen

2014-05-01

The challenges of detecting hazardous biological materials are manifold: Such material has to be discriminated from other substances in various natural surroundings. The detection sensitivity should be extremely high. As living material may reproduce itself, already one single bacterium may represent a high risk. Of course, identification should be quite fast with a low false alarm rate. Up to now, there is no single technique to solve this problem. Point sensors may collect material and identify it, but the problems of fast identification and especially of appropriate positioning of local collectors are sophisticated. On the other hand, laser based standoff detection may instantaneously provide the information of some accidental spillage of material by detecting the generated thin cloud. LIF technique may classify but hardly identify the substance. A solution can be the use of LIF technique in a first step to collect primary data and - if necessary- followed by utilizing these data for an optimized positioning of point sensors. We perform studies on an open air laser test range at distances between 20 and 135 m applying LIF technique to detect and classify aerosols. In order to employ LIF capability, we use a laser source emitting two wavelengths alternatively, 280 and 355 nm, respectively. Moreover, the time dependence of fluorescence spectra is recorded by a gated intensified CCD camera. Signal processing is performed by dedicated software for spectral pattern recognition. The direct comparison of all results leads to a basic classification of the various compounds.

Detection of fecal residue on poultry carcasses by laser induced fluorescence imaging techniques

USDA-ARS?s Scientific Manuscript database

The potential use of laser-induced fluorescence imaging techniques was investigated for the detection of diluted fecal matters from various parts of the digestive tract, including colon, ceca, small intestine, and duodenum, on poultry carcasses. One of the challenges for using fluorescence imaging f...

Planar temperature measurement in compressible flows using laser-induced iodine fluorescence

NASA Technical Reports Server (NTRS)

Hartfield, Roy J., Jr.; Hollo, Steven D.; Mcdaniel, James C.

1991-01-01

A laser-induced iodine fluorescence technique that is suitable for the planar measurement of temperature in cold nonreacting compressible air flows is investigated analytically and demonstrated in a known flow field. The technique is based on the temperature dependence of the broadband fluorescence from iodine excited by the 514-nm line of an argon-ion laser. Temperatures ranging from 165 to 245 K were measured in the calibration flow field. This technique makes complete, spatially resolved surveys of temperature practical in highly three-dimensional, low-temperature compressible flows.

Laser Induced Fluorescence Studies of NO Kinetics in Short Pulse Air and Air-Fuel Nonequilibrium Discharges

NASA Astrophysics Data System (ADS)

Lempert, Walter; Uddi, Mruthunjaya; Adamovich, Igor

2008-10-01

Laser Induced Fluorescence is used to measure absolute NO concentrations in air, methane-air, and ethylene-air non-equilibrium plasmas, as a function of time after initiation of a single 25 nsec discharge pulse. Peak NO density in air at 60 torr is ˜8.10^12 cm-3 occurring at ˜500 μs after the pulse, with decay time of ˜16.5 msec. Peak NO atom mole fraction in methane-air at φ=0.5 is approximately equal to that in pure air with similar rise and decay rate. In φ = 0.5 ethylene-air, the rise and decay times are also comparable to air and methane--air, but peak NO concentration is a factor of ˜2.5 lower. Spontaneous emission measurements show that N2(C) and NO (A) decay in ˜25ns and ˜2.5μs, respectively. Kinetic modeling calculations incorporating Boltzmann solver for EEDF, and electron impact and full air species kinetics, complemented with the GRI Mech 3.0 hydrocarbon oxidation mechanism, are compared with the experimental data using three different mechanisms. It is concluded that processes involving long lifetime (˜100 μsec) meta-stable states, such as N2 (X,v) and O2(b^1σ), which are formed by quenching of the metastable N2 (A) state by ground state O2, play a dominant role in NO formation.

Measurements of KrF laser-induced O2 fluorescence in high-temperature atmospheric air

NASA Technical Reports Server (NTRS)

Grinstead, Jay H.; Laufer, Gabriel; Mcdaniel, James C., Jr.

1993-01-01

Conditions for obtaining laser-induced O2 fluorescence using a tunable KrF laser has been determined theoretically and experimentally. With this laser source, O2 rotational temperature measurement is possible even in the absence of vibrational equilibrium. Temperature measurement using a two-line excitation scheme has been demonstrated in a high-temperature atmospheric-air furnace. A measurement uncertainty of 10.7 percent for the temperature range 1325-1725 K was realized. At atmospheric pressure, O2 LIF measurements are possible for air temperatures above 1250 K. Interference from OH fluorescence in reacting flows can be avoided by the proper selection of O2 transitions. Depletion of the ground state population by the incident laser is negligible for intensities below 7.5 x 10 to the 6th W/sq cm/per cm.

Predictive Techniques for Spacecraft Cabin Air Quality Control

NASA Technical Reports Server (NTRS)

Perry, J. L.; Cromes, Scott D. (Technical Monitor)

2001-01-01

As assembly of the International Space Station (ISS) proceeds, predictive techniques are used to determine the best approach for handling a variety of cabin air quality challenges. These techniques use equipment offgassing data collected from each ISS module before flight to characterize the trace chemical contaminant load. Combined with crew metabolic loads, these data serve as input to a predictive model for assessing the capability of the onboard atmosphere revitalization systems to handle the overall trace contaminant load as station assembly progresses. The techniques for predicting in-flight air quality are summarized along with results from early ISS mission analyses. Results from groundbased analyses of in-flight air quality samples are compared to the predictions to demonstrate the technique's relative conservatism.

LIFES: Laser Induced Fluorescence and Environmental Sensing. [remote sensing technique for marine environment

NASA Technical Reports Server (NTRS)

Houston, W. R.; Stephenson, D. G.; Measures, R. M.

1975-01-01

A laboratory investigation has been conducted to evaluate the detection and identification capabilities of laser induced fluorescence as a remote sensing technique for the marine environment. The relative merits of fluorescence parameters including emission and excitation profiles, intensity and lifetime measurements are discussed in relation to the identification of specific targets of the marine environment including crude oils, refined petroleum products, fish oils and algae. Temporal profiles displaying the variation of lifetime with emission wavelength have proven to add a new dimension of specificity and simplicity to the technique.

A "turn-on" fluorescent sensor for ozone detection in ambient air using protein-directed gold nanoclusters.

PubMed

Wu, Di; Qi, Wenjing; Liu, Chun; Zhang, Qing

2017-04-01

A "turn-on" fluorescent sensor for ozone using bovine serum albumin-directed gold nanoclusters (BSA-Au NCs) via energy transfer was developed. The spectral overlap of fluorescent spectrum of BSA-Au NCs with absorption spectrum of indigo carmine (IDS) was utilized. Ozone cleaves C = C bond of IDS and suppresses energy transfer from BSA-Au NCs to IDS. Therefore, this proposed fluorescent sensor is a "turn-on" detection motif. It is the first application of fluorescent nanoclusters in sensitively detecting ozone from 0.2 to 12 μM with the limit of detection of 35 nM (the volume of 500 μL, 1.68 ppb). The proposed fluorescent sensor for ozone is more sensitive and faster (within 2 min) than most methods and is with good selectivity for ozone detection against other reactive oxygen species, reactive nitrogen, or metallic ions. Besides, the proposed method is also utlized in ozone detection in ambient air by monitoring 1 h (60 min) in Qijiang district in Chongqing city. The average of concentration of ozone in ambient air ranges from 44.97 to 52.85 μg/m 3 . The results are compared with the automatic monitoring data provided by Qijiang Environmental Monitoring Station and the relative deviations range, respectively, from 2.1 to 5.6%, which suggests that it is a promising fluorescent sensor for ozone in ambient air. This study not only develops a new model of energy transfer motif using BSA-Au NCs as donor and IDS as acceptor but also expands the application of BSA-Au NCs in environmental science. Graphical abstract A "turn-on" fluorescent sensor for ozone detection using bovine serum albumin-directed gold nanoclusters (BSA-Au NCs) via energy transfer is developed. It is the first time to utilize spectral overlap of fluorescent spectrum of BSA-Au NCs with absorption spectrum of indigo carmine and to achieve fast, sensitive, and selective ozone detection with a limit of detection of down to 35 nM (the volume of 500 μL, 1.68 ppb).

Fluorescence lifetime technique for surgical imaging, guidance and augmented reality (Conference Presentation)

NASA Astrophysics Data System (ADS)

Marcu, Laura

2017-02-01

The surgeon's limited ability to accurately delineate the tumor margin during surgical interventions is one key challenge in clinical management of cancer. New methods for guiding tumor resection decisions are needed. Numerous studies have shown that tissue autofluorescence properties have the potential to asses biochemical features associates with distinct pathologies in tissue and to distinguish various cancers from normal tissues. However, despite these promising reports, autofluorescence techniques were sparsely adopted in clinical settings. Moreover, when adopted they were primarily used for pre-operative diagnosis rather than guiding interventions. To address this need, we have researched and engineered instrumentation that utilizes label-free fluorescence lifetime contrast to characterize tissue biochemical features in vivo in patients and methodologies conducive to real-time (few seconds) diagnosis of tissue pathologies during surgical procedures. This presentation overviews clinically-compatible multispectral fluorescence lifetime imaging techniques developed in our laboratory and their ability to operate as stand-alone tools, integrated in a biopsy needle and in conjunction with the da Vinci surgical robot. We present pre-clinical and clinical studies in patients that demonstrate the potential of these techniques for intraoperative assessment of brain tumors and head and neck cancer. Current results demonstrate that intrinsic fluorescence signals can provide useful contrast for delineation distinct types of tissues including tumors intraoperatively. Challenges and solutions in the clinical implementation of these techniques are discussed.

Cleansing technique using high-velocity steam-air micromist jet spray.

PubMed

Fukuda, Koichi; Ishihara, Masayuki; Murakami, Kaoru; Nakamura, Shingo; Sato, Yoko; Kuwabara, Masahiro; Fujita, Masanori; Kiyosawa, Tomoharu; Yokoe, Hidetaka

2017-10-01

Application of a high-velocity steam-air micromist jet spray (HVS-AMJS; micromist average diameter: 2.4 μm) for cleansing the skin is proposed. Low-pressure steam is mixed with compressed air (pH 6.5) in a nozzle, and then sprayed at a pressure of ≦0.25 MPa and a velocity of ≧0.34 m/s on the skin or surface of material located approximately 5-10 cm from the nozzle. The temperature on the sprayed surface and water flow rate could be controlled between 42 °C and 46 °C and at approximately 50 mL/min, respectively. Compared with ultrasonic cleansing with tap water and rubbing with only tap water, the HVS-AMJS successfully removed fluorescent lotion covering pieces of wood and significantly reduced both the number of coliforms and the total viable counts on pieces of wood and gauze. Furthermore, the HVS-AMJS effectively removed oily ink from the skin of hairless rats, and temporarily elevated the skin temperature and blood flow, indicating massage effects. The striking characteristics of this cleansing technique using HVS-AMJS are not only its ability to remove microbes and residue without using any chemicals or detergents but also its massage effects.

Survey of air cargo forecasting techniques

NASA Technical Reports Server (NTRS)

Kuhlthan, A. R.; Vermuri, R. S.

1978-01-01

Forecasting techniques currently in use in estimating or predicting the demand for air cargo in various markets are discussed with emphasis on the fundamentals of the different forecasting approaches. References to specific studies are cited when appropriate. The effectiveness of current methods is evaluated and several prospects for future activities or approaches are suggested. Appendices contain summary type analyses of about 50 specific publications on forecasting, and selected bibliographies on air cargo forecasting, air passenger demand forecasting, and general demand and modalsplit modeling.

The Fluorescent-Oil Film Method and Other Techniques for Boundary-Layer Flow Visualization

NASA Technical Reports Server (NTRS)

Loving, Donald L.; Katzoff, S.

1959-01-01

A flow-visualization technique, known as the fluorescent-oil film method, has been developed which appears to be generally simpler and to require less experience and development of technique than previously published methods. The method is especially adapted to use in the large high-powered wind tunnels which require considerable time to reach the desired test conditions. The method consists of smearing a film of fluorescent oil over a surface and observing where the thickness is affected by the shearing action of the boundary layer. These films are detected and identified, and their relative thicknesses are determined by use of ultraviolet light. Examples are given of the use of this technique. Other methods that show promise in the study of boundary-layer conditions are described. These methods include the use of a temperature-sensitive fluorescent paint and the use of a radiometer that is sensitive to the heat radiation from a surface. Some attention is also given to methods that can be used with a spray apparatus in front of the test model.

Visual air quality simulation techniques

NASA Astrophysics Data System (ADS)

Molenar, John V.; Malm, William C.; Johnson, Christopher E.

Visual air quality is primarily a human perceptual phenomenon beginning with the transfer of image-forming information through an illuminated, scattering and absorbing atmosphere. Visibility, especially the visual appearance of industrial emissions or the degradation of a scenic view, is the principal atmospheric characteristic through which humans perceive air pollution, and is more sensitive to changing pollution levels than any other air pollution effect. Every attempt to quantify economic costs and benefits of air pollution has indicated that good visibility is a highly valued and desired environmental condition. Measurement programs can at best approximate the state of the ambient atmosphere at a few points in a scenic vista viewed by an observer. To fully understand the visual effect of various changes in the concentration and distribution of optically important atmospheric pollutants requires the use of aerosol and radiative transfer models. Communication of the output of these models to scientists, decision makers and the public is best done by applying modern image-processing systems to generate synthetic images representing the modeled air quality conditions. This combination of modeling techniques has been under development for the past 15 yr. Initially, visual air quality simulations were limited by a lack of computational power to simplified models depicting Gaussian plumes or uniform haze conditions. Recent explosive growth in low cost, high powered computer technology has allowed the development of sophisticated aerosol and radiative transfer models that incorporate realistic terrain, multiple scattering, non-uniform illumination, varying spatial distribution, concentration and optical properties of atmospheric constituents, and relative humidity effects on aerosol scattering properties. This paper discusses these improved models and image-processing techniques in detail. Results addressing uniform and non-uniform layered haze conditions in both

Measurement of Ambient Air Motion of D. I. Gasoline Spray by LIF-PIV

NASA Astrophysics Data System (ADS)

Yamakawa, Masahisa; Isshiki, Seiji; Yoshizaki, Takuo; Nishida, Keiya

Ambient air velocity distributions in and around a D. I. gasoline spray were measured using a combination of LIF and PIV techniques. A rhodamine and water solution was injected into ambient air to disperse the fine fluorescent liquid particles used as tracers. A fuel spray was injected into the fluorescent tracer cloud and was illuminated by an Nd: YAG laser light sheet (532nm). The scattered light from the spray droplets and tracers was cut off by a high-pass filter (>560nm). As the fluorescence (>600nm) was transmitted through the high-pass filter, the tracer images were captured using a CCD camera and the ambient air velocity distribution could be obtained by PIV based on the images. This technique was applied to a D. I. gasoline spray. The ambient air flowed up around the spray and entered into the tail of the spray. Furthermore, the relative velocity between the spray and ambient air was investigated.

Compressed Air/Vacuum Transportation Techniques

NASA Astrophysics Data System (ADS)

Guha, Shyamal

2011-03-01

General theory of compressed air/vacuum transportation will be presented. In this transportation, a vehicle (such as an automobile or a rail car) is powered either by compressed air or by air at near vacuum pressure. Four version of such transportation is feasible. In all versions, a ``c-shaped'' plastic or ceramic pipe lies buried a few inches under the ground surface. This pipe carries compressed air or air at near vacuum pressure. In type I transportation, a vehicle draws compressed air (or vacuum) from this buried pipe. Using turbine or reciprocating air cylinder, mechanical power is generated from compressed air (or from vacuum). This mechanical power transferred to the wheels of an automobile (or a rail car) drives the vehicle. In type II-IV transportation techniques, a horizontal force is generated inside the plastic (or ceramic) pipe. A set of vertical and horizontal steel bars is used to transmit this force to the automobile on the road (or to a rail car on rail track). The proposed transportation system has following merits: virtually accident free; highly energy efficient; pollution free and it will not contribute to carbon dioxide emission. Some developmental work on this transportation will be needed before it can be used by the traveling public. The entire transportation system could be computer controlled.

Injectant mole-fraction imaging in compressible mixing flows using planar laser-induced iodine fluorescence

NASA Technical Reports Server (NTRS)

Hartfield, Roy J., Jr.; Abbitt, John D., III; Mcdaniel, James C.

1989-01-01

A technique is described for imaging the injectant mole-fraction distribution in nonreacting compressible mixing flow fields. Planar fluorescence from iodine, seeded into air, is induced by a broadband argon-ion laser and collected using an intensified charge-injection-device array camera. The technique eliminates the thermodynamic dependence of the iodine fluorescence in the compressible flow field by taking the ratio of two images collected with identical thermodynamic flow conditions but different iodine seeding conditions.

A fluorescent imaging technique for quantifying spray deposits on plant leaves

USDA-ARS?s Scientific Manuscript database

Because of the unique characteristics of electrostatically-charged sprays, use of traditional methods to quantify deposition from these sprays has been challenging. A new fluorescent imaging technique was developed to quantify spray deposits from electrostatically-charged sprays on natural plant lea...

Techniques for Forecasting Air Passenger Traffic

NASA Technical Reports Server (NTRS)

Taneja, N.

1972-01-01

The basic techniques of forecasting the air passenger traffic are outlined. These techniques can be broadly classified into four categories: judgmental, time-series analysis, market analysis and analytical. The differences between these methods exist, in part, due to the degree of formalization of the forecasting procedure. Emphasis is placed on describing the analytical method.

Effects of fungal species, cultivation time, growth substrate, and air exposure velocity on the fluorescence properties of airborne fungal spores.

PubMed

Saari, S; Mensah-Attipoe, J; Reponen, T; Veijalainen, A M; Salmela, A; Pasanen, P; Keskinen, J

2015-12-01

Real-time bioaerosol monitoring is possible with fluorescence based instruments. This study provides information on major factors that can affect the fluorescence properties of airborne fungal spores. Two fluorescence-based bioaerosol detectors, BioScout, and ultraviolet aerodynamic particle sizer (UVAPS), were used to study fluorescent particle fractions (FPFs) of released spores of three fungal species (Aspergillus versicolor, Cladosporium cladosporioides, and Penicillium brevicompactum). Two culture media (agar and gypsum board), three ages of the culture (one week, one month, and four months), and three aerosolization air velocities (5, 15, and 27 m/s) were tested. The results showed that the FPF values for spores released from gypsum were typically lower than for those released from agar indicating that poor nutrient substrate produces spores with lower amounts of fluorescent compounds. The results also showed higher FPF values with lower air velocities in aerosolization. This indicates that easily released fully developed spores have more fluorescent compounds compared to forcibly extracted non-matured spores. The FPFs typically were lower with older samples. The FPF results between the two instruments were similar, except with four-month-old samples. The results can be utilized in field measurements of fungal spores to estimate actual concentrations and compare different instruments with fluorescence-based devices as well as in instrument calibration and testing in laboratory conditions. Fluorescence-based instruments are the only choice for real-time detection of fungal spores at the moment. In general, all fluorescence-based bioaerosol instruments are tested against known bacterial and fungal spores in laboratory conditions. This study showed that fungal species, growth substrate, age of culture, and air current exposure rate have an effect on detection efficiency of fungal spores in the fluorescence-based instruments. Therefore, these factors should be

Monitoring of nanoclay-protein adsorption isotherms via fluorescence techniques.

PubMed

Felbeck, Tom; Moss, Sebastian; Botas, Alexandre M P; Lezhnina, Marina M; Ferreira, Rute A S; Carlos, Luís D; Kynast, Ulrich H

2017-09-01

The investigation of nanoparticles and their interaction with bio-macromolecules have become an important issue; the widely discussed protein corona around nanoparticles and their biological fate in general have drawn particular attention. Here, we focus on nanoclay dispersions and the use of solvatochromic fluorescent dyes (Dansyl and Coumarin 153) for monitoring the interaction with two model proteins, bovine serum albumin and β-lactoglobulin. On one hand, these dyes are poorly emissive in water, but experience a boost in their fluorescence when adsorbed into the hydrophobic domains of proteins. On the other hand, (nano)clays and clay minerals have previously been investigated in terms of their individual protein adsorption isotherms and their usefulness for the solubilization of water-insoluble dyes into an aqueous environment. In the following, we have combined all three individual parts (nanoclay, fluorophore and protein) in dispersions in a wide range of concentration ratios to systematically study the various adsorption processes via fluorescence techniques. In order to clarify the extent of dye diffusion and adsorption-desorption equilibria in the investigations, nanoclay hybrids with an adsorbed dye (Coumarin 153) and a covalently conjugated dye (Dansyl) were compared. The results suggest that the fluorescence progression of protein titration curves correlate with the amount of protein adsorbed, matching their reported adsorption isotherms on hectorite clays. Furthermore, experimental data on the protein monolayer formation around the nanoclays could be extracted due to only minor alterations of the dispersions' optical quality and transparency. In this manner, a fluorescence-based monitor for the formation of the globular protein layer around the nanoclay was realized. Copyright © 2017 Elsevier B.V. All rights reserved.

New approaches to the analysis of complex samples using fluorescence lifetime techniques and organized media

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hertz, P.R.

Fluorescence spectroscopy is a highly sensitive and selective tool for the analysis of complex systems. In order to investigate the efficacy of several steady state and dynamic techniques for the analysis of complex systems, this work focuses on two types of complex, multicomponent samples: petrolatums and coal liquids. It is shown in these studies dynamic, fluorescence lifetime-based measurements provide enhanced discrimination between complex petrolatum samples. Additionally, improved quantitative analysis of multicomponent systems is demonstrated via incorporation of organized media in coal liquid samples. This research provides the first systematic studies of (1) multifrequency phase-resolved fluorescence spectroscopy for dynamic fluorescence spectralmore » fingerprinting of complex samples, and (2) the incorporation of bile salt micellar media to improve accuracy and sensitivity for characterization of complex systems. In the petroleum studies, phase-resolved fluorescence spectroscopy is used to combine spectral and lifetime information through the measurement of phase-resolved fluorescence intensity. The intensity is collected as a function of excitation and emission wavelengths, angular modulation frequency, and detector phase angle. This multidimensional information enhances the ability to distinguish between complex samples with similar spectral characteristics. Examination of the eigenvalues and eigenvectors from factor analysis of phase-resolved and steady state excitation-emission matrices, using chemometric methods of data analysis, confirms that phase-resolved fluorescence techniques offer improved discrimination between complex samples as compared with conventional steady state methods.« less

Application of fluorescently labeled tracer technique for detection of natural active macromolecules in Chinese medicine.

PubMed

Zeng, Qiao-Hui; Zhang, Xue-Wu; Xu, Kai-Peng; Jiang, Jian-Guo

2014-02-01

Active substances in traditional Chinese Medicine (TCM) contain not only a variety of small molecules, but also many other macromolecules (TCMMs), such as proteins, peptides and polysaccharides. Active TCMM can achieve good therapeutic effects by regulating the body's overall function with lower side effects. This review summarized the literatures published in recent years on the application of fluorescently labeled tracer technique for detection of natural active macromolecules in TCM. Classified by fluorescent markers, applications of fluorescein, rhodamine, and quantum dots (QDs) in TCMM active tracer are reviewed, and the methods and principles of TCMM fluorescent marker are illustrated. Studies on active TCMMs and their action mechanism are quite difficult due to a multitarget, multicomponent, and multipath system of TCM. However, the development of fluorescently labeled active tracer technique (FLATT) provides this research with new tools. Traditional fluorescent markers have many deficiencies, such as easily quenched, short luminous cycle, and intrinsic toxicity. Relatively, FLATT has many obvious advantages, and its application in TCMM is still at the early stage. In order to improve the overall level of fluorescence labeling in TCMM active tracer, the improvement on FLATT's detection sensitivity and biological affinity is urgent and critical to allow study of these interesting molecules.

Detection of Biomass in New York City Aerosols: Light Scattering and Optical Fluorescence Techniques

NASA Astrophysics Data System (ADS)

Niebauer, M.; Alimova, A.; Katz, A.; Xu, M.; Rudolph, E.; Steiner, J.; Alfano, R. R.

2005-12-01

Optical spectroscopy is an ideal method for detecting bacteria and spores in real time. Optical fluorescence spectroscopy examination of New York City aerosols is used to quantify the mass of bacteria spores present in air masses collected at 14 liters/minute onto silica fiber filters, and on silica fiber ribbons using an Environmental Beta Attenuation Monitor manufactured by MetOne Instruments configured for the PM2.5 fraction. Dipicolinic acid (DPA), a molecule found primarily in bacterial spores, is the most characteristic component of spores in trial experiments on over 200 collected aerosol samples. DPA is extracted from the spores using a heat bath and chelated with Terbium. The DPA:Tb is detected by measuring its characteristic fluorescence with emission bands at 490, 545 and 585 nm for 270 nm excitation. Light scattering also measures the size distribution for a number of a variety of bacteria - Bacillus subtilis (rod shaped), Staphylococcus aureus (spherical) and Pseudomonas aeruginosa (short rods) establishing that optical techniques satisfactorily distinguish populations based on their variable morphology. Size and morphology are obtained by applying a variation of the Gaussian Ray Approximation theory of anomalous diffraction theory to an analysis of the transmission spectra in the range of 0.4 to 1.0 microns. In test experiments, the refractive index of the inner spore core of Bacillus subtilis decreases from 1.51 to 1.39 while the spore radius enlarges from 0.38 to 0.6 micrometers. Optical determinations are verified by oil-immersion techniques and by scanning electron microscope measurements. Characterization of spores, germinating spore materials, and bacteria is considered vital to tracing bacteria in the environment, for the development of life-detection systems for planetary exploration, monitoring pathogens in environmental systems, and for the preparation of anti-terrorism strategies.

A measurement of time-averaged aerosol optical depth using air-showers observed in stereo by HiRes

NASA Astrophysics Data System (ADS)

High Resolution Fly'S Eye Collaboration; Abbasi, R. U.; Abu-Zayyad, T.; Amann, J. F.; Archbold, G.; Atkins, R.; Belov, K.; Belz, J. W.; Benzvi, S.; Bergman, D. R.; Boyer, J. H.; Cannon, C. T.; Cao, Z.; Connolly, B. M.; Fedorova, Y.; Finley, C. B.; Hanlon, W. F.; Hoffman, C. M.; Holzscheiter, M. H.; Hughes, G. A.; Hüntemeyer, P.; Jui, C. C. H.; Kirn, M. A.; Knapp, B. C.; Loh, E. C.; Manago, N.; Mannel, E. J.; Martens, K.; Matthews, J. A. J.; Matthews, J. N.; O'Neill, A.; Reil, K.; Roberts, M. D.; Schnetzer, S. R.; Seman, M.; Sinnis, G.; Smith, J. D.; Sokolsky, P.; Song, C.; Springer, R. W.; Stokes, B. T.; Thomas, S. B.; Thomson, G. B.; Tupa, D.; Westerhoff, S.; Wiencke, L. R.; Zech, A.

2006-03-01

Air fluorescence measurements of cosmic ray energy must be corrected for attenuation of the atmosphere. In this paper, we show that the air-showers themselves can yield a measurement of the aerosol attenuation in terms of optical depth, time-averaged over extended periods. Although the technique lacks statistical power to make the critical hourly measurements that only specialized active instruments can achieve, we note the technique does not depend on absolute calibration of the detector hardware, and requires no additional equipment beyond the fluorescence detectors that observe the air showers. This paper describes the technique, and presents results based on analysis of 1258 air-showers observed in stereo by the High Resolution Fly’s Eye over a four year span.

Preparation and application of new fluorescein-labeled fumonisins B1 in fluorescence polarization analysis technique

USDA-ARS?s Scientific Manuscript database

Objective: To prepare a new fluorescent tracer against common mycotoxins such as fumonisin B1 in order to replace 6-(4,6-Dichlorotriazinyl) aminofluorescein (6-DTAF), an expensive marker, and to develop a technique for quick detection of fumonisin B1 based on the principle of fluorescence polarizati...

An optical clearing technique for plant tissues allowing deep imaging and compatible with fluorescence microscopy.

PubMed

Warner, Cherish A; Biedrzycki, Meredith L; Jacobs, Samuel S; Wisser, Randall J; Caplan, Jeffrey L; Sherrier, D Janine

2014-12-01

We report on a nondestructive clearing technique that enhances transmission of light through specimens from diverse plant species, opening unique opportunities for microscope-enabled plant research. After clearing, plant organs and thick tissue sections are amenable to deep imaging. The clearing method is compatible with immunocytochemistry techniques and can be used in concert with common fluorescent probes, including widely adopted protein tags such as GFP, which has fluorescence that is preserved during the clearing process. © 2014 American Society of Plant Biologists. All Rights Reserved.

Ambient measurements of biological aerosol particles near Killarney, Ireland: a comparison between real-time fluorescence and microscopy techniques

NASA Astrophysics Data System (ADS)

Healy, D. A.; Huffman, J. A.; O'Connor, D. J.; Pöhlker, C.; Pöschl, U.; Sodeau, J. R.

2014-02-01

Primary biological aerosol particles (PBAP) can contribute significantly to the coarse particle burden in many environments, may thus influence climate and precipitation systems as cloud nuclei, and can spread disease to humans, animals, and plants. Measurements of PBAP in natural environments taken at high time- and size- resolution are, however, sparse and so large uncertainties remain in the role that biological particles play in the Earth system. In this study two commercial real-time fluorescence particle sensors and a Sporewatch single-stage particle impactor were operated continuously from 2 August to 2 September 2010 at a rural sampling location in Killarney National Park in south western Ireland. A cascade impactor was operated periodically to collect size-resolved particles during exemplary periods. Here we report the first ambient comparison of the waveband integrated bioaerosol sensor (WIBS-4) with the ultraviolet aerodynamic particle sizer (UV-APS) and also compare these real-time fluorescence techniques with results of fluorescence and optical microscopy of impacted samples. Both real-time instruments showed qualitatively similar behaviour, with increased fluorescent bioparticle concentrations at night when relative humidity was highest and temperature was lowest. The fluorescent particle number from the FL3 channel of the WIBS-4 and from the UV-APS were strongly correlated and dominated by a 3 μm mode in the particle size distribution. The WIBS FL2 channel exhibited particle modes at approx. 1 and 3 μm, and each were correlated with the concentration of fungal spores commonly observed in air samples collected at the site (ascospores, basidiospores, Ganoderma spp.). The WIBS FL1 channel exhibited variable multi-modal distributions turning into a broad featureless single mode after averaging and exhibited poor correlation with fungal spore concentrations, which may be due to the detection of bacterial and non-biological fluorescent particles

Holographic techniques for cellular fluorescence microscopy

NASA Astrophysics Data System (ADS)

Kim, Myung K.

2017-04-01

We have constructed a prototype instrument for holographic fluorescence microscopy (HFM) based on self-interference incoherent digital holography (SIDH) and demonstrate novel imaging capabilities such as differential 3D fluorescence microscopy and optical sectioning by compressive sensing.

Development of a fluorescence lidar for biological aerosol detection in the air

NASA Astrophysics Data System (ADS)

He, Tingyao; Rao, Zhimin

2017-10-01

In order to investigate biological aerosols in the air, a fluorescence lidar has being developed at Laser Radar Center of Remote Sensing of Atmosphere, Xi'an University of Technology. The fluorescence lidar is constructed with a pulsed Nd:YAG laser, employing at based harmonic (1064 nm), second harmonic (532 nm) and fourth harmonic (266 nm) simultaneously, with a repetition rate of 10 Hz. A 250 mm diameter custom telescope is used to collect optical spectra ranging from 260-1100 nm. In the Infrared detection, an avalanche diode (APD) is used, and two photomultiplier tubes (PMTs) for two linear orthogonal polarization detection at a wavelength of 532 nm. Range-resolved fluorescence signals are collected in 32 channels of compound PMT sensor coupled with Czerny-Turner spectrograph. Based on the current configurations, we performed a series of numerical simulations to estimate the maximal detectable ranges and the minimal detectable concentrations of biological aerosols with various conditions. With a relative error of less than 10%, simulated results show that the system is able to monitor biological aerosols within detected distances of 1.3 km and of 2.0 km at daytime and nighttime, respectively. The developing fluorescence lidar is also capable to identify a minimum concentration of bio-aerosols at about 150 particles;L-1 with daytime operation and 100 particles;L-1 with nighttime at a distance of about 0.1 km. We truly believe that the fluorescence lidar could be spread in the field of remote sensing of biological aerosols in the near future.

Effect of Clouds on Apertures of Space-based Air Fluorescence Detectors

NASA Technical Reports Server (NTRS)

Sokolsky, P.; Krizmanic, J.

2003-01-01

Space-based ultra-high-energy cosmic ray detectors observe fluorescence light from extensive air showers produced by these particles in the troposphere. Clouds can scatter and absorb this light and produce systematic errors in energy determination and spectrum normalization. We study the possibility of using IR remote sensing data from MODIS and GOES satellites to delimit clear areas of the atmosphere. The efficiency for detecting ultra-high-energy cosmic rays whose showers do not intersect clouds is determined for real, night-time cloud scenes. We use the MODIS SST cloud mask product to define clear pixels for cloud scenes along the equator and use the OWL Monte Carlo to generate showers in the cloud scenes. We find the efficiency for cloud-free showers with closest approach of three pixels to a cloudy pixel is 6.5% exclusive of other factors. We conclude that defining a totally cloud-free aperture reduces the sensitivity of space-based fluorescence detectors to unacceptably small levels.

New techniques for fluorescence background rejection in microscopy and endoscopy

NASA Astrophysics Data System (ADS)

Ventalon, Cathie

2009-03-01

Confocal microscopy is a popular technique in the bioimaging community, mainly because it provides optical sectioning. However, its standard implementation requires 3-dimensional scanning of focused illumination throughout the sample. Efficient non-scanning alternatives have been implemented, among which the simple and well-established incoherent structured illumination microscopy (SIM) [1]. We recently proposed a similar technique, called Dynamic Speckle Illumination (DSI) microscopy, wherein the incoherent grid illumination pattern is replaced with a coherent speckle illumination pattern from a laser, taking advantage of the fact that speckle contrast is highly maintained in a scattering media, making the technique well adapted to tissue imaging [2]. DSI microscopy relies on the illumination of a sample with a sequence of dynamic speckle patterns and an image processing algorithm based only on an a priori knowledge of speckle statistics. The choice of this post-processing algorithm is crucial to obtain a good sectioning strength: in particular, we developed a novel post-processing algorithm based one wavelet pre-filtering of the raw images and obtained near-confocal fluorescence sectioning in a mouse brain labeled with GFP, with a good image quality maintained throughout a depth of ˜100 μm [3]. In the purpose of imaging fluorescent tissue at higher depth, we recently applied structured illumination to endoscopy. We used a similar set-up wherein the illumination pattern (a one-dimensional grid) is transported to the sample with an imaging fiber bundle with miniaturized objective and the fluorescence image is collected through the same bundle. Using a post-processing algorithm similar to the one previously described [3], we obtained high-quality images of a fluorescein-labeled rat colonic mucosa [4], establishing the potential of our endomicroscope for bioimaging applications. [4pt] Ref: [0pt] [1] M. A. A. Neil et al, Opt. Lett. 22, 1905 (1997) [0pt] [2] C

Ambient air contamination: Characterization and detection techniques

NASA Technical Reports Server (NTRS)

Nulton, C. P.; Silvus, H. S.

1985-01-01

Techniques to characterize and detect sources of ambient air contamination are described. Chemical techniques to identify indoor contaminants are outlined, they include gas chromatography, or colorimetric detection. Organics generated from indoor materials at ambient conditions and upon combustion are characterized. Piezoelectric quartz crystals are used as precision frequency determining elements in electronic oscillators.

Fluorescence exclusion: A simple versatile technique to calculate cell volumes and local heights (Conference Presentation)

NASA Astrophysics Data System (ADS)

Thouvenin, Olivier; Fink, Mathias; Boccara, A. Claude

2017-02-01

Understanding volume regulation during mitosis is technically challenging. Indeed, a very sensitive non invasive imaging over time scales ranging from seconds to hours and over large fields is required. Therefore, Quantitative Phase Imaging (QPI) would be a perfect tool for such a project. However, because of asymmetric protein segregation during mitosis, an efficient separation of the refractive index and the height in the phase signal is required. Even though many strategies to make such a separation have been developed, they usually are difficult to implement, have poor sensitivity, or cannot be performed in living cells, or in a single shot. In this paper, we will discuss the use of a new technique called fluorescence exclusion to perform volume measurements. By coupling such technique with a simultaneous phase measurement, we were also able to recover the refractive index inside the cells. Fluorescence exclusion is a versatile and powerful technique that allows the volume measurement of many types of cells. A fluorescent dye, which cannot penetrate inside the cells, is mixed with the external medium in a confined environment. Therefore, the fluorescent signal depends on the inverse of the object's height. We could demonstrate both experimentally and theoretically that fluorescence exclusion can accurately measure cell volumes, even for cells much higher than the depth of focus of the objective. A local accurate height and RI measurement can also be obtained for smaller cells. We will also discuss the way to optimize the confinement of the observation chamber, either mechanically or optically.

Air Coupled Acoustic Thermography (ACAT) Inspection Technique

NASA Technical Reports Server (NTRS)

Zalameda, Joseph; Winfree, William P.; Yost, William T.

2007-01-01

The scope of this effort is to determine the viability of a new heating technique using a noncontact acoustic excitation source. Because of low coupling between air and the structure, a synchronous detection method is employed. Any reduction in the out of plane stiffness improves the acoustic coupling efficiency and as a result, defective areas have an increase in temperature relative to the surrounding area. Hence a new measurement system, based on air-coupled acoustic energy and synchronous detection is presented. An analytical model of a clamped circular plate is given, experimentally tested, and verified. Repeatability confirms the technique with a measurement uncertainty of plus or minus 6.2 percent. The range of frequencies used was 800-2,000 Hertz. Acoustic excitation and consequent thermal detection of flaws in a helicopter blade is examined and results indicate that air coupled acoustic excitation enables the detection of core damage in sandwich honeycomb structures.

A Fluorescent Marking and Re-count Technique Using the Invasive Earthworm, Pontoscolex corethrurus (Annelida: Oligochaeta)

Treesearch

Grizelle Gonzalez; Elianid Espinoza; Zhigang Liu; Xiaoming Zou

2006-01-01

We used a fluorescence technique to mark and re-count the invasive earthworm, Pontoscolex corethrurus from PVC tubes established in a forest and a recently abandoned pasture in Puerto Rico to test the effects of the labeling treatment on earthworm population survival over time. A fluorescent marker was injected into the earthworms in the middle third section of the...

Fluorescence particle detector for real-time quantification of viable organisms in air

NASA Astrophysics Data System (ADS)

Luoma, Greg; Cherrier, Pierre P.; Piccioni, Marc; Tanton, Carol; Herz, Steve; DeFreez, Richard K.; Potter, Michael; Girvin, Kenneth L.; Whitney, Ronald

2002-02-01

The ability to detect viable organisms in air in real time is important in a number of applications. Detecting high levels of airborne organisms in hospitals can prevent post-operative infections and the spread of diseases. Monitoring levels of airborne viable organisms in pharmaceutical facilities can ensure safe production of drugs or vaccines. Monitoring airborne bacterial levels in meat processing plants can help to prevent contamination of food products. Monitoring the level of airborne organisms in bio-containment facilities can ensure that proper procedures are being followed. Finally, detecting viable organisms in real time is a key to defending against biological agent attacks. This presentation describes the development and performance of a detector, based on fluorescence particle counting technology, where an ultraviolet laser is used to count particles by light scattering and elicit fluorescence from specific biomolecules found only in living organisms. The resulting detector can specifically detect airborne particles containing living organisms from among the large majority of other particles normally present in air. Efforts to develop the core sensor technology, focusing on integrating an UV laser with a specially designed particle-counting cell will be highlighted. The hardware/software used to capture the information from the sensor, provide an alarm in the presence of an unusual biological aerosol content will also be described. Finally, results from experiments to test the performance of the detector will be presented.

Nitric oxide density measurements in air and air/fuel nanosecond pulse discharges by laser induced fluorescence

NASA Astrophysics Data System (ADS)

Uddi, M.; Jiang, N.; Adamovich, I. V.; Lempert, W. R.

2009-04-01

Laser induced fluorescence is used to measure absolute nitric oxide concentrations in air, methane-air and ethylene-air non-equilibrium plasmas, as a function of time after initiation of a single pulse, 20 kV peak voltage, 25 ns pulse duration discharge. A mixture of NO and nitrogen with known composition (4.18 ppm NO) is used for calibration. Peak NO density in air at 60 Torr, after a single pulse, is ~8 × 1012 cm-3 (~4.14 ppm) occurring at ~250 µs after the pulse, with decay time of ~16.5 ms. Peak NO atom mole fraction in a methane-air mixture with equivalence ratio of phiv = 0.5 is found to be approximately equal to that in air, with approximately the same rise and decay rate. In an ethylene-air mixture (also with equivalence ratio of phiv = 0.5), the rise and decay times are comparable to air and methane-air, but the peak NO concentration is reduced by a factor of approximately 2.5. Spontaneous emission measurements show that excited electronic states N2(C 3Π) and NO(A 2Σ) in air at P = 60 Torr decay within ~20 ns and ~1 µs, respectively. Kinetic modelling calculations incorporating air plasma kinetics complemented with the GRI Mech 3.0 hydrocarbon oxidation mechanism are compared with the experimental data using three different NO production mechanisms. It is found that NO concentration rise after the discharge pulse is much faster than predicted by Zel'dovich mechanism reactions, by two orders of magnitude, but much slower compared with reactions of electronically excited nitrogen atoms and molecules, also by two orders of magnitude. It is concluded that processes involving long lifetime (~100 µs) metastable states, such as N2(X 1Σ,v) and O2(b 1Σ), formed by quenching of the metastable N2(A 3Σ) state by ground electronic state O2, may play a dominant role in NO formation. NO decay, in all cases, is found to be dominated by the reverse Zel'dovich reaction, NO + O → N + O2, as well as by conversion into NO2 in a reaction of NO with ozone.

Simple and direct method for detecting phosphorus in air at normal pressure and temperature using a combination of LIBS and LIFS techniques

NASA Astrophysics Data System (ADS)

Al-Jeffery, Mohammad O.; Kondou, H.; Belenkevitch, Alexander; Azzeer, Abdallah M.

2002-05-01

The Environmental Protection Agency (EAP) designated phosphorus as hazardous material; it is flammable and poisonous. Phosphorus attacks the respiratory system, liver, kidneys, jaw, teeth, blood, eyes, and skin. Phosphorus is an element that has a high detection limit when using laser-induced breakdown spectroscopy (LIBS) techniques. In order to improve on detection limits, laser-induced fluorescence spectroscopy (LIFS) has been proposed, as an extension to LIBS. The ultimate goal of this work is to use the combined LIBS & LIFS techniques to detect the presence of phosphorus in air and to measure its level. In order to provide 'proof-of-concept' results, the sample used for our experiment was prepared using the 'igniting' strip of a safety match box. The spectrally and temporally resolved detection of the specific atomic emission revealed analytical information about the elemental composition of the sample. A tunable Ti: sapphire laser, at the resonance wavelength of 253.4 nm, was then used to probe the plume by exciting the phosphorus element and we measured the fluorescence from the atoms at 213.62 nm and 214.91 nm. The whole experiment was carried out in a few minutes. We have thus demonstrated for the first time, to our knowledge, the use of LIBS and LIFS in air quality monitoring and in particular for phosphorus detection.

Review of Fluorescence-Based Velocimetry Techniques to Study High-Speed Compressible Flows

NASA Technical Reports Server (NTRS)

Bathel, Brett F.; Johansen, Criag; Inman, Jennifer A.; Jones, Stephen B.; Danehy, Paul M.

2013-01-01

This paper reviews five laser-induced fluorescence-based velocimetry techniques that have been used to study high-speed compressible flows at NASA Langley Research Center. The techniques discussed in this paper include nitric oxide (NO) molecular tagging velocimetry (MTV), nitrogen dioxide photodissociation (NO2-to-NO) MTV, and NO and atomic oxygen (O-atom) Doppler-shift-based velocimetry. Measurements of both single-component and two-component velocity have been performed using these techniques. This paper details the specific application and experiment for which each technique has been used, the facility in which the experiment was performed, the experimental setup, sample results, and a discussion of the lessons learned from each experiment.

Control Techniques for Particulate Air Pollutants.

ERIC Educational Resources Information Center

National Air Pollution Control Administration (DHEW), Washington, DC.

Included is a comprehensive review of the approaches commonly recommended for controlling the sources of particulate air pollution. Not all possible combinations of control techniques that might bring about more stringent control of each individual source are reviewed. The many agricultural, commercial, domestic, industrial, and municipal…

Time-Domain Fluorescence Lifetime Imaging Techniques Suitable for Solid-State Imaging Sensor Arrays

PubMed Central

Li, David Day-Uei; Ameer-Beg, Simon; Arlt, Jochen; Tyndall, David; Walker, Richard; Matthews, Daniel R.; Visitkul, Viput; Richardson, Justin; Henderson, Robert K.

2012-01-01

We have successfully demonstrated video-rate CMOS single-photon avalanche diode (SPAD)-based cameras for fluorescence lifetime imaging microscopy (FLIM) by applying innovative FLIM algorithms. We also review and compare several time-domain techniques and solid-state FLIM systems, and adapt the proposed algorithms for massive CMOS SPAD-based arrays and hardware implementations. The theoretical error equations are derived and their performances are demonstrated on the data obtained from 0.13 μm CMOS SPAD arrays and the multiple-decay data obtained from scanning PMT systems. In vivo two photon fluorescence lifetime imaging data of FITC-albumin labeled vasculature of a P22 rat carcinosarcoma (BD9 rat window chamber) are used to test how different algorithms perform on bi-decay data. The proposed techniques are capable of producing lifetime images with enough contrast. PMID:22778606

Intrinsic Tryptophan Fluorescence in the Detection and Analysis of Proteins: A Focus on Förster Resonance Energy Transfer Techniques

PubMed Central

Ghisaidoobe, Amar B. T.; Chung, Sang J.

2014-01-01

Förster resonance energy transfer (FRET) occurs when the distance between a donor fluorophore and an acceptor is within 10 nm, and its application often necessitates fluorescent labeling of biological targets. However, covalent modification of biomolecules can inadvertently give rise to conformational and/or functional changes. This review describes the application of intrinsic protein fluorescence, predominantly derived from tryptophan (λEX ∼ 280 nm, λEM ∼ 350 nm), in protein-related research and mainly focuses on label-free FRET techniques. In terms of wavelength and intensity, tryptophan fluorescence is strongly influenced by its (or the protein’s) local environment, which, in addition to fluorescence quenching, has been applied to study protein conformational changes. Intrinsic Förster resonance energy transfer (iFRET), a recently developed technique, utilizes the intrinsic fluorescence of tryptophan in conjunction with target-specific fluorescent probes as FRET donors and acceptors, respectively, for real time detection of native proteins. PMID:25490136

SIL-STED microscopy technique enhancing super-resolution of fluorescence microscopy

NASA Astrophysics Data System (ADS)

Park, No-Cheol; Lim, Geon; Lee, Won-sup; Moon, Hyungbae; Choi, Guk-Jong; Park, Young-Pil

2017-08-01

We have characterized a new type STED microscope which combines a high numerical aperture (NA) optical head with a solid immersion lens (SIL), and we call it as SIL-STED microscope. The advantage of a SIL-STED microscope is that its high NA of the SIL makes it superior to a general STED microscope in lateral resolution, thus overcoming the optical diffraction limit at the macromolecular level and enabling advanced super-resolution imaging of cell surface or cell membrane structure and function Do. This study presents the first implementation of higher NA illumination in a STED microscope limiting the fluorescence lateral resolution to about 40 nm. The refractive index of the SIL which is made of material KTaO3 is about 2.23 and 2.20 at a wavelength of 633 nm and 780 nm which are used for excitation and depletion in STED imaging, respectively. Based on the vector diffraction theory, the electric field focused by the SILSTED microscope is numerically calculated so that the numerical results of the point dispersion function of the microscope and the expected resolution could be analyzed. For further investigation, fluorescence imaging of nano size fluorescent beads is fulfilled to show improved performance of the technique.

Instantaneous temperature field measurements using planar laser-induced fluorescence.

PubMed

Seitzman, J M; Kychakoff, G; Hanson, R K

1985-09-01

A single-pulse, laser-induced-fluorescence diagnostic for the measurement of two-dimensional temperature fields in combustion flows is described. The method uses sheet illumination from a tunable laser to excite planar laserinduced fluorescence in a stable tracer molecule, seeded at constant mole fraction into the flow field. The temporal resolution of this technique is determined by the laser pulse length. Experimental results are presented for a rodstabilized, premixed methane-air flame, using the Q(1) (22) line of the nitric oxide A(2) Sigma(+) (v = 0) ? X(2)II((1/2))(v = 0) transition (lambda approximately 225.6 nm).

The indirect fluorescent antibody technique as a method for detecting antibodies in aborted fetuses.

PubMed Central

Miller, R B; Wilkie, B N

1979-01-01

In this investigation the indirect fluorescent antibody technique was used to titrate antibodies in bovine sera to parainfluenza 3, infectious bovine rhinotracheitis virus and bovine viral diarrhea virus. These results were compared to those determined on the same samples by hemagglutination inhibition for parainfluenza 3 virus and serum neutralization for bovine virus diarrhea and infectious bovine rhinotracheitis virus. The results of the serological methods agreed closely. The indirect fluorescent antibody technique is a rapid and sensitive method for detecting antibodies and the procedure lends itself to use in diagnostic laboratories. In addition to the above viruses the presence or absence of antibodies to bovine coronavirus and bovine adenovirus 3 were determined by the indirect fluorescent antibody technique in thoracic fluids from 100 aborted fetuses and 50 nonaborted fetuses. Results on these samples were not compared to hemagglutination inhibition or serum neutralization as the condition of fluid samples from aborted fetuses renders interpretation of such tests unreliable. Antibodies to one or more viruses were detected in 30 of the 100 aborted fetuses and in seven of the 50 nonaborted fetuses. Antibodies to more than one agent were detected in eleven of the 100 aborted and in one of the 50 nonaborted fetuses. Reasons for this occurrence and application of the test in determination of causes of abortion are discussed. PMID:226243

Time-resolved fluorescence spectroscopy for chemical sensors

NASA Astrophysics Data System (ADS)

Draxler, Sonja; Lippitsch, Max E.

1996-07-01

A family of sensors is presented with fluorescence decay-time measurements used as the sensing technique. The concept is to take a single fluorophore with a suitably long fluorescence decay time as the basic building block for numerous different sensors. Analyte recognition can be performed by different functional groups that are necessary for selective interaction with the analyte. To achieve this, the principle of excited-state electron transfer is applied with pyrene as the fluorophore. Therefore the same instrumentation based on a small, ambient air-nitrogen laser and solid-state electronics can be used to measure different analytes, for example, oxygen, pH, carbon dioxide, potassium, ammonium, lead, cadmium, zinc, and phosphate.

Fluorescence hyperspectral imaging technique for the foreign substance detection on fresh-cut lettuce

USDA-ARS?s Scientific Manuscript database

Nondestructive methods based on fluorescence hyperspectral imaging (HSI) techniques were developed in order to detect worms on fresh-cut lettuce. The optimal wavebands for detecting worms on fresh-cut lettuce were investigated using the one-way ANOVA analysis and correlation analysis. The worm detec...

UV-fluorescence spectroscopic technique in the diagnosis of breast, ovarian, uterus, and cervix cancer

NASA Astrophysics Data System (ADS)

Das, Bidyut B.; Glassman, Wenling S.; Alfano, Robert R.; Cleary, Joseph; Prudente, R.; Celmer, Edward J.; Lubicz, Stephanie

1991-06-01

Malignant breast tumors can be separated from benign and normal tissues using uv-fluorescence spectroscopic technique. Using the same method one can also distinguish cancerous tissues from noncancerous ones in case of cervix, uterus and ovary.

Shot noise limited detection of OH using the technique of laser induced fluorescence

NASA Technical Reports Server (NTRS)

Bakalyar, D. M.; Davis, L. I., Jr.; Guo, C.; James, J. V.; Kakos, S.; Morris, P. T.; Wang, C. C.

1984-01-01

Nearly shot-noise limited detection of OH using the technique of laser-induced fluorescence is reported. A LIDAR configuration is used to excite fluorescence in a large volume and a narrow-bandwidth interference filter provides spectral discrimination. This arrangement alleviates the effect of ozone interference and facilitates image processing at relatively close distances. The detection limit is determined mainly by the shot-noise of the solar background. Ground-based measurements in Dearborn indicate a detection limit of better than 1 x 10 to the 6th power OH/cubic cm over a forty-minute acquisition period. Under favorable conditions, a comparable detection limit was also observed for airborne measurements.

Novel air-injection technique to locate the medial cut end of lacerated canaliculus.

PubMed

Liu, Bingqian; Li, Yonghao; Long, Chongde; Wang, Zhonghao; Liang, Xuanwei; Ge, Jian; Wang, Zhichong

2013-12-01

Locating the medial cut end of the severed canaliculus is the most difficult aspect of canalicular repair, especially in patients with more medial laceration, severe oedema, persistent errhysis and a narrow canaliculus. Irrigation is a widely used technique to identify the cut end; however, we found that air injected through the intact canaliculus with a straight needle failed to reflux when the common canaliculus or lacrimal sac was not blocked. We describe a simple, safe and efficient air-injection technique to identify the medial cut edge of a lacerated canaliculus. In this method, we initially submersed the medial canthus under normal saline, then injected filtered air through the intact canaliculus using a side port stainless steel probe with a closed round tip. The tip was designed to block the common canaliculus to form a relatively closed system. The efficiency of this novel air-injection technique was equivalent to the traditional technique but does not require the cooperation of the patient to blow air. Using this technique, the medial cut end was successfully identified by locating the air-bubble exit within minutes in 19 cases of mono-canalicular laceration without any complication.

Status of air-shower measurements with sparse radio arrays

NASA Astrophysics Data System (ADS)

Schröder, Frank G.

2017-03-01

This proceeding gives a summary of the current status and open questions of the radio technique for cosmic-ray air showers, assuming that the reader is already familiar with the principles. It includes recent results of selected experiments not present at this conference, e.g., LOPES and TREND. Current radio arrays like AERA or Tunka-Rex have demonstrated that areas of several km2 can be instrumented for reasonable costs with antenna spacings of the order of 200m. For the energy of the primary particle such sparse antenna arrays can already compete in absolute accuracy with other precise techniques, like the detection of air-fluorescence or air-Cherenkov light. With further improvements in the antenna calibration, the radio detection might become even more accurate. For the atmospheric depth of the shower maximum, Xmax, currently only the dense array LOFAR features a precision similar to the fluorescence technique, but analysis methods for the radio measurement of Xmax are still under development. Moreover, the combination of radio and muon measurements is expected to increase the accuracy of the mass composition, and this around-the-clock recording is not limited to clear nights as are the light-detection methods. Consequently, radio antennas will be a valuable add-on for any air shower array targeting the energy range above 100 PeV.

Technique for Increasing the Selectivity of the Method of Laser Fragmentation/Laser-Induced Fluorescence

NASA Astrophysics Data System (ADS)

Bobrovnikov, S. M.; Gorlov, E. V.; Zharkov, V. I.

2018-05-01

A technique for increasing the selectivity of the method of detecting high-energy materials (HEMs) based on laser fragmentation of HEM molecules with subsequent laser excitation of fluorescence of the characteristic NO fragments from the first vibrational level of the ground state is suggested.

A simple and sensitive fluorescence method for the determination of trace ozone in air using acridine red as a probe.

PubMed

Liu, Qingye; Lin, Chenyin; Zhang, Xinghui; Wen, Guiqing; Liang, Aihui

2014-12-01

The ozone in an air sample was trapped by H3 BO3 -LK solution to produce iodine (I2) that interacted with excess I(-) to form I3(-). In pH 4.0 acetate buffer solutions, the I3(-) reacted with acridine red to form acridine red-I3 ion association particles that resulted in the fluorescence peak decreased at 553 nm. The decreased value ΔF553 nm is linear to the O3 concentration in the range 0.08-53.3 × 10(-6) mol/L, with a detection limit of 4 × 10(-8) mol/L. This fluorescence method was used to determine ozone in air samples, and the results were in agreement with that of indigo carmine spectrophotometry. Copyright © 2014 John Wiley & Sons, Ltd.

75 FR 59180 - Approval and Promulgation of Air Quality Implementation Plans; Maryland; Control Technique...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-09-27

... Promulgation of Air Quality Implementation Plans; Maryland; Control Technique Guidelines for Paper, Film, and... Technique Guidelines (CTG) for paper, film, and foil coatings. These amendments will reduce volatile organic... Promulgation of Air Quality Implementation Plans; Maryland; Control Technique Guidelines for Paper, Film, and...

Open-air multispectral fluorescence-guided surgery platform for intraoperative detection of malignant tissue under ambient lighting conditions

NASA Astrophysics Data System (ADS)

Behrooz, Ali; Vasquez, Kristine O.; Waterman, Peter; Meganck, Jeff; Peterson, Jeffrey D.; Miller, Peter; Kempner, Joshua

2017-02-01

Intraoperative resection of tumors currently relies upon the surgeon's ability to visually locate and palpate tumor nodules. Undetected residual malignant tissue often results in the need for additional treatment or surgical intervention. The Solaris platform is a multispectral open-air fluorescence imaging system designed for translational fluorescence-guided surgery. Solaris supports video-rate imaging in four fixed fluorescence channels ranging from visible to near infrared, and a multispectral channel equipped with a liquid crystal tunable filter (LCTF) for multispectral image acquisition (520-620 nm). Identification of tumor margins using reagents emitting in the visible spectrum (400-650 nm), such as fluorescein isothiocyanate (FITC), present challenges considering the presence of auto-fluorescence from tissue and food in the gastrointestinal (GI) tract. To overcome this, Solaris acquires LCTF-based multispectral images, and by applying an automated spectral unmixing algorithm to the data, separates reagent fluorescence from tissue and food auto-fluorescence. The unmixing algorithm uses vertex component analysis to automatically extract the primary pure spectra, and resolves the reagent fluorescent signal using non-negative least squares. For validation, intraoperative in vivo studies were carried out in tumor-bearing rodents injected with FITC-dextran reagent that is primarily residing in malignant tissue 24 hours post injection. In the absence of unmixing, fluorescence from tumors is not distinguishable from that of surrounding tissue. Upon spectral unmixing, the FITC-labeled malignant regions become well defined and detectable. The results of these studies substantiate the multispectral power of Solaris in resolving FITC-based agent signal in deep tumor masses, under ambient and surgical light, and enhancing the ability to surgically resect them.

Fluorescence Imaging Reveals Surface Contamination

NASA Technical Reports Server (NTRS)

Schirato, Richard; Polichar, Raulf

1992-01-01

In technique to detect surface contamination, object inspected illuminated by ultraviolet light to make contaminants fluoresce; low-light-level video camera views fluorescence. Image-processing techniques quantify distribution of contaminants. If fluorescence of material expected to contaminate surface is not intense, tagged with low concentration of dye.

Chemical reactivation of quenched fluorescent protein molecules enables resin-embedded fluorescence microimaging

PubMed Central

Xiong, Hanqing; Zhou, Zhenqiao; Zhu, Mingqiang; Lv, Xiaohua; Li, Anan; Li, Shiwei; Li, Longhui; Yang, Tao; Wang, Siming; Yang, Zhongqin; Xu, Tonghui; Luo, Qingming; Gong, Hui; Zeng, Shaoqun

2014-01-01

Resin embedding is a well-established technique to prepare biological specimens for microscopic imaging. However, it is not compatible with modern green-fluorescent protein (GFP) fluorescent-labelling technique because it significantly quenches the fluorescence of GFP and its variants. Previous empirical optimization efforts are good for thin tissue but not successful on macroscopic tissue blocks as the quenching mechanism remains uncertain. Here we show most of the quenched GFP molecules are structurally preserved and not denatured after routine embedding in resin, and can be chemically reactivated to a fluorescent state by alkaline buffer during imaging. We observe up to 98% preservation in yellow-fluorescent protein case, and improve the fluorescence intensity 11.8-fold compared with unprocessed samples. We demonstrate fluorescence microimaging of resin-embedded EGFP/EYFP-labelled tissue block without noticeable loss of labelled structures. This work provides a turning point for the imaging of fluorescent protein-labelled specimens after resin embedding. PMID:24886825

An intercomparison of nitric oxide measurement techniques

NASA Technical Reports Server (NTRS)

Hoell, J. M., Jr.; Gregory, G. L.; Mcdougal, D. S.; Carroll, M. A.; Mcfarland, M.; Ridley, B. A.; Davis, D. D.; Bradshaw, J.; Rodgers, M. O.; Torres, A. L.

1985-01-01

Results from an intercomparison of techniques to measure tropospheric levels of nitric oxide (NO) are discussed. The intercomparison was part of the National Aeronautics and Space Administration's Global Tropospheric Experiment and was conducted at Wallops Island, VA, in July 1983. Instruments intercompared included a laser-induced fluorescence system and two chemiluminescence instruments. The intercomparisons were performed with ambient air at NO mixing ratios ranging from 10 to 60 pptv and NO-enriched ambient air at mixing ratios from 20 to 170 pptv. All instruments sampled from a common manifold. The techniques exhibited a high degree of correlation among themselves and with changes in the NO mixing ratio. Agreement among the three techniques was placed at approximately + or - 30 percent. Within this level of agreement, no artifacts or species interferences were identified.

Air Quality Forecasting through Different Statistical and Artificial Intelligence Techniques

NASA Astrophysics Data System (ADS)

Mishra, D.; Goyal, P.

2014-12-01

Urban air pollution forecasting has emerged as an acute problem in recent years because there are sever environmental degradation due to increase in harmful air pollutants in the ambient atmosphere. In this study, there are different types of statistical as well as artificial intelligence techniques are used for forecasting and analysis of air pollution over Delhi urban area. These techniques are principle component analysis (PCA), multiple linear regression (MLR) and artificial neural network (ANN) and the forecasting are observed in good agreement with the observed concentrations through Central Pollution Control Board (CPCB) at different locations in Delhi. But such methods suffers from disadvantages like they provide limited accuracy as they are unable to predict the extreme points i.e. the pollution maximum and minimum cut-offs cannot be determined using such approach. Also, such methods are inefficient approach for better output forecasting. But with the advancement in technology and research, an alternative to the above traditional methods has been proposed i.e. the coupling of statistical techniques with artificial Intelligence (AI) can be used for forecasting purposes. The coupling of PCA, ANN and fuzzy logic is used for forecasting of air pollutant over Delhi urban area. The statistical measures e.g., correlation coefficient (R), normalized mean square error (NMSE), fractional bias (FB) and index of agreement (IOA) of the proposed model are observed in better agreement with the all other models. Hence, the coupling of statistical and artificial intelligence can be use for the forecasting of air pollutant over urban area.

Remote sensing of OH in the atmosphere using the technique of laser-induced fluorescence

NASA Technical Reports Server (NTRS)

Wang, C. C.

1983-01-01

The use of a laser-induced fluorescence technique for the sensitive measurement of the atmospheric hydroxyl radical is discussed. Results of laboratory studies of the fluorescence and other spectroscopic properties of OH which allow the calculation of OH concentrations from the returned signals for various altitudes, water vapor contents and temperatures are presented. The experimental setup used for airborne OH measurements is then described, with particular attention given to the use of a telescope for excitation and light collection in a coaxial configuration and the periodic tuning of the exciting radiation necessary to obtain an OH signal in the presence of strong solar and nonresonant fluorescence backgrounds. The best detection limit obtained to date with the system is noted to be about 700,000 OH/cu cm, and it is expected that, with planned improvements in detection and tuning schemes, limits in the neighborhood of 1,000,000 OH/cu cm will be achieved routinely.

Air pollution source identification

NASA Technical Reports Server (NTRS)

Fordyce, J. S.

1975-01-01

Techniques for air pollution source identification are reviewed, and some results obtained with them are evaluated. Described techniques include remote sensing from satellites and aircraft, on-site monitoring, and the use of injected tracers and pollutants themselves as tracers. The use of a large number of trace elements in ambient airborne particulate matter as a practical means of identifying sources is discussed in detail. Sampling and analysis techniques are described, and it is shown that elemental constituents can be related to specific source types such as those found in the earth's crust and those associated with specific industries. Source identification sytems are noted which utilize charged particle X-ray fluorescence analysis of original field data.

Development of a New Time-Resolved Laser-Induced Fluorescence Technique

NASA Astrophysics Data System (ADS)

Durot, Christopher; Gallimore, Alec

2012-10-01

We are developing a time-resolved laser-induced fluorescence (LIF) technique to interrogate the ion velocity distribution function (VDF) of EP thruster plumes down to the microsecond time scale. Better measurements of dynamic plasma processes will lead to improvements in simulation and prediction of thruster operation and erosion. We present the development of the new technique and results of initial tests. Signal-to-noise ratio (SNR) is often a challenge for LIF studies, and it is only more challenging for time-resolved measurements since a lock-in amplifier cannot be used with a long time constant. The new system uses laser modulation on the order of MHz, which enables the use of electronic filtering and phase-sensitive detection to improve SNR while preserving time-resolved information. Statistical averaging over many cycles to further improve SNR is done in the frequency domain. This technique can have significant advantages, including (1) larger spatial maps enabled by shorter data acquisition time and (2) the ability to average data without creating a phase reference by modifying the thruster operating condition with a periodic cutoff in discharge current, which can modify the ion velocity distribution.

Ultrafast Method for the Analysis of Fluorescence Lifetime Imaging Microscopy Data Based on the Laguerre Expansion Technique

PubMed Central

Jo, Javier A.; Fang, Qiyin; Marcu, Laura

2007-01-01

We report a new deconvolution method for fluorescence lifetime imaging microscopy (FLIM) based on the Laguerre expansion technique. The performance of this method was tested on synthetic and real FLIM images. The following interesting properties of this technique were demonstrated. 1) The fluorescence intensity decay can be estimated simultaneously for all pixels, without a priori assumption of the decay functional form. 2) The computation speed is extremely fast, performing at least two orders of magnitude faster than current algorithms. 3) The estimated maps of Laguerre expansion coefficients provide a new domain for representing FLIM information. 4) The number of images required for the analysis is relatively small, allowing reduction of the acquisition time. These findings indicate that the developed Laguerre expansion technique for FLIM analysis represents a robust and extremely fast deconvolution method that enables practical applications of FLIM in medicine, biology, biochemistry, and chemistry. PMID:19444338

Evaluation of the photocatalytic activity of Ln3+-TiO2 nanomaterial using fluorescence technique for real wastewater treatment.

PubMed

Saif, M; Aboul-Fotouh, S M K; El-Molla, S A; Ibrahim, M M; Ismail, L F M

2014-07-15

Evaluation the photocatalytic activity of different Ln(3+) modified TiO2 nanomaterials using fluorescence based technique has rarely been reported. In the present work, xmol Ln(3+) modified TiO2 nanomaterials (Ln = Nd(3+), Sm(3+), Eu(3+), Gd(3+), Dy(3+) and Er(3+) ions; x = 0.005, 0.008, 0.01, 0.02 and 0.03) were synthesized by sol-gel method and characterized using different advanced techniques. The photocatalytic efficiency of the modified TiO2 expressed in the charge carrier separation and OH radicals formation were assigned using TiO2 fluorescence quenching and fluorescence probe methods, respectively. The obtained fluorescence measurements confirm that doping treatment significantly decreases the electron-hole recombination probability in the obtained Ln(3+)/TiO2. Moreover, the rate of OH radicals formation is increased by doping. The highly active nanoparticles (0.02Gd(3+)/TiO2 and 0.01Eu(3+)/TiO2) were applied for industrial wastewater treatment using solar radiation as a renewable energy source. Copyright © 2014 Elsevier B.V. All rights reserved.

Compressed air injection technique to standardize block injection pressures.

PubMed

Tsui, Ban C H; Li, Lisa X Y; Pillay, Jennifer J

2006-11-01

Presently, no standardized technique exists to monitor injection pressures during peripheral nerve blocks. Our objective was to determine if a compressed air injection technique, using an in vitro model based on Boyle's law and typical regional anesthesia equipment, could consistently maintain injection pressures below a 1293 mmHg level associated with clinically significant nerve injury. Injection pressures for 20 and 30 mL syringes with various needle sizes (18G, 20G, 21G, 22G, and 24G) were measured in a closed system. A set volume of air was aspirated into a saline-filled syringe and then compressed and maintained at various percentages while pressure was measured. The needle was inserted into the injection port of a pressure sensor, which had attached extension tubing with an injection plug clamped "off". Using linear regression with all data points, the pressure value and 99% confidence interval (CI) at 50% air compression was estimated. The linearity of Boyle's law was demonstrated with a high correlation, r = 0.99, and a slope of 0.984 (99% CI: 0.967-1.001). The net pressure generated at 50% compression was estimated as 744.8 mmHg, with the 99% CI between 729.6 and 760.0 mmHg. The various syringe/needle combinations had similar results. By creating and maintaining syringe air compression at 50% or less, injection pressures will be substantially below the 1293 mmHg threshold considered to be an associated risk factor for clinically significant nerve injury. This technique may allow simple, real-time and objective monitoring during local anesthetic injections while inherently reducing injection speed.

Feasibility of measuring temperature and density fluctuations in air using laser-induced O2 fluorescence

NASA Technical Reports Server (NTRS)

Massey, G. A.; Lemon, C. J.

1984-01-01

A tunable line-narrowed ArF laser can selectively excite several rotation al lines of the Schumann-Runge band system of O2 in air. The resulting ultraviolet fluorescence can be monitored at 90 deg to the laser beam axis, permitting space and time resolved observation of density and temperature fluctuations in turbulence. Experiments and calculations show that + or - 1 K, + or - 1 percent density, 1 cu mm spatial, and 1 microsecond temporal resolution can be achieved simultaneously under some conditions.

An Intelligent Harmonic Synthesis Technique for Air-Gap Eccentricity Fault Diagnosis in Induction Motors

NASA Astrophysics Data System (ADS)

Li, De Z.; Wang, Wilson; Ismail, Fathy

2017-11-01

Induction motors (IMs) are commonly used in various industrial applications. To improve energy consumption efficiency, a reliable IM health condition monitoring system is very useful to detect IM fault at its earliest stage to prevent operation degradation, and malfunction of IMs. An intelligent harmonic synthesis technique is proposed in this work to conduct incipient air-gap eccentricity fault detection in IMs. The fault harmonic series are synthesized to enhance fault features. Fault related local spectra are processed to derive fault indicators for IM air-gap eccentricity diagnosis. The effectiveness of the proposed harmonic synthesis technique is examined experimentally by IMs with static air-gap eccentricity and dynamic air-gap eccentricity states under different load conditions. Test results show that the developed harmonic synthesis technique can extract fault features effectively for initial IM air-gap eccentricity fault detection.

Transient Fluorescence Spectroscopy and laser induced fluorescence lifetimes of terbium doped dipicolinic acid

NASA Astrophysics Data System (ADS)

Makoui, Anali

We have investigated the use of deep UV laser induced fluorescence for the sensitive detection and spectroscopic lifetime studies of terbium doped dipicolinic acid (DPA-Tb) and used this to study the optical characteristics of DPA which is a chemical surrounding most bacterial spores. Background absorption spectra, fluorescence spectra, and Excitation Emission Matrix (EEM) spectra were made of the DPA-Tb complex, using both fixed 266 nm wavelength and tunable (220 nm--280 nm) UV laser excitations. Of importance, the fluorescence lifetimes of the four main fluorescence peaks (488 nm, 543 nm, 581 nm, and 618 nm) of the DPA-Tb complex have been measured for the first time to our knowledge. The lifetimes of all the fluorescing lines have been measured as a function of DPA-Tb concentration, solvent pH, and solvent composition, including that for the weakest fluorescing line of DPA-Tb at 618 nm. In addition, a new spectroscopic lifetime measurement technique, which we call "Transient Fluorescence Spectroscopy", was developed. In this technique, a weak, quasi-CW, amplitude modulated UV laser (8.5 kHz) was used to measure the lifetimes of the fluorescence lines, and yields insight into energy transfer and excitation lifetimes within the system. This technique is especially useful when a high power laser is not either available or not suitable. In the latter case, this would be when a high power pulsed deep-UV laser could produce bleaching or destruction of the biological specimen. In addition, this technique simulated the excitation and fluorescence emission of the DPA-Tb using a 4-level energy model, and solved the dynamic transient rate equations to predict the temporal behavior of the DPA-Tb emitted fluorescence. Excellent agreement between the experiments and the simulation were found. This technique has the potential to provide a more accurate value for the fluorescence lifetime values. In addition, with the use of asymmetric excitation waveforms, the dynamic

A Fluorescence Recovery After Photobleaching (FRAP) Technique for the Measurement of Solute Transport Across Surfactant-Laden Interfaces

NASA Technical Reports Server (NTRS)

Browne, Edward P.; Hatton, T. Alan

1996-01-01

The technique of Fluorescence Recovery After Photobleaching (FRAP) has been applied to the measurement of interfacial transport in two-phase systems. FRAP exploits the loss of fluorescence exhibited by certain fluorophores when over-stimulated (photobleached), so that a two-phase system, originally at equilibrium, can be perturbed without disturbing the interface by strong light from an argon-ion laser and its recovery monitored by a microscope-mounted CCD camera as it relaxes to a new equilibrium. During this relaxation, the concentration profiles of the probe solute are measured on both sides of the interface as a function of time, yielding information about the transport characteristics of the system. To minimize the size of the meniscus between the two phases, a photolithography technique is used to selectively treat the glass walls of the cell in which the phases are contained. This allows concentration measurements to be made very close to the interface and increases the sensitivity of the FRAP technique.

A Planar-Fluorescence Imaging Technique for Studying Droplet-Turbulence Interactions in Vaporizing Sprays

NASA Technical Reports Server (NTRS)

Santavicca, Dom A.; Coy, E.

1990-01-01

Droplet turbulence interactions directly affect the vaporization and dispersion of droplets in liquid sprays and therefore play a major role in fuel oxidizer mixing in liquid fueled combustion systems. Proper characterization of droplet turbulence interactions in vaporizing sprays require measurement of droplet size velocity and size temperature correlations. A planar, fluorescence imaging technique is described which is being developed for simultaneously measuring the size, velocity, and temperature of individual droplets in vaporizing sprays. Preliminary droplet size velocity correlation measurements made with this technique are presented. These measurements are also compared to and show very good agreement with measurements made in the same spray using a phase Doppler particle analyzer.

Temperature measurement in a compressible flow field using laser-induced iodine fluorescence

NASA Technical Reports Server (NTRS)

Fletcher, D. G.; Mcdaniel, J. C.

1987-01-01

The thermometric capability of a two-line fluorescence technique using iodine seed molecules in air is investigated analytically and verified experimentally in a known steady compressible flow field. Temperatures ranging from 165 to 295 K were measured in the flowfield using two iodine transitions accessed with a 30-GHz dye-laser scan near 543 nm. The effect of pressure broadening on temperature measurement is evaluated.

Ultrafast fluorescence upconversion technique and its applications to proteins.

PubMed

Chosrowjan, Haik; Taniguchi, Seiji; Tanaka, Fumio

2015-08-01

The basic principles and main characteristics of the ultrafast time-resolved fluorescence upconversion technique (conventional and space-resolved), including requirements for nonlinear crystals, mixing spectral bandwidth, acceptance angle, etc., are presented. Applications to flavoproteins [wild-type (WT) FMN-binding protein and its W32Y, W32A, E13R, E13K, E13Q and E13T mutants] and photoresponsive proteins [WT photoactive yellow protein and its R52Q mutant in solution and as single crystals] are demonstrated. For flavoproteins, investigations elucidating the effects of ionic charges on ultrafast electron transfer (ET) dynamics are summarized. It is shown that replacement of the ionic amino acid Glu13 and the resulting modification of the electrostatic charge distribution in the protein chromphore-binding pocket substantially alters the ultrafast fluorescence quenching dynamics and ET rate in FMN-binding protein. It is concluded that, together with donor-acceptor distances, electrostatic interactions between ionic photoproducts and other ionic groups in the proteins are important factors influencing the ET rates. In WT photoactive yellow protein and the R52Q mutant, ultrafast photoisomerization dynamics of the chromophore (deprotonated trans-p-coumaric acid) in liquid and crystal phases are investigated. It is shown that the primary dynamics in solution and single-crystal phases are quite similar; hence, the photocycle dynamics and structural differences observed at longer time scales arise mostly from the structural restraints imposed by the crystal lattice rigidity versus the flexibility in solution. © 2014 FEBS.

High-Collection-Efficiency Fluorescence Detection Cell

NASA Technical Reports Server (NTRS)

Hanisco, Thomas; Cazorla, Maria; Swanson, Andrew

2013-01-01

A new fluorescence cell has been developed for the laser induced fluorescence (LIF) detection of formaldehyde. The cell is used to sample a flow of air that contains trace concentrations of formaldehyde. The cell provides a hermetically sealed volume in which a flow of air containing formaldehyde can be illuminated by a laser. The cell includes the optics for transmitting the laser beam that is used to excite the formaldehyde and for collecting the resulting fluorescence. The novelty of the cell is its small size and simple design that provides a more robust and cheaper alternative to the state of the art. Despite its simplicity, the cell provides the same sensitivity to detection as larger, more complicated cells.

Agricultural pest monitoring using fluorescence lidar techniques. Feasibility study

NASA Astrophysics Data System (ADS)

Mei, L.; Guan, Z. G.; Zhou, H. J.; Lv, J.; Zhu, Z. R.; Cheng, J. A.; Chen, F. J.; Löfstedt, C.; Svanberg, S.; Somesfalean, G.

2012-03-01

The fluorescence of different types of planthopper ( Hemiptera) and moth ( Lepidoptera), which constitute important Chinese agricultural pests, was investigated both in situ in a laboratory setting and remotely using a fluorescence light detection and ranging (lidar) system operating at a range of about 50 m. The natural autofluorescence of different species, as well as the fluorescence from insects that had been dusted with fluorescent dye powder for identification were studied. Autofluorescence spectra of both moths and planthoppers show a maximum intensity peak around 450 nm. Bleaching upon long-time laser illumination was modest and did not affect the shape of the spectrum. A single dyed rice planthopper, a few mm in size, could be detected at 50 m distance by using the fluorescence lidar system. By employing various marking dyes, different types of agricultural pest could be determined. We suggest that lidar may be used in studies of migration and movement of pest insects, including studies of their behavior in the vicinity of pheromone traps and in pheromone-treated fields.

High resolution x-ray fluorescence spectroscopy - a new technique for site- and spin-selectivity

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wang, Xin

1996-12-01

X-ray spectroscopy has long been used to elucidate electronic and structural information of molecules. One of the weaknesses of x-ray absorption is its sensitivity to all of the atoms of a particular element in a sample. Through out this thesis, a new technique for enhancing the site- and spin-selectivity of the x-ray absorption has been developed. By high resolution fluorescence detection, the chemical sensitivity of K emission spectra can be used to identify oxidation and spin states; it can also be used to facilitate site-selective X-ray Absorption Near Edge Structure (XANES) and site-selective Extended X-ray Absorption Fine Structure (EXAFS). Themore » spin polarization in K fluorescence could be used to generate spin selective XANES or spin-polarized EXAFS, which provides a new measure of the spin density, or the nature of magnetic neighboring atoms. Finally, dramatic line-sharpening effects by the combination of absorption and emission processes allow observation of structure that is normally unobservable. All these unique characters can enormously simplify a complex x-ray spectrum. Applications of this novel technique have generated information from various transition-metal model compounds to metalloproteins. The absorption and emission spectra by high resolution fluorescence detection are interdependent. The ligand field multiplet model has been used for the analysis of K{alpha} and K{beta} emission spectra. First demonstration on different chemical states of Fe compounds has shown the applicability of site selectivity and spin polarization. Different interatomic distances of the same element in different chemical forms have been detected using site-selective EXAFS.« less

First Results of Using a UVTron Flame Sensor to Detect Alpha-Induced Air Fluorescence in the UVC Wavelength Range.

PubMed

Crompton, Anita J; Gamage, Kelum A A; Bell, Steven; Wilson, Andrew P; Jenkins, Alex; Trivedi, Divyesh

2017-11-29

In this work, a robust stand-off alpha detection method using the secondary effects of alpha radiation has been sought. Alpha particles ionise the surrounding atmosphere as they travel. Fluorescence photons produced as a consequence of this can be used to detect the source of the alpha emissions. This paper details experiments carried out to detect this fluorescence, with the focus on photons in the ultraviolet C (UVC) wavelength range (180-280 nm). A detector, UVTron R9533 (Hamamatsu, 325-6, Sunayama-cho, Naka-ku, Hamamatsu City, Shizuoka Pref., 430-8587, Japan), designed to detect the UVC emissions from flames for fire alarm purposes, was tested in various gas atmospheres with a 210 Po alpha source to determine if this could provide an avenue for stand-off alpha detection. The results of the experiments show that this detector is capable of detecting alpha-induced air fluorescence in normal indoor lighting conditions, as the interference from daylight and artificial lighting is less influential on this detection system which operates below the UVA and UVB wavelength ranges (280-315 nm and 315-380 nm respectively). Assuming a standard 1 r 2 drop off in signal, the limit of detection in this configuration can be calculated to be approximately 240 mm, well beyond the range of alpha-particles in air, which indicates that this approach could have potential for stand-off alpha detection. The gas atmospheres tested produced an increase in the detector count, with xenon having the greatest effect with a measured 52% increase in the detector response in comparison to the detector response in an air atmosphere. This type of alpha detection system could be operated at a distance, where it would potentially provide a more cost effective, safer, and faster solution in comparison with traditional alpha detection methods to detect and characterise alpha contamination in nuclear decommissioning and security applications.

Development of a measurement system for nitrate radical and dinitrogen pentoxide using a thermal conversion/laser-induced fluorescence technique

NASA Astrophysics Data System (ADS)

Matsumoto, Jun; Kosugi, Naohiro; Imai, Hidekazu; Kajii, Yoshizumi

2005-06-01

An instrument for measuring atmospheric nitrate radical (NO3) and dinitrogen pentoxide (N2O5) has been developed by a thermal conversion/laser-induced fluorescence (TC/LIF) technique. N2O5 is thermally decomposed and converted to NO3, which is measured by laser-induced fluorescence. In situ, fast-response, sensitive measurement of NO3/N2O5 is expected by use of LIF. In detecting NO3, dual-wavelength excitation at 622.96 and 618.81nm was adopted to remove potential interference and to guarantee high selectivity. A high-power dye laser system was used as the source of excitation light. To measure ambient air directly, the NO3 detection cell was placed on the rooftop. The laser beam was guided by an optical fiber into the excitation cell. Transmittance of the laser beam was 80% for a 10m long fiber. To calibrate the instrument, the series of thermal decomposition of N2O5 and the gas phase titration of NO3 by NO were conducted. NO3 reduction by adding NO was also applied to determine accurately the zero points of the detector. After optimization of conditions such as gate timing in photon counting and the settings of the N2O5 converter, the present detection limits of NO3 and N2O5 were determined to be 4 and 6pptv, respectively, for the integration time of 10min (signal-to-noise ratio=1). It was confirmed that the interference of NO2 on N2O5 detection is negligible, but can be significant for NO3 measurement when NO2 concentration is extremely high (>100ppbv ). Measurement of N2O5 in ambient air was made in the urban area of Tokyo, Japan. Observed data demonstrated the capacity of the TC/LIF instrument with a powerful dye laser and a single-path excitation cell for ambient measurements. In this article, we focus on the instrumentation and characterization.

Planetary Surface Analysis Using Fast Laser Spectroscopic Techniques: Combined Microscopic Raman, LIBS, and Fluorescence Spectroscopy

NASA Astrophysics Data System (ADS)

Blacksberg, J.; Rossman, G. R.; Maruyama, Y.; Charbon, E.

2011-12-01

In situ exploration of planetary surfaces has to date required multiple techniques that, when used together, yield important information about their formation histories and evolution. We present a time-resolved laser spectroscopic technique that could potentially collect complementary sets of data providing information on mineral structure, composition, and hydration state. Using a picosecond-scale pulsed laser and a fast time-resolved detector we can simultaneously collect spectra from Raman, Laser Induced Breakdown Spectroscopy (LIBS), and fluorescence emissions that are separated in time due to the unique decay times of each process. The use of a laser with high rep rate (40 KHz) and low pulse energy (1 μJ/pulse) allows us to rapidly collect high signal to noise Raman spectra while minimizing sample damage. Increasing the pulse energy by about an order of magnitude creates a microscopic plasma near the surface and enables the collection of LIBS spectra at an unusually high rep rate and low pulse energy. Simultaneously, broader fluorescence peaks can be detected with lifetimes varying from nanosecond to microsecond. We will present Raman, LIBS, and fluorescence spectra obtained on natural mineral samples such as sulfates, clays, pyroxenes and carbonates that are of interest for Mars mineralogy. We demonstrate this technique using a photocathode-based streak camera detector as well as a newly-developed solid state Single Photon Avalanche Diode (SPAD) sensor array based on Complementary Metal-Oxide Semiconductor (CMOS) technology. We will discuss the impact of system design and detector choice on science return of a potential planetary surface mission, with a specific focus on size, weight, power, and complexity. The research described here was carried out at the Jet Propulsion Laboratory, California Institute of Technology, under a contract with the National Aeronautics and Space Administration (NASA).

Direct fluorescent antibody technique for the detection of bacterial kidney disease in paraffin-embedded tissues

USGS Publications Warehouse

Ochiai, T.; Yasutake, W.T.; Gould, R.W.

1985-01-01

The direct fluorescent antibody technique (FAT) was successfully used to detect the causative agent of bacterial kidney disease (BKD), Renibacterium salmoninarum, in Bouin's solution flexed and paraffinembedded egg and tissue sections. This method is superior to gram stain and may be particularly useful in detecting the BKD organism in fish with low-grade infection.

UV red fluorescence of Eubacterium lentum.

PubMed Central

Mosca, A; Strong, C A; Finegold, S M

1993-01-01

Twenty-nine clinical isolates of Eubacterium lentum and two type species were evaluated for the ability to fluoresce under UV light. Twenty-one of the 29 isolates and both of the reference strains showed orange-to-red fluorescence. This fluorescence did not require blood or hemin in the culture media and did not fade upon air exposure. The fluorescent pigment, after extraction by 1 N NaOH, showed peak excitation at a wavelength of around 400 nm. The capacity of E. lentum to produce fluorescence may be a useful and time-sparing laboratory aid for its identification. PMID:8463378

Fluorescence lifetime in cardiovascular diagnostics

NASA Astrophysics Data System (ADS)

Marcu, Laura

2010-01-01

We review fluorescence lifetime techniques including time-resolved laser-induced fluorescence spectroscopy (TR-LIFS) and fluorescence lifetime imaging microscopy (FLIM) instrumentation and associated methodologies that allow for characterization and diagnosis of atherosclerotic plaques. Emphasis is placed on the translational research potential of TR-LIFS and FLIM and on determining whether intrinsic fluorescence signals can be used to provide useful contrast for the diagnosis of high-risk atherosclerotic plaque. Our results demonstrate that these techniques allow for the discrimination of important biochemical features involved in atherosclerotic plaque instability and rupture and show their potential for future intravascular applications.

Fluorescence lifetime in cardiovascular diagnostics.

PubMed

Marcu, Laura

2010-01-01

We review fluorescence lifetime techniques including time-resolved laser-induced fluorescence spectroscopy (TR-LIFS) and fluorescence lifetime imaging microscopy (FLIM) instrumentation and associated methodologies that allow for characterization and diagnosis of atherosclerotic plaques. Emphasis is placed on the translational research potential of TR-LIFS and FLIM and on determining whether intrinsic fluorescence signals can be used to provide useful contrast for the diagnosis of high-risk atherosclerotic plaque. Our results demonstrate that these techniques allow for the discrimination of important biochemical features involved in atherosclerotic plaque instability and rupture and show their potential for future intravascular applications.

Fluorescence lifetime in cardiovascular diagnostics

PubMed Central

Marcu, Laura

2010-01-01

We review fluorescence lifetime techniques including time-resolved laser-induced fluorescence spectroscopy (TR-LIFS) and fluorescence lifetime imaging microscopy (FLIM) instrumentation and associated methodologies that allow for characterization and diagnosis of atherosclerotic plaques. Emphasis is placed on the translational research potential of TR-LIFS and FLIM and on determining whether intrinsic fluorescence signals can be used to provide useful contrast for the diagnosis of high-risk atherosclerotic plaque. Our results demonstrate that these techniques allow for the discrimination of important biochemical features involved in atherosclerotic plaque instability and rupture and show their potential for future intravascular applications. PMID:20210432

Experimental and numerical investigation of a scaled-up passive micromixer using fluorescence technique

NASA Astrophysics Data System (ADS)

Fan, Yanfeng; Hassan, Ibrahim

2010-09-01

The present paper investigates experimentally and numerically a scaled-up micromixer that combines the mixing principles of focusing/diverging and flow split-and-recombine. The micromixer consists of two units called “cross” and “omega”, which are similar to a zigzag structure. The total length is 199.5 mm with a depth of 3 mm. Fluorescence technique is used in the present study for local quantitative measurements of concentration. Two syringe pumps are used to supply the working fluids at two inlets. The testing range of Reynolds number is at 1 ≤ Re ≤ 50. The results of the experiment, obtained by fluorescence technique, are supported by the mixing visualization. The experimental results show that the mixing efficiency decreases at Re ≤ 10 and increases at Re ≥ 10. This is caused by the change in mixing mechanism from mass-diffusion domination to mass-convection domination. After five cells, the mixing efficiency reaches to 70% at Re = 50. The computational fluid dynamics is applied to assist in the understanding of fluid characteristics in channels. The simulation has a good agreement with the experiment. Based on the simulation results, vortices are observed in the channels at high Re, which could stretch and fold the fluids to enhance the effect of mass-convection on mixing. This design has the potential to be developed for micromixers with high flow rates.

Advanced fluorescence microscopy techniques for the life sciences

PubMed Central

Aguib, Yasmine; Yacoub, Magdi H.

The development of super-resolved fluorescence microscopy, for which the Nobel Prize was awarded in 2014, has been a topic of interest to physicists and biologists alike. It is inevitable that numerous questions in biomedical research cannot be answered by means other than direct observation. In this review, advances to fluorescence microscopy are covered in a widely accessible fashion to facilitate its use in decisions related to its acquisition and utilization in biomedical research. PMID:29043264

Invited Review Article: Imaging techniques for harmonic and multiphoton absorption fluorescence microscopy

PubMed Central

Carriles, Ramón; Schafer, Dawn N.; Sheetz, Kraig E.; Field, Jeffrey J.; Cisek, Richard; Barzda, Virginijus; Sylvester, Anne W.; Squier, Jeffrey A.

2009-01-01

We review the current state of multiphoton microscopy. In particular, the requirements and limitations associated with high-speed multiphoton imaging are considered. A description of the different scanning technologies such as line scan, multifoci approaches, multidepth microscopy, and novel detection techniques is given. The main nonlinear optical contrast mechanisms employed in microscopy are reviewed, namely, multiphoton excitation fluorescence, second harmonic generation, and third harmonic generation. Techniques for optimizing these nonlinear mechanisms through a careful measurement of the spatial and temporal characteristics of the focal volume are discussed, and a brief summary of photobleaching effects is provided. Finally, we consider three new applications of multiphoton microscopy: nonlinear imaging in microfluidics as applied to chemical analysis and the use of two-photon absorption and self-phase modulation as contrast mechanisms applied to imaging problems in the medical sciences. PMID:19725639

Two-dimensional fluorescence correlation spectroscopy: resolution of fluorescence of tryptophan residues in horse heart myoglobin.

PubMed

Nakashima, Kenichi; Yuda, Kazuki; Ozaki, Yukihiro; Noda, Isao

2003-11-01

Generalized two-dimensional (2D) fluorescence correlation spectroscopy has been used to resolve fluorescence of two tryptophan (Trp) residues in horse heart myoglobin. Fluorescence quenching is employed as a perturbation mode for causing intensity changes in the fluorescence (quenching perturbation). Two kinds of quenchers, iodide ion and acrylamide, are used for inducing fluorescence intensity change. This technique works because the Trp residue located at the 7th position (W7) is known to be easily accessible to the quencher, whereas that located at the 14th position (W14) is not. By this technique, the fluorescence spectra of the two Trp residues were clearly resolved. From asynchronous maps, it was also shown that the quenching of W7 fluorescence is brought about prior to the quenching of W14 fluorescence. This result is consistent with the structure of horse heart myoglobin that was proposed earlier. Furthermore, it was elucidated that the present 2D analysis is not interfered with by Raman bands of the solvents, which sometimes brings difficulty into conventional fluorescence analysis.

Optimal fluorescence waveband determination for detecting defect cherry tomatoes using fluorescence excitation-emission matrix

USDA-ARS?s Scientific Manuscript database

A multi-spectral fluorescence imaging technique was used to detect defect cherry tomatoes. The fluorescence excitation and emission matrix was used to measure for defects, sound surface, and stem areas to determine the optimal fluorescence excitation and emission wavelengths for discrimination. Two-...

Herbal Extract Incorporated Nanofiber Fabricated by an Electrospinning Technique and its Application to Antimicrobial Air Filtration.

PubMed

Choi, Jeongan; Yang, Byeong Joon; Bae, Gwi-Nam; Jung, Jae Hee

2015-11-18

Recently, with the increased attention to indoor air quality, antimicrobial air filtration techniques have been studied widely to inactivate hazardous airborne microorganisms effectively. In this study, we demonstrate herbal extract incorporated (HEI) nanofibers synthesized by an electrospinning technique and their application to antimicrobial air filtration. As an antimicrobial herbal material, an ethanolic extract of Sophora flavescens, which exhibits great antibacterial activity against pathogens, was mixed with the polymer solution for the electrospinning process. We measured various characteristics of the synthesized HEI nanofibers, such as fiber morphology, fiber size distribution, and thermal stability. For application of the electrospun HEI nanofibers, we made highly effective air filters with 99.99% filtration efficiency and 99.98% antimicrobial activity against Staphylococcus epidermidis. The pressure drop across the HEI nanofiber air filter was 4.75 mmH2O at a face air velocity of 1.79 cm/s. These results will facilitate the implementation of electrospun HEI nanofiber techniques to control air quality and protect against hazardous airborne microorganisms.

Road traffic air and noise pollution exposure assessment - A review of tools and techniques.

PubMed

Khan, Jibran; Ketzel, Matthias; Kakosimos, Konstantinos; Sørensen, Mette; Jensen, Steen Solvang

2018-09-01

Road traffic induces air and noise pollution in urban environments having negative impacts on human health. Thus, estimating exposure to road traffic air and noise pollution (hereafter, air and noise pollution) is important in order to improve the understanding of human health outcomes in epidemiological studies. The aims of this review are (i) to summarize current practices of modelling and exposure assessment techniques for road traffic air and noise pollution (ii) to highlight the potential of existing tools and techniques for their combined exposure assessment for air and noise together with associated challenges, research gaps and priorities. The study reviews literature about air and noise pollution from urban road traffic, including other relevant characteristics such as the employed dispersion models, Geographic Information System (GIS)-based tool, spatial scale of exposure assessment, study location, sample size, type of traffic data and building geometry information. Deterministic modelling is the most frequently used assessment technique for both air and noise pollution of short-term and long-term exposure. We observed a larger variety among air pollution models as compared to the applied noise models. Correlations between air and noise pollution vary significantly (0.05-0.74) and are affected by several parameters such as traffic attributes, building attributes and meteorology etc. Buildings act as screens for the dispersion of pollution, but the reduction effect is much larger for noise than for air pollution. While, meteorology has a greater influence on air pollution levels as compared to noise, although also important for noise pollution. There is a significant potential for developing a standard tool to assess combined exposure of traffic related air and noise pollution to facilitate health related studies. GIS, due to its geographic nature, is well established and has a significant capability to simultaneously address both exposures. Copyright �

Time-resolved resonance fluorescence spectroscopy for study of chemical reactions in laser-induced plasmas.

PubMed

Liu, Lei; Deng, Leimin; Fan, Lisha; Huang, Xi; Lu, Yao; Shen, Xiaokang; Jiang, Lan; Silvain, Jean-François; Lu, Yongfeng

2017-10-30

Identification of chemical intermediates and study of chemical reaction pathways and mechanisms in laser-induced plasmas are important for laser-ablated applications. Laser-induced breakdown spectroscopy (LIBS), as a promising spectroscopic technique, is efficient for elemental analyses but can only provide limited information about chemical products in laser-induced plasmas. In this work, time-resolved resonance fluorescence spectroscopy was studied as a promising tool for the study of chemical reactions in laser-induced plasmas. Resonance fluorescence excitation of diatomic aluminum monoxide (AlO) and triatomic dialuminum monoxide (Al 2 O) was used to identify these chemical intermediates. Time-resolved fluorescence spectra of AlO and Al 2 O were used to observe the temporal evolution in laser-induced Al plasmas and to study their formation in the Al-O 2 chemistry in air.

Ultraviolet-Visible and Fluorescence Spectroscopy Techniques Are Important Diagnostic Tools during the Progression of Atherosclerosis: Diet Zinc Supplementation Retarded or Delayed Atherosclerosis

PubMed Central

Abdelhalim, Mohamed Anwar K.; Moussa, Sherif A. Abdelmottaleb; AL-Mohy, Yanallah Hussain

2013-01-01

Background. In this study, we examined whether UV-visible and fluorescence spectroscopy techniques detect the progression of atherosclerosis in serum of rabbits fed on high-cholesterol diet (HCD) and HCD supplemented with zinc (HCD + Zn) compared with the control. Methods. The control rabbits group was fed on 100 g/day of normal diet. The HCD group was fed on Purina Certified Rabbit Chow supplemented with 1.0% cholesterol plus 1.0% olive oil (100 g/day) for the same period. The HCD + Zn group was fed on normal Purina Certified Rabbit Chow plus 1.0% cholesterol and 1.0% olive oil supplemented with 470 ppm Zn for the same feeding period. UV-visible and fluorescence spectroscopy and biochemistry in Rabbit's blood serum and blood hematology were measured in Rabbit's blood. Results. We found that the fluorescent peak of HCD shifted toward UV-visible wavelength compared with the control using fluorescent excitation of serum at 192 nm. In addition, they showed that supplementation of zinc (350 ppm) restored the fluorescent peak closely to the control. By using UV-visible spectroscopy approach, we found that the peak absorbance of HCD (about 280 nm) was higher than that of control and that zinc supplementation seemed to decrease the absorbance. Conclusions. This study demonstrates that ultraviolet-visible and fluorescence spectroscopy techniques can be applied as noninvasive techniques on a sample blood serum for diagnosing or detecting the progression of atherosclerosis. The Zn supplementation to rabbits fed on HCD delays or retards the progression of atherosclerosis. Inducing anemia in rabbits fed on HCD delays the progression of atherosclerosis. PMID:24350281

GAS CHROMATOGRAPHIC TECHNIQUES FOR THE MEASUREMENT OF ISOPRENE IN AIR

EPA Science Inventory

The chapter discusses gas chromatographic techniques for measuring isoprene in air. Such measurement basically consists of three parts: (1) collection of sufficient sample volume for representative and accurate quantitation, (2) separation (if necessary) of isoprene from interfer...

In Situ Live-Cell Nucleus Fluorescence Labeling with Bioinspired Fluorescent Probes.

PubMed

Ding, Pan; Wang, Houyu; Song, Bin; Ji, Xiaoyuan; Su, Yuanyuan; He, Yao

2017-08-01

Fluorescent imaging techniques for visualization of nuclear structure and function in live cells are fundamentally important for exploring major cellular events. The ideal cellular labeling method is capable of realizing label-free, in situ, real-time, and long-term nucleus labeling in live cells, which can fully obtain the nucleus-relative information and effectively alleviate negative effects of alien probes on cellular metabolism. However, current established fluorescent probes-based strategies (e.g., fluorescent proteins-, organic dyes-, fluorescent organic/inorganic nanoparticles-based imaging techniques) are unable to simultaneously realize label-free, in situ, long-term, and real-time nucleus labeling, resulting in inevitable difficulties in fully visualizing nuclear structure and function in live cells. To this end, we present a type of bioinspired fluorescent probes, which are highly efficacious for in situ and label-free tracking of nucleus in long-term and real-time manners. Typically, the bioinspired polydopamine (PDA) nanoparticles, served as fluorescent probes, can be readily synthesized in situ within live cell nucleus without any further modifications under physiological conditions (37 °C, pH ∼7.4). Compared with other conventional nuclear dyes (e.g., propidium iodide (PI), Hoechst), superior spectroscopic properties (e.g., quantum yield of ∼35.8% and high photostability) and low cytotoxicity of PDA-based probes enable long-term (e.g., 3 h) fluorescence tracking of nucleus. We also demonstrate the generality of this type of bioinspired fluorescent probes in different cell lines and complex biological samples.

Implementation of Satellite Techniques in the Air Transport

NASA Astrophysics Data System (ADS)

Fellner, Andrzej; Jafernik, Henryk

2016-06-01

The article shows process of the implementation satellite systems in Polish aviation which contributed to accomplishment Performance-Based Navigation (PBN) concept. Since 1991 authors have introduced Satellite Navigation Equipment in Polish Air Forces. The studies and researches provide to the Polish Air Force alternative approaches, modernize their navigation and landing systems and achieve compatibility with systems of the North Atlantic Treaty Organization (NATO) and International Civil Aviation Organization (ICAO). Acquired experience, conducted military tests and obtained results enabled to take up work scientifically - research in the environment of the civil aviation. Therefore in 2008 there has been launched cooperation with Polish Air Navigation Services Agency (PANSA). Thanks to cooperation, there have been compiled and fulfilled three fundamental international projects: EGNOS APV MIELEC (EGNOS Introduction in European Eastern Region - APV Mielec), HEDGE (Helicopters Deploy GNSS in Europe), SHERPA (Support ad-Hoc to Eastern Region Pre-operational in GNSS). The successful completion of these projects enabled implementation 21 procedures of the RNAV GNSS final approach at Polish airports, contributing to the implementation of PBN in Poland as well as ICAO resolution A37-11. Results of conducted research which served for the implementation of satellite techniques in the air transport constitute the meaning of this material.

Identification Of Natural Dyes On Archaeological Textile Objects Using Laser Induced Fluorescent Technique

DOE Office of Scientific and Technical Information (OSTI.GOV)

Abdel-Kareem, O.; Eltokhy, A.; Harith, M. A.

2011-09-22

This study aims to evaluate the use of Laser Fluorescent as a non-destructive technique for identification of natural dyes on archaeological textile objects. In this study wool textile samples were dyed with 10 natural dyes such as cochineal, cutch, henna, indigo, Lac, madder, safflower, saffron, sumac and turmeric. These dyes common present on archaeological textile objects to be used as standard dyed textile samples. These selected natural dyes will be used as known references that can be used a guide to identify unknown archaeological dyes. The dyed textile samples were investigated with laser radiation in different wavelengths to detect themore » best wavelengths for identification each dye. This study confirms that Laser Florescent is very useful and a rapid technique can be used as a non-destructive technique for identification of natural dyes on archaeological textile objects. The results obtained with this study can be a guide for all conservators in identification of natural organic dyes on archaeological textile objects.« less

Identification Of Natural Dyes On Archaeological Textile Objects Using Laser Induced Fluorescent Technique

NASA Astrophysics Data System (ADS)

Abdel-Kareem, O.; Eltokhy, A.; Harith, M. A.

2011-09-01

This study aims to evaluate the use of Laser Fluorescent as a non-destructive technique for identification of natural dyes on archaeological textile objects. In this study wool textile samples were dyed with 10 natural dyes such as cochineal, cutch, henna, indigo, Lac, madder, safflower, saffron, sumac and turmeric. These dyes common present on archaeological textile objects to be used as standard dyed textile samples. These selected natural dyes will be used as known references that can be used a guide to identify unknown archaeological dyes. The dyed textile samples were investigated with laser radiation in different wavelengths to detect the best wavelengths for identification each dye. This study confirms that Laser Florescent is very useful and a rapid technique can be used as a non-destructive technique for identification of natural dyes on archaeological textile objects. The results obtained with this study can be a guide for all conservators in identification of natural organic dyes on archaeological textile objects.

Identification of Scleractinian Coral Recruits Using Fluorescent Censusing and DNA Barcoding Techniques

PubMed Central

Hsu, Chia-Min; de Palmas, Stéphane; Kuo, Chao-Yang; Denis, Vianney; Chen, Chaolun Allen

2014-01-01

The identification of coral recruits has been problematic due to a lack of definitive morphological characters being available for higher taxonomic resolution. In this study, we tested whether fluorescent detection of coral recruits used in combinations of different DNA-barcoding markers (cytochrome oxidase I gene [COI], open reading frame [ORF], and nuclear Pax-C intron [PaxC]) could be useful for increasing the resolution of coral spat identification in ecological studies. One hundred and fifty settlement plates were emplaced at nine sites on the fringing reefs of Kenting National Park in southern Taiwan between April 2011 and September 2012. A total of 248 living coral spats and juveniles (with basal areas ranging from 0.21 to 134.57 mm2) were detected on the plates with the aid of fluorescent light and collected for molecular analyses. Using the COI DNA barcoding technique, 90.3% (224/248) of coral spats were successfully identified into six genera, including Acropora, Isopora, Montipora, Pocillopora, Porites, and Pavona. PaxC further separated I. cuneata and I. palifera of Isopora from Acropora, and ORF successfully identified the species of Pocillopora (except P. meandrina and P. eydouxi). Moreover, other cnidarian species such as actinarians, zoanthids, and Millepora species were visually found using fluorescence and identified by COI DNA barcoding. This combination of existing approaches greatly improved the taxonomic resolution of early coral life stages, which to date has been mainly limited to the family level based on skeletal identification. Overall, this study suggests important improvements for the identification of coral recruits in ecological studies. PMID:25211345

Identification of scleractinian coral recruits using fluorescent censusing and DNA barcoding techniques.

PubMed

Hsu, Chia-Min; de Palmas, Stéphane; Kuo, Chao-Yang; Denis, Vianney; Chen, Chaolun Allen

2014-01-01

The identification of coral recruits has been problematic due to a lack of definitive morphological characters being available for higher taxonomic resolution. In this study, we tested whether fluorescent detection of coral recruits used in combinations of different DNA-barcoding markers (cytochrome oxidase I gene [COI], open reading frame [ORF], and nuclear Pax-C intron [PaxC]) could be useful for increasing the resolution of coral spat identification in ecological studies. One hundred and fifty settlement plates were emplaced at nine sites on the fringing reefs of Kenting National Park in southern Taiwan between April 2011 and September 2012. A total of 248 living coral spats and juveniles (with basal areas ranging from 0.21 to 134.57 mm(2)) were detected on the plates with the aid of fluorescent light and collected for molecular analyses. Using the COI DNA barcoding technique, 90.3% (224/248) of coral spats were successfully identified into six genera, including Acropora, Isopora, Montipora, Pocillopora, Porites, and Pavona. PaxC further separated I. cuneata and I. palifera of Isopora from Acropora, and ORF successfully identified the species of Pocillopora (except P. meandrina and P. eydouxi). Moreover, other cnidarian species such as actinarians, zoanthids, and Millepora species were visually found using fluorescence and identified by COI DNA barcoding. This combination of existing approaches greatly improved the taxonomic resolution of early coral life stages, which to date has been mainly limited to the family level based on skeletal identification. Overall, this study suggests important improvements for the identification of coral recruits in ecological studies.

Binding of caffeine with caffeic acid and chlorogenic acid using fluorescence quenching, UV/vis and FTIR spectroscopic techniques.

PubMed

Belay, Abebe; Kim, Hyung Kook; Hwang, Yoon-Hwae

2016-03-01

The interactions of caffeine (CF) with chlorogenic acid (CGA) and caffeic acid (CFA) were investigated by fluorescence quenching, UV/vis and Fourier transform infrared (FTIR) spectroscopic techniques. The results of the study indicated that the fluorescence quenching between caffeine and hydroxycinnamic acids could be rationalized in terms of static quenching or the formation of non-fluorescent CF-CFA and CF-CGA complexes. From fluorescence quenching spectral analysis, the quenching constant (KSV), quenching rate constant (kq), number of binding sites (n), thermodynamic properties and conformational changes of the interaction were determined. The quenching constants (KSV) between CF and CGA, CFA are 1.84 × 10(4) and 1.04 × 10(4) L/mol at 298 K and their binding site n is ~ 1. Thermodynamic parameters determined using the Van't Hoff equation indicated that hydrogen bonds and van der Waal's forces have a major role in the reaction of caffeine with caffeic acid and chlorogenic acid. The 3D fluorescence, UV/vis and FTIR spectra also showed that the binding of CF with CFA and CGA induces conformational changes in CFA and CGA. Copyright © 2015 John Wiley & Sons, Ltd.

First Results of Using a UVTron Flame Sensor to Detect Alpha-Induced Air Fluorescence in the UVC Wavelength Range

PubMed Central

Crompton, Anita J.; Wilson, Andrew P.; Jenkins, Alex; Trivedi, Divyesh

2017-01-01

In this work, a robust stand-off alpha detection method using the secondary effects of alpha radiation has been sought. Alpha particles ionise the surrounding atmosphere as they travel. Fluorescence photons produced as a consequence of this can be used to detect the source of the alpha emissions. This paper details experiments carried out to detect this fluorescence, with the focus on photons in the ultraviolet C (UVC) wavelength range (180–280 nm). A detector, UVTron R9533 (Hamamatsu, 325-6, Sunayama-cho, Naka-ku, Hamamatsu City, Shizuoka Pref., 430-8587, Japan), designed to detect the UVC emissions from flames for fire alarm purposes, was tested in various gas atmospheres with a 210Po alpha source to determine if this could provide an avenue for stand-off alpha detection. The results of the experiments show that this detector is capable of detecting alpha-induced air fluorescence in normal indoor lighting conditions, as the interference from daylight and artificial lighting is less influential on this detection system which operates below the UVA and UVB wavelength ranges (280–315 nm and 315–380 nm respectively). Assuming a standard 1r2 drop off in signal, the limit of detection in this configuration can be calculated to be approximately 240 mm, well beyond the range of alpha-particles in air, which indicates that this approach could have potential for stand-off alpha detection. The gas atmospheres tested produced an increase in the detector count, with xenon having the greatest effect with a measured 52% increase in the detector response in comparison to the detector response in an air atmosphere. This type of alpha detection system could be operated at a distance, where it would potentially provide a more cost effective, safer, and faster solution in comparison with traditional alpha detection methods to detect and characterise alpha contamination in nuclear decommissioning and security applications. PMID:29186051

New developments of X-ray fluorescence imaging techniques in laboratory

NASA Astrophysics Data System (ADS)

Tsuji, Kouichi; Matsuno, Tsuyoshi; Takimoto, Yuki; Yamanashi, Masaki; Kometani, Noritsugu; Sasaki, Yuji C.; Hasegawa, Takeshi; Kato, Shuichi; Yamada, Takashi; Shoji, Takashi; Kawahara, Naoki

2015-11-01

X-ray fluorescence (XRF) analysis is a well-established analytical technique with a long research history. Many applications have been reported in various fields, such as in the environmental, archeological, biological, and forensic sciences as well as in industry. This is because XRF has a unique advantage of being a nondestructive analytical tool with good precision for quantitative analysis. Recent advances in XRF analysis have been realized by the development of new x-ray optics and x-ray detectors. Advanced x-ray focusing optics enables the making of a micro x-ray beam, leading to micro-XRF analysis and XRF imaging. A confocal micro-XRF technique has been applied for the visualization of elemental distributions inside the samples. This technique was applied for liquid samples and for monitoring chemical reactions such as the metal corrosion of steel samples in the NaCl solutions. In addition, a principal component analysis was applied for reducing the background intensity in XRF spectra obtained during XRF mapping, leading to improved spatial resolution of confocal micro-XRF images. In parallel, the authors have proposed a wavelength dispersive XRF (WD-XRF) imaging spectrometer for a fast elemental imaging. A new two dimensional x-ray detector, the Pilatus detector was applied for WD-XRF imaging. Fast XRF imaging in 1 s or even less was demonstrated for Euro coins and industrial samples. In this review paper, these recent advances in laboratory-based XRF imaging, especially in a laboratory setting, will be introduced.

Correlative Electron and Fluorescence Microscopy of Magnetotactic Bacteria in Liquid: Toward In Vivo Imaging

PubMed Central

Woehl, Taylor J.; Kashyap, Sanjay; Firlar, Emre; Perez-Gonzalez, Teresa; Faivre, Damien; Trubitsyn, Denis; Bazylinski, Dennis A.; Prozorov, Tanya

2014-01-01

Magnetotactic bacteria biomineralize ordered chains of uniform, membrane-bound magnetite or greigite nanocrystals that exhibit nearly perfect crystal structures and species-specific morphologies. Transmission electron microscopy (TEM) is a critical technique for providing information regarding the organization of cellular and magnetite structures in these microorganisms. However, conventional TEM can only be used to image air-dried or vitrified bacteria removed from their natural environment. Here we present a correlative scanning TEM (STEM) and fluorescence microscopy technique for imaging viable cells of Magnetospirillum magneticum strain AMB-1 in liquid using an in situ fluid cell TEM holder. Fluorescently labeled cells were immobilized on microchip window surfaces and visualized in a fluid cell with STEM, followed by correlative fluorescence imaging to verify their membrane integrity. Notably, the post-STEM fluorescence imaging indicated that the bacterial cell wall membrane did not sustain radiation damage during STEM imaging at low electron dose conditions. We investigated the effects of radiation damage and sample preparation on the bacteria viability and found that approximately 50% of the bacterial membranes remained intact after an hour in the fluid cell, decreasing to ~30% after two hours. These results represent a first step toward in vivo studies of magnetite biomineralization in magnetotactic bacteria. PMID:25358460

Correlative Electron and Fluorescence Microscopy of Magnetotactic Bacteria in Liquid: Toward In Vivo Imaging

DOE PAGES

Woehl, Taylor J.; Kashyap, Sanjay; Firlar, Emre; ...

2014-10-31

Magnetotactic bacteria biomineralize ordered chains of uniform, membrane-bound magnetite or greigite nanocrystals that exhibit nearly perfect crystal structures and species-specific morphologies. Transmission electron microscopy (TEM) is a critical technique for providing information regarding the organization of cellular and magnetite structures in these microorganisms. However, conventional TEM can only be used to image air-dried or vitrified bacteria removed from their natural environment. Here we present a correlative scanning TEM (STEM) and fluorescence microscopy technique for imaging viable cells of Magnetospirillum magneticum strain AMB-1 in liquid using an in situ fluid cell TEM holder. Fluorescently labeled cells were immobilized on microchip windowmore » surfaces and visualized in a fluid cell with STEM, followed by correlative fluorescence imaging to verify their membrane integrity. Notably, the post-STEM fluorescence imaging indicated that the bacterial cell wall membrane did not sustain radiation damage during STEM imaging at low electron dose conditions. We investigated the effects of radiation damage and sample preparation on the bacteria viability and found that approximately 50% of the bacterial membranes remained intact after an hour in the fluid cell, decreasing to ~30% after two hours. These results represent a first step toward in vivo studies of magnetite biomineralization in magnetotactic bacteria.« less

Fluorescence detection of small gastrointestinal tumours: principles, technique, first clinical experience.

PubMed

Orth, K; Russ, D; Steiner, R; Beger, H G

2000-11-01

Photodynamic therapy (PDT) is a form of cancer treatment based on the selective accumulation of a photosensitizer in neoplastic tissue. The fluorescent properties of a photosensitizer permit diagnostic localization of primary tumours and/or metastasis. Occult lesions are hard to detect and can easily be missed during routine laparoscopy. Fluorescence observation offers additional optical information and the ability to detect these occult tumours. Clinically, we used 5-aminolevulinic acid for peritoneal staining and tumour demarcation via tumour-specific fluorescence induced by protoporphyrin IX. For laparoscopic observations, a "D-Light" system was used; the conventional white light source was equipped with an optical blocking filter that transmits at the excitation wavelength (380-450 nm) and blocks all other parts of the spectrum. With the aid of a suitable observation filter, the relevant fluorescence was detectable. With the help of this fluorescence we increased the capacity to detect occult tumours, that were missed with white-light observation (9/26). In the gastrointestinal tract, we used a krypton laser at 405 nm for PP IX fluorescence induction. Although there were high sensitivity rates for neoplasms (81% peritoneal carcinomas, 60% gastric cancer), no exact histopathological statement could be achieved at because of false-positive fluorescence, mainly caused by inflammation (6/32). Current clinical goals and the future perspectives of photodynamic diagnostic are discussed.

Measurement of the flux of ultra high energy cosmic rays by the stereo technique

NASA Astrophysics Data System (ADS)

High Resolution Fly'S Eye Collaboration; Abbasi, R. U.; Abu-Zayyad, T.; Al-Seady, M.; Allen, M.; Amann, J. F.; Archbold, G.; Belov, K.; Belz, J. W.; Bergman, D. R.; Blake, S. A.; Brusova, O. A.; Burt, G. W.; Cannon, C.; Cao, Z.; Deng, W.; Fedorova, Y.; Findlay, J.; Finley, C. B.; Gray, R. C.; Hanlon, W. F.; Hoffman, C. M.; Holzscheiter, M. H.; Hughes, G.; Hüntemeyer, P.; Ivanov, D.; Jones, B. F.; Jui, C. C. H.; Kim, K.; Kirn, M. A.; Loh, E. C.; Maestas, M. M.; Manago, N.; Marek, L. J.; Martens, K.; Matthews, J. A. J.; Matthews, J. N.; Moore, S. A.; O'Neill, A.; Painter, C. A.; Perera, L.; Reil, K.; Riehle, R.; Roberts, M. D.; Rodriguez, D.; Sasaki, M.; Schnetzer, S. R.; Scott, L. M.; Sinnis, G.; Smith, J. D.; Snow, R.; Sokolsky, P.; Springer, R. W.; Stokes, B. T.; Stratton, S. R.; Thomas, J. R.; Thomas, S. B.; Thomson, G. B.; Tupa, D.; Wiencke, L. R.; Zech, A.; Zhang, B. K.; Zhang, X.; Zhang, Y.; High Resolution Fly's Eye Collaboration

2009-08-01

The High Resolution Fly’s Eye (HiRes) experiment has measured the flux of ultrahigh energy cosmic rays using the stereoscopic air fluorescence technique. The HiRes experiment consists of two detectors that observe cosmic ray showers via the fluorescence light they emit. HiRes data can be analyzed in monocular mode, where each detector is treated separately, or in stereoscopic mode where they are considered together. Using the monocular mode the HiRes collaboration measured the cosmic ray spectrum and made the first observation of the Greisen-Zatsepin-Kuzmin cutoff. In this paper we present the cosmic ray spectrum measured by the stereoscopic technique. Good agreement is found with the monocular spectrum in all details.

Fluorescence-Assisted Gamma Spectrometry for Surface Contamination Analysis

NASA Astrophysics Data System (ADS)

Ihantola, Sakari; Sand, Johan; Perajarvi, Kari; Toivonen, Juha; Toivonen, Harri

2013-02-01

A fluorescence-based alpha-gamma coincidence spectrometry approach has been developed for the analysis of alpha-emitting radionuclides. The thermalization of alpha particles in air produces UV light, which in turn can be detected over long distances. The simultaneous detection of UV and gamma photons allows detailed gamma analyses of a single spot of interest even in highly active surroundings. Alpha particles can also be detected indirectly from samples inside sealed plastic bags, which minimizes the risk of cross-contamination. The position-sensitive alpha-UV-gamma coincidence technique reveals the presence of alpha emitters and identifies the nuclides ten times faster than conventional gamma spectrometry.

Quantification of OH and HO2 radicals during the low-temperature oxidation of hydrocarbons by Fluorescence Assay by Gas Expansion technique

PubMed Central

Blocquet, Marion; Schoemaecker, Coralie; Amedro, Damien; Herbinet, Olivier; Battin-Leclerc, Frédérique; Fittschen, Christa

2013-01-01

•OH and •HO2 radicals are known to be the key species in the development of ignition. A direct measurement of these radicals under low-temperature oxidation conditions (T = 550–1,000 K) has been achieved by coupling a technique named fluorescence assay by gas expansion, an experimental technique designed for the quantification of these radicals in the free atmosphere, to a jet-stirred reactor, an experimental device designed for the study of low-temperature combustion chemistry. Calibration allows conversion of relative fluorescence signals to absolute mole fractions. Such radical mole fraction profiles will serve as a benchmark for testing chemical models developed to improve the understanding of combustion processes. PMID:24277836

Determination of phosphorus in steel by the combined technique of laser induced breakdown spectrometry with laser induced fluorescence spectrometry

NASA Astrophysics Data System (ADS)

Kondo, Hiroyuki; Hamada, Naoya; Wagatsuma, Kazuaki

2009-09-01

Laser induced breakdown spectrometry (LIBS) combined with laser induced fluorescence spectrometry (LIFS) has been applied for detection of trace-level phosphorus in steel. The plasma induced by irradiation of Nd:YAG laser pulse for ablation was illuminated by the 3rd harmonic of Ti:Sapphire laser tuned to one of the resonant lines for phosphorus in the wavelength region of 253-256 nm. An excitation line for phosphorus was selected to give the highest signal-to-noise ratio. Fluorescence signals, P213.62 and P214.91 nm, were observed with high selectivity at the contents as low as several tens µg g - 1 . Fluorescence intensities were in a good linear correlation with the contents. Fluorescence intensity ratio of a collisionally assisted line (213.62 nm) to a direct transition line (214.91 nm) was discussed in terms of the analytical conditions and experimental results were compared with a calculation based on rate equations. Since the fluorescence signal light in the wavelength range longer than 200 nm can be transmitted relatively easily, even through fiber optics of moderate length, LIBS/LIFS would be a versatile technique in on-site applications for the monitoring of phosphorus contents in steel.

Biological applications of confocal fluorescence polarization microscopy

NASA Astrophysics Data System (ADS)

Bigelow, Chad E.

Fluorescence polarization microscopy is a powerful modality capable of sensing changes in the physical properties and local environment of fluorophores. In this thesis we present new applications for the technique in cancer diagnosis and treatment and explore the limits of the modality in scattering media. We describe modifications to our custom-built confocal fluorescence microscope that enable dual-color imaging, optical fiber-based confocal spectroscopy and fluorescence polarization imaging. Experiments are presented that indicate the performance of the instrument for all three modalities. The limits of confocal fluorescence polarization imaging in scattering media are explored and the microscope parameters necessary for accurate polarization images in this regime are determined. A Monte Carlo routine is developed to model the effect of scattering on images. Included in it are routines to track the polarization state of light using the Mueller-Stokes formalism and a model for fluorescence generation that includes sampling the excitation light polarization ellipse, Brownian motion of excited-state fluorophores in solution, and dipole fluorophore emission. Results from this model are compared to experiments performed on a fluorophore-embedded polymer rod in a turbid medium consisting of polystyrene microspheres in aqueous suspension. We demonstrate the utility of the fluorescence polarization imaging technique for removal of contaminating autofluorescence and for imaging photodynamic therapy drugs in cell monolayers. Images of cells expressing green fluorescent protein are extracted from contaminating fluorescein emission. The distribution of meta-tetrahydroxypheny1chlorin in an EMT6 cell monolayer is also presented. A new technique for imaging enzyme activity is presented that is based on observing changes in the anisotropy of fluorescently-labeled substrates. Proof-of-principle studies are performed in a model system consisting of fluorescently labeled bovine

An objective isobaric/isentropic technique for upper air analysis

NASA Technical Reports Server (NTRS)

Mancuso, R. L.; Endlich, R. M.; Ehernberger, L. J.

1981-01-01

An objective meteorological analysis technique is presented whereby both horizontal and vertical upper air analyses are performed. The process used to interpolate grid-point values from the upper-air station data is the same as for grid points on both an isobaric surface and a vertical cross-sectional plane. The nearby data surrounding each grid point are used in the interpolation by means of an anisotropic weighting scheme, which is described. The interpolation for a grid-point potential temperature is performed isobarically; whereas wind, mixing-ratio, and pressure height values are interpolated from data that lie on the isentropic surface that passes through the grid point. Two versions (A and B) of the technique are evaluated by qualitatively comparing computer analyses with subjective handdrawn analyses. The objective products of version A generally have fair correspondence with the subjective analyses and with the station data, and depicted the structure of the upper fronts, tropopauses, and jet streams fairly well. The version B objective products correspond more closely to the subjective analyses, and show the same strong gradients across the upper front with only minor smoothing.

Compressed air injection technique to standardize block injection pressures : [La technique d'injection d'air comprimé pour normaliser les pressions d'injection d'un blocage nerveux].

PubMed

Tsui, Ban C H; Li, Lisa X Y; Pillay, Jennifer J

2006-11-01

Presently, no standardized technique exists to monitor injection pressures during peripheral nerve blocks. Our objective was to determine if a compressed air injection technique, using an in vitro model based on Boyle's law and typical regional anesthesia equipment, could consistently maintain injection pressures below a 1293 mmHg level associated with clinically significant nerve injury. Injection pressures for 20 and 30 mL syringes with various needle sizes ( 18G, 20G, 21 G, 22G, and 24G) were measured in a closed system. A set volume of air was aspirated into a saline-filled syringe and then compressed and maintained at various percentages while pressure was measured. The needle was inserted into the injection port of a pressure sensor, which had attached extension tubing with an injection plug clamped "off". Using linear regression with all data points, the pressure value and 99% confidence interval (CI) at 50% air compression was estimated. The linearity of Boyle's law was demonstrated with a high correlation, r = 0.99, and a slope of 0.984 (99% CI: 0.967-1.001). The net pressure generated at 50% compression was estimated as 744.8 mmHg, with the 99% CI between 729.6 and 760.0 mmHg. The various syringe/needle combinations had similar results. By creating and maintaining syringe air compression at 50% or less, injection pressures will be substantially below the 1293 mmHg threshold considered to be an associated risk factor for clinically significant nerve injury. This technique may allow simple, real-time and objective monitoring during local anesthetic injections while inherently reducing injection speed. Présentement, aucune technique normalisée ne permet de vérifier les pressions d'injection pendant les blocages nerveux périphériques. Nous voulions vérifier si une technique d'injection d'air comprimé, utilisant un modèle in vitro fondé sur la loi de Boyle et du matériel propre à l'anesthésie régionale, pouvait maintenir avec régularité les

Demonstration of plant fluorescence by imaging technique and Intelligent FluoroSensor

NASA Astrophysics Data System (ADS)

Lenk, Sándor; Gádoros, Patrik; Kocsányi, László; Barócsi, Attila

2015-10-01

Photosynthesis is a process that converts carbon-dioxide into organic compounds, especially into sugars, using the energy of sunlight. The absorbed light energy is used mainly for photosynthesis initiated at the reaction centers of chlorophyll-protein complexes, but part of it is lost as heat and chlorophyll fluorescence. Therefore, the measurement of the latter can be used to estimate the photosynthetic activity. The basic method, when illuminating intact leaves with strong light after a dark adaptation of at least 20 minutes resulting in a transient change of fluorescence emission of the fluorophore chlorophyll-a called `Kautsky effect', is demonstrated by an imaging setup. The experimental kit includes a high radiant blue LED and a CCD camera (or a human eye) equipped with a red transmittance filter to detect the changing fluorescence radiation. However, for the measurement of several fluorescence parameters, describing the plant physiological processes in detail, the variation of several excitation light sources and an adequate detection method are needed. Several fluorescence induction protocols (e.g. traditional Kautsky, pulse amplitude modulated and excitation kinetic), are realized in the Intelligent FluoroSensor instrument. Using it, students are able to measure different plant fluorescence induction curves, quantitatively determine characteristic parameters and qualitatively interpret the measured signals.

Photobleaching correction in fluorescence microscopy images

NASA Astrophysics Data System (ADS)

Vicente, Nathalie B.; Diaz Zamboni, Javier E.; Adur, Javier F.; Paravani, Enrique V.; Casco, Víctor H.

2007-11-01

Fluorophores are used to detect molecular expression by highly specific antigen-antibody reactions in fluorescence microscopy techniques. A portion of the fluorophore emits fluorescence when irradiated with electromagnetic waves of particular wavelengths, enabling its detection. Photobleaching irreversibly destroys fluorophores stimulated by radiation within the excitation spectrum, thus eliminating potentially useful information. Since this process may not be completely prevented, techniques have been developed to slow it down or to correct resulting alterations (mainly, the decrease in fluorescent signal). In the present work, the correction by photobleaching curve was studied using E-cadherin (a cell-cell adhesion molecule) expression in Bufo arenarum embryos. Significant improvements were observed when applying this simple, inexpensive and fast technique.

Survey of statistical techniques used in validation studies of air pollution prediction models

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bornstein, R D; Anderson, S F

1979-03-01

Statistical techniques used by meteorologists to validate predictions made by air pollution models are surveyed. Techniques are divided into the following three groups: graphical, tabular, and summary statistics. Some of the practical problems associated with verification are also discussed. Characteristics desired in any validation program are listed and a suggested combination of techniques that possesses many of these characteristics is presented.

Laser-Stimulated Fluorescence in Paleontology

PubMed Central

Kaye, Thomas G.; Falk, Amanda R.; Pittman, Michael; Sereno, Paul C.; Burnham, David A.; Gong, Enpu; Xu, Xing; Wang, Yinan

2015-01-01

Fluorescence using ultraviolet (UV) light has seen increased use as a tool in paleontology over the last decade. Laser-stimulated fluorescence (LSF) is a next generation technique that is emerging as a way to fluoresce paleontological specimens that remain dark under typical UV. A laser’s ability to concentrate very high flux rates both at the macroscopic and microscopic levels results in specimens fluorescing in ways a standard UV bulb cannot induce. Presented here are five paleontological case histories that illustrate the technique across a broad range of specimens and scales. Novel uses such as back-lighting opaque specimens to reveal detail and detection of specimens completely obscured by matrix are highlighted in these examples. The recent cost reductions in medium-power short wavelength lasers and use of standard photographic filters has now made this technique widely accessible to researchers. This technology has the potential to automate multiple aspects of paleontology, including preparation and sorting of microfossils. This represents a highly cost-effective way to address paleontology's preparatory bottleneck. PMID:26016843

Fluorescent techniques for discovery and characterization of phosphopantetheinyl transferase inhibitors

PubMed Central

Kosa, Nicolas M.; Foley, Timothy L.; Burkart, Michael D.

2016-01-01

Phosphopantetheinyl transferase (E.C. 2.7.8.-) activates biosynthetic pathways that synthesize both primary and secondary metabolites in bacteria. Inhibitors of these enzymes have the potential to serve as antibiotic compounds that function through a unique mode of action and possess clinical utility. Here we report a direct and continuous assay for this enzyme class based upon monitoring polarization of a fluorescent phosphopantetheine analog as it is transferred from a low molecular weight coenzyme A substrate to higher molecular weight protein acceptor. We demonstrate the utility of this method for the biochemical characterization of phosphopantetheinyl transferase Sfp, a canonical representative from this class. We also establish the portability of this technique to other homologs by adapting the assay to function with the human phosphopantetheinyl transferase, a target for which a microplate detection method does not currently exist. Comparison of these targets provides a basis to predict therapeutic index of inhibitor candidates and offers a valuable characterization of enzyme activity. PMID:24192555

Fluorescence from polystyrene - Photochemical processes in polymeric systems, 7

NASA Technical Reports Server (NTRS)

Gupta, M. C.; Gupta, A.

1983-01-01

Results are presented for measurements of the fluorescence spectra of polystyrene in dilute solution and in pure solid films. It is determined that a major potential source of experimental error is the concurrent photooxidative degradation in air which may obscure fluorescence emission from monomeric sites in solid films at 25 C. The fluorescence spectra of oriented films are evaluated in terms of the monomer to excimer fluorescence intensity ratio and the excimer 'red shift'. The monomer to excimer fluorescence intensity ratio is determined to be significantly higher in fluid solution than in solid film.

Multispectral fluorescence imaging technique for discrimination of cucumber (Cucumis Sativus) seed viability

USDA-ARS?s Scientific Manuscript database

In this study, we developed a nondestructive method for discriminating viable cucumber (Cucumis sativus) seeds based on hyperspectral fluorescence imaging. The fluorescence spectra of cucumber seeds in the 420–700 nm range were extracted from hyperspectral fluorescence images obtained using 365 nm u...

Widefield fluorescence sectioning with HiLo microscopy.

PubMed

Mertz, Jerome; Lim, Daryl; Chu, Kengyeh K; Bozinovic, Nenad; Ford, Timothy

2009-01-01

HiLo microscopy is a widefield fluorescence imaging technique that provides depth discrimination by combining two images, one with non-uniform illumination and one with uniform illumination. We discuss the theory of this technique and a variety of practical implementations in brain-tissue imaging and fluorescence endomicroscopy.

Shot noise limited detection of OH using the technique of laser-induced fluorescence

NASA Technical Reports Server (NTRS)

Bakalyar, D. M.; Davis, L. I., Jr.; Guo, C.; James, J. V.; Wang, C. C.; Kakos, S.; Morris, P. T.

1984-01-01

Nearly shot-noise limited detection of OH using the technique of laser-induced fluorescence is reported. A LIDAR configuration is used to excite fluoresence in a large volume and a narrow-bandwidth interference filter provides spectral discrimination. This arrangement alleviates the effect of ozone interference and facilitates image processing at relatively close distances. The detection limit is determined mainly by the short-noise of the solar background. Ground-based measurements in Dearborn indicate a detection limit of better than 1 x 10 to the 6th power OH/cubic cm over a forty-minute acquisition period. Under favorable conditions, a comparable detection limit was also observed for airborne measurements.

Visualization of oxygen transfer across the air-water interface using a fluorescence oxygen visualization method.

PubMed

Lee, Minhee

2002-04-01

Oxygen concentration fields in a water body were visualized by the fluorescence oxygen visualization (FOV) method. Pyrenebutyric acid (PBA) was used as a fluorescent indicator of oxygen, and an intensive charge coupled-device (ICCD) camera as an image detector. Sequential images (over 2000 images) of the oxygen concentration field around the surface water of the tank (1 x 1 x 0.75 m3) were produced during the 3 h experiment. From image processing, the accurate pathway of oxygen-rich, cold water at the water surface was also visualized. The amount of oxygen transferred through the air-water interface during the experiment was measured and the oxygen transfer coefficient (K(L)) was determined as 0.22 m/d, which was much higher than that is expected in molecular diffusion. Results suggest that vertical penetration of cold water was the main pathway of oxygen in the water body in the tank. The average velocity of cold water penetrating downward in water body was also measured from consecutive images and the value was 0.3-0.6 mm/s. The FOV method used in this research should have wide application in experimental fluid mechanics and can also provide a phenomenological description of oxygen transfer under physically realizable natural conditions in lakes and reservoirs.

Laser-induced fluorescence imaging of bacteria

NASA Astrophysics Data System (ADS)

Hilton, Peter J.

1998-12-01

This paper outlines a method for optically detecting bacteria on various backgrounds, such as meat, by imaging their laser induced auto-fluorescence response. This method can potentially operate in real-time, which is many times faster than current bacterial detection methods, which require culturing of bacterial samples. This paper describes the imaging technique employed whereby a laser spot is scanned across an object while capturing, filtering, and digitizing the returned light. Preliminary results of the bacterial auto-fluorescence are reported and plans for future research are discussed. The results to date are encouraging with six of the eight bacterial strains investigated exhibiting auto-fluorescence when excited at 488 nm. Discrimination of these bacterial strains against red meat is shown and techniques for reducing background fluorescence discussed.

Air data position-error calibration using state reconstruction techniques

NASA Technical Reports Server (NTRS)

Whitmore, S. A.; Larson, T. J.; Ehernberger, L. J.

1984-01-01

During the highly maneuverable aircraft technology (HiMAT) flight test program recently completed at NASA Ames Research Center's Dryden Flight Research Facility, numerous problems were experienced in airspeed calibration. This necessitated the use of state reconstruction techniques to arrive at a position-error calibration. For the HiMAT aircraft, most of the calibration effort was expended on flights in which the air data pressure transducers were not performing accurately. Following discovery of this problem, the air data transducers of both aircraft were wrapped in heater blankets to correct the problem. Additional calibration flights were performed, and from the resulting data a satisfactory position-error calibration was obtained. This calibration and data obtained before installation of the heater blankets were used to develop an alternate calibration method. The alternate approach took advantage of high-quality inertial data that was readily available. A linearized Kalman filter (LKF) was used to reconstruct the aircraft's wind-relative trajectory; the trajectory was then used to separate transducer measurement errors from the aircraft position error. This calibration method is accurate and inexpensive. The LKF technique has an inherent advantage of requiring that no flight maneuvers be specially designed for airspeed calibrations. It is of particular use when the measurements of the wind-relative quantities are suspected to have transducer-related errors.

Modern developments in electron-beam fluorescence

NASA Technical Reports Server (NTRS)

Cattolica, Robert J.

1991-01-01

Recent developments in the area of electron-beam fluorescence are discussed with special attention given to the experience in the use of the electron-beam fluorescence in flight research. A new measurement approach, called electron-photon fluorescence (EPF), is described, and it is shown that EPF offers the potential of overcoming some of the disadvantages of electron-beam fluorescence in high-density flows. Examples of using the EPF technique are presented.

Photocontrollable Fluorescent Proteins for Superresolution Imaging

PubMed Central

Shcherbakova, Daria M.; Sengupta, Prabuddha; Lippincott-Schwartz, Jennifer; Verkhusha, Vladislav V.

2014-01-01

Superresolution fluorescence microscopy permits the study of biological processes at scales small enough to visualize fine subcellular structures that are unresolvable by traditional diffraction-limited light microscopy. Many superresolution techniques, including those applicable to live cell imaging, utilize genetically encoded photocontrollable fluorescent proteins. The fluorescence of these proteins can be controlled by light of specific wavelengths. In this review, we discuss the biochemical and photophysical properties of photocontrollable fluorescent proteins that are relevant to their use in superresolution microscopy. We then describe the recently developed photoactivatable, photoswitchable, and reversibly photoswitchable fluorescent proteins, and we detail their particular usefulness in single-molecule localization–based and nonlinear ensemble–based superresolution techniques. Finally, we discuss recent applications of photocontrollable proteins in superresolution imaging, as well as how these applications help to clarify properties of intracellular structures and processes that are relevant to cell and developmental biology, neuroscience, cancer biology and biomedicine. PMID:24895855

X-ray fluorescence measurements of dissolved gas and cavitation

DOE PAGES

Duke, Daniel J.; Kastengren, Alan L.; Swantek, Andrew B.; ...

2016-09-28

The dynamics of dissolved gas and cavitation are strongly coupled, yet these phenomena are difficult to measure in-situ. Both create voids in the fluid that can be difficult to distinguish. In this paper, we present an application of X-ray fluorescence in which liquid density and total noncondensible gas concentration (both dissolved and nucleated) are simultaneously measured. The liquid phase is doped with 400 ppm of a bromine tracer, and dissolved air is removed and substituted with krypton. Fluorescent emission at X-ray wavelengths is simultaneously excited from the Br and Kr with a focused monochromatic X-ray beam from a synchrotron source.more » We measure the flow in a cavitating nozzle 0.5 mm in diameter. From Br fluorescence, total displacement of the liquid is measured. From Kr fluorescence, the mass fraction of both dissolved and nucleated gas is measured. Volumetric displacement of liquid due to both cavitation and gas precipitation can be separated through estimation of the local equilibrium dissolved mass fraction. The uncertainty in the line of sight projected densities of the liquid and gas phases is 4–6 %. The high fluorescence yields and energies of Br and Kr allow small mass fractions of gas to be measured, down to 10 -5, with an uncertainty of 8 %. Finally, these quantitative measurements complement existing optical diagnostic techniques and provide new insight into the diffusion of gas into cavitation bubbles, which can increase their internal density, pressure and lifetimes by orders of magnitude.« less

Fluorescent optical liquid level sensor

DOEpatents

Weiss, Jonathan D.

2001-01-01

A liquid level sensor comprising a transparent waveguide containing fluorescent material that is excited by light of a first wavelength and emits at a second, longer wavelength. The upper end of the waveguide is connected to a light source at the first wavelength through a beveled portion of the waveguide such that the input light is totally internally reflected within the waveguide above an air/liquid interface in a tank but is transmitted into the liquid below this interface. Light is emitted from the fluorescent material only in those portions of the waveguide that are above the air/liquid interface, to be collected at the upper end of the waveguide by a detector that is sensitive only to the second wavelength. As the interface moves down in the tank, the signal strength from the detector will increase.

Measurement of resistance to solute transport across surfactant-laden interfaces using a Fluorescence Recovery After Photobleaching (FRAP) technique

NASA Technical Reports Server (NTRS)

Browne, Edward P.; Nivaggioli, Thierry; Hatton, T. Alan

1994-01-01

A noninvasive fluorescence recovery after photobleaching (FRAP) technique is under development to measure interfacial transport in two phase systems without disturbing the interface. The concentration profiles of a probe solute are measured in both sides of the interface by argon-ion laser, and the system relaxation is then monitored by a microscope-mounted CCD camera.

Air/fuel ratio visualization in a diesel spray

NASA Astrophysics Data System (ADS)

Carabell, Kevin David

1993-01-01

To investigate some features of high pressure diesel spray ignition, we have applied a newly developed planar imaging system to a spray in an engine-fed combustion bomb. The bomb is designed to give flow characteristics similar to those in a direct injection diesel engine yet provide nearly unlimited optical access. A high pressure electronic unit injector system with on-line manually adjustable main and pilot injection features was used. The primary scalar of interest was the local air/fuel ratio, particularly near the spray plumes. To make this measurement quantitative, we have developed a calibration LIF technique. The development of this technique is the key contribution of this dissertation. The air/fuel ratio measurement was made using biacetyl as a seed in the air inlet to the engine. When probed by a tripled Nd:YAG laser the biacetyl fluoresces, with a signal proportional to the local biacetyl concentration. This feature of biacetyl enables the fluorescent signal to be used as as indicator of local fuel vapor concentration. The biacetyl partial pressure was carefully controlled, enabling estimates of the local concentration of air and the approximate local stoichiometry in the fuel spray. The results indicate that the image quality generated with this method is sufficient for generating air/fuel ratio contours. The processes during the ignition delay have a marked effect on ignition and the subsequent burn. These processes, vaporization and pre-flame kinetics, very much depend on the mixing of the air and fuel. This study has shown that poor mixing and over-mixing of the air and fuel will directly affect the type of ignition. An optimal mixing arrangement exists and depends on the swirl ratio in the engine, the number of holes in the fuel injector and the distribution of fuel into a pilot and main injection. If a short delay and a diffusion burn is desired, the best mixing parameters among those surveyed would be a high swirl ratio, a 4-hole nozzle and a

DIRECT AND INDIRECT FLUORESCENT-ANTIBODY TECHNIQUES FOR THE PSITTACOSIS-LYMPHOGRANULOMA VENEREUM-TRACHOMA GROUP OF AGENTS1

PubMed Central

Ross, Martin R.; Borman, Earle K.

1963-01-01

Ross, Martin R. (Connecticut State Department of Health, Hartford) and Earle K. Borman. Direct and indirect fluorescent-antibody techniques for the psittacosis-lymphogranuloma venereum-trachoma group of agents. J. Bacteriol. 85:851–858. 1963.—Direct and indirect fluorescent-antibody (FA) techniques were developed for the detection of group antigen in infected tissue cultures and the titration of group antibody in human antiserum. The growth of the agent of meningopneumonitis (MP) in mouse embryo lung cell monolayers was followed by infectivity and complement-fixing (CF) antigen titrations, and cytological examination of FA stained cultures. Although infectivity and CF antigen reached a peak at 2 days and remained constant for an additional 3 days, only cells tested 2 to 3 days after infection were suitable for FA staining with labeled anti-MP serum because of excessive artifacts in the older cultures. Fluorescein isothiocyanate-labeled rooster and guinea pig anti-MP serums and human antipsittacosis serums were titrated in direct FA and hemagglutination-inhibition (HI) tests. The rooster conjugate showed brighter staining and higher antibody titers than the guinea pig or human conjugates and was more effective in detecting minimal amounts of virus antigen. FA staining reactions with 1 and 2 units of labeled rooster serum were inhibited by unlabeled rooster serum but clear-cut inhibition with human antipsittacosis serum could not be demonstrated. The indirect FA technique was successfully used for the titration of group antibody in human serum. A comparison of the indirect FA, HI, and CF tests showed the indirect FA technique to be intermediate in sensitivity between the HI and CF tests. None of the three tests showed significant cross reactions with human serums reactive for influenza A and B; parainfluenza 1, 2, and 3; respiratory syncytial virus; Q fever; or the primary atypical pneumonia agent. PMID:14044954

Fluorescence calibration method for single-particle aerosol fluorescence instruments

NASA Astrophysics Data System (ADS)

Shipley Robinson, Ellis; Gao, Ru-Shan; Schwarz, Joshua P.; Fahey, David W.; Perring, Anne E.

2017-05-01

Real-time, single-particle fluorescence instruments used to detect atmospheric bioaerosol particles are increasingly common, yet no standard fluorescence calibration method exists for this technique. This gap limits the utility of these instruments as quantitative tools and complicates comparisons between different measurement campaigns. To address this need, we have developed a method to produce size-selected particles with a known mass of fluorophore, which we use to calibrate the fluorescence detection of a Wideband Integrated Bioaerosol Sensor (WIBS-4A). We use mixed tryptophan-ammonium sulfate particles to calibrate one detector (FL1; excitation = 280 nm, emission = 310-400 nm) and pure quinine particles to calibrate the other (FL2; excitation = 280 nm, emission = 420-650 nm). The relationship between fluorescence and mass for the mixed tryptophan-ammonium sulfate particles is linear, while that for the pure quinine particles is nonlinear, likely indicating that not all of the quinine mass contributes to the observed fluorescence. Nonetheless, both materials produce a repeatable response between observed fluorescence and particle mass. This procedure allows users to set the detector gains to achieve a known absolute response, calculate the limits of detection for a given instrument, improve the repeatability of the instrumental setup, and facilitate intercomparisons between different instruments. We recommend calibration of single-particle fluorescence instruments using these methods.

Proximal sensing of plant-pathogen interactions in spring barley with three fluorescence techniques.

PubMed

Leufen, Georg; Noga, Georg; Hunsche, Mauricio

2014-06-24

In the last years fluorescence spectroscopy has come to be viewed as an essential approach in key research fields of applied plant sciences. However, the quantity and particularly the quality of information produced by different equipment might vary considerably. In this study we investigate the potential of three optical devices for the proximal sensing of plant-pathogen interactions in four genotypes of spring barley. For this purpose, the fluorescence lifetime, the image-resolved multispectral fluorescence and selected indices of a portable multiparametric fluorescence device were recorded at 3, 6, and 9 days after inoculation (dai) from healthy leaves as well as from leaves inoculated with powdery mildew (Blumeria graminis) or leaf rust (Puccinia hordei). Genotype-specific responses to pathogen infections were revealed already at 3 dai by higher fluorescence mean lifetimes in the spectral range from 410 to 560 nm in the less susceptible varieties. Noticeable pathogen-induced modifications were also revealed by the 'Blue-to-Far-Red Fluorescence Ratio' and the 'Simple Fluorescence Ratio'. Particularly in the susceptible varieties the differences became more evident in the time-course of the experiment i.e., following the pathogen development. The relevance of the blue and green fluorescence to exploit the plant-pathogen interaction was demonstrated by the multispectral fluorescence imaging system. As shown, mildewed leaves were characterized by exceptionally high blue fluorescence, contrasting the values observed in rust inoculated leaves. Further, we confirm that the intensity of green fluorescence depends on the pathogen infection and the stage of disease development; this information might allow a differentiation of both diseases. Moreover, our results demonstrate that the detection area might influence the quality of the information, although it had a minor impact only in the current study. Finally, we highlight the relevance of different excitation

75 FR 59084 - Approval and Promulgation of Air Quality Implementation Plans; Maryland; Control Technique...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-09-27

... Promulgation of Air Quality Implementation Plans; Maryland; Control Technique Guidelines for Paper, Film, and... from paper, film, and foil coatings. Specifically, Maryland is amending its regulations by adopting the requirements of EPA's Control Technique Guidelines (CTG) for Paper, Film, and Foil Coatings. These amendments...

Assessing Photosynthesis by Fluorescence Imaging

ERIC Educational Resources Information Center

Saura, Pedro; Quiles, Maria Jose

2011-01-01

This practical paper describes a novel fluorescence imaging experiment to study the three processes of photochemistry, fluorescence and thermal energy dissipation, which compete during the dissipation of excitation energy in photosynthesis. The technique represents a non-invasive tool for revealing and understanding the spatial heterogeneity in…

Copper ion sensing with fluorescent electrospun nanofibers.

PubMed

Ongun, Merve Zeyrek; Ertekin, Kadriye; Gocmenturk, Mustafa; Ergun, Yavuz; Suslu, Aslıhan

2012-05-01

In this work, the use of electrospun nanofibrous materials as highly responsive fluorescence quenching-based copper sensitive chemosensor is reported. Poly(methyl methacrylate) and ethyl cellulose were used as polymeric support materials. Sensing slides were fabricated by electrospinning technique. Copper sensors based on the change in the fluorescence signal intensity of fluoroionophore; N'-3-(4-(dimethylamino phenly)allylidene)isonicotinohydrazide. The sensor slides exhibited high sensitivities due to the high surface area of the nanofibrous membrane structures. The preliminary results of Stern-Volmer analysis show that the sensitivities of electrospun nanofibrous membranes to detect Cu(II) ions are 6-20-fold higher than those of the continuous thin films. By this way we obtained linear calibration plots for Cu(II) ions in the concentration range of 10(-12)-10(-5)M. The response times of the sensing slides were less than 1 min. Stability of the employed ionophore in the matrix materials was excellent and when stored in the ambient air of the laboratory there was no significant drift in signal intensity after 6 months. Our stability tests are still in progress. Copyright © 2012 Elsevier B.V. All rights reserved.

Constraining Microwave Emission from Extensive Air Showers via the MIDAS Experiment

NASA Astrophysics Data System (ADS)

Richardson, Matthew; Privitera, Paolo

2017-01-01

Ultra high energy cosmic rays (UHECRs) are accelerated by the most energetic processes in the universe. Upon entering Earth’s atmosphere they produce particle showers known as extensive air showers (EASs). Observatories like the Pierre Auger Observatory sample the particles and light produced by the EASs through large particle detector arrays or nitrogen fluorescence detectors to ascertain the fundamental properties of UHECRs. The large sample of high quality data provided by the Pierre Auger Observatory can be attributed to the hybrid technique which utilizes the two aforementioned techniques simultaneously; however, the limitation of only being able to observe nitrogen fluorescence from EASs on clear moonless nights yields a limited 10% duty cycle for the hybrid technique. One proposal for providing high quality data at increased statistics is the observation of isotropic microwave emission from EASs, as such emission would be observed with a 100% duty cycle. Measurements of microwave emission from laboratory air plasmas conducted by Gorham et al. (2008) produced promising results indicating that the microwave emission should be observable using inexpensive detectors. The Microwave Detection of Air Showers (MIDAS) experiment was built at the University of Chicago to characterize the isotropic microwave emission from EASs and has collected 359 days of observational data at the location of the Pierre Auger experiment. We have performed a time coincidence analysis between this data and data from Pierre Auger and we report a null result. This result places stringent limits on microwave emission from EASs and demonstrates that the laboratory measurements of Gorham et al. (2008) are not applicable to EASs, thus diminishing the feasibility of using isotropic microwave emission to detect EASs.

The effect of the air-blowing step on the technique sensitivity of four different adhesive systems.

PubMed

Spreafico, Diego; Semeraro, Stefano; Mezzanzanica, Dario; Re, Dino; Gagliani, Massimo; Tanaka, Toru; Sano, Hidehiko; Sidhu, Sharanbir K

2006-03-01

To evaluate the technique sensitivity of four different adhesive systems using different air-blowing pressure. Four adhesive systems were employed: Clearfil SE Bond [SE] (Kuraray, Japan), G-Bond [GB] (GC Corporation, Japan), Adper Prompt L-Pop [LP] (3M ESPE, USA) and an experimental adhesive, SSB-200 [SSB] (Kuraray, Japan). Twenty-four extracted molars were used. After grinding the coronal enamel surface, the teeth were divided into two equal groups. The first group's teeth were randomly assigned for bonding with the different adhesives using gentle air-blowing (g). For the teeth of the second group, the four adhesive systems were applied using strong air-blowing (s). After storage overnight in 37 degrees C water, the bonded specimens were sectioned into sticks (1 mm x 1 mm wide), which were subjected to microtensile bond strength testing (microTBS) at a crosshead speed of 1 mm/min. The load at failure of each specimen was recorded and the data were analyzed by one-way ANOVA and Tukey HSD tests. The surfaces of the fractured specimens were observed using SEM to determine the failure mode. The results of the microTBS test showed that the highest bond strengths tended to be with SE for both gentle and strong air-blowing, and the significantly lowest for SSB with strong air streaming. Comparing the two techniques, significant differences were noted only for SSB-200 (P < 0.05). For each material, the SEM evaluation did not show distinct differences in the nature of the fractures between the two techniques, except for SSB-200. The adhesives tested are not technique sensitive, except SSB-200, with regards to the air-blowing step.

Macroscopic fluorescence imaging: a novel technique to monitor retention and distribution of injected microspheres in an experimental model of ischemic heart failure.

PubMed

Martens, Andreas; Rojas, Sebastian V; Baraki, Hassina; Rathert, Christian; Schecker, Natalie; Hernandez, Sara Rojas; Schwanke, Kristin; Zweigerdt, Robert; Martin, Ulrich; Saito, Shunsuke; Haverich, Axel; Kutschka, Ingo

2014-01-01

The limited effectiveness of cardiac cell therapy has generated concern regarding its clinical relevance. Experimental studies show that cell retention and engraftment are low after injection into ischemic myocardium, which may restrict therapy effectiveness significantly. Surgical aspects and mechanical loss are suspected to be the main culprits behind this phenomenon. As current techniques of monitoring intramyocardial injections are complex and time-consuming, the aim of the study was to develop a fast and simple model to study cardiac retention and distribution following intramyocardial injections. For this purpose, our main hypothesis was that macroscopic fluorescence imaging could adequately serve as a detection method for intramyocardial injections. A total of 20 mice underwent ligation of the left anterior descending artery (LAD) for myocardial infarction. Fluorescent microspheres with cellular dimensions were used as cell surrogates. Particles (5 × 10(5)) were injected into the infarcted area of explanted resting hearts (Ex vivo myocardial injetions EVMI, n = 10) and in vivo into beating hearts (In vivo myocardial injections IVMI, n = 10). Microsphere quantification was performed by fluorescence imaging of explanted organs. Measurements were repeated after a reduction to homogenate dilutions. Cardiac microsphere retention was 2.78 × 10(5) ± 0.31 × 10(5) in the EVMI group. In the IVMI group, cardiac retention of microspheres was significantly lower (0.74 × 10(5) ± 0.18 × 10(5); p<0.05). Direct fluorescence imaging revealed venous drainage through the coronary sinus, resulting in a microsphere accumulation in the left (0.90 × 10(5) ± 0.20 × 10(5)) and the right (1.07 × 10(5) ± 0.17 × 10(5)) lung. Processing to homogenates involved further particle loss (p<0.05) in both groups. We developed a fast and simple direct fluorescence imaging method for biodistribution analysis which enabled the quantification of fluorescent microspheres after

Demonstration of lipofuscin and Nissl bodies in crystal violet stained sections using a fluorescence technique or pyronin Y stain.

PubMed

Terr, L I

1986-09-01

This paper presents two simple, reliable methods for identification of lipofuscin and Nissl bodies in the same section. One method shows that lipofuscin stained with crystal violet retains its ability to fluoresce and can be observed under the fluorescence microscope after the stain has faded. Fading is accompanied by a gradual increase in the intensity of the fluorescence and is complete in about 5 min. Exciting illumination from this part of the spectrum also substantially fades staining of other autofluorescing tissue elements, such as lipids. Nonfluorescing structures, such as Nissl bodies, remain stained. By changing from transillumination with tungsten light to epifluorescent illumination and vice versa, both types of structures--Nissl bodies and lipofuscin--can be identified in the same section. The second technique uses pyronin Y for staining Nissl bodies in preparations previously stained with crystal violet. Nissl bodies are stained pink but lipofuscin remains violet. Lipofuscin in these sections also remains autofluorescent after the crystal violet stain has faded under violet or near-UV light.

Fluorescence quantum yield of carbon dioxide for quantitative UV laser-induced fluorescence in high-pressure flames

NASA Astrophysics Data System (ADS)

Lee, T.; Bessler, W. G.; Yoo, J.; Schulz, C.; Jeffries, J. B.; Hanson, R. K.

2008-11-01

The fluorescence quantum yield for ultraviolet laser-induced fluorescence of CO2 is determined for selected excitation wavelengths in the range 215-250 nm. Wavelength-resolved laser-induced fluorescence (LIF) spectra of CO2, NO, and O2 are measured in the burned gases of a laminar CH4/air flame ( φ=0.9 and 1.1) at 20 bar with additional NO seeded into the flow. The fluorescence spectra are fit to determine the relative contribution of the three species to infer an estimate of fluorescence quantum yield for CO2 that ranges from 2-8×10-6 depending on temperature and excitation wavelength with an estimated uncertainty of ±0.5×10-6. The CO2 fluorescence signal increases linearly with gas pressure for flames with constant CO2 mole fraction for the 10 to 60 bar range, indicating that collisional quenching is not an important contributor to the CO2 fluorescence quantum yield. Spectral simulation calculations are used to choose two wavelengths for excitation of CO2, 239.34 and 242.14 nm, which minimize interference from LIF of NO and O2. Quantitative LIF images of CO2 are demonstrated using these two excitation wavelengths and the measured fluorescence quantum yield.

The Use of Sensory Analysis Techniques to Assess the Quality of Indoor Air.

PubMed

Lewkowska, Paulina; Dymerski, Tomasz; Gębicki, Jacek; Namieśnik, Jacek

2017-01-02

The quality of indoor air is one of the significant elements that influences people's well-being and health inside buildings. Emissions of pollutants, which may cause odor nuisance, are the main reason for people's complaints regarding the quality of indoor air. As a result, it is necessary to perform tests aimed at identifying the sources of odors inside buildings. The article contains basic information on the characteristics of the sources of indoor air pollution and the influence of the odor detection threshold on people's health and comfort. An attempt was also made to classify and use sensory analysis techniques to perform tests of the quality of indoor air, which would enable identification of sensory experience and would allow for indication of the degree of their intensity.

Open-source do-it-yourself multi-color fluorescence smartphone microscopy

PubMed Central

Sung, Yulung; Campa, Fernando; Shih, Wei-Chuan

2017-01-01

Fluorescence microscopy is an important technique for cellular and microbiological investigations. Translating this technique onto a smartphone can enable particularly powerful applications such as on-site analysis, on-demand monitoring, and point-of-care diagnostics. Current fluorescence smartphone microscope setups require precise illumination and imaging alignment which altogether limit its broad adoption. We report a multi-color fluorescence smartphone microscope with a single contact lens-like add-on lens and slide-launched total-internal-reflection guided illumination for three common tasks in investigative fluorescence microscopy: autofluorescence, fluorescent stains, and immunofluorescence. The open-source, simple and cost-effective design has the potential for do-it-yourself fluorescence smartphone microscopy. PMID:29188104

Simultaneous off-axis multiplexed holography and regular fluorescence microscopy of biological cells.

PubMed

Nygate, Yoav N; Singh, Gyanendra; Barnea, Itay; Shaked, Natan T

2018-06-01

We present a new technique for obtaining simultaneous multimodal quantitative phase and fluorescence microscopy of biological cells, providing both quantitative phase imaging and molecular specificity using a single camera. Our system is based on an interferometric multiplexing module, externally positioned at the exit of an optical microscope. In contrast to previous approaches, the presented technique allows conventional fluorescence imaging, rather than interferometric off-axis fluorescence imaging. We demonstrate the presented technique for imaging fluorescent beads and live biological cells.

Terahertz-Radiation-Enhanced Emission of Fluorescence from Gas Plasma

NASA Astrophysics Data System (ADS)

Liu, Jingle; Zhang, X.-C.

2009-12-01

We report the study of femtosecond laser-induced air plasma fluorescence under the illumination of terahertz (THz) pulses. Semiclassical modeling and experimental verification indicate that time-resolved THz radiation-enhanced emission of fluorescence is dominated by the electron kinetics and the electron-impact excitation of gas molecules or ions. We demonstrate that the temporal waveform of the THz field could be retrieved from the transient enhanced fluorescence, making omnidirectional, coherent detection available for THz time-domain spectroscopy.

The use of a modified technique to reduce radioactive air contamination in aerosol lung ventilation imaging.

PubMed

Avison, M; Hart, G

2001-06-01

The aim of this study was to reduce airborne contamination resulting from the use of aerosols in lung ventilation scintigraphy. Lung ventilation imaging is frequently performed with 99mTc-diethylenetriaminepentaacetate aerosol (DTPA), derived from a commercial nebuliser. Airborne contamination is a significant problem with this procedure; it results in exposure of staff to radiation and can reduce gamma camera performance when the ventilation is performed in the camera room. We examined the level of airborne contamination resulting from the standard technique with one of the most popular nebuliser kits and tested a modification which significantly reduced airborne contamination. Air contamination was measured while ventilating 122 patients. The modified technique reduced air contamination by a mean value of 64% (p = 0.028) compared with the standard control technique. Additionally, differences in contamination were examined when a mask or mouthpiece was used as well as differences between operators. A simplified method of monitoring air contamination is presented using a commonly available surface contamination monitor. The index so derived was proportional to air contamination (r = 0.88). The problems and regulations associated with airborne contamination are discussed.

Excitation/Detection Strategies for OH Planar Laser-Induced Fluorescence Measurements in the Presence of Interfering Fuel Signal and Absorption Effects

NASA Technical Reports Server (NTRS)

Heath, Christopher M.; Anderson, Robert C.; Hicks, Yolanda R.

2011-01-01

Planar laser-induced fluorescence (PLIF) excitation/detection methods have been applied to obtain spatial distributions of the hydroxyl [OH] reacting intermediary and hydrocarbon [HC] primary species in laminar and turbulent combustion reactions. In this report, broadband and narrowband excitation/filtering techniques are explored to identify an optimal experimental configuration yielding significant fluorescent signal with low absorption losses. The combustion environments analyzed include 1) a laminar non-premixed methane/air flame and 2) a turbulent, non-premixed Jet-A/air fueled flame within a lean flame tube combustor. Hydrocarbon-based fuel and OH were excited via the R1 (1), R1(10) and R2(7) transitions of the A(sup 2)Epsilon(+) X(sup 2)pi(1,0) band using a broadband Nd:YAG pumped optical parametric oscillator (OPO) and narrowband Nd:YAG/dye laser with ultraviolet frequency extension (UVX) package. Variables tested for influence on fluorescent signal and absorption characteristics were excitation line, laser energy, exciting linewidth, combustion reactants, and test flow conditions. Results are intended to guide the transition from a dye/UVX laser to an OPO system for performing advanced diagnostics of low-emission combustion concepts.

A flight investigation of oscillating air forces: Equipment and technique

NASA Technical Reports Server (NTRS)

Reed, W. H., III

1975-01-01

The equipment and techniques are described which are to be used in a project aimed at measuring oscillating air forces and dynamic aeroelastic response of a swept wing airplane at high subsonic speeds. Electro-hydraulic inertia type shakers installed in the wing tips will excite various elastic airplane modes while the related oscillating chordwise pressures at two spanwise wing stations and the wing mode shapes are recorded on magnetic tape. The data reduction technique, following the principle of a wattmeter harmonic analyzer employed by Bratt, Wight, and Tilly, utilizes magnetic tape and high speed electronic multipliers to record directly the real and imaginary components of oscillatory data signals relative to a simple harmonic reference signal. Through an extension of this technique an automatic flight-flutter-test data analyzer is suggested in which vector plots of mechanical admittance or impedance would be plotted during the flight test.

pH-Mediated Fluorescent Polymer Particles and Gel from Hyperbranched Polyethylenimine and the Mechanism of Intrinsic Fluorescence.

PubMed

Liu, Shi Gang; Li, Na; Ling, Yu; Kang, Bei Hua; Geng, Shuo; Li, Nian Bing; Luo, Hong Qun

2016-02-23

We report that fluorescence properties and morphology of hyperbranched polyethylenimine (hPEI) cross-linked with formaldehyde are highly dependent on the pH values of the cross-linking reaction. Under acidic and neutral conditions, water-soluble fluorescent copolymer particles (CPs) were produced. However, under basic conditions, white gels with weak fluorescence emission would be obtained. The water-soluble hPEI-formaldehyde (hPEI-F) CPs show strong intrinsic fluorescence without the conjugation to any classical fluorescent agents. By the combination of spectroscopy and microscopy techniques, the mechanism of fluorescence emission was discussed. We propose that the intrinsic fluorescence originates from the formation of a Schiff base in the cross-linking process between hPEI and formaldehyde. Schiff base bonds are the fluorescence-emitting moieties, and the compact structure of hPEI-F CPs plays an important role in their strong fluorescence emission. The exploration on fluorescence mechanism may provide a new strategy to prepare fluorescent polymer particles. In addition, the investigation shows that the hPEI-F CPs hold potential as a fluorescent probe for the detection of copper ions in aqueous media.

Fluorescence optical imaging in anticancer drug delivery.

PubMed

Etrych, Tomáš; Lucas, Henrike; Janoušková, Olga; Chytil, Petr; Mueller, Thomas; Mäder, Karsten

2016-03-28

In the past several decades, nanosized drug delivery systems with various targeting functions and controlled drug release capabilities inside targeted tissues or cells have been intensively studied. Understanding their pharmacokinetic properties is crucial for the successful transition of this research into clinical practice. Among others, fluorescence imaging has become one of the most commonly used imaging tools in pre-clinical research. The development of increasing numbers of suitable fluorescent dyes excitable in the visible to near-infrared wavelengths of the spectrum has significantly expanded the applicability of fluorescence imaging. This paper focuses on the potential applications and limitations of non-invasive imaging techniques in the field of drug delivery, especially in anticancer therapy. Fluorescent imaging at both the cellular and systemic levels is discussed in detail. Additionally, we explore the possibility for simultaneous treatment and imaging using theranostics and combinations of different imaging techniques, e.g., fluorescence imaging with computed tomography. Copyright © 2016 Elsevier B.V. All rights reserved.

qF-SSOP: real-time optical property corrected fluorescence imaging

PubMed Central

Valdes, Pablo A.; Angelo, Joseph P.; Choi, Hak Soo; Gioux, Sylvain

2017-01-01

Fluorescence imaging is well suited to provide image guidance during resections in oncologic and vascular surgery. However, the distorting effects of tissue optical properties on the emitted fluorescence are poorly compensated for on even the most advanced fluorescence image guidance systems, leading to subjective and inaccurate estimates of tissue fluorophore concentrations. Here we present a novel fluorescence imaging technique that performs real-time (i.e., video rate) optical property corrected fluorescence imaging. We perform full field of view simultaneous imaging of tissue optical properties using Single Snapshot of Optical Properties (SSOP) and fluorescence detection. The estimated optical properties are used to correct the emitted fluorescence with a quantitative fluorescence model to provide quantitative fluorescence-Single Snapshot of Optical Properties (qF-SSOP) images with less than 5% error. The technique is rigorous, fast, and quantitative, enabling ease of integration into the surgical workflow with the potential to improve molecular guidance intraoperatively. PMID:28856038

Field Observations of Bioaerosols: What We've Learned from Fluorescence, Genetic, and Microscopic Techniques (Invited)

NASA Astrophysics Data System (ADS)

Huffman, J. A.; Fröhlich-Nowoisky, J.; Després, V. R.; Elbert, W.; Sinha, B.; Andreae, M. O.; Pöschl, U.

2009-12-01

Biogenic aerosols are ubiquitous in the Earth’s atmosphere, influencing atmospheric chemistry and physics, the biosphere, climate, and public health. They play an important role in the spread of biological organisms, and they can cause or enhance human, animal, and plant diseases. Moreover, they can initiate the formation of clouds and precipitation as cloud condensation and ice nuclei (CCN, IN). Primary biogenic aerosol particles (PBAP) such as pollen, fungal spores, and bacteria are emitted directly from the biosphere to the atmosphere. Microscopic investigations have shown that PBAP account for up to ~30% of fine and up to ~70% of coarse particulate matter in rural and rain forest air, and the estimates of PBA emissions range from ~60 Tg a-1 of fine particles up to ~1000 Tg a-1 of total particulate matter. Fungal spores account for a large proportion of PBA with typical number and mass concentrations of ~104 m-3 and ~1 μg m-3 in continental boundary layer air and estimated global emissions of the order of ~50 Tg a-1 and 200 m-2 s-1, respectively [1]. The actual abundance, variability and diversity of PBAP are still poorly understood and quantified, however. By measuring fluorescence at excitation and emission wavelengths specific to viable cells, online techniques with time resolution of minutes are able to detect fluorescent biological aerosol particles (FBAP), which represent a lower limit for the actual abundance of coarse (> 1 μm) PBAP [2]. Continuous sampling (1 - 4 months) was performed at various locations including pristine rain forest, rural and polluted urban sites. Each study exhibited a similar average particle number distribution dominated by a peak at ~3 μm, with coarse FBAP concentrations of the order of ~5x104 m-3 and ~1 μg m-3. Recent advances in the DNA analysis and molecular genetic characterization of aerosol filter samples yield new information about the sources and composition of PBA and provide new insight into regional and global

Fluorescence lifetime imaging ophthalmoscopy.

PubMed

Dysli, Chantal; Wolf, Sebastian; Berezin, Mikhail Y; Sauer, Lydia; Hammer, Martin; Zinkernagel, Martin S

2017-09-01

Imaging techniques based on retinal autofluorescence have found broad applications in ophthalmology because they are extremely sensitive and noninvasive. Conventional fundus autofluorescence imaging measures fluorescence intensity of endogenous retinal fluorophores. It mainly derives its signal from lipofuscin at the level of the retinal pigment epithelium. Fundus autofluorescence, however, can not only be characterized by the spatial distribution of the fluorescence intensity or emission spectrum, but also by a characteristic fluorescence lifetime function. The fluorescence lifetime is the average amount of time a fluorophore remains in the excited state following excitation. Fluorescence lifetime imaging ophthalmoscopy (FLIO) is an emerging imaging modality for in vivo measurement of lifetimes of endogenous retinal fluorophores. Recent reports in this field have contributed to our understanding of the pathophysiology of various macular and retinal diseases. Within this review, the basic concept of fluorescence lifetime imaging is provided. It includes technical background information and correlation with in vitro measurements of individual retinal metabolites. In a second part, clinical applications of fluorescence lifetime imaging and fluorescence lifetime features of selected retinal diseases such as Stargardt disease, age-related macular degeneration, choroideremia, central serous chorioretinopathy, macular holes, diabetic retinopathy, and retinal artery occlusion are discussed. Potential areas of use for fluorescence lifetime imaging ophthalmoscopy will be outlined at the end of this review. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

8-Anilino-1-naphthalenesulfonate/Layered Double Hydroxide Ultrathin Films: Small Anion Assembly and Its Potential Application as a Fluorescent Biosensor.

PubMed

Zhang, Ping; Li, Ling; Zhao, Yun; Tian, Zeyun; Qin, Yumei; Lu, Jun

2016-09-06

The fluorescent dye 8-anilino-1-naphthalenesulfonate (ANS) is a widely used fluorescent probe molecule for biochemistry analysis. This paper reported the fabrication of ANS/layered double hydroxide nanosheets (ANS/LDH)n ultrathin films (UTFs) via the layer-by-layer small anion assembly technique based on electrostatic interaction and two possible weak interactions: hydrogen-bond and induced electrostatic interactions between ANS and positive-charged LDH nanosheets. The obtained UTFs show a long-range-ordered periodic layered stacking structure and weak fluorescence in dry air or water, but it split into three narrow strong peaks in a weak polarity environment induced by the two-dimensional (2D) confinement effect of the LDH laminate; the fluorescence intensity increases with decreasing the solvent polarity, concomitant with the blue shift of the emission peaks, which show good sensoring reversibility. Meanwhile, the UTFs exhibit selective fluorescence enhancement to the bovine serum albumin (BSA)-like protein biomolecules, and the rate of fluorescence enhancement with the protein concentration is significantly different with the different protein aggregate states. The (ANS/LDH)n UTF has the potential to be a novel type of biological flourescence sensor material.

Some Secrets of Fluorescent Proteins: Distinct Bleaching in Various Mounting Fluids and Photoactivation of Cyan Fluorescent Proteins at YFP-Excitation

PubMed Central

Malkani, Naila; Schmid, Johannes A.

2011-01-01

Background The use of spectrally distinct variants of green fluorescent protein (GFP) such as cyan or yellow mutants (CFP and YFP, respectively) is very common in all different fields of life sciences, e.g. for marking specific proteins or cells or to determine protein interactions. In the latter case, the quantum physical phenomenon of fluorescence resonance energy transfer (FRET) is exploited by specific microscopy techniques to visualize proximity of proteins. Methodology/Principal Findings When we applied a commonly used FRET microscopy technique - the increase in donor (CFP)-fluorescence after bleaching of acceptor fluorophores (YFP), we obtained good signals in live cells, but very weak signals for the same samples after fixation and mounting in commercial microscopy mounting fluids. This observation could be traced back to much faster bleaching of CFP in these mounting media. Strikingly, the opposite effect of the mounting fluid was observed for YFP and also for other proteins such as Cerulean, TFP or Venus. The changes in photostability of CFP and YFP were not caused by the fixation but directly dependent on the mounting fluid. Furthermore we made the interesting observation that the CFP-fluorescence intensity increases by about 10 - 15% after illumination at the YFP-excitation wavelength – a phenomenon, which was also observed for Cerulean. This photoactivation of cyan fluorescent proteins at the YFP-excitation can cause false-positive signals in the FRET-microscopy technique that is based on bleaching of a yellow FRET acceptor. Conclusions/Significance Our results show that photostability of fluorescent proteins differs significantly for various media and that CFP bleaches significantly faster in commercial mounting fluids, while the opposite is observed for YFP and some other proteins. Moreover, we show that the FRET microscopy technique that is based on bleaching of the YFP is prone to artifacts due to photoactivation of cyan fluorescent proteins

Some secrets of fluorescent proteins: distinct bleaching in various mounting fluids and photoactivation of cyan fluorescent proteins at YFP-excitation.

PubMed

Malkani, Naila; Schmid, Johannes A

2011-04-07

The use of spectrally distinct variants of green fluorescent protein (GFP) such as cyan or yellow mutants (CFP and YFP, respectively) is very common in all different fields of life sciences, e.g. for marking specific proteins or cells or to determine protein interactions. In the latter case, the quantum physical phenomenon of fluorescence resonance energy transfer (FRET) is exploited by specific microscopy techniques to visualize proximity of proteins. When we applied a commonly used FRET microscopy technique--the increase in donor (CFP)-fluorescence after bleaching of acceptor fluorophores (YFP), we obtained good signals in live cells, but very weak signals for the same samples after fixation and mounting in commercial microscopy mounting fluids. This observation could be traced back to much faster bleaching of CFP in these mounting media. Strikingly, the opposite effect of the mounting fluid was observed for YFP and also for other proteins such as Cerulean, TFP or Venus. The changes in photostability of CFP and YFP were not caused by the fixation but directly dependent on the mounting fluid. Furthermore we made the interesting observation that the CFP-fluorescence intensity increases by about 10-15% after illumination at the YFP-excitation wavelength--a phenomenon, which was also observed for Cerulean. This photoactivation of cyan fluorescent proteins at the YFP-excitation can cause false-positive signals in the FRET-microscopy technique that is based on bleaching of a yellow FRET acceptor. Our results show that photostability of fluorescent proteins differs significantly for various media and that CFP bleaches significantly faster in commercial mounting fluids, while the opposite is observed for YFP and some other proteins. Moreover, we show that the FRET microscopy technique that is based on bleaching of the YFP is prone to artifacts due to photoactivation of cyan fluorescent proteins under these conditions.

Adaptive optical fluorescence microscopy.

PubMed

Ji, Na

2017-03-31

The past quarter century has witnessed rapid developments of fluorescence microscopy techniques that enable structural and functional imaging of biological specimens at unprecedented depth and resolution. The performance of these methods in multicellular organisms, however, is degraded by sample-induced optical aberrations. Here I review recent work on incorporating adaptive optics, a technology originally applied in astronomical telescopes to combat atmospheric aberrations, to improve image quality of fluorescence microscopy for biological imaging.

Characterization of Air Plane Soot Surrogates using Raman spectroscopy and laser ablation techniques

NASA Astrophysics Data System (ADS)

Chazallon, Bertrand; Ortega, Ismael Kenneth; Ikhenazene, Raouf; Pirim, Claire; Carpentier, Yvain; Irimiea, Cornelia; Focsa, Cristian; Ouf, François-Xavier

2016-04-01

Aviation alters the composition of the atmosphere globally and can thus drive climate change and ozone depletion [1]. Aircraft exhaust plumes contain species (gases and soot particles) produced by the combustion of kerosene with ambient air in the combustion chamber of the engine. Soot particles emitted by air-planes produce persistent contrails in the upper troposphere in ice-supersaturated air masses that contribute to cloudiness and impact the radiative properties of the atmosphere. These aerosol-cloud interactions represent one of the largest sources of uncertainty in global climate models [2]. Though the formation of atmospheric ice particles has been studied for many years [3], there are still numerous opened questions on nucleation properties of soot particles [4], as the ice nucleation experiments showed a large spread in results depending on the nucleation mode chosen and origin of the soot produced. The reasons behind these discrepancies reside in the different physico-chemical properties (composition, structure) of soot particles produced in different conditions, e.g., with respect to fuel or combustion techniques. In this work, we use Raman microscopy (514 and 785 nm excitation wavelengths) and ablation techniques (Secondary Ions Mass Spectrometry, and Laser Desorption Mass Spectrometry) to characterize soot particle surrogates produced from a CAST generator (propane fuel, four different global equivalence ratios). They are produced as analogues of air-plane soot collected at different engine regimes (PowerJet SaM-146 turbofan) simulating a landing and take-off (LTO) cycle (MERMOSE project (http://mermose.onera.fr/)) [6]. The spectral parameters of the first-order Raman bands of these soot samples are analyzed using a de-convolution approach described by Sadezky et al. (2005) [5]. A systematic Raman analysis is carried out to select a number of parameters (laser wavelength, irradiance at sample, exposure time) that will alter the sample and the

Catheter-based time-gated near-infrared fluorescence/OCT imaging system

NASA Astrophysics Data System (ADS)

Lu, Yuankang; Abran, Maxime; Cloutier, Guy; Lesage, Frédéric

2018-02-01

We developed a new dual-modality intravascular imaging system based on fast time-gated fluorescence intensity imaging and spectral domain optical coherence tomography (SD-OCT) for the purpose of interventional detection of atherosclerosis. A pulsed supercontinuum laser was used for fluorescence and OCT imaging. A double-clad fiber (DCF)- based side-firing catheter was designed and fabricated to have a 23 μm spot size at a 2.2 mm working distance for OCT imaging. Its single-mode core is used for OCT, while its inner cladding transports fluorescence excitation light and collects fluorescent photons. The combination of OCT and fluorescence imaging was achieved by using a DCF coupler. For fluorescence detection, we used a time-gated technique with a novel single-photon avalanche diode (SPAD) working in an ultra-fast gating mode. A custom-made delay chip was integrated in the system to adjust the delay between the excitation laser pulse and the SPAD gate-ON window. This technique allowed to detect fluorescent photons of interest while rejecting most of the background photons, thus leading to a significantly improved signal to noise ratio (SNR). Experiments were carried out in turbid media mimicking tissue with an indocyanine green (ICG) inclusion (1 mM and 100 μM) to compare the time-gated technique and the conventional continuous detection technique. The gating technique increased twofold depth sensitivity, and tenfold SNR at large distances. The dual-modality imaging capacity of our system was also validated with a silicone-based tissue-mimicking phantom.

Longitudinal in vivo two-photon fluorescence imaging

PubMed Central

Crowe, Sarah E.; Ellis-Davies, Graham C.R.

2014-01-01

Fluorescence microscopy is an essential technique for the basic sciences, especially biomedical research. Since the invention of laser scanning confocal microscopy in 1980s, that enabled imaging both fixed and living biological tissue with three-dimensional precision, high-resolution fluorescence imaging has revolutionized biological research. Confocal microscopy, by its very nature, has one fundamental limitation. Due to the confocal pinhole, deep tissue fluorescence imaging is not practical. In contrast (no pun intended), two-photon fluorescence microscopy allows, in principle, the collection of all emitted photons from fluorophores in the imaged voxel, dramatically extending our ability to see deep into living tissue. Since the development of transgenic mice with genetically encoded fluorescent protein in neocortical cells in 2000, two-photon imaging has enabled the dynamics of individual synapses to be followed for up to two years. Since the initial landmark contributions to this field in 2002, the technique has been used to understand how neuronal structure are changed by experience, learning and memory and various diseases. Here we provide a basic summary of the crucial elements that are required for such studies, and discuss many applications of longitudinal two-photon fluorescence microscopy that have appeared since 2002. PMID:24214350

Laser induced fluorescence technique for environmental applications

NASA Astrophysics Data System (ADS)

Utkin, Andrei B.; Felizardo, Rui; Gameiro, Carla; Matos, Ana R.; Cartaxana, Paulo

2014-08-01

We discuss the development of laser induced fluorescence sensors and their application in the evaluation of water pollution and physiological status of higher plants and algae. The sensors were built on the basis of reliable and robust solid-state Nd:YAG lasers. They demonstrated good efficiency in: i) detecting and characterizing oil spills and dissolved organic matter; ii) evaluating the impact of stress on higher plants (cork oak, maritime pine, and genetically modified Arabidopsis); iii) tracking biomass changes in intertidal microphytobenthos; and iv) mapping macroalgal communities in the Tagus Estuary.

Luminescence and fluorescence of essential oils. Fluorescence imaging in vivo of wild chamomile oil.

PubMed

Boschi, F; Fontanella, M; Calderan, L; Sbarbati, A

2011-06-16

Essential oils are currently of great importance to pharmaceutical companies, cosmetics producers and manufacturers of veterinary products. They are found in perfumes, creams, bath products, and household cleaning substances, and are used for flavouring food and drinks. It is well known that some of them act on the respiratory apparatus. The increasing interest in optical imaging techniques and the development of related technologies have made possible the investigation of the optical properties of several compounds. Luminescent properties of essential oils have not been extensively investigated. We evaluated the luminescent and fluorescent emissions of several essential oils, in order to detect them in living organisms by exploiting their optical properties. Some fluorescent emission data were high enough to be detected in dermal treatments. Consequently, we demonstrated how the fluorescent signal can be monitored for at least three hours on the skin of living mice treated with wild chamomile oil. The results encourage development of this technique to investigate the properties of drugs and cosmetics containing essential oils.

Air-coupled ultrasound: a novel technique for monitoring the curing of thermosetting matrices.

PubMed

Lionetto, Francesca; Tarzia, Antonella; Maffezzoli, Alfonso

2007-07-01

A custom-made, air-coupled ultrasonic device was applied to cure monitoring of thick samples (7-10 mm) of unsaturated polyester resin at room temperature. A key point was the optimization of the experimental setup in order to propagate compression waves during the overall curing reaction by suitable placement of the noncontact transducers, placed on the same side of the test material, in the so-called pitch-catch configuration. The progress of polymerization was monitored through the variation of the time of flight of the propagating longitudinal waves. The exothermic character of the polymerization was taken into account by correcting the measured value of time of flight with that one in air, obtained by sampling the air velocity during the experiment. The air-coupled ultrasonic results were compared with those obtained from conventional contact ultrasonic measurements. The good agreement between the air-coupled ultrasonic results and those obtained by the rheological analysis demonstrated the reliability of air-coupled ultrasound in monitoring the changes of viscoelastic properties at gelation and vitrification. The position of the transducers on the same side of the sample makes this technique suitable for on-line cure monitoring during several composite manufacturing technologies.

Fluorescence background subtraction technique for hybrid fluorescence molecular tomography/x-ray computed tomography imaging of a mouse model of early stage lung cancer.

PubMed

Ale, Angelique; Ermolayev, Vladimir; Deliolanis, Nikolaos C; Ntziachristos, Vasilis

2013-05-01

The ability to visualize early stage lung cancer is important in the study of biomarkers and targeting agents that could lead to earlier diagnosis. The recent development of hybrid free-space 360-deg fluorescence molecular tomography (FMT) and x-ray computed tomography (XCT) imaging yields a superior optical imaging modality for three-dimensional small animal fluorescence imaging over stand-alone optical systems. Imaging accuracy was improved by using XCT information in the fluorescence reconstruction method. Despite this progress, the detection sensitivity of targeted fluorescence agents remains limited by nonspecific background accumulation of the fluorochrome employed, which complicates early detection of murine cancers. Therefore we examine whether x-ray CT information and bulk fluorescence detection can be combined to increase detection sensitivity. Correspondingly, we research the performance of a data-driven fluorescence background estimator employed for subtraction of background fluorescence from acquisition data. Using mice containing known fluorochromes ex vivo, we demonstrate the reduction of background signals from reconstructed images and sensitivity improvements. Finally, by applying the method to in vivo data from K-ras transgenic mice developing lung cancer, we find small tumors at an early stage compared with reconstructions performed using raw data. We conclude with the benefits of employing fluorescence subtraction in hybrid FMT-XCT for early detection studies.

Bioaerosol detection and classification using dual excitation wavelength laser-induced fluorescence

NASA Astrophysics Data System (ADS)

Jonsson, Per; Wästerby, Pär.; Gradmark, Per-Åke; Hedborg, Julia; Larsson, Anders; Landström, Lars

2015-05-01

We present results obtained by a detection system designed to measure laser-induced fluorescence from individual aerosol particles using dual excitation wavelengths. The aerosol is sampled from ambient air and via a 1 mm diameter nozzle, surrounded by a sheath air flow, confined into a particle beam. A continuous wave blue laser at 404 nm is focused on the aerosol beam and two photomultiplier tubes monitor the presence of individual particles by simultaneous measuring the scattered light and any induced fluorescence. When a particle is present in the detection volume, a laser pulse is triggered from an ultraviolet laser at 263 nm and the corresponding fluorescence spectrum is acquired with a spectrometer based on a diffraction grating and a 32 channel photomultiplier tube array with single-photon sensitivity. The spectrometer measures the fluorescence spectra in the wavelength region from 250 to 800 nm. In the present report, data were measured on different monodisperse reference aerosols, simulants of biological warfare agents, and different interference aerosol particles, e.g. pollen. In the analysis of the experimental data, i.e., the time-resolved scattered and fluorescence signals from 404 nm c.w. light excitation and the fluorescence spectra obtained by a pulsed 263 nm laser source, we use multivariate data analysis methods to classify each individual aerosol particle.

Laser induced fluorescence in algae: A new technique for remote detection

NASA Technical Reports Server (NTRS)

Friedman, E. J.; Hickman, G. D.

1972-01-01

Measurements of the absorption and fluorescence spectra were obtained for four various types of marine and fresh water algae using a pulsed N2/Ne dye laser as the source of excitation. The absorption maxima for the algae ranged from 420 to 675 nm, while their fluorescent spectra ranged from 580 to 685 nm. It appears feasible that various algal species can be identified by detection of their fluorescent signatures using a tunable laser as the excitation source. However, if one is concerned only with detection of chlorophyll a, the optimum excitation is approximately 600 + 50 nm while detection is at 685 nm. An analysis of both calculations and laboratory results indicates that it should be feasible to measure chlorophyll a in concentrations as low as 1.0 mg/m3 using a 100 kW peak pulsed laser from an altitude of 500 meters.

Characterization of uranium bearing material using x-ray fluorescence and direct gamma-rays measurement techniques

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mujaini, M., E-mail: madihah@uniten.edu.my; Chankow, N.; Yusoff, M. Z.

2016-01-22

Uranium ore can be easily detected due to various gamma-ray energies emitted from uranium daughters particularly from {sup 238}U daughters such as {sup 214}Bi, {sup 214}Pb and {sup 226}Ra. After uranium is extracted from uranium ore, only low energy gamma-rays emitted from {sup 235}U may be detected if the detector is placed in close contact to the specimen. In this research, identification and characterization of uranium bearing materials is experimentally investigated using direct measurement of gamma-rays from {sup 235}U in combination with the x-ray fluorescence (XRF) technique. Measurement of gamma-rays can be conducted by using high purity germanium (HPGe) detectormore » or cadmium telluride (CdTe) detector while a {sup 57}Coradioisotope-excited XRF spectrometer using CdTe detector is used for elemental analysis. The proposed technique was tested with various uranium bearing specimens containing natural, depleted and enriched uranium in both metallic and powder forms.« less

Comparison of Techniques for Non-Intrusive Fuel Drop Size Measurements in a Subscale Gas Turbine Combustor

NASA Technical Reports Server (NTRS)

Zaller, Michelle; Anderson, Robert C.; Hicks, Yolanda R.; Locke, Randy J.

1999-01-01

In aviation gas turbine combustors, many factors, such as the degree and extent of fuel/air mixing and fuel vaporization achieved prior to combustion, influence the formation of undesirable pollutants. To assist in analyzing the extent of fuel/air mixing, flow visualization techniques have been used to interrogate the fuel distributions during subcomponent tests of lean-burning fuel injectors. Discrimination between liquid and vapor phases of the fuel was determined by comparing planar laser-induced fluorescence (PLIF) images, elastically-scattered light images, and phase/Doppler interferometer measurements. Estimates of Sauter mean diameters are made by ratioing PLIF and Mie scattered intensities for various sprays, and factors affecting the accuracy of these estimates are discussed. Mie calculations of absorption coefficients indicate that the fluorescence intensities of individual droplets are proportional to their surface areas, instead of their volumes, due to the high absorbance of the liquid fuel for the selected excitation wavelengths.

Fundamental limits to superresolution fluorescence microscopy

NASA Astrophysics Data System (ADS)

Small, Alex

2013-02-01

Superresolution fluorescence microscopy techniques such as PALM, STORM, STED, and Structured Illumination Microscopy (SIM) enable imaging of live cells at nanometer resolution. The common theme in all of these techniques is that the diffraction limit is circumvented by controlling the states of fluorescent molecules. Although the samples are labeled very densely (i.e. with spacing much smaller than the Airy distance), not all of the molecules are emitting at the same time. Consequently, one does not encounter overlapping blurs. In the deterministic techniques (STED, SIM) the achievable resolution scales as the wavelength of light divided by the square root of the intensity of a beam used to control the fluorescent state. In the stochastic techniques (PALM, STORM), the achievable resolution scales as the wavelength of light divided by the square root of the number of photons collected. Although these limits arise from very different mechanisms (parabolic beam profiles for STED and SIM, statistics for PALM and STORM), in all cases the resolution scales inversely with the square root of a measure of the number of photons used in the experiment. We have developed a proof that this relationship between resolution and photon count is universal to techniques that control the states of fluorophores using classical light. Our proof encompasses linear and nonlinear optics, as well as computational post-processing techniques for extracting information beyond the diffraction limit. If there are techniques that can achieve a more efficient relationship between resolution and photon count, those techniques will require light exhibiting non-classical correlations.

[Simultaneous determination of multiple elements in airborne particulate samples by X-ray fluorescence spectrometry].

PubMed

Takada, T; Hitosugi, M; Kadowaki, T; Kudo, M

1983-07-01

An energy dispersive X-ray fluorescence spectrometer (EDX) has been applied to determine multielements in the workplace air. The standards for X-ray fluorescence analysis were prepared by the chelate precipitation method on polyvinyl chloride (PVC) membrane filter. And, the specimens were prepared to deposit various metal compounds of different chemical forms by the suspension method on PVC membrane filter, and they were determined with EDX and atomic absorption spectrometer (AAS). The results obtained were as follows. Though there is a difference by each element, an amount less than 3 microgram/cm2 per unit area makes it possible to undergo multielement analysis, that is, is has no influence on fine particle effect (particle size; under 5 microns). Then, effects of the X-ray intensity by different chemical forms are negligible. At the presence the neighboring element and other elements this technique showed greater precision by carrying out on corrective treatment, etc. The coefficient of variation of this technique was in the range of 2.5-6.5% at DDTC-Cu of 0.5-5.0 micrograms/cm2, with the limit of detection for As : 0.002 microgram/cm2, Zn : 0.003 microgram/cm2, Pb : 0.003 microgram/cm2, Cu : 0.004 microgram/cm2, Ni : 0.003 microgram/cm2, Fe : 0.005 microgram/cm2, Mn : 0.008 microgram/cm2, Cr : 0.013 microgram/cm2, respectively. Aerosols collected at the workplace were analyzed with EDX and AAS, and the obtained results showed good agreement with such regression line as y = 1.04 chi + 0.04, the coefficient of correlation being r = 0.995. From these results, this technique was found to be a very excellent method for monitoring of multielements in the workplace air.

Fluorescence characterization of clinically-important bacteria.

PubMed

Dartnell, Lewis R; Roberts, Tom A; Moore, Ginny; Ward, John M; Muller, Jan-Peter

2013-01-01

Healthcare-associated infections (HCAI/HAI) represent a substantial threat to patient health during hospitalization and incur billions of dollars additional cost for subsequent treatment. One promising method for the detection of bacterial contamination in a clinical setting before an HAI outbreak occurs is to exploit native fluorescence of cellular molecules for a hand-held, rapid-sweep surveillance instrument. Previous studies have shown fluorescence-based detection to be sensitive and effective for food-borne and environmental microorganisms, and even to be able to distinguish between cell types, but this powerful technique has not yet been deployed on the macroscale for the primary surveillance of contamination in healthcare facilities to prevent HAI. Here we report experimental data for the specification and design of such a fluorescence-based detection instrument. We have characterized the complete fluorescence response of eleven clinically-relevant bacteria by generating excitation-emission matrices (EEMs) over broad wavelength ranges. Furthermore, a number of surfaces and items of equipment commonly present on a ward, and potentially responsible for pathogen transfer, have been analyzed for potential issues of background fluorescence masking the signal from contaminant bacteria. These include bedside handrails, nurse call button, blood pressure cuff and ward computer keyboard, as well as disinfectant cleaning products and microfiber cloth. All examined bacterial strains exhibited a distinctive double-peak fluorescence feature associated with tryptophan with no other cellular fluorophore detected. Thus, this fluorescence survey found that an emission peak of 340nm, from an excitation source at 280nm, was the cellular fluorescence signal to target for detection of bacterial contamination. The majority of materials analysed offer a spectral window through which bacterial contamination could indeed be detected. A few instances were found of potential problems

Fluorescence technique for on-line monitoring of state of hydrogen-producing microorganisms

DOEpatents

Seibert, Michael [Lakewood, CO; Makarova, Valeriya [Golden, CO; Tsygankov, Anatoly A [Pushchino, RU; Rubin, Andrew B [Moscow, RU

2007-06-12

In situ fluorescence method to monitor state of sulfur-deprived algal culture's ability to produce H.sub.2 under sulfur depletion, comprising: a) providing sulfur-deprived algal culture; b) illuminating culture; c) measuring onset of H.sub.2 percentage in produced gas phase at multiple times to ascertain point immediately after anerobiosis to obtain H.sub.2 data as function of time; and d) determining any abrupt change in three in situ fluorescence parameters; i) increase in F.sub.t (steady-state level of chlorophyll fluorescence in light adapted cells); ii) decrease in F.sub.m', (maximal saturating light induced fluorescence level in light adapted cells); and iii) decrease in .DELTA.F/F.sub.m'=(F.sub.m'-F.sub.t)/F.sub.m' (calculated photochemical activity of photosystem II (PSII) signaling full reduction of plastoquinone pool between PSII and PSI, which indicates start of anaerobic conditions that induces synthesis of hydrogenase enzyme for subsequent H.sub.2 production that signal oxidation of plastoquinone pool asmain factor to regulate H.sub.2 under sulfur depletion.

Miniaturized side-viewing imaging probe for fluorescence lifetime imaging (FLIM): validation with fluorescence dyes, tissue structural proteins and tissue specimens

PubMed Central

Elson, D S; Jo, J A

2007-01-01

We report a side viewing fibre-based endoscope that is compatible with intravascular imaging and fluorescence lifetime imaging microscopy (FLIM). The instrument has been validated through testing with fluorescent dyes and collagen and elastin powders using the Laguerre expansion deconvolution technique to calculate the fluorescence lifetimes. The instrument has also been tested on freshly excised unstained animal vascular tissues. PMID:19503759

Fluorescence lifetime imaging of optically levitated aerosol: a technique to quantitatively map the viscosity of suspended aerosol particles.

PubMed

Fitzgerald, C; Hosny, N A; Tong, H; Seville, P C; Gallimore, P J; Davidson, N M; Athanasiadis, A; Botchway, S W; Ward, A D; Kalberer, M; Kuimova, M K; Pope, F D

2016-08-21

We describe a technique to measure the viscosity of stably levitated single micron-sized aerosol particles. Particle levitation allows the aerosol phase to be probed in the absence of potentially artefact-causing surfaces. To achieve this feat, we combined two laser based techniques: optical trapping for aerosol particle levitation, using a counter-propagating laser beam configuration, and fluorescent lifetime imaging microscopy (FLIM) of molecular rotors for the measurement of viscosity within the particle. Unlike other techniques used to measure aerosol particle viscosity, this allows for the non-destructive probing of viscosity of aerosol particles without interference from surfaces. The well-described viscosity of sucrose aerosol, under a range of relative humidity conditions, is used to validate the technique. Furthermore we investigate a pharmaceutically-relevant mixture of sodium chloride and salbutamol sulphate under humidities representative of in vivo drug inhalation. Finally, we provide a methodology for incorporating molecular rotors into already levitated particles, thereby making the FLIM/optical trapping technique applicable to real world aerosol systems, such as atmospheric aerosols and those generated by pharmaceutical inhalers.

An evolutionary outlook of air traffic flow management techniques

NASA Astrophysics Data System (ADS)

Kistan, Trevor; Gardi, Alessandro; Sabatini, Roberto; Ramasamy, Subramanian; Batuwangala, Eranga

2017-01-01

In recent years Air Traffic Flow Management (ATFM) has become pertinent even in regions without sustained overload conditions caused by dense traffic operations. Increasing traffic volumes in the face of constrained resources has created peak congestion at specific locations and times in many areas of the world. Increased environmental awareness and economic drivers have combined to create a resurgent interest in ATFM as evidenced by a spate of recent ATFM conferences and workshops mediated by official bodies such as ICAO, IATA, CANSO the FAA and Eurocontrol. Significant ATFM acquisitions in the last 5 years include South Africa, Australia and India. Singapore, Thailand and Korea are all expected to procure ATFM systems within a year while China is expected to develop a bespoke system. Asia-Pacific nations are particularly pro-active given the traffic growth projections for the region (by 2050 half of all air traffic will be to, from or within the Asia-Pacific region). National authorities now have access to recently published international standards to guide the development of national and regional operational concepts for ATFM, geared to Communications, Navigation, Surveillance/Air Traffic Management and Avionics (CNS+A) evolutions. This paper critically reviews the field to determine which ATFM research and development efforts hold the best promise for practical technological implementations, offering clear benefits both in terms of enhanced safety and efficiency in times of growing air traffic. An evolutionary approach is adopted starting from an ontology of current ATFM techniques and proceeding to identify the technological and regulatory evolutions required in the future CNS+A context, as the aviation industry moves forward with a clearer understanding of emerging operational needs, the geo-political realities of regional collaboration and the impending needs of global harmonisation.

Single-molecule fluorescence microscopy review: shedding new light on old problems

PubMed Central

Shashkova, Sviatlana

2017-01-01

Fluorescence microscopy is an invaluable tool in the biosciences, a genuine workhorse technique offering exceptional contrast in conjunction with high specificity of labelling with relatively minimal perturbation to biological samples compared with many competing biophysical techniques. Improvements in detector and dye technologies coupled to advances in image analysis methods have fuelled recent development towards single-molecule fluorescence microscopy, which can utilize light microscopy tools to enable the faithful detection and analysis of single fluorescent molecules used as reporter tags in biological samples. For example, the discovery of GFP, initiating the so-called ‘green revolution’, has pushed experimental tools in the biosciences to a completely new level of functional imaging of living samples, culminating in single fluorescent protein molecule detection. Today, fluorescence microscopy is an indispensable tool in single-molecule investigations, providing a high signal-to-noise ratio for visualization while still retaining the key features in the physiological context of native biological systems. In this review, we discuss some of the recent discoveries in the life sciences which have been enabled using single-molecule fluorescence microscopy, paying particular attention to the so-called ‘super-resolution’ fluorescence microscopy techniques in live cells, which are at the cutting-edge of these methods. In particular, how these tools can reveal new insights into long-standing puzzles in biology: old problems, which have been impossible to tackle using other more traditional tools until the emergence of new single-molecule fluorescence microscopy techniques. PMID:28694303

Instantaneous three-dimensional visualization of concentration distributions in turbulent flows with crossed-plane laser-induced fluorescence imaging

NASA Astrophysics Data System (ADS)

Hoffmann, A.; Zimmermann, F.; Scharr, H.; Krömker, S.; Schulz, C.

2005-01-01

A laser-based technique for measuring instantaneous three-dimensional species concentration distributions in turbulent flows is presented. The laser beam from a single laser is formed into two crossed light sheets that illuminate the area of interest. The laser-induced fluorescence (LIF) signal emitted from excited species within both planes is detected with a single camera via a mirror arrangement. Image processing enables the reconstruction of the three-dimensional data set in close proximity to the cutting line of the two light sheets. Three-dimensional intensity gradients are computed and compared to the two-dimensional projections obtained from the two directly observed planes. Volume visualization by digital image processing gives unique insight into the three-dimensional structures within the turbulent processes. We apply this technique to measurements of toluene-LIF in a turbulent, non-reactive mixing process of toluene and air and to hydroxyl (OH) LIF in a turbulent methane-air flame upon excitation at 248 nm with a tunable KrF excimer laser.

Fluorescence correlation spectroscopy: principles and applications.

PubMed

Bacia, Kirsten; Haustein, Elke; Schwille, Petra

2014-07-01

Fluorescence correlation spectroscopy (FCS) is used to study the movements and the interactions of biomolecules at extremely dilute concentrations, yielding results with good spatial and temporal resolutions. Using a number of technical developments, FCS has become a versatile technique that can be used to study a variety of sample types and can be advantageously combined with other methods. Unlike other fluorescence-based techniques, the analysis of FCS data is not based on the average intensity of the fluorescence emission but examines the minute intensity fluctuations caused by spontaneous deviations from the mean at thermal equilibrium. These fluctuations can result from variations in local concentrations owing to molecular mobility or from characteristic intermolecular or intramolecular reactions of fluorescently labeled biomolecules present at low concentrations. Here, we provide a basic introduction to FCS, including its technical development and theoretical basis, experimental setup of an FCS system, adjustment of a setup, data acquisition, and analysis of FCS measurements. Finally, the application of FCS to the study of lipid bilayer membranes and to living cells is discussed. © 2014 Cold Spring Harbor Laboratory Press.

Polyester Fabric's Fluorescent Dyeing in Supercritical Carbon Dioxide and its Fluorescence Imaging.

PubMed

Xiong, Xiaoqing; Xu, Yanyan; Zheng, Laijiu; Yan, Jun; Zhao, Hongjuan; Zhang, Juan; Sun, Yanfeng

2017-03-01

As one of the most important coumarin-like dyes, disperse fluorescent Yellow 82 exhibits exceptionally large two-photon effects. Here, it was firstly introduced into the supercritical CO 2 dyeing polyester fabrics in this work. Results of the present work showed that the dyeing parameters such as the dyeing time, pressure and temperature had remarkable influences on the color strength of fabrics. The optimized dyeing condition in supercritical CO 2 dyeing has been proposed that the dyeing time was 60 min; the pressure was 25 MPa and the temperature was 120 °C. As a result, acceptable products were obtained with the wash and rub fastness rating at 5 or 4-5. The polyester fabrics dyed with fluorescent dyes can be satisfied for the requirement of manufacturing warning clothing. Importantly, the confocal microscopy imaging technology was successfully introduced into textile fields to observe the distribution and fluorescence intensity of disperse fluorescent Yellow 82 on polyester fabrics. As far as we know, this is the first report about supercritical CO 2 dyeing polyester fabrics based on disperse fluorescent dyes. It will be very helpful for the further design of new fluorescent functional dyes suitable for supercritical CO 2 dyeing technique.

Study of fluorescence quenching of Barley α-amylase

NASA Astrophysics Data System (ADS)

Bakkialakshmi, S.; Shanthi, B.; Bhuvanapriya, T.

2012-05-01

The fluorescence quenching of Barley α-amylase by acrylamide and succinimide has been studied in water using steady-state and time-resolved fluorescence techniques. The steady-state fluorescence quenching technique has been performed in three different pHs (i.e., 6, 7 and 8) of water. Ground state and excited state binding constants (Kg &Ke) have been calculated. From the calculated binding constants (Kg &Ke) the free energy changes for the ground (ΔGg) and excited (ΔGe) states have been calculated and are presented in tables. UV and FTIR spectra have also been recorded to prove the binding of Barley α-amylase with acrylamide and succinimide.

Detecting fluorescence hot-spots using mosaic maps generated from multimodal endoscope imaging

NASA Astrophysics Data System (ADS)

Yang, Chenying; Soper, Timothy D.; Seibel, Eric J.

2013-03-01

Fluorescence labeled biomarkers can be detected during endoscopy to guide early cancer biopsies, such as high-grade dysplasia in Barrett's Esophagus. To enhance intraoperative visualization of the fluorescence hot-spots, a mosaicking technique was developed to create full anatomical maps of the lower esophagus and associated fluorescent hot-spots. The resultant mosaic map contains overlaid reflectance and fluorescence images. It can be used to assist biopsy and document findings. The mosaicking algorithm uses reflectance images to calculate image registration between successive frames, and apply this registration to simultaneously acquired fluorescence images. During this mosaicking process, the fluorescence signal is enhanced through multi-frame averaging. Preliminary results showed that the technique promises to enhance the detectability of the hot-spots due to enhanced fluorescence signal.

Laser-induced fluorescence spectroscopy in tissue local necrosis detection

NASA Astrophysics Data System (ADS)

Cip, Ondrej; Buchta, Zdenek; Lesundak, Adam; Randula, Antonin; Mikel, Bretislav; Lazar, Josef; Veverkova, Lenka

2014-03-01

The recent effort leads to reliable imaging techniques which can help to a surgeon during operations. The fluorescence spectroscopy was selected as very useful online in vivo imaging method to organics and biological materials analysis. The presented work scopes to a laser induced fluorescence spectroscopy technique to detect tissue local necrosis in small intestine surgery. In first experiments, we tested tissue auto-fluorescence technique but a signal-to-noise ratio didn't express significant results. Then we applied a contrast dye - IndoCyanine Green (ICG) which absorbs and emits wavelengths in the near IR. We arranged the pilot experimental setup based on highly coherent extended cavity diode laser (ECDL) used for stimulating of some critical areas of the small intestine tissue with injected ICG dye. We demonstrated the distribution of the ICG exciter with the first file of shots of small intestine tissue of a rabbit that was captured by high sensitivity fluorescent cam.

Single molecule fluorescence imaging as a technique for barium tagging in neutrinoless double beta decay

NASA Astrophysics Data System (ADS)

Jones, B. J. P.; McDonald, A. D.; Nygren, D. R.

2016-12-01

Background rejection is key to success for future neutrinoless double beta decay experiments. To achieve sensitivity to effective Majorana lifetimes of ~ 1028 years, backgrounds must be controlled to better than 0.1 count per ton per year, beyond the reach of any present technology. In this paper we propose a new method to identify the birth of the barium daughter ion in the neutrinoless double beta decay of 136Xe. The method adapts Single Molecule Fluorescent Imaging, a technique from biochemistry research with demonstrated single ion sensitivity. We explore possible SMFI dyes suitable for the problem of barium ion detection in high pressure xenon gas, and develop a fiber-coupled sensing system with which we can detect the presence of bulk Ba++ ions remotely. We show that our sensor produces signal-to-background ratios as high as 85 in response to Ba++ ions when operated in aqueous solution. We then describe the next stage of this R&D program, which will be to demonstrate chelation and fluorescence in xenon gas. If a successful barium ion tag can be developed using SMFI adapted for high pressure xenon gas detectors, the first essentially zero background, ton-scale neutrinoless double beta decay technology could be realized.

Fluorescence Studies of Lysozyme Nucleation

NASA Technical Reports Server (NTRS)

Pusey, Marc L.; Smith, Lori

1998-01-01

Fluorescence is one of the most powerful tools available for the study of macromolecules. For example, fluorescence can be used to study self association through methods such as anisotropy (the rotational rate of the molecule in solution), quenching (the accessibility of a bound probe to the bulk solution), and resonance energy transfer (measurement of the distance between two species). Fluorescence can also be used to study the local environment of the probe molecules, and the changes in that environment which accompany crystal nucleation and growth. However fluorescent techniques have been very much underutilized in macromolecular growth studies. One major advantage is that the fluorescent species generally must be at low concentration, typically ca 10-5 to 10-6 M. Thus one can study a very wide range of solution conditions, ranging from very high to very low protein concentration, he latter of which are not readily accessible to scattering techniques. We have prepared a number of fluorescent derivatives of chicken egg white lysozyme (CEWL). Fluorescent probes have been attached to two different sites, ASP 101 and the N-terrninal amine, with a sought for use in different lines of study. Preliminary resonance energy transfer studies have been -carried out using pyrene acetic acid (Ex 340 mn, Em 376 nm) lysozyme as a donor and cascade blue (Ex 377 run, Em 423 nm) labeled lysozyme as an acceptor. The emission of both the pyrene and cascade blue probes was followed as a function of the salt protein concentrations. The data show an increase in cascade blue and a concomitant decrease in the pyrene fluorescence as either the salt or protein concentrations are increased, suggesting that the two species are approaching each other close enough for resonance energy transfer to occur. This data can be analyzed to measure the distance between the probe molecules and, knowing their locations on the protein molecule their distances from and orientations with respect to each

Capture of Fluorescence Decay Times by Flow Cytometry

PubMed Central

Naivar, Mark A.; Jenkins, Patrick; Freyer, James P.

2012-01-01

In flow cytometry, the fluorescence decay time of an excitable species has been largely underutilized and is not likely found as a standard parameter on any imaging cytometer, sorting, or analyzing system. Most cytometers lack fluorescence lifetime hardware mainly owing to two central issues. Foremost, research and development with lifetime techniques has lacked proper exploitation of modern laser systems, data acquisition boards, and signal processing techniques. Secondly, a lack of enthusiasm for fluorescence lifetime applications in cells and with bead-based assays has persisted among the greater cytometry community. In this unit, we describe new approaches that address these issues and demonstrate the simplicity of digitally acquiring fluorescence relaxation rates in flow. The unit is divided into protocol and commentary sections in order to provide a most comprehensive discourse on acquiring the fluorescence lifetime with frequency-domain methods. The unit covers (i) standard fluorescence lifetime acquisition (protocol-based) with frequency-modulated laser excitation, (ii) digital frequency-domain cytometry analyses, and (iii) interfacing fluorescence lifetime measurements onto sorting systems. Within the unit is also a discussion on how digital methods are used for aliasing in order to harness higher frequency ranges. Also, a final discussion is provided on heterodyning and processing of waveforms for multi-exponential decay extraction. PMID:25419263

Time-resolved multicolor two-photon excitation fluorescence microscopy of cells and tissues

NASA Astrophysics Data System (ADS)

Zheng, Wei

2014-11-01

Multilabeling which maps the distribution of different targets is an indispensable technique in many biochemical and biophysical studies. Two-photon excitation fluorescence (TPEF) microscopy of endogenous fluorophores combining with conventional fluorescence labeling techniques such as genetically encoded fluorescent protein (FP) and fluorescent dyes staining could be a powerful tool for imaging living cells. However, the challenge is that the excitation and emission wavelength of these endogenous fluorophores and fluorescent labels are very different. A multi-color ultrafast source is required for the excitation of multiple fluorescence molecules. In this study, we developed a two-photon imaging system with excitations from the pump femtosecond laser and the selected supercontinuum generated from a photonic crystal fiber (PCF). Multiple endogenous fluorophores, fluorescent proteins and fluorescent dyes were excited in their optimal wavelengths simultaneously. A time- and spectral-resolved detection system was used to record the TPEF signals. This detection technique separated the TPEF signals from multiple sources in time and wavelength domains. Cellular organelles such as nucleus, mitochondria, microtubule and endoplasmic reticulum, were clearly revealed in the TPEF images. The simultaneous imaging of multiple fluorophores of cells will greatly aid the study of sub-cellular compartments and protein localization.

Photon and neutron interrogation techniques for chemical explosives detection in air cargo: A critical review

DOE Office of Scientific and Technical Information (OSTI.GOV)

Runkle, Robert C.; White, Timothy A.; Miller, Erin A.

Scanning cargo transported via aircraft ("air cargo") for explosive threats is a problem that, at present, lacks a comprehensive technical solution. While explosives detection in the baggage-scanning domain has a rich history that sheds light on potential solutions for air cargo, baggage scanning differs in several ways and thus one cannot look to the present array of technologies. Some contemporary solutions, like trace analysis, are not readily applied to cargo due to sampling challenges while the larger geometry of air cargo makes others less effective. This review article examines an array of interrogation techniques using photons and neutrons as incidentmore » particles. We first present a summary of the signatures and observables explosives provide and review how they have been exploited in baggage scanning. Following this is a description of the challenges posed by the air cargo application space. After considering interrogation sources, methods focused on transmission imaging, sub-surface examination and elemental characterization are described. It is our goal to shed light on the technical promise of each method while largely deferring questions that revolve around footprint, safety and conduct of operations. Our overarching intent is that a comprehensive understanding of potential techniques will foster development of a comprehensive solution.« less

Particle Image Velocimetry Applications Using Fluorescent Dye-Doped Particles

NASA Technical Reports Server (NTRS)

Petrosky, Brian J.; Maisto, Pietro; Lowe, K. Todd; Andre, Matthieu A.; Bardet, Philippe M.; Tiemsin, Patsy I.; Wohl, Christopher J.; Danehy, Paul M.

2015-01-01

Polystyrene latex sphere particles are widely used to seed flows for velocimetry techniques such as Particle Image Velocimetry (PIV) and Laser Doppler Velocimetry (LDV). These particles may be doped with fluorescent dyes such that signals spectrally shifted from the incident laser wavelength may be detected via Laser Induced Fluorescence (LIF). An attractive application of the LIF signal is achieving velocimetry in the presence of strong interference from laser scatter, opening up new research possibilities very near solid surfaces or at liquid/gas interfaces. Additionally, LIF signals can be used to tag different fluid streams to study mixing. While fluorescence-based PIV has been performed by many researchers for particles dispersed in water flows, the current work is among the first in applying the technique to micron-scale particles dispersed in a gas. A key requirement for such an application is addressing potential health hazards from fluorescent dyes; successful doping of Kiton Red 620 (KR620) has enabled the use of this relatively safe dye for fluorescence PIV for the first time. In this paper, basic applications proving the concept of PIV using the LIF signal from KR620-doped particles are exhibited for a free jet and a twophase flow apparatus. Results indicate that while the fluorescence PIV techniques are roughly 2 orders of magnitude weaker than Mie scattering, they provide a viable method for obtaining data in flow regions previously inaccessible via standard PIV. These techniques have the potential to also complement Mie scattering signals, for example in multi-stream and/or multi-phase experiments.

Luminescence and fluorescence of essential oils. Fluorescence imaging in vivo of wild chamomile oil

PubMed Central

Boschi, F.; Fontanella, M.; Calderan, L.; Sbarbati, A.

2011-01-01

Essential oils are currently of great importance to pharmaceutical companies, cosmetics producers and manufacturers of veterinary products. They are found in perfumes, creams, bath products, and household cleaning substances, and are used for flavouring food and drinks. It is well known that some of them act on the respiratory apparatus. The increasing interest in optical imaging techniques and the development of related technologies have made possible the investigation of the optical properties of several compounds. Luminescent properties of essential oils have not been extensively investigated. We evaluated the luminescent and fluorescent emissions of several essential oils, in order to detect them in living organisms by exploiting their optical properties. Some fluorescent emission data were high enough to be detected in dermal treatments. Consequently, we demonstrated how the fluorescent signal can be monitored for at least three hours on the skin of living mice treated with wild chamomile oil. The results encourage development of this technique to investigate the properties of drugs and cosmetics containing essential oils. PMID:22193298

Beyond radio-displacement techniques for Identification of CB1 Ligands: The First Application of a Fluorescence-quenching Assay

PubMed Central

Bruno, Agostino; Lembo, Francesca; Novellino, Ettore; Stornaiuolo, Mariano; Marinelli, Luciana

2014-01-01

Cannabinoid type 1 Receptor (CB1) belongs to the GPCR family and it has been targeted, so far, for the discovery of drugs aimed at the treatment of neuropathic pain, nausea, vomit, and food intake disorders. Here, we present the development of the first fluorescent assay enabling the measurement of kinetic binding constants for CB1orthosteric ligands. The assay is based on the use of T1117, a fluorescent analogue of AM251. We prove that T1117 binds endogenous and recombinant CB1 receptors with nanomolar affinity. Moreover, T1117 binding to CB1 is sensitive to the allosteric ligand ORG27569 and thus it is applicable to the discovery of new allosteric drugs. The herein presented assay constitutes a sustainable valid alternative to the expensive and environmental impacting radiodisplacement techniques and paves the way for an easy, fast and cheap high-throughput drug screening toward CB1 for identification of new orthosteric and allosteric modulators. PMID:24441508

Stereoscopic Planar Laser-Induced Fluorescence Imaging at 500 kHz

NASA Technical Reports Server (NTRS)

Medford, Taylor L.; Danehy, Paul M.; Jones, Stephen B.; Jiang, N.; Webster, M.; Lempert, Walter; Miller, J.; Meyer, T.

2011-01-01

A new measurement technique for obtaining time- and spatially-resolved image sequences in hypersonic flows is developed. Nitric-oxide planar laser-induced fluorescence (NO PLIF) has previously been used to investigate transition from laminar to turbulent flow in hypersonic boundary layers using both planar and volumetric imaging capabilities. Low flow rates of NO were typically seeded into the flow, minimally perturbing the flow. The volumetric imaging was performed at a measurement rate of 10 Hz using a thick planar laser sheet that excited NO fluorescence. The fluorescence was captured by a pair of cameras having slightly different views of the flow. Subsequent stereoscopic reconstruction of these images allowed the three-dimensional flow structures to be viewed. In the current paper, this approach has been extended to 50,000 times higher repetition rates. A laser operating at 500 kHz excites the seeded NO molecules, and a camera, synchronized with the laser and fitted with a beam-splitting assembly, acquires two separate images of the flow. The resulting stereoscopic images provide three-dimensional flow visualizations at 500 kHz for the first time. The 200 ns exposure time in each frame is fast enough to freeze the flow while the 500 kHz repetition rate is fast enough to time-resolve changes in the flow being studied. This method is applied to visualize the evolving hypersonic flow structures that propagate downstream of a discrete protuberance attached to a flat plate. The technique was demonstrated in the NASA Langley Research Center s 31-Inch Mach 10 Air Tunnel facility. Different tunnel Reynolds number conditions, NO flow rates and two different cylindrical protuberance heights were investigated. The location of the onset of flow unsteadiness, an indicator of transition, was observed to move downstream during the tunnel runs, coinciding with an increase in the model temperature.

Diode laser-based thermometry using two-line atomic fluorescence of indium and gallium

NASA Astrophysics Data System (ADS)

Borggren, Jesper; Weng, Wubin; Hosseinnia, Ali; Bengtsson, Per-Erik; Aldén, Marcus; Li, Zhongshan

2017-12-01

A robust and relatively compact calibration-free thermometric technique using diode lasers two-line atomic fluorescence (TLAF) for reactive flows at atmospheric pressures is investigated. TLAF temperature measurements were conducted using indium and, for the first time, gallium atoms as temperature markers. The temperature was measured in a multi-jet burner running methane/air flames providing variable temperatures ranging from 1600 to 2000 K. Indium and gallium were found to provide a similar accuracy of 2.7% and precision of 1% over the measured temperature range. The reliability of the TLAF thermometry was further tested by performing simultaneous rotational CARS measurements in the same experiments.

Trends in fluorescence imaging and related techniques to unravel biological information.

PubMed

Haustein, Elke; Schwille, Petra

2007-09-01

Optical microscopy is among the most powerful tools that the physical sciences have ever provided biology. It is indispensable for basic lab work, as well as for cutting edge research, as the visual monitoring of life processes still belongs to the most compelling evidences for a multitude of biomedical applications. Along with the rapid development of new probes and methods for the analysis of laser induced fluorescence, optical microscopy over past years experienced a vast increase of both new techniques and novel combinations of established methods to study biological processes with unprecedented spatial and temporal precision. On the one hand, major technical advances have significantly improved spatial resolution. On the other hand, life scientists are moving toward three- and even four-dimensional cell biology and biophysics involving time as a crucial coordinate to quantitatively understand living specimen. Monitoring the whole cell or tissue in real time, rather than producing snap-shot-like two-dimensional projections, will enable more physiological and, thus, more clinically relevant experiments, whereas an increase in temporal resolution facilitates monitoring fast nonperiodic processes as well as the quantitative analysis of characteristic dynamics.

Trends in fluorescence imaging and related techniques to unravel biological information

PubMed Central

Haustein, Elke; Schwille, Petra

2007-01-01

Optical microscopy is among the most powerful tools that the physical sciences have ever provided biology. It is indispensable for basic lab work, as well as for cutting edge research, as the visual monitoring of life processes still belongs to the most compelling evidences for a multitude of biomedical applications. Along with the rapid development of new probes and methods for the analysis of laser induced fluorescence, optical microscopy over past years experienced a vast increase of both new techniques and novel combinations of established methods to study biological processes with unprecedented spatial and temporal precision. On the one hand, major technical advances have significantly improved spatial resolution. On the other hand, life scientists are moving toward three- and even four-dimensional cell biology and biophysics involving time as a crucial coordinate to quantitatively understand living specimen. Monitoring the whole cell or tissue in real time, rather than producing snap-shot-like two-dimensional projections, will enable more physiological and, thus, more clinically relevant experiments, whereas an increase in temporal resolution facilitates monitoring fast nonperiodic processes as well as the quantitative analysis of characteristic dynamics. PMID:19404444

Development of a particle-trap preconcentration-soft ionization mass spectrometric technique for the quantification of mercury halides in air.

PubMed

Deeds, Daniel A; Ghoshdastidar, Avik; Raofie, Farhad; Guérette, Élise-Andrée; Tessier, Alain; Ariya, Parisa A

2015-01-01

Measurement of oxidized mercury, Hg(II), in the atmosphere poses a significant analytical challenge as Hg(II) is present at ultra-trace concentrations (picograms per cubic meter air). Current technologies are sufficiently sensitive to measure the total Hg present as Hg(II) but cannot determine the chemical speciation of Hg(II). We detail here the development of a soft ionization mass spectrometric technique coupled with preconcentration onto nano- or microparticle-based traps prior to analysis for the measurement of mercury halides in air. The current methodology has comparable detection limits (4-11 pg m(-3)) to previously developed techniques for the measurement of total inorganic mercury in air while allowing for the identification of HgX2 in collected samples. Both mercury chloride and mercury bromide have been sporadically detected in Montreal urban and indoor air using atmospheric pressure chemical ionization-mass spectrometry (APCI-MS). We discuss limitations and advantages of the current technique and discuss potential avenues for future research including quantitative trace measurements of a larger range of mercury compounds.

Histological methods to determine blood flow distribution with fluorescent microspheres.

PubMed

Luchtel, D L; Boykin, J C; Bernard, S L; Glenny, R W

1998-11-01

We evaluated several histological methods and determined their advantages and disadvantages for histological studies of tissues and organs perfused with fluorescent microspheres. Microspheres retained their fluorescence in 7-10 microm serial sections with a change in the antimedium from toluene when samples were fixed in formalin and embedded in paraffin. Several antimedia allowed both wax infiltration of tissue and preservation of microsphere fluorescence. Histoclear II was the best substitute for toluene. When samples were fixed in formalin and embedded in glycol methacrylate, thinner (3-5 microm) sections provided greater histological detail but had fewer microspheres per section. Air dried lung tissue followed by Vibratome sectioning provided thick sections (100 microm) that facilitated rapid survey of large volumes of tissue for microspheres but limited histological detail, and the air drying procedure was restricted to lung tissue. Samples fixed in formalin followed by Vibratome sectioning of unembedded tissue provided better histological detail of lung tissue and was also useful for other organs. These sections were more difficult to handle and to mount on slides compared to air dried tissue, whereas fixed tissue embedded in gelatin provided better tissue support for Vibratome sectioning. Rapid freezing followed by cryo-microtome sectioning resulted in frozen sections that were relatively difficult to handle compared to embedded or unembedded tissue; they also deteriorated relatively rapidly with time. Paraffin sections were stained with hematoxylin and eosin or with aqueous methyl green, although tissue autofluorescence by itself was usually sufficient to identify histological features. Methacrylate sections quenched tissue autofluorescence, and Lee's stain or Richardson's stain were used for staining sections. Toluene based mountants such as Cytoseal quenched fluorescence, particularly the red fluorescent microspheres. Aqueous based mountants such as

Estimation of the Invisible Energy in Extensive Air Showers with the Data Collected by the Pierre Auger Observatory

NASA Astrophysics Data System (ADS)

Mariazzi, Analisa

The determination of the energy of primary cosmic rays from their extensive air showers using the fluorescence technique requires an estimation of the energy carried away by particles reaching the ground that do not deposit all their energy in the atmosphere. This estimation is typically made using Monte Carlo simulations and depends on the assumed primary particle mass and on model predictions for hadron-air collisions at high energies. In this work we review the method that the Pierre Auger Collaboration uses to obtain the invisible energy directly from hybrid events measured simultaneously with the fluorescence and the surface detectors of the Pierre Auger Observatory. As a corroboration of these results, a new method for the determination of the invisible energy using an independent data set is also presented. Both methods agree within systematic uncertainties, reducing significantly the biases related to differences between the high energy hadronic interaction models and data.

Potential of Fluorescence Imaging Techniques To Monitor Mutagenic PAH Uptake by Microalga

PubMed Central

2015-01-01

Benzo[a]pyrene (BaP), a polycyclic aromatic hydrocarbon (PAH), is one of the major environmental pollutants that causes mutagenesis and cancer. BaP has been shown to accumulate in phytoplankton and zooplankton. We have studied the localization and aggregation of BaP in Chlorella sp., a microalga that is one of the primary producers in the food chain, using fluorescence confocal microscopy and fluorescence lifetime imaging microscopy with the phasor approach to characterize the location and the aggregation of BaP in the cell. Our results show that BaP accumulates in the lipid bodies of Chlorella sp. and that there is Förster resonance energy transfer between BaP and photosystems of Chlorella sp., indicating the close proximity of the two molecular systems. The lifetime of BaP fluorescence was measured to be 14 ns in N,N-dimethylformamide, an average of 7 ns in Bold’s basal medium, and 8 ns in Chlorella cells. Number and brightness analysis suggests that BaP does not aggregate inside Chlorella sp. (average brightness = 5.330), while it aggregates in the supernatant. In Chlorella grown in sediments spiked with BaP, in 12 h the BaP uptake could be visualized using fluorescence microscopy. PMID:25020149

Ultrafast fluorescence spectroscopy via upconversion applications to biophysics.

PubMed

Xu, Jianhua; Knutson, Jay R

2008-01-01

This chapter reviews basic concepts of nonlinear fluorescence upconversion, a technique whose temporal resolution is essentially limited only by the pulse width of the ultrafast laser. Design aspects for upconversion spectrophotofluorometers are discussed, and a recently developed system is described. We discuss applications in biophysics, particularly the measurement of time-resolved fluorescence spectra of proteins (with subpicosecond time resolution). Application of this technique to biophysical problems such as dynamics of tryptophan, peptides, proteins, and nucleic acids is reviewed.

New Approaches in Soil Organic Matter Fluorescence; A Solid Phase Fluorescence Approach

NASA Astrophysics Data System (ADS)

Bowman, M. M.; Sanclements, M.; McKnight, D. M.

2017-12-01

Fluorescence spectroscopy is a well-established technique to investigate the composition of organic matter in aquatic systems and is increasingly applied to soil organic matter (SOM). Current methods require that SOM be extracted into a liquid prior to analysis by fluorescence spectroscopy. Soil extractions introduce an additional layer of complexity as the composition of the organic matter dissolved into solution varies based upon the selected extractant. Water is one of the most commonly used extractant, but only extracts the water-soluble fraction of the SOM with the insoluble soil organic matter fluorescence remaining in the soil matrix. We propose the use of solid phase fluorescence on whole soils as a potential tool to look at the composition of organic matter without the extraction bias and gain a more complete understand of the potential for fluorescence as a tool in terrestrial studies. To date, the limited applications of solid phase fluorescence have ranged from food and agriculture to pharmaceutical with no clearly defined methods and limitations available. We are aware of no other studies that use solid phase fluorescence and thus no clear methods to look at SOM across a diverse set of soil types and ecosystems. With this new approach to fluorescence spectroscopy there are new challenges, such as blank correction, inner filter effect corrections, and sample preparation. This work outlines a novel method for analyzing soil organic matter using solid phase fluorescence across a wide range of soils collected from the National Ecological Observatory Network (NEON) eco-domains. This method has shown that organic matter content in soils must be diluted to 2% to reduce backscattering and oversaturation of the detector in forested soils. In mineral horizons (A) there is observed quenching of the humic-like organic matter, which is likely a result of organo-mineral complexation. Finally, we present preliminary comparisons between solid and liquid phase

Study of interaction of hypericin and its pharmaceutical preparation by fluorescence techniques.

PubMed

Liu, Jun; Saw, Constance Lay Lay; Olivo, Malini; Sudhaharan, Thankiah; Ahmed, Sohail; Heng, Paul Wan Sia; Wohland, Thorsten

2009-01-01

We report the detection of interactions between a photosensitizer, hypericin (HY), and its solvent system prepared with a formulation additive, polyvinylpyrrolidone (PVP), a commonly used pharmaceutical excipient. Fluorescence correlation spectroscopy (FCS) and fluorescence lifetime imaging microscopy (FLIM) were used to study aggregation and binding of HY in the presence of PVP. Digitized fluorescence endoscopic imaging (DFEI) was used to study the effect of the pharmaceutical formulation in the in vivo tumor implanted chick chorioallantoic membrane (CAM) model. The results presented reveal the coordination of HY-PVP binding, HY disaggregation in the presence of PVP, and strengthened HY tumor uptake selectivity. PVP is thus suggested as a potential adjuvant to previously investigated N-methyl pyrrolidone (NMP) in the HY delivery system as well as a replacement for the conventionally used albumin in the HY bladder instillation fluids preparation for clinical use.

Selective plane illumination microscopy (SPIM) with time-domain fluorescence lifetime imaging microscopy (FLIM) for volumetric measurement of cleared mouse brain samples

NASA Astrophysics Data System (ADS)

Funane, Tsukasa; Hou, Steven S.; Zoltowska, Katarzyna Marta; van Veluw, Susanne J.; Berezovska, Oksana; Kumar, Anand T. N.; Bacskai, Brian J.

2018-05-01

We have developed an imaging technique which combines selective plane illumination microscopy with time-domain fluorescence lifetime imaging microscopy (SPIM-FLIM) for three-dimensional volumetric imaging of cleared mouse brains with micro- to mesoscopic resolution. The main features of the microscope include a wavelength-adjustable pulsed laser source (Ti:sapphire) (near-infrared) laser, a BiBO frequency-doubling photonic crystal, a liquid chamber, an electrically focus-tunable lens, a cuvette based sample holder, and an air (dry) objective lens. The performance of the system was evaluated with a lifetime reference dye and micro-bead phantom measurements. Intensity and lifetime maps of three-dimensional human embryonic kidney (HEK) cell culture samples and cleared mouse brain samples expressing green fluorescent protein (GFP) (donor only) and green and red fluorescent protein [positive Förster (fluorescence) resonance energy transfer] were acquired. The results show that the SPIM-FLIM system can be used for sample sizes ranging from single cells to whole mouse organs and can serve as a powerful tool for medical and biological research.

Recommendations for fluorescence instrument qualification: the new ASTM Standard Guide.

PubMed

DeRose, Paul C; Resch-Genger, Ute

2010-03-01

Aimed at improving quality assurance and quantitation for modern fluorescence techniques, ASTM International (ASTM) is about to release a Standard Guide for Fluorescence, reviewed here. The guide's main focus is on steady state fluorometry, for which available standards and instrument characterization procedures are discussed along with their purpose, suitability, and general instructions for use. These include the most relevant instrument properties needing qualification, such as linearity and spectral responsivity of the detection system, spectral irradiance reaching the sample, wavelength accuracy, sensitivity or limit of detection for an analyte, and day-to-day performance verification. With proper consideration of method-inherent requirements and limitations, many of these procedures and standards can be adapted to other fluorescence techniques. In addition, procedures for the determination of other relevant fluorometric quantities including fluorescence quantum yields and fluorescence lifetimes are briefly introduced. The guide is a clear and concise reference geared for users of fluorescence instrumentation at all levels of experience and is intended to aid in the ongoing standardization of fluorescence measurements.

Portal vein territory identification using indocyanine green fluorescence imaging: Technical details and short-term outcomes.

PubMed

Kobayashi, Yuta; Kawaguchi, Yoshikuni; Kobayashi, Kosuke; Mori, Kazuhiro; Arita, Junichi; Sakamoto, Yoshihiro; Hasegawa, Kiyoshi; Kokudo, Norihiro

2017-12-01

Portal vein (PV) territory identification during liver resection may be performed using indocyanine green (ICG) fluorescence imaging technique. However, the technical details of the fluorescence staining technique have not been fully elucidated. This study was performed to demonstrate the technical details of PV territory identification using fluorescence imaging and evaluates the short-term outcomes. From 2011 to 2015, 105 underwent liver resection at the University of Tokyo Hospital with one of the following fluorescence staining techniques by transhepatic PV injection or intravenous injection of ICG: single staining (n = 36), multiple staining (n = 31), counterstaining (n = 22), negative staining (n = 13), or paradoxical negative staining (n = 3). The PV territory was identified as a region with fluorescence or a defect of fluorescence using one of the five staining techniques. ICG was administered by transhepatic PV injection in all but the negative staining technique, which employed intravenous injection. No adverse events associated with the ICG administration occurred. The mortality, postoperative total morbidity, and the major complication (Clavien-Dindo grade ≥III) rates were 0.0%, 14.3%, and 7.6%. We have demonstrated the technical details of five types of fluorescence staining techniques. These techniques are safe to perform and facilitate clear visualization of the PV territory in real time, enhancing the efficacy of anatomical removal of such territories. © 2017 Wiley Periodicals, Inc.

Fluorescence Characterization of Clinically-Important Bacteria

PubMed Central

Dartnell, Lewis R.; Roberts, Tom A.; Moore, Ginny; Ward, John M.; Muller, Jan-Peter

2013-01-01

Healthcare-associated infections (HCAI/HAI) represent a substantial threat to patient health during hospitalization and incur billions of dollars additional cost for subsequent treatment. One promising method for the detection of bacterial contamination in a clinical setting before an HAI outbreak occurs is to exploit native fluorescence of cellular molecules for a hand-held, rapid-sweep surveillance instrument. Previous studies have shown fluorescence-based detection to be sensitive and effective for food-borne and environmental microorganisms, and even to be able to distinguish between cell types, but this powerful technique has not yet been deployed on the macroscale for the primary surveillance of contamination in healthcare facilities to prevent HAI. Here we report experimental data for the specification and design of such a fluorescence-based detection instrument. We have characterized the complete fluorescence response of eleven clinically-relevant bacteria by generating excitation-emission matrices (EEMs) over broad wavelength ranges. Furthermore, a number of surfaces and items of equipment commonly present on a ward, and potentially responsible for pathogen transfer, have been analyzed for potential issues of background fluorescence masking the signal from contaminant bacteria. These include bedside handrails, nurse call button, blood pressure cuff and ward computer keyboard, as well as disinfectant cleaning products and microfiber cloth. All examined bacterial strains exhibited a distinctive double-peak fluorescence feature associated with tryptophan with no other cellular fluorophore detected. Thus, this fluorescence survey found that an emission peak of 340nm, from an excitation source at 280nm, was the cellular fluorescence signal to target for detection of bacterial contamination. The majority of materials analysed offer a spectral window through which bacterial contamination could indeed be detected. A few instances were found of potential problems

A simple semi-empirical technique for apportioning the impact of roadways on air quality in an urban neighbourhood

NASA Astrophysics Data System (ADS)

Elangasinghe, M. A.; Dirks, K. N.; Singhal, N.; Costello, S. B.; Longley, I.; Salmond, J. A.

2014-02-01

Air pollution from the transport sector has a marked effect on human health, so isolating the pollutant contribution from a roadway is important in understanding its impact on the local neighbourhood. This paper proposes a novel technique based on a semi-empirical air pollution model to quantify the impact from a roadway on the air quality of a local neighbourhood using ambient records of a single air pollution monitor. We demonstrate the proposed technique using a case study, in which we quantify the contribution from a major highway with respect to the local background concentration in Auckland, New Zealand. Comparing the diurnal variation of the model-separated background contribution with real measurements from a site upwind of the highway shows that the model estimates are reliable. Amongst all of the pollutants considered, the best estimations of the background were achieved for nitrogen oxides. Although the multi-pronged approach worked well for predominantly vehicle-related pollutants, it could not be used effectively to isolate emissions of PM10 due to the complex and less predictable influence of natural sources (such as marine aerosols). The proposed approach is useful in situations where ambient records from an upwind background station are not available (as required by other techniques) and is potentially transferable to situations such as intersections and arterial roads. Applying this technique to longer time series could help to understand the changes in pollutant concentrations from the road and background sources for different emission scenarios, for different years or seasons. Modelling results also show the potential of such a hybrid semi-empirical models to contribute to our understanding of the physical parameters determining air quality and to validate emissions inventory data.

Hyperspectral small animal fluorescence imaging: spectral selection imaging

NASA Astrophysics Data System (ADS)

Leavesley, Silas; Jiang, Yanan; Patsekin, Valery; Hall, Heidi; Vizard, Douglas; Robinson, J. Paul

2008-02-01

Molecular imaging is a rapidly growing area of research, fueled by needs in pharmaceutical drug-development for methods for high-throughput screening, pre-clinical and clinical screening for visualizing tumor growth and drug targeting, and a growing number of applications in the molecular biology fields. Small animal fluorescence imaging employs fluorescent probes to target molecular events in vivo, with a large number of molecular targeting probes readily available. The ease at which new targeting compounds can be developed, the short acquisition times, and the low cost (compared to microCT, MRI, or PET) makes fluorescence imaging attractive. However, small animal fluorescence imaging suffers from high optical scattering, absorption, and autofluorescence. Much of these problems can be overcome through multispectral imaging techniques, which collect images at different fluorescence emission wavelengths, followed by analysis, classification, and spectral deconvolution methods to isolate signals from fluorescence emission. We present an alternative to the current method, using hyperspectral excitation scanning (spectral selection imaging), a technique that allows excitation at any wavelength in the visible and near-infrared wavelength range. In many cases, excitation imaging may be more effective at identifying specific fluorescence signals because of the higher complexity of the fluorophore excitation spectrum. Because the excitation is filtered and not the emission, the resolution limit and image shift imposed by acousto-optic tunable filters have no effect on imager performance. We will discuss design of the imager, optimizing the imager for use in small animal fluorescence imaging, and application of spectral analysis and classification methods for identifying specific fluorescence signals.

Spirally-patterned pinhole arrays for long-term fluorescence cell imaging.

PubMed

Koo, Bon Ung; Kang, YooNa; Moon, SangJun; Lee, Won Gu

2015-11-07

Fluorescence cell imaging using a fluorescence microscope is an extensively used technique to examine the cell nucleus, internal structures, and other cellular molecules with fluorescence response time and intensity. However, it is difficult to perform high resolution cell imaging for a long period of time with this technique due to necrosis and apoptosis depending on the type and subcellular location of the damage caused by phototoxicity. A large number of studies have been performed to resolve this problem, but researchers have struggled to meet the challenge between cellular viability and image resolution. In this study, we employ a specially designed disc to reduce cell damage by controlling total fluorescence exposure time without deterioration of the image resolution. This approach has many advantages such as, the apparatus is simple, cost-effective, and easily integrated into the optical pathway through a conventional fluorescence microscope.

Directed molecular evolution to design advanced red fluorescent proteins.

PubMed

Subach, Fedor V; Piatkevich, Kiryl D; Verkhusha, Vladislav V

2011-11-29

Fluorescent proteins have become indispensable imaging tools for biomedical research. Continuing progress in fluorescence imaging, however, requires probes with additional colors and properties optimized for emerging techniques. Here we summarize strategies for development of red-shifted fluorescent proteins. We discuss possibilities for knowledge-based rational design based on the photochemistry of fluorescent proteins and the position of the chromophore in protein structure. We consider advances in library design by mutagenesis, protein expression systems and instrumentation for high-throughput screening that should yield improved fluorescent proteins for advanced imaging applications.

Air versus saline in the loss of resistance technique for identification of the epidural space.

PubMed

Antibas, Pedro L; do Nascimento Junior, Paulo; Braz, Leandro G; Vitor Pereira Doles, João; Módolo, Norma S P; El Dib, Regina

2014-07-18

The success of epidural anaesthesia depends on correct identification of the epidural space. For several decades, the decision of whether to use air or physiological saline during the loss of resistance technique for identification of the epidural space has been governed by the personal experience of the anaesthesiologist. Epidural block remains one of the main regional anaesthesia techniques. It is used for surgical anaesthesia, obstetrical analgesia, postoperative analgesia and treatment of chronic pain and as a complement to general anaesthesia. The sensation felt by the anaesthesiologist from the syringe plunger with loss of resistance is different when air is compared with saline (fluid). Frequently fluid allows a rapid change from resistance to non-resistance and increased movement of the plunger. However, the ideal technique for identification of the epidural space remains unclear. • To evaluate the efficacy and safety of both air and saline in the loss of resistance technique for identification of the epidural space.• To evaluate complications related to the air or saline injected. We searched the Cochrane Central Register of Controlled Trials (CENTRAL) (2013, Issue 9), MEDLINE, EMBASE and the Latin American and Caribbean Health Science Information Database (LILACS) (from inception to September 2013). We applied no language restrictions. The date of the most recent search was 7 September 2013. We included randomized controlled trials (RCTs) and quasi-randomized controlled trials (quasi-RCTs) on air and saline in the loss of resistance technique for identification of the epidural space. Two review authors independently assessed trial quality and extracted data. We included in the review seven studies with a total of 852 participants. The methodological quality of the included studies was generally ranked as showing low risk of bias in most domains, with the exception of one study, which did not mask participants. We were able to include data from 838

A Preliminary Study of Krypton Laser-Induced Fluorescence

DTIC Science & Technology

2010-07-01

Induced Fluorescence 5b. GRANT NUMBER 5c. PROGRAM ELEMENT NUMBER 6. AUTHOR(S) William A. Hargus, Jr. (AFRL/RZSS) 5d. PROJECT NUMBER R 5e. TASK...replacement for xenon. This study examines the potential applications of laser-induced fluorescence as a plasma diagnostic technique for Kr I and Kr...II. Candidate electronic transitions are examined to determine their suitability for successful routine application of laser-induced fluorescence

Optical calibration of the Auger fluorescence telescopes

NASA Astrophysics Data System (ADS)

Matthews, John A. J.

2003-02-01

The Pierre Auger Observatory is optimized to study the cosmic ray spectrum in the region of the Greisen-Zatsepin-Kuz'min (GZK) cutoff, i.e.cosmic rays with energies of ~1020eV. Cosmic rays are detected as extensive air showers. To measure these showers each Auger site combines a 3000sq-km ground array with air fluorescence telescopes into a hybrid detector. Our design choice is motivated by the heightened importance of the energy scale, and related systematic uncertainties in shower energies, for experiments investigating the GZK cutoff. This paper focuses on the optical calibration of the Auger fluorescence telescopes. The optical calibration is done three independent ways: an absolute end-to-end calibration using a uniform, calibrated intensity, light-source at the telescope entrance aperture, a component by component calibration using both laboratory and in-situ measurements, and Rayleigh scattered light from external laser beams. The calibration concepts and related instrumentation are summarized. Results from the 5-month engineering array test are presented.

Doppler ultrasound surveillance in deep tunneling compressed-air work with Trimix breathing: bounce dive technique compared to saturation-excursion technique.

PubMed

Vellinga, T P van Rees; Sterk, W; de Boer, A G E M; van der Beek, A J; Verhoeven, A C; van Dijk, F J H

2008-01-01

The Western Scheldt Tunneling Project in The Netherlands provided a unique opportunity to evaluate two deep-diving techniques with Doppler ultrasound surveillance. Divers used the bounce diving techniques for repair and maintenance of the TBM. The tunnel boring machine jammed at its deepest depth. As a result the work time was not sufficient. The saturation diving technique was developed and permitted longer work time at great depth. Thirty-one divers were involved in this project. Twenty-three divers were examined using Doppler ultrasound. Data analysis addressed 52 exposures to Trimix at 4.6-4.8 bar gauge using the bounce technique and 354 exposures to Trimix at 4.0-6.9 bar gauge on saturation excursions. No decompression incidents occurred with either technique during the described phase of the project. Doppler ultrasound revealed that the bubble loads assessed in both techniques were generally low. We find out, that despite longer working hours, shorter decompression times and larger physical workloads, the saturation-excursion technique was associated with significant lower bubble grades than in the bounce technique using Doppler Ultrasound. We conclude that the saturation-excursion technique with Trimix is a good option for deep and long exposures in caisson work. The Doppler technique proved valuable, and it should be incorporated in future compressed-air work.

77 FR 58953 - Approval and Promulgation of Air Quality Implementation Plans; Delaware; Control Technique...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-09-25

... Promulgation of Air Quality Implementation Plans; Delaware; Control Technique Guidelines for Plastic Parts... categories: Plastic Parts, Metal Furniture, Large Appliances, and Miscellaneous Metal Parts. EPA is approving... Compounds, sections 2.0 ``Definitions,'' 12.0 ``Surface Coating of Plastic Parts,'' 19.0 ``Coating of Metal...

NaOCl degradation of a HEMA-free all-in-one adhesive bonded to enamel and dentin following two air-blowing techniques.

PubMed

De Munck, Jan; Ermis, R Banu; Koshiro, Kenichi; Inoue, Satoshi; Ikeda, Takatsumi; Sano, Hidehiko; Van Landuyt, Kirsten L; Van Meerbeek, Bart

2007-01-01

Phase-separation within HEMA-free all-in-one dental adhesives may result in the entrapment of droplets within the adhesive resin. Strongly air-blowing prior to polymerization, can remove most of these droplets. The objective of this study was to evaluate the effect these droplets may have on the resistance of the adhesive-tooth interface to NaOCl degradation. The micro-tensile bond strength (microTBS) to enamel and dentin was determined when a HEMA-free all-in-one adhesive was applied either following a mild or strong air-blowing technique. The bonds were also exposed to an aqueous sodium hypochlorite (NaOCl) solution for 1h, following a recently introduced methodology to mimic in vivo bond degradation. This study revealed that strong air-blowing of the adhesive only resulted in a significantly higher micro-tensile bond strength (microTBS) to dentin, but not to enamel. Likewise, NaOCl only reduced the microTBS to dentin for both the mild and strong air-blowing technique, but again not the microTBS to enamel. Failure analysis by SEM clearly revealed that strong air-blowing is less effective in droplet removal when the adhesive was applied in small and narrow class-I cavities, as compared to when it was applied to flat surfaces. NaOCl did preferentially dissolve the hybrid layer at dentin, and more for the mild than for the strong air-blowing technique. A strong air-blowing procedure resulted in a more NaOCl-resistant hybrid layer, so that it can be concluded that a HEMA-free one-step adhesive definitely benefits from a strong air-blowing technique.

Evaluation of OH laser-induced fluorescence techniques for supersonic combustion diagnostics

NASA Technical Reports Server (NTRS)

Quagliaroli, T. M.; Laufer, G.; Krauss, R. H.; Mcdaniel, J. C., Jr.

1992-01-01

The limitations on application of dye laser and narrowband tunable KrF excimer laser systems to planar OH fluorescence measurements in supersonic combustion test facilities are examined. Included in the analysis are effects of collisional quenching, beam absorption, fluorescence trapping, and signal strengths on achievable measurement accuracy using several excitation and detection options for either of the two laser systems. Dye-based laser systems are found to be the method of choice for imaging OH concentrations less than 10 exp 15 per cu cm, while the KrF based systems provide significant reduction in measurement ambiguity for concentrations in excess of 10 exp 15 per cu cm.

Ultrafast Fluorescence Spectroscopy via Upconversion: Applications to Biophysics

PubMed Central

Xu, Jianhua; Knutson, Jay R.

2012-01-01

This chapter reviews basic concepts of nonlinear fluorescence upconversion, a technique whose temporal resolution is essentially limited only by the pulse width of the ultrafast laser. Design aspects for upconversion spectrophotofluorometers are discussed, and a recently developed system is described. We discuss applications in biophysics, particularly the measurement of time-resolved fluorescence spectra of proteins (with subpicosecond time resolution). Application of this technique to biophysical problems such as dynamics of tryptophan, peptides, proteins, and nucleic acids is reviewed. PMID:19152860

Fluorescence fluctuation spectroscopy for clinical applications

NASA Astrophysics Data System (ADS)

Olson, Eben

Fluorescence correlation spectroscopy (FCS) and the related techniques of brightness analysis have become standard tools in biological and biophysical research. By analyzing the statistics of fluorescence emitted from a restricted volume, a number of parameters including concentrations, diffusion coefficients and chemical reaction rates can be determined. The single-molecule sensitivity, spectral selectivity, small sample volume and non-perturbative measurement mechanism of FCS make it an excellent technique for the study of molecular interactions. However, its adoption outside of the research laboratory has been limited. Potential reasons for this include the cost and complexity of the required apparatus. In this work, the application of fluorescence fluctuation analysis to several clinical problems is considered. Optical designs for FCS instruments which reduce the cost and increase alignment tolerance are presented. Brightness analysis of heterogenous systems, with application to the characterization of protein aggregates and multimer distributions, is considered. Methods for FCS-based assays of two clinically relevant proteins, von Willebrand factor and haptoglobin, are presented as well.

Fluorescence imaging of angiogenesis in green fluorescent protein-expressing tumors

NASA Astrophysics Data System (ADS)

Yang, Meng; Baranov, Eugene; Jiang, Ping; Li, Xiao-Ming; Wang, Jin W.; Li, Lingna; Yagi, Shigeo; Moossa, A. R.; Hoffman, Robert M.

2002-05-01

The development of therapeutics for the control of tumor angiogenesis requires a simple, reliable in vivo assay for tumor-induced vascularization. For this purpose, we have adapted the orthotopic implantation model of angiogenesis by using human and rodent tumors genetically tagged with Aequorea victoria green fluorescent protein (GFP) for grafting into nude mice. Genetically-fluorescent tumors can be readily imaged in vivo. The non-luminous induced capillaries are clearly visible against the bright tumor fluorescence examined either intravitally or by whole-body luminance in real time. Fluorescence shadowing replaces the laborious histological techniques for determining blood vessel density. High-level GFP-expressing tumor cell lines made it possible to acquire the high-resolution real-time fluorescent optical images of angiogenesis in both primary tumors and their metastatic lesions in various human and rodent tumor models by means of a light-based imaging system. Intravital images of angiogenesis onset and development were acquired and quantified from a GFP- expressing orthotopically-growing human prostate tumor over a 19-day period. Whole-body optical imaging visualized vessel density increasing linearly over a 20-week period in orthotopically-growing, GFP-expressing human breast tumor MDA-MB-435. Vessels in an orthotopically-growing GFP- expressing Lewis lung carcinoma tumor were visualized through the chest wall via a reversible skin flap. These clinically-relevant angiogenesis mouse models can be used for real-time in vivo evaluation of agents inhibiting or promoting tumor angiogenesis in physiological micro- environments.

Contribution of glue layer into epidermis sample fluorescence dynamics

NASA Astrophysics Data System (ADS)

Salomatina, Elena V.; Chernova, Svetlana P.; Pravdin, Alexander B.

2000-04-01

In this work, the temporal behavior of autofluorescence of epidermis samples under UV-irradiation has ben studied. The samples were prepared using surface epidermis stripping technique. Fluorescence spectra and kinetic curves of fluorescence intensity have been obtained. It has been concluded that the glue composition used allows the measurement of epidermis fluorescence dynamics with the first 60 min of experiment.

Development of a neutral embedding resin for optical imaging of fluorescently labeled biological tissue

NASA Astrophysics Data System (ADS)

Zhou, Hongfu; Gang, Yadong; Chen, Shenghua; Wang, Yu; Xiong, Yumiao; Li, Longhui; Yin, Fangfang; Liu, Yue; Liu, Xiuli; Zeng, Shaoqun

2017-10-01

Plastic embedding is widely applied in light microscopy analyses. Previous studies have shown that embedding agents and related techniques can greatly affect the quality of biological tissue embedding and fluorescent imaging. Specifically, it is difficult to preserve endogenous fluorescence using currently available acidic commercial embedding resins and related embedding techniques directly. Here, we developed a neutral embedding resin that improved the green fluorescent protein (GFP), yellow fluorescent protein (YFP), and DsRed fluorescent intensity without adjusting the pH value of monomers or reactivating fluorescence in lye. The embedding resin had a high degree of polymerization, and its fluorescence preservation ratios for GFP, YFP, and DsRed were 126.5%, 155.8%, and 218.4%, respectively.

Laser-induced fluorescence in the detection of esophageal carcinoma

NASA Astrophysics Data System (ADS)

Wang, Kenneth K.; Gutta, Kumar; Laukka, Mark A.; Densmore, John

1995-01-01

Laser induced fluorescence (LIF) is a technique which can perform an 'optical biopsy' of gastrointestinal mucosa. LIF was performed in resected specimens using a pulsed N2-laser coupled fiberoptically to a probe. Fluorescence was measured using a 0.2 meter spectroscope with an intensified photodiode array. Measurements were made on fresh (<30 minutes after resection) esophageal specimens containing normal mucosa, Barrett's esophagus, and adenocarcinoma. Each tissue section was examined using an optical probe consisting of a central fiber for delivering the excitation energy and a 6 fiber bundle surrounding the central fiber for detection of the fluorescence. An excitation wavelength of 337 nm was used which generated 3-ns pulses while fluorescence intensities were acquired from 300-800 nm. Spectra were obtained from each section in a standardized fashion and background spectra subtracted. Fluorescence readings were taken from 54 normal esophageal sections and 32 sections of adenocarcinoma. A fluorescence index obtained from the tumor sections was 0.68+/- 0.01 compared with 0.51+/- 0.01 for the normal sections (p<0.001). Using a discriminant value of 0.65, this technique had a sensitivity of 81% and a specificity of 100% for detection of malignant tissue. The positive predictive value was 100% and the negative predictive value was 90% for an overall accuracy of 93%. LIF is a promising technique which has the capability of distinguishing normal versus malignant tissue in the esophagus with good accuracy.

Activatable fluorescent probes in fluorescence-guided surgery: Practical considerations.

PubMed

Mochida, Ai; Ogata, Fusa; Nagaya, Tadanobu; Choyke, Peter L; Kobayashi, Hisataka

2018-02-15

Fluorescence-guided imaging during surgery is a promising technique that is increasingly used to aid surgeons in identifying sites of tumor and surgical margins. Of the two types of fluorescent probes, always-on and activatable, activatable probes are preferred because they produce higher target-to-background ratios, thus improving sensitivity compared with always-on probes that must contend with considerable background signal. There are two types of activatable probes: 1) enzyme-reactive probes that are normally quenched but can be activated after cleavage by cancer-specific enzymes (activity-based probes) and 2) molecular-binding probes which use cancer targeting moieties such as monoclonal antibodies to target receptors found in abundance on cancers and are activated after internalization and lysosomal processing (binding-based probes). For fluorescence-guided intraoperative surgery, enzyme-reactive probes are superior because they can react quickly, require smaller dosages especially for topical applications, have limited side effects, and have favorable pharmacokinetics. Enzyme-reactive probes are easier to use, fit better into existing work flows in the operating room and have minimal toxicity. Although difficult to prove, it is assumed that the guidance provided to surgeons by these probes results in more effective surgeries with better outcomes for patients. In this review, we compare these two types of activatable fluorescent probes for their ease of use and efficacy. Published by Elsevier Ltd.

Synchronous fluorescence spectroscopy for analysis of wine and wine distillates

NASA Astrophysics Data System (ADS)

Andreeva, Ya.; Borisova, E.; Genova, Ts.; Zhelyazkova, Al.; Avramov, L.

2015-01-01

Wine and brandies are multicomponent systems and conventional fluorescence techniques, relying on recording of single emission or excitation spectra, are often insufficient. In such cases synchronous fluorescence spectra can be used for revealing the potential of the fluorescence techniques. The technique is based on simultaneously scanning of the excitation and emission wavelength with constant difference (Δλ) maintained between them. In this study the measurements were made using FluoroLog3 spectrofluorimeter (HORIBA Jobin Yvon, France) and collected for excitation and emission in the wavelength region 220 - 700 nm using wavelength interval Δλ from 10 to 100 nm in 10 nm steps. This research includes the results obtained for brandy and red wine samples. Fluorescence analysis takes advantage in the presence of natural fluorophores in wines and brandies, such as gallic, vanillic, p-coumaric, syringic, ferulic acid, umbelliferone, scopoletin and etc. Applying of synchronous fluorescence spectroscopy for analysis of these types of alcohols allows us to estimate the quality of wines and also to detect adulteration of brandies like adding of a caramel to wine distillates for imitating the quality of the original product aged in oak casks.

78 FR 34306 - Approval and Promulgation of Air Quality Implementation Plans: North Carolina; Control Techniques...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-06-07

... Promulgation of Air Quality Implementation Plans: North Carolina; Control Techniques Guidelines and Reasonably Available Control Technology AGENCY: Environmental Protection Agency (EPA). ACTION: Proposed rule. SUMMARY... Carolina's commitment associated with the conditional approval of its reasonably available control...

Food-Growing, Air- And Water-Cleaning Module

NASA Technical Reports Server (NTRS)

Sauer, R. L.; Scheld, H. W.; Mafnuson, J. W.

1988-01-01

Apparatus produces fresh vegetables and removes pollutants from air. Hydroponic apparatus performs dual function of growing fresh vegetables and purifying air and water. Leafy vegetables rooted in granular growth medium grow in light of fluorescent lamps. Air flowing over leaves supplies carbon dioxide and receives fresh oxygen from them. Adaptable to production of food and cleaning of air and water in closed environments as in underwater research stations and submarines.

An air flow sensor for neonatal mechanical ventilation applications based on a novel fiber-optic sensing technique

DOE Office of Scientific and Technical Information (OSTI.GOV)

Battista, L.; Sciuto, S. A.; Scorza, A.

2013-03-15

In this work, a simple and low-cost air flow sensor, based on a novel fiber-optic sensing technique has been developed for monitoring air flows rates supplied by a neonatal ventilator to support infants in intensive care units. The device is based on a fiber optic sensing technique allowing (a) the immunity to light intensity variations independent by measurand and (b) the reduction of typical shortcomings affecting all biomedical fields (electromagnetic interference and patient electrical safety). The sensing principle is based on the measurement of transversal displacement of an emitting fiber-optic cantilever due to action of air flow acting on it;more » the fiber tip displacement is measured by means of a photodiode linear array, placed in front of the entrance face of the emitting optical fiber in order to detect its light intensity profile. As the measurement system is based on a detection of the illumination pattern, and not on an intensity modulation technique, it results less sensitive to light intensity fluctuation independent by measurand than intensity-based sensors. The considered technique is here adopted in order to develop two different configurations for an air flow sensor suitable for the measurement of air flow rates typically occurring during mechanical ventilation of newborns: a mono-directional and a bi-directional transducer have been proposed. A mathematical model for the air flow sensor is here proposed and a static calibration of two different arrangements has been performed: a measurement range up to 3.00 Multiplication-Sign 10{sup -4} m{sup 3}/s (18.0 l/min) for the mono-directional sensor and a measurement range of {+-}3.00 Multiplication-Sign 10{sup -4} m{sup 3}/s ({+-}18.0 l/min) for the bi-directional sensor are experimentally evaluated, according to the air flow rates normally encountered during tidal breathing of infants with a mass lower than 10 kg. Experimental data of static calibration result in accordance with the proposed

An air flow sensor for neonatal mechanical ventilation applications based on a novel fiber-optic sensing technique.

PubMed

Battista, L; Sciuto, S A; Scorza, A

2013-03-01

In this work, a simple and low-cost air flow sensor, based on a novel fiber-optic sensing technique has been developed for monitoring air flows rates supplied by a neonatal ventilator to support infants in intensive care units. The device is based on a fiber optic sensing technique allowing (a) the immunity to light intensity variations independent by measurand and (b) the reduction of typical shortcomings affecting all biomedical fields (electromagnetic interference and patient electrical safety). The sensing principle is based on the measurement of transversal displacement of an emitting fiber-optic cantilever due to action of air flow acting on it; the fiber tip displacement is measured by means of a photodiode linear array, placed in front of the entrance face of the emitting optical fiber in order to detect its light intensity profile. As the measurement system is based on a detection of the illumination pattern, and not on an intensity modulation technique, it results less sensitive to light intensity fluctuation independent by measurand than intensity-based sensors. The considered technique is here adopted in order to develop two different configurations for an air flow sensor suitable for the measurement of air flow rates typically occurring during mechanical ventilation of newborns: a mono-directional and a bi-directional transducer have been proposed. A mathematical model for the air flow sensor is here proposed and a static calibration of two different arrangements has been performed: a measurement range up to 3.00 × 10(-4) m(3)∕s (18.0 l∕min) for the mono-directional sensor and a measurement range of ±3.00 × 10(-4) m(3)∕s (±18.0 l∕min) for the bi-directional sensor are experimentally evaluated, according to the air flow rates normally encountered during tidal breathing of infants with a mass lower than 10 kg. Experimental data of static calibration result in accordance with the proposed theoretical model: for the mono

An air flow sensor for neonatal mechanical ventilation applications based on a novel fiber-optic sensing technique

NASA Astrophysics Data System (ADS)

Battista, L.; Sciuto, S. A.; Scorza, A.

2013-03-01

In this work, a simple and low-cost air flow sensor, based on a novel fiber-optic sensing technique has been developed for monitoring air flows rates supplied by a neonatal ventilator to support infants in intensive care units. The device is based on a fiber optic sensing technique allowing (a) the immunity to light intensity variations independent by measurand and (b) the reduction of typical shortcomings affecting all biomedical fields (electromagnetic interference and patient electrical safety). The sensing principle is based on the measurement of transversal displacement of an emitting fiber-optic cantilever due to action of air flow acting on it; the fiber tip displacement is measured by means of a photodiode linear array, placed in front of the entrance face of the emitting optical fiber in order to detect its light intensity profile. As the measurement system is based on a detection of the illumination pattern, and not on an intensity modulation technique, it results less sensitive to light intensity fluctuation independent by measurand than intensity-based sensors. The considered technique is here adopted in order to develop two different configurations for an air flow sensor suitable for the measurement of air flow rates typically occurring during mechanical ventilation of newborns: a mono-directional and a bi-directional transducer have been proposed. A mathematical model for the air flow sensor is here proposed and a static calibration of two different arrangements has been performed: a measurement range up to 3.00 × 10-4 m3/s (18.0 l/min) for the mono-directional sensor and a measurement range of ±3.00 × 10-4 m3/s (±18.0 l/min) for the bi-directional sensor are experimentally evaluated, according to the air flow rates normally encountered during tidal breathing of infants with a mass lower than 10 kg. Experimental data of static calibration result in accordance with the proposed theoretical model: for the mono-directional configuration, the

Fluorescent speckle microscopy of microtubules: how low can you go?

PubMed

Waterman-Storer, C M; Salmon, E D

1999-12-01

Fluorescent speckle microscopy (FSM) is a new technique for visualizing the movement, assembly, and turnover of macromolecular assemblies like the cytoskeleton in living cells. In this method, contrast is created by coassembly of a small fraction of fluorescent subunits in a pool of unlabeled subunits. Random variation in association creates a nonuniform "fluorescent speckle" pattern. Fluorescent speckle movements in time-lapse recordings stand out to the eye and can be measured. Because fluorescent speckles represent fiduciary marks on the polymer lattice, FSM provides the opportunity for the first time to see the 2- and 3-dimensional trajectories of lattice movements within large arrays of polymers as well as identifying sites of assembly and disassembly of individual polymers. The technique works with either microinjection of fluorescently labeled subunits or expression of subunits ligated to green fluorescent protein (GFP). We have found for microtubules assembled in vitro that speckles containing one fluorophore can be detected and recorded using a conventional wide-field epi-fluorescence light microscope and digital imaging with a low noise cooled CCD camera. In living cells, optimal speckle contrast occurs at fractions of labeled tubulin of approximately 0.1-0.5% where the fluorescence of each speckle corresponds to one to seven fluorophores per resolvable unit (approximately 0.27 microm) in the microscope. This small fraction of labeled subunits significantly reduces out-of-focus fluorescence and greatly improves visibility of fluorescently labeled structures and their dynamics in thick regions of living cells.

Intercomparison of four different in-situ techniques for ambient formaldehyde measurements in urban air

NASA Astrophysics Data System (ADS)

Hak, C.; Pundt, I.; Trick, S.; Kern, C.; Platt, U.; Dommen, J.; Ordóñez, C.; Prévôt, A. S. H.; Junkermann, W.; Astorga-Lloréns, C.; Larsen, B. R.; Mellqvist, J.; Strandberg, A.; Yu, Y.; Galle, B.; Kleffmann, J.; Lörzer, J. C.; Braathen, G. O.; Volkamer, R.

2005-11-01

Results from an intercomparison of several currently used in-situ techniques for the measurement of atmospheric formaldehyde (CH2O) are presented. The measurements were carried out at Bresso, an urban site in the periphery of Milan (Italy) as part of the FORMAT-I field campaign. Eight instruments were employed by six independent research groups using four different techniques: Differential Optical Absorption Spectroscopy (DOAS), Fourier Transform Infra Red (FTIR) interferometry, the fluorimetric Hantzsch reaction technique (five instruments) and a chromatographic technique employing C18-DNPH-cartridges (2,4-dinitrophenylhydrazine). White type multi-reflection systems were employed for the optical techniques in order to avoid spatial CH2O gradients and ensure the sampling of nearly the same air mass by all instruments. Between 23 and 31 July 2002, up to 13 ppbv of CH2O were observed. The concentrations lay well above the detection limits of all instruments. The formaldehyde concentrations determined with DOAS, FTIR and the Hantzsch instruments were found to agree within ±11%, with the exception of one Hantzsch instrument, which gave systematically higher values. The two hour integrated samples by DNPH yielded up to 25% lower concentrations than the data of the continuously measuring instruments averaged over the same time period. The consistency between the DOAS and the Hantzsch method was better than during previous intercomparisons in ambient air with slopes of the regression line not significantly differing from one. The differences between the individual Hantzsch instruments could be attributed in part to the calibration standards used. Possible systematic errors of the methods are discussed.

Intercomparison of four different in-situ techniques for ambient formaldehyde measurements in urban air

NASA Astrophysics Data System (ADS)

Hak, C.; Pundt, I.; Kern, C.; Platt, U.; Dommen, J.; Ordóñez, C.; Prévôt, A. S. H.; Junkermann, W.; Astorga-Lloréns, C.; Larsen, B. R.; Mellqvist, J.; Strandberg, A.; Yu, Y.; Galle, B.; Kleffmann, J.; Lörzer, J. C.; Braathen, G. O.; Volkamer, R.

2005-05-01

Results from an intercomparison of several currently used in-situ techniques for the measurement of atmospheric formaldehyde (CH2O) are presented. The measurements were carried out at Bresso, an urban site in the periphery of Milan (Italy) as part of the FORMAT-I field campaign. Eight instruments were employed by six independent research groups using four different techniques: Differential Optical Absorption Spectroscopy (DOAS), Fourier Transform Infra Red (FTIR) interferometry, the fluorimetric Hantzsch reaction technique (five instruments) and a chromatographic technique employing C18-DNPH-cartridges (2,4-dinitrophenylhydrazine). White type multi-reflection systems were employed for the optical techniques in order to avoid spatial CH2O gradients and ensure the sampling of nearly the same air mass by all instruments. Between 23 and 31 July 2002, up to 13 ppbv of CH2O were observed. The concentrations lay well above the detection limits of all instruments. The formaldehyde concentrations determined with DOAS, FTIR and the Hantzsch instruments were found to agree within ±11%, with the exception of one Hantzsch instrument, which gave systematically higher values. The two hour integrated samples by DNPH yielded up to 25% lower concentrations than the data of the continuously measuring instruments averaged over the same time period. The consistency between the DOAS and the Hantzsch method was better than during previous intercomparisons in ambient air with slopes of the regression line not significantly differing from one. The differences between the individual Hantzsch instruments could be attributed in part to the calibration standards used. Possible systematic errors of the methods are discussed.

Preparation and Characterization of Fluorescent SiO2 Microspheres

NASA Astrophysics Data System (ADS)

Xu, Cui; Zhang, Hao; Guan, Ruifang

2018-01-01

Fluorescent compound without typical fluorophores was synthesized with citric acid (CA) and aminopropyltriethoxysilane (APTS) firstly, and then it was grafted to the surface of the prepared SiO2 microspheres by chemical reaction. The fluorescent SiO2 microspheres with good fluorescent properties were obtained by optimizing the reaction conditions. And the morphology and structure of the fluorescent SiO2 microspheres have been characterized by scanning electron microscopy (SEM) and fourier transform infrared (FTIR) spectroscopy. The results showed that the preparation of fluorescent SiO2 microspheres have good monodispersity and narrow particle size distribution. Moreover, the fluorescent SiO2 microspheres can be applied to detect Fe3+ in aqueous solution, prepare fluorescent SiO2 rubber, and have potential to be applied in the fluorescent labeling and fingerprint appearing technique fields.

Application of cell-surface engineering for visualization of yeast in bread dough: development of a fluorescent bio-imaging technique in the mixing process of dough.

PubMed

Maeda, Tatsuro; Shiraga, Seizaburo; Araki, Tetsuya; Ueda, Mitsuyoshi; Yamada, Masaharu; Takeya, Koji; Sagara, Yasuyuki

2009-07-01

Cell-surface engineering (Ueda et al., 2000) has been applied to develop a novel technique to visualize yeast in bread dough. Enhanced green fluorescent protein (EGFP) was bonded to the surface of yeast cells, and 0.5% EGFP yeasts were mixed into the dough samples at four different mixing stages. The samples were placed on a cryostat at -30 degrees C and sliced at 10 microm. The sliced samples were observed at an excitation wavelength of 480 nm and a fluorescent wavelength of 520 nm. The results indicated that the combination of the EGFP-displayed yeasts, rapid freezing, and cryo-sectioning made it possible to visualize 2-D distribution of yeast in bread dough to the extent that the EGFP yeasts could be clearly distinguished from the auto-fluorescent background of bread dough.

A new technique for preliminary estimates of TRU activity on air sample filters and radiological smears.

PubMed

Hayes, Robert

2004-10-01

In most nuclear facilities, fixed air samplers and sometimes portable air samplers are used where some probability of a release exists but is not expected, and so the added expense and effort of using a continuous air monitor is not deemed justified. When a release is suspected, naturally occurring radioactive material buildup on the filter typically prevents any quantitative measurements within the first day or so. Likewise, outdoor air measurements suffer from the same limitations (such as those taken during the Los Alamos fires) and so any rapid quantifiable measurements of fixed air sampler/portable air sampler filters which are technically defendable (even though conservative) are of use. The technique presented here is only intended for use in routine health physics survey applications and does not presently appear to be appropriate for sub pico Curie activity determinations. This study evaluates the utility of using a portable continuous air monitor as an alpha spectrometer to make transuranic activity determinations of samples using both the built in algorithm for air monitoring and a simple region of interest analysis. All samples evaluated were from air sample filters taken using a portable air sampler. Samples were taken over many months to quantify effects from natural variation in radon progeny activity distributions.

Fluorescence spectroscopy for diagnosis of squamous intraepithelial lesions of the cervix.

PubMed

Mitchell, M F; Cantor, S B; Ramanujam, N; Tortolero-Luna, G; Richards-Kortum, R

1999-03-01

To calculate receiver operating characteristic (ROC) curves for fluorescence spectroscopy in order to measure its performance in the diagnosis of squamous intraepithelial lesions (SILs) and to compare these curves with those for other diagnostic methods: colposcopy, cervicography, speculoscopy, Papanicolaou smear screening, and human papillomavirus (HPV) testing. Data from our previous clinical study were used to calculate ROC curves for fluorescence spectroscopy. Curves for other techniques were calculated from other investigators' reports. To identify these, a MEDLINE search for articles published from 1966 to 1996 was carried out, using the search terms "colposcopy," "cervicoscopy," "cervicography," "speculoscopy," "Papanicolaou smear," "HPV testing," "fluorescence spectroscopy," and "polar probe" in conjunction with the terms "diagnosis," "positive predictive value," "negative predictive value," and "receiver operating characteristic curve." We found 270 articles, from which articles were selected if they reported results of studies involving high-disease-prevalence populations, reported findings of studies in which colposcopically directed biopsy was the criterion standard, and included sufficient data for recalculation of the reported sensitivities and specificities. We calculated ROC curves for fluorescence spectroscopy using Bayesian and neural net algorithms. A meta-analytic approach was used to calculate ROC curves for the other techniques. Areas under the curves were calculated. Fluorescence spectroscopy using the neural net algorithm had the highest area under the ROC curve, followed by fluorescence spectroscopy using the Bayesian algorithm, followed by colposcopy, the standard diagnostic technique. Cervicography, Papanicolaou smear screening, and HPV testing performed comparably with each other but not as well as fluorescence spectroscopy and colposcopy. Fluorescence spectroscopy performs better than colposcopy and other techniques in the diagnosis of

Inducible fluorescent speckle microscopy

PubMed Central

Aguiar, Paulo; Belsley, Michael; Maiato, Helder

2016-01-01

The understanding of cytoskeleton dynamics has benefited from the capacity to generate fluorescent fiducial marks on cytoskeleton components. Here we show that light-induced imprinting of three-dimensional (3D) fluorescent speckles significantly improves speckle signal and contrast relative to classic (random) fluorescent speckle microscopy. We predict theoretically that speckle imprinting using photobleaching is optimal when the laser energy and fluorophore responsivity are related by the golden ratio. This relation, which we confirm experimentally, translates into a 40% remaining signal after speckle imprinting and provides a rule of thumb in selecting the laser power required to optimally prepare the sample for imaging. This inducible speckle imaging (ISI) technique allows 3D speckle microscopy to be performed in readily available libraries of cell lines or primary tissues expressing fluorescent proteins and does not preclude conventional imaging before speckle imaging. As a proof of concept, we use ISI to measure metaphase spindle microtubule poleward flux in primary cells and explore a scaling relation connecting microtubule flux to metaphase duration. PMID:26783303

Inducible fluorescent speckle microscopy.

PubMed

Pereira, António J; Aguiar, Paulo; Belsley, Michael; Maiato, Helder

2016-01-18

The understanding of cytoskeleton dynamics has benefited from the capacity to generate fluorescent fiducial marks on cytoskeleton components. Here we show that light-induced imprinting of three-dimensional (3D) fluorescent speckles significantly improves speckle signal and contrast relative to classic (random) fluorescent speckle microscopy. We predict theoretically that speckle imprinting using photobleaching is optimal when the laser energy and fluorophore responsivity are related by the golden ratio. This relation, which we confirm experimentally, translates into a 40% remaining signal after speckle imprinting and provides a rule of thumb in selecting the laser power required to optimally prepare the sample for imaging. This inducible speckle imaging (ISI) technique allows 3D speckle microscopy to be performed in readily available libraries of cell lines or primary tissues expressing fluorescent proteins and does not preclude conventional imaging before speckle imaging. As a proof of concept, we use ISI to measure metaphase spindle microtubule poleward flux in primary cells and explore a scaling relation connecting microtubule flux to metaphase duration. © 2016 Pereira et al.

Doppler-shifted fluorescence imaging of velocity fields in supersonic reacting flows

NASA Technical Reports Server (NTRS)

Allen, M. G.; Davis, S. J.; Kessler, W. J.; Sonnenfroh, D. M.

1992-01-01

The application of Doppler-shifted fluorescence imaging of velocity fields in supersonic reacting flows is analyzed. Focussing on fluorescence of the OH molecule in typical H2-air Scramjet flows, the effects of uncharacterized variations in temperature, pressure, and collisional partner composition across the measurement plane are examined. Detailed measurements of the (1,0) band OH lineshape variations in H2-air combustions are used, along with single-pulse and time-averaged measurements of an excimer-pumped dye laser, to predict the performance of a model velocimeter with typical Scramjet flow properties. The analysis demonstrates the need for modification and control of the laser bandshape in order to permit accurate velocity measurements in the presence of multivariant flow properties.

Methods for Broadband Spectral Analysis: Intrinsic Fluorescence Temperature Sensing as an Example.

PubMed

Zhang, Weiwei; Wang, Guoyao; Baxter, Greg W; Collins, Stephen F

2017-06-01

A systematic study was performed on the temperature-dependent fluorescence of (Ba,Sr) 2 SiO 4 :Eu 2+ . The barycenter and extended intensity ratio techniques were proposed to characterize the broadband fluorescence spectra. These techniques and other known methods (listed below) were employed and compared in the fluorescent temperature sensing experiment. Multiple sensing functions were obtained using the behaviors of: (1) the barycenter location of the emission band; (2) the emission bandwidth; and (3) the ratio of intensities at different wavelengths in the emission band, respectively. The barycenter technique was not limited by the spectrometer resolution and worked well while the peak location method failed. All the sensing functions were based on the intrinsic characteristics of the fluorescence of the phosphor and demonstrated nearly linear relationships with temperature in the measuring range. The multifunctional temperature-sensing abilities of the phosphor can be applied in a point thermometer or thermal mapping. The new techniques were validated successfully for characterizing various spectra.

Fluorescent imaging of cancerous tissues for targeted surgery

PubMed Central

Bu, Lihong; Shen, Baozhong; Cheng, Zhen

2014-01-01

To maximize tumor excision and minimize collateral damage is the primary goal of cancer surgery. Emerging molecular imaging techniques have to “image-guided surgery” developing into “molecular imaging-guided surgery”, which is termed “targeted surgery” in this review. Consequently, the precision of surgery can be advanced from tissue-scale to molecule-scale, enabling “targeted surgery” to be a component of “targeted therapy”. Evidence from numerous experimental and clinical studies has demonstrated significant benefits of fluorescent imaging in targeted surgery with preoperative molecular diagnostic screening. Fluorescent imaging can help to improve intraoperative staging and enable more radical cytoreduction, detect obscure tumor lesions in special organs, highlight tumor margins, better map lymph node metastases, and identify important normal structures intraoperatively. Though limited tissue penetration of fluorescent imaging and tumor heterogeneity are two major hurdles for current targeted surgery, multimodality imaging and multiplex imaging may provide potential solutions to overcome these issues, respectively. Moreover, though many fluorescent imaging techniques and probes have been investigated, targeted surgery remains at a proof-of-principle stage. The impact of fluorescent imaging on cancer surgery will likely be realized through persistent interdisciplinary amalgamation of research in diverse fields. PMID:25064553

Handheld Fluorescence Microscopy based Flow Analyzer.

PubMed

Saxena, Manish; Jayakumar, Nitin; Gorthi, Sai Siva

2016-03-01

Fluorescence microscopy has the intrinsic advantages of favourable contrast characteristics and high degree of specificity. Consequently, it has been a mainstay in modern biological inquiry and clinical diagnostics. Despite its reliable nature, fluorescence based clinical microscopy and diagnostics is a manual, labour intensive and time consuming procedure. The article outlines a cost-effective, high throughput alternative to conventional fluorescence imaging techniques. With system level integration of custom-designed microfluidics and optics, we demonstrate fluorescence microscopy based imaging flow analyzer. Using this system we have imaged more than 2900 FITC labeled fluorescent beads per minute. This demonstrates high-throughput characteristics of our flow analyzer in comparison to conventional fluorescence microscopy. The issue of motion blur at high flow rates limits the achievable throughput in image based flow analyzers. Here we address the issue by computationally deblurring the images and show that this restores the morphological features otherwise affected by motion blur. By further optimizing concentration of the sample solution and flow speeds, along with imaging multiple channels simultaneously, the system is capable of providing throughput of about 480 beads per second.

Frequently asked questions about in vivo chlorophyll fluorescence: practical issues.

PubMed

Kalaji, Hazem M; Schansker, Gert; Ladle, Richard J; Goltsev, Vasilij; Bosa, Karolina; Allakhverdiev, Suleyman I; Brestic, Marian; Bussotti, Filippo; Calatayud, Angeles; Dąbrowski, Piotr; Elsheery, Nabil I; Ferroni, Lorenzo; Guidi, Lucia; Hogewoning, Sander W; Jajoo, Anjana; Misra, Amarendra N; Nebauer, Sergio G; Pancaldi, Simonetta; Penella, Consuelo; Poli, DorothyBelle; Pollastrini, Martina; Romanowska-Duda, Zdzislawa B; Rutkowska, Beata; Serôdio, João; Suresh, Kancherla; Szulc, Wiesław; Tambussi, Eduardo; Yanniccari, Marcos; Zivcak, Marek

2014-11-01

The aim of this educational review is to provide practical information on the hardware, methodology, and the hands on application of chlorophyll (Chl) a fluorescence technology. We present the paper in a question and answer format like frequently asked questions. Although nearly all information on the application of Chl a fluorescence can be found in the literature, it is not always easily accessible. This paper is primarily aimed at scientists who have some experience with the application of Chl a fluorescence but are still in the process of discovering what it all means and how it can be used. Topics discussed are (among other things) the kind of information that can be obtained using different fluorescence techniques, the interpretation of Chl a fluorescence signals, specific applications of these techniques, and practical advice on different subjects, such as on the length of dark adaptation before measurement of the Chl a fluorescence transient. The paper also provides the physiological background for some of the applied procedures. It also serves as a source of reference for experienced scientists.

Single-Shot Optical Sectioning Using Two-Color Probes in HiLo Fluorescence Microscopy

PubMed Central

Muro, Eleonora; Vermeulen, Pierre; Ioannou, Andriani; Skourides, Paris; Dubertret, Benoit; Fragola, Alexandra; Loriette, Vincent

2011-01-01

We describe a wide-field fluorescence microscope setup which combines HiLo microscopy technique with the use of a two-color fluorescent probe. It allows one-shot fluorescence optical sectioning of thick biological moving sample which is illuminated simultaneously with a flat and a structured pattern at two different wavelengths. Both homogenous and structured fluorescence images are spectrally separated at detection and combined similarly with the HiLo microscopy technique. We present optically sectioned full-field images of Xenopus laevis embryos acquired at 25 images/s frame rate. PMID:21641327

Microencapsulated Fluorescent Dye Penetrant.

DTIC Science & Technology

1979-07-01

Microencapsulated fluorescent dye pentrant materials were evaluated for feasibility as a technique to detect cracks on metal surfaces when applied as...a free flowing dry powder. Various flourescent dye solutions in addition to a commercial penetrant (Zyglo ZL-30) were microencapsulated and tested on

Integrating fluorescence and interactance measurements to improve apple maturity assessment

NASA Astrophysics Data System (ADS)

Noh, Hyun Kwon; Lu, Renfu

2006-10-01

Fluorescence and reflectance (or interactance) are promising techniques for measuring fruit quality and condition. Our previous research showed that a hyperspectral imaging technique integrating fluorescence and reflectance could improve predictions of selected quality parameters compared to single sensing techniques. The objective of this research was to use a low cost spectrometer for rapid acquisition of fluorescence and interactance spectra from apples and develop an algorithm integrating the two types of data for predicting skin and flesh color, fruit firmness, starch index, soluble solids content, and titratable acid. Experiments were performed to measure UV light induced transient fluorescence and interactance spectra from 'Golden Delicious' apples that were harvested over a period of four weeks during the 2005 harvest season. Standard destructive tests were performed to measure maturity parameters from the apples. Principal component (PC) analysis was applied to the interactance and fluorescence data. A back-propagation feedforward neural network with the inputs of PC data was used to predict individual maturity parameters. Interactance mode was consistently better than fluorescence mode in predicting the maturity parameters. Integrating interactance and fluorescence improved predictions of all parameters except flesh chroma; values of the correlation coefficient for firmness, soluble solids content, starch index, and skin and flesh hue were 0.77, 0.77, 0.89, 0.99, and 0.96 respectively, with the corresponding standard errors of 6.93 N, 0.90%, 0.97 g/L, 0.013 rad, and 0.013 rad. These results represented 4.1% to 23.5% improvements in terms of standard error, in comparison with the better results from the two single sensing methods. Integrating interactance and fluorescence can better assess apple maturity and quality.

Tomographic imaging of OH laser-induced fluorescence in laminar and turbulent jet flames

NASA Astrophysics Data System (ADS)

Li, Tao; Pareja, Jhon; Fuest, Frederik; Schütte, Manuel; Zhou, Yihui; Dreizler, Andreas; Böhm, Benjamin

2018-01-01

In this paper a new approach for 3D flame structure diagnostics using tomographic laser-induced fluorescence (Tomo-LIF) of the OH radical was evaluated. The approach combined volumetric illumination with a multi-camera detection system of eight views. Single-shot measurements were performed in a methane/air premixed laminar flame and in a non-premixed turbulent methane jet flame. 3D OH fluorescence distributions in the flames were reconstructed using the simultaneous multiplicative algebraic reconstruction technique. The tomographic measurements were compared and validated against results of OH-PLIF in the laminar flame. The effects of the experimental setup of the detection system and the size of the volumetric illumination on the quality of the tomographic reconstructions were evaluated. Results revealed that the Tomo-LIF is suitable for volumetric reconstruction of flame structures with acceptable spatial resolution and uncertainty. It was found that the number of views and their angular orientation have a strong influence on the quality and accuracy of the tomographic reconstruction while the illumination volume thickness influences mainly the spatial resolution.

Dielectric controlled excited state relaxation pathways of a representative push-pull stilbene: a mechanistic study using femtosecond fluorescence up-conversion technique.

PubMed

Rafiq, Shahnawaz; Sen, Pratik

2013-02-28

Femtosecond fluorescence up-conversion technique was employed to reinvestigate the intriguing dependence of fluorescence quantum yield of trans-4-dimethylamino-4(')-nitrostilbene (DNS) on dielectric properties of the media. In polar solvents, such as methanol and acetonitrile, the two time components of the fluorescence transients were assigned to intramolecular charge transfer (ICT) dynamics and to the depletion of the ICT state to the ground state via internal conversion along the torsional coordinate of nitro moiety. The viscosity independence of the first time component indicates the absence of any torsional coordinate in the charge transfer process. In slightly polar solvent (carbon tetrachloride) the fluorescence transients show a triple exponential behavior. The first time component was assigned to the formation of the ICT state on a 2 ps time scale. Second time component was assigned to the relaxation of the ICT state via two torsion controlled channels. First channel involves the torsional motion about the central double bond leading to the trans-cis isomerization via a conical intersection or avoided crossing. The other channel contributing to the depopulation of ICT state involves the torsional coordinates of dimethylanilino and∕or nitrophenyl moieties and leads to the formation of a conformationally relaxed state, which subsequently relaxes back to the ground state radiatively, and is responsible for the high fluorescence quantum yield of DNS in slightly polar solvents such as carbon tetrachloride, toluene, etc. The excited singlet state which is having a dominant π-π∗ character may also decay via intersystem crossing to the n-π∗ triplet manifold and thus accounts for the observed triplet yield of the molecule in slightly polar solvents.

Dielectric controlled excited state relaxation pathways of a representative push-pull stilbene: A mechanistic study using femtosecond fluorescence up-conversion technique

NASA Astrophysics Data System (ADS)

Rafiq, Shahnawaz; Sen, Pratik

2013-02-01

Femtosecond fluorescence up-conversion technique was employed to reinvestigate the intriguing dependence of fluorescence quantum yield of trans-4-dimethylamino-4'-nitrostilbene (DNS) on dielectric properties of the media. In polar solvents, such as methanol and acetonitrile, the two time components of the fluorescence transients were assigned to intramolecular charge transfer (ICT) dynamics and to the depletion of the ICT state to the ground state via internal conversion along the torsional coordinate of nitro moiety. The viscosity independence of the first time component indicates the absence of any torsional coordinate in the charge transfer process. In slightly polar solvent (carbon tetrachloride) the fluorescence transients show a triple exponential behavior. The first time component was assigned to the formation of the ICT state on a 2 ps time scale. Second time component was assigned to the relaxation of the ICT state via two torsion controlled channels. First channel involves the torsional motion about the central double bond leading to the trans-cis isomerization via a conical intersection or avoided crossing. The other channel contributing to the depopulation of ICT state involves the torsional coordinates of dimethylanilino and/or nitrophenyl moieties and leads to the formation of a conformationally relaxed state, which subsequently relaxes back to the ground state radiatively, and is responsible for the high fluorescence quantum yield of DNS in slightly polar solvents such as carbon tetrachloride, toluene, etc. The excited singlet state which is having a dominant π-π* character may also decay via intersystem crossing to the n-π* triplet manifold and thus accounts for the observed triplet yield of the molecule in slightly polar solvents.

Monitoring biological aerosols using UV fluorescence

NASA Astrophysics Data System (ADS)

Eversole, Jay D.; Roselle, Dominick; Seaver, Mark E.

1999-01-01

An apparatus has been designed and constructed to continuously monitor the number density, size, and fluorescent emission of ambient aerosol particles. The application of fluorescence to biological particles suspended in the atmosphere requires laser excitation in the UV spectral region. In this study, a Nd:YAG laser is quadrupled to provide a 266 nm wavelength to excite emission from single micrometer-sized particles in air. Fluorescent emission is used to continuously identify aerosol particles of biological origin. For calibration, biological samples of Bacillus subtilis spores and vegetative cells, Esherichia coli, Bacillus thuringiensis and Erwinia herbicola vegetative cells were prepared as suspensions in water and nebulized to produce aerosols. Detection of single aerosol particles, provides elastic scattering response as well as fluorescent emission in two spectral bands simultaneously. Our efforts have focuses on empirical characterization of the emission and scattering characteristics of various bacterial samples to determine the feasibility of optical discrimination between different cell types. Preliminary spectroscopic evidence suggest that different samples can be distinguished as separate bio-aerosol groups. In addition to controlled sample results, we will also discuss the most recent result on the effectiveness of detection outdoor releases and variations in environmental backgrounds.

Quantitative, spectrally-resolved intraoperative fluorescence imaging

PubMed Central

Valdés, Pablo A.; Leblond, Frederic; Jacobs, Valerie L.; Wilson, Brian C.; Paulsen, Keith D.; Roberts, David W.

2012-01-01

Intraoperative visual fluorescence imaging (vFI) has emerged as a promising aid to surgical guidance, but does not fully exploit the potential of the fluorescent agents that are currently available. Here, we introduce a quantitative fluorescence imaging (qFI) approach that converts spectrally-resolved data into images of absolute fluorophore concentration pixel-by-pixel across the surgical field of view (FOV). The resulting estimates are linear, accurate, and precise relative to true values, and spectral decomposition of multiple fluorophores is also achieved. Experiments with protoporphyrin IX in a glioma rodent model demonstrate in vivo quantitative and spectrally-resolved fluorescence imaging of infiltrating tumor margins for the first time. Moreover, we present images from human surgery which detect residual tumor not evident with state-of-the-art vFI. The wide-field qFI technique has broad implications for intraoperative surgical guidance because it provides near real-time quantitative assessment of multiple fluorescent biomarkers across the operative field. PMID:23152935

Exciplex Fluorescence Systems for Two-Phase Visualization.

NASA Astrophysics Data System (ADS)

Kim, J.-U.; Golding, B.; Schock, H. J.; Nocera, D. G.; Keller, P.

1996-03-01

We report the development of diagnostic chemical systems for vapor-liquid visualization based on an exciplex (excited state complex) formed between dimethyl- or diethyl-substituted aniline and trimethyl-substituted naphthalenes. Quantum yields of individual monomers were measured and the exciplex emission spectra as well as fluorescence quenching mechanisms were analyzed. Quenching occurs by both static and dynamic mechanisms. Among the many formulations investigated in this study, a system consisting of 7% 1,4,6-trimethylnaphthalene (1,4,6-TMN) and 5% N,N-dimethylaniline (DMA) in 88% isooctane exciplex was found to be useful for the laser- induced fluorescence technique. The technique is expected to find application in observing mixture formation in diesel or spark ignition engines with spectrally well-separated fluorescence images obtained from the monomer and exciplex constituents dissolved in the gasoline fuel. *Supported by NSF MRSEC DMR-9400417 and the Center for Fundamental Materials Research.

Fluorescence microscopy for the characterization of structural integrity

NASA Technical Reports Server (NTRS)

Street, Kenneth W.; Leonhardt, Todd A.

1991-01-01

The absorption characteristics of light and the optical technique of fluorescence microscopy for enhancing metallographic interpretation are presented. Characterization of thermally sprayed coatings by optical microscopy suffers because of the tendency for misidentification of the microstructure produced by metallographic preparation. Gray scale, in bright field microscopy, is frequently the only means of differentiating the actual structural details of porosity, cracking, and debonding of coatings. Fluorescence microscopy is a technique that helps to distinguish the artifacts of metallographic preparation (pullout, cracking, debonding) from the microstructure of the specimen by color contrasting structural differences. Alternative instrumentation and the use of other dye systems are also discussed. The combination of epoxy vacuum infiltration with fluorescence microscopy to verify microstructural defects is an effective means to characterize advanced materials and to assess structural integrity.

Development of a robust field technique to quantify the air-void distribution in fresh concrete.

DOT National Transportation Integrated Search

2013-07-01

In order to make concrete frost durable it is common to provide a small and well distributed air void system. Current measuring techniques require weeks to complete on hardened and polished samples of concrete. This report presents the results of a n...

Clinical application of indocyanine green-fluorescence imaging during hepatectomy

PubMed Central

Ishizawa, Takeaki; Saiura, Akio

2016-01-01

In hepatobiliary surgery, the fluorescence and bile excretion of indocyanine green (ICG) can be used for real-time visualization of biological structure. Fluorescence cholangiography is used to obtain fluorescence images of the bile ducts following intrabiliary injection of 0.025−0.5 mg/mL ICG or intravenous injection of 2.5 mg ICG. Recently, the latter technique has been used in laparoscopic/robotic cholecystectomy. Intraoperative fluorescence imaging can be used to identify subcapsular hepatic tumors. Primary and secondary hepatic malignancy can be identified by intraoperative fluorescence imaging using preoperative intravenous injection of ICG through biliary excretion disorders that exist in cancerous tissues of hepatocellular carcinoma (HCC) and in non-cancerous hepatic parenchyma around adenocarcinoma foci. Intraoperative fluorescence imaging may help detect tumors to be removed, especially during laparoscopic hepatectomy, in which visual inspection and palpation are limited, compared with open surgery. Fluorescence imaging can also be used to identify hepatic segments. Boundaries of hepatic segments can be visualized following injection of 0.25−2.5 mg/mL ICG into the portal veins or by intravenous injection of 2.5 mg ICG following closure of the proximal portal pedicle toward hepatic regions to be removed. These techniques enable identification of hepatic segments before hepatectomy and during parenchymal transection for anatomic resection. Advances in imaging systems will increase the use of fluorescence imaging as an intraoperative navigation tool that can enhance the safety and accuracy of open and laparoscopic/robotic hepatobiliary surgery. PMID:27500144

Tuning Fluorescence Direction with Plasmonic Metal–Dielectric– Metal Substrates

PubMed Central

Choudhury, Sharmistha Dutta; Badugu, Ramachandram; Nowaczyk, Kazimierz; Ray, Krishanu; Lakowicz, Joseph R.

2013-01-01

Controlling the emission properties of fluorophores is essential for improving the performance of fluorescence-based techniques in modern biochemical research, medical diagnosis, and sensing. Fluorescence emission is isotropic in nature, which makes it difficult to capture more than a small fraction of the total emission. Metal– dielectric–metal (MDM) substrates, discussed in this Letter, convert isotropic fluorescence into beaming emission normal to the substrate. This improves fluorescence collection efficiency and also opens up new avenues for a wide range of fluorescence-based applications. We suggest that MDM substrates can be readily adapted for multiple uses, such as in microarray formats, for directional fluorescence studies of multiple probes or for molecule-specific sensing with a high degree of spatial control over the fluorescence emission. SECTION: Physical Processes in Nanomaterials and Nanostructures PMID:24013521

Using soft computing techniques to predict corrected air permeability using Thomeer parameters, air porosity and grain density

NASA Astrophysics Data System (ADS)

Nooruddin, Hasan A.; Anifowose, Fatai; Abdulraheem, Abdulazeez

2014-03-01

Soft computing techniques are recently becoming very popular in the oil industry. A number of computational intelligence-based predictive methods have been widely applied in the industry with high prediction capabilities. Some of the popular methods include feed-forward neural networks, radial basis function network, generalized regression neural network, functional networks, support vector regression and adaptive network fuzzy inference system. A comparative study among most popular soft computing techniques is presented using a large dataset published in literature describing multimodal pore systems in the Arab D formation. The inputs to the models are air porosity, grain density, and Thomeer parameters obtained using mercury injection capillary pressure profiles. Corrected air permeability is the target variable. Applying developed permeability models in recent reservoir characterization workflow ensures consistency between micro and macro scale information represented mainly by Thomeer parameters and absolute permeability. The dataset was divided into two parts with 80% of data used for training and 20% for testing. The target permeability variable was transformed to the logarithmic scale as a pre-processing step and to show better correlations with the input variables. Statistical and graphical analysis of the results including permeability cross-plots and detailed error measures were created. In general, the comparative study showed very close results among the developed models. The feed-forward neural network permeability model showed the lowest average relative error, average absolute relative error, standard deviations of error and root means squares making it the best model for such problems. Adaptive network fuzzy inference system also showed very good results.

The Fluorescent Antibody Technique in the Diagnosis of Equine Rhinopneumonitis Virus Abortion

PubMed Central

Smith, I. M.; Girard, A.; Corner, A. H.; Mitchell, D.

1972-01-01

Using two known positive equine viral rhinopneumonitis (EVR) sera, conjugates were prepared with fluorescein isothiocyanate and tested for specificity using EVR infected tissue culture cells. The conjugate was then applied to selected tissues from 32 aborted fetuses and foals submitted during a natural outbreak of EVR. Antigen was detected in various tissues by immunofluorescence in 20 cases (62.5%). In 24 cases bovine fetal kidney cell monolayers were inoculated with a pool of lung and liver and EVR virus was isolated from 15 (62.5%). Histological examination of various tissues from 29 cases resulted in the diagnosis of EVR in 19 (65.5%), based upon the presence of focal areas of necrosis and intranuclear inclusion bodies. Correlation of results was not obtained in two cases. One was diagnosed positive histologically and negative on fluorescence, the other was negative histologically and by virus isolation but showed fluorescence. The distribution of fluorescence in various infected fetal tissues indicated that the combined examination of lung and thymus gland was most likely to provide a positive diagnosis. ImagesFig. 1.Fig. 2. PMID:4114979

Fluorescence microscopy: A tool to study autophagy

NASA Astrophysics Data System (ADS)

Rai, Shashank; Manjithaya, Ravi

2015-08-01

Autophagy is a cellular recycling process through which a cell degrades old and damaged cellular components such as organelles and proteins and the degradation products are reused to provide energy and building blocks. Dysfunctional autophagy is reported in several pathological situations. Hence, autophagy plays an important role in both cellular homeostasis and diseased conditions. Autophagy can be studied through various techniques including fluorescence based microscopy. With the advancements of newer technologies in fluorescence microscopy, several novel processes of autophagy have been discovered which makes it an essential tool for autophagy research. Moreover, ability to tag fluorescent proteins with sub cellular targets has enabled us to evaluate autophagy processes in real time under fluorescent microscope. In this article, we demonstrate different aspects of autophagy in two different model organisms i.e. yeast and mammalian cells, with the help of fluorescence microscopy.

Detection of intaoral lesions using a fluorescence camera

NASA Astrophysics Data System (ADS)

Thoms, Michael

2006-02-01

Optical methods for the detection of carious lesions, calculus and plaque have the advantage of being minimally invasive. The use of endogeneous fluorescence markers like porphyrins could simplify the application of fluorescence techniques in the dental practice. It is known that porphyrins are produced by some of the bacterial species that are present in the oral cavity. Since porphyrins have an excitation band at about 400nm they have the potential to be used as fluorescent markers of locations in the oral cavity where the production of bacteria is out of the limits of healthy regions. Further, modern and efficient GaN-based semiconductor diodes emit light in this spectral range and thus make the implementation of fluorescence sensors with excitation at this wavelength easy. Carious lesions, calculus and plaque have been measured using a self build fluorescence camera using GaN-diodes for illumination at 405nm. Further, emission spectra under this excitation were recorded. For the latter purpose freshly extracted teeth were used. It has been found that already in the case of an initial carious lesion red porphyrin-fluorescence is emitted whereas it is absent in healthy enamel. In already brown coloured carious lesions the emission bands of porphyrin are present but the observed overall fluorescence intensity is lower, probably due to the absorption of the fluorescence by the carious defect itself. In dental calculus, dental plaque and subgingival concrements porphyrin originated luminescence was found as well. Since in these cases the emission spectra differ slightly it can be concluded that they originate from different types of porphyrins and thus also from different bacteria. These results show that this fluorescence technique can be a promising method to diagnose carious lesions, calculus and plaque.

Quantum dots versus organic fluorophores in fluorescent deep-tissue imaging--merits and demerits.

PubMed

Bakalova, Rumiana; Zhelev, Zhivko; Gadjeva, Veselina

2008-12-01

The use of fluorescence in deep-tissue imaging is rapidly expanding in last several years. The progress in fluorescent molecular probes and fluorescent imaging techniques gives an opportunity to detect single cells and even molecular targets in live organisms. The highly sensitive and high-speed fluorescent molecular sensors and detection devices allow the application of fluorescence in functional imaging. With the development of novel bright fluorophores based on nanotechnologies and 3D fluorescence scanners with high spatial and temporal resolution, the fluorescent imaging has a potential to become an alternative of the other non-invasive imaging techniques as magnetic resonance imaging, positron-emission tomography, X-ray, computing tomography. The fluorescent imaging has also a potential to give a real map of human anatomy and physiology. The current review outlines the advantages of fluorescent nanoparticles over conventional organic dyes in deep-tissue imaging in vivo and defines the major requirements to the "perfect fluorophore". The analysis proceeds from the basic principles of fluorescence and major characteristics of fluorophores, light-tissue interactions, and major limitations of fluorescent deep-tissue imaging. The article is addressed to a broad readership - from specialists in this field to university students.

Minimizing brain shift during functional neurosurgical procedures - a simple burr hole technique that can decrease CSF loss and intracranial air.

PubMed

Coenen, V A; Abdel-Rahman, A; McMaster, J; Bogod, N; Honey, C R

2011-11-01

Exact stereotactic placement of deep brain stimulation electrodes during functional stereotactic neurosurgical procedures can be impeded by intraoperative brain shift. Brain shift has been shown to correlate with the amount of intracranial (subdural) air detected on early postoperative imaging studies. We report a simple burr hole technique that reduces the loss of cerebrospinal fluid (CSF) and has the potential to significantly reduce the amount of postoperative intracranial air. A total of 16 patients were studied with half (group 2) receiving the burr hole technique designed to seal the CSF space and thereby reducing CSF loss. The other 8 patients (group 1) received the standard burr hole technique. The 2 groups were of similar age, gender, diagnosis (Parkinson's disease, n=14; cervical dystonia n=2), and surgical targets. All patients received bilateral electrodes either in the subthalamic nucleus (STN, n=14) or in the globus pallidum internus (GPi, n=2) avoiding transventricular trajectories. Early postoperative 3-dimensional computed tomography (3D CT) was used to check for possible bleeding, DBS lead location, and the amount of intracranial air. Intracranial air was assessed manually in a volumetric slice-by-slice approach in the individual postoperative CT and the groups compared by t-test. Group 2 showed significantly lower postoperative intracranial air volumes (4.86 ± 4.35cc) as compared to group 1 (27.59 ± 17.80 cc, p=0.0083*). The duration of surgery, however, was significantly longer for group 1 (435 ± 56.05 min) as compared to group 2 (316 ± 34.79 min,p=0.00015*).The time span between the conclusion of the operation and postoperative 3DCT was similar for both groups. This new and simple burr hole technique was associated with a significant reduction in postoperative intracranial air. Reduction of intracranial air will ultimately reduce brain shift. That total operation time does not influence intracranial air is discussed as well as the

Mercury in Environmental and Biological Samples Using Online Combustion with Sequential Atomic Absorption and Fluorescence Measurements: A Direct Comparison of Two Fundamental Techniques in Spectrometry

ERIC Educational Resources Information Center

Cizdziel, James V.

2011-01-01

In this laboratory experiment, students quantitatively determine the concentration of an element (mercury) in an environmental or biological sample while comparing and contrasting the fundamental techniques of atomic absorption spectrometry (AAS) and atomic fluorescence spectrometry (AFS). A mercury analyzer based on sample combustion,…

Binding of fluorescent acridine dyes acridine orange and 9-aminoacridine to hemoglobin: Elucidation of their molecular recognition by spectroscopy, calorimetry and molecular modeling techniques.

PubMed

Chatterjee, Sabyasachi; Kumar, Gopinatha Suresh

2016-06-01

The molecular interaction between hemoglobin (HHb), the major human heme protein, and the acridine dyes acridine orange (AO) and 9-aminoacridine (9AA) was studied by various spectroscopic, calorimetric and molecular modeling techniques. The dyes formed stable ground state complex with HHb as revealed from spectroscopic data. Temperature dependent fluorescence data showed the strength of the dye-protein complexation to be inversely proportional to temperature and the fluorescence quenching was static in nature. The binding-induced conformational change in the protein was investigated using circular dichroism, synchronous fluorescence, 3D fluorescence and FTIR spectroscopy results. Circular dichroism data also quantified the α-helicity change in hemoglobin due to the binding of acridine dyes. Calorimetric studies revealed the binding to be endothermic in nature for both AO and 9AA, though the latter had higher affinity, and this was also observed from spectroscopic data. The binding of both dyes was entropy driven. pH dependent fluorescence studies revealed the existence of electrostatic interaction between the protein and dye molecules. Molecular modeling studies specified the binding site and the non-covalent interactions involved in the association. Overall, the results revealed that a small change in the acridine chromophore leads to remarkable alteration in the structural and thermodynamic aspects of binding to HHb. Copyright © 2016 Elsevier B.V. All rights reserved.

Single-shot optical sectioning using two-color probes in HiLo fluorescence microscopy.

PubMed

Muro, Eleonora; Vermeulen, Pierre; Ioannou, Andriani; Skourides, Paris; Dubertret, Benoit; Fragola, Alexandra; Loriette, Vincent

2011-06-08

We describe a wide-field fluorescence microscope setup which combines HiLo microscopy technique with the use of a two-color fluorescent probe. It allows one-shot fluorescence optical sectioning of thick biological moving sample which is illuminated simultaneously with a flat and a structured pattern at two different wavelengths. Both homogenous and structured fluorescence images are spectrally separated at detection and combined similarly with the HiLo microscopy technique. We present optically sectioned full-field images of Xenopus laevis embryos acquired at 25 images/s frame rate. Copyright © 2011 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Digitally synthesized beat frequency-multiplexed fluorescence lifetime spectroscopy

PubMed Central

Chan, Jacky C. K.; Diebold, Eric D.; Buckley, Brandon W.; Mao, Sien; Akbari, Najva; Jalali, Bahram

2014-01-01

Frequency domain fluorescence lifetime imaging is a powerful technique that enables the observation of subtle changes in the molecular environment of a fluorescent probe. This technique works by measuring the phase delay between the optical emission and excitation of fluorophores as a function of modulation frequency. However, high-resolution measurements are time consuming, as the excitation modulation frequency must be swept, and faster low-resolution measurements at a single frequency are prone to large errors. Here, we present a low cost optical system for applications in real-time confocal lifetime imaging, which measures the phase vs. frequency spectrum without sweeping. Deemed Lifetime Imaging using Frequency-multiplexed Excitation (LIFE), this technique uses a digitally-synthesized radio frequency comb to drive an acousto-optic deflector, operated in a cat’s-eye configuration, to produce a single laser excitation beam modulated at multiple beat frequencies. We demonstrate simultaneous fluorescence lifetime measurements at 10 frequencies over a bandwidth of 48 MHz, enabling high speed frequency domain lifetime analysis of single- and multi-component sample mixtures. PMID:25574449

A portable time-domain LED fluorimeter for nanosecond fluorescence lifetime measurements

NASA Astrophysics Data System (ADS)

Wang, Hongtao; Qi, Ying; Mountziaris, T. J.; Salthouse, Christopher D.

2014-05-01

Fluorescence lifetime measurements are becoming increasingly important in chemical and biological research. Time-domain lifetime measurements offer fluorescence multiplexing and improved handling of interferers compared with the frequency-domain technique. In this paper, an all solid-state, filterless, and highly portable light-emitting-diode based time-domain fluorimeter (LED TDF) is reported for the measurement of nanosecond fluorescence lifetimes. LED based excitation provides more wavelengths options compared to laser diode based excitation, but the excitation is less effective due to the uncollimated beam, less optical power, and longer latency in state transition. Pulse triggering and pre-bias techniques were implemented in our LED TDF to improve the peak optical power to over 100 mW. The proposed pulsing circuit achieved an excitation light fall time of less than 2 ns. Electrical resetting technique realized a time-gated photo-detector to remove the interference of the excitation light with fluorescence. These techniques allow the LED fluorimeter to accurately measure the fluorescence lifetime of fluorescein down to concentration of 0.5 μM. In addition, all filters required in traditional instruments are eliminated for the non-attenuated excitation/emission light power. These achievements make the reported device attractive to biochemical laboratories seeking for highly portable lifetime detection devices for developing sensors based on fluorescence lifetime changes. The device was initially validated by measuring the lifetimes of three commercial fluorophores and comparing them with reported lifetime data. It was subsequently used to characterize a ZnSe quantum dot based DNA sensor.

A portable time-domain LED fluorimeter for nanosecond fluorescence lifetime measurements

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wang, Hongtao; Salthouse, Christopher D., E-mail: salthouse@ecs.umass.edu; Center for Personalized Health Monitoring, University of Massachusetts, Amherst, Massachusetts 01003

2014-05-15

Fluorescence lifetime measurements are becoming increasingly important in chemical and biological research. Time-domain lifetime measurements offer fluorescence multiplexing and improved handling of interferers compared with the frequency-domain technique. In this paper, an all solid-state, filterless, and highly portable light-emitting-diode based time-domain fluorimeter (LED TDF) is reported for the measurement of nanosecond fluorescence lifetimes. LED based excitation provides more wavelengths options compared to laser diode based excitation, but the excitation is less effective due to the uncollimated beam, less optical power, and longer latency in state transition. Pulse triggering and pre-bias techniques were implemented in our LED TDF to improve themore » peak optical power to over 100 mW. The proposed pulsing circuit achieved an excitation light fall time of less than 2 ns. Electrical resetting technique realized a time-gated photo-detector to remove the interference of the excitation light with fluorescence. These techniques allow the LED fluorimeter to accurately measure the fluorescence lifetime of fluorescein down to concentration of 0.5 μM. In addition, all filters required in traditional instruments are eliminated for the non-attenuated excitation/emission light power. These achievements make the reported device attractive to biochemical laboratories seeking for highly portable lifetime detection devices for developing sensors based on fluorescence lifetime changes. The device was initially validated by measuring the lifetimes of three commercial fluorophores and comparing them with reported lifetime data. It was subsequently used to characterize a ZnSe quantum dot based DNA sensor.« less

Modulated Chlorophyll "a" Fluorescence: A Tool for Teaching Photosynthesis

ERIC Educational Resources Information Center

Marques da Silva, Jorge; Bernardes da Silva, Anabela; Padua, Mario

2007-01-01

"In vivo" chlorophyll "a" fluorescence is a key technique in photosynthesis research. The recent release of a low cost, commercial, modulated fluorometer enables this powerful technology to be used in education. Modulated chlorophyll a fluorescence measurement "in vivo" is here proposed as a tool to demonstrate basic…

Note: On-chip multifunctional fluorescent-magnetic Janus helical microswimmers

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hwang, G., E-mail: gilgueng.hwang@lpn.cnrs.fr; Decanini, D.; Leroy, L.

Microswimmers integrated into microfluidic devices that are capable of self-illumination through fluorescence could revolutionize many aspects of technology, especially for biological applications. Few illumination and propulsion techniques of helical microswimmers inside microfluidic channels have been demonstrated. This paper presents the fabrication, detachment, and magnetic propulsions of multifunctional fluorescent-magnetic helical microswimmers integrated inside microfluidics. The fabrication process is based on two-photon laser lithography to pattern 3-D nanostructures from fluorescent photoresist coupled with conventional microfabrication techniques for magnetic thin film deposition by shadowing. After direct integration inside a microfluidic device, injected gas bubble allows gentle detachment of the integrated helical microswimmers whosemore » magnetic propulsion can then be directly applied inside the microfluidic channel using external electromagnetic coil setup. With their small scale, fluorescence, excellent resistance to liquid/gas surface tension, and robust propulsion capability inside the microfluidic channel, the microswimmers can be used as high-resolution and large-range mobile micromanipulators inside microfluidic channels.« less

Laser-induced fluorescence of space-exposed polyurethane

NASA Technical Reports Server (NTRS)

Hill, Ralph H., Jr.

1993-01-01

The object of this work was to utilize laser-induced fluorescence technique to characterize several samples of space-exposed polyurethane. These samples were flown on the Long Duration Exposure Facility (LDEF), which was in a shuttle-like orbit for nearly 6 years. Because of our present work to develop laser-induced-fluorescence inspection techniques for polymers, space-exposed samples and controls were lent to us for evaluation. These samples had been attached to the outer surface of LDEF; therefore, they were subjected to thermal cycling, solar ultraviolet radiation, vacuum, and atomic oxygen. It is well documented that atomic oxygen and ultraviolet exposure have detrimental effects on many polymers. This was a unique opportunity to make measurements on material that had been naturally degraded by an unusual environment. During our past work, data have come from artificially degraded samples and generally have demonstrated a correlation between laser-induced fluorescence and tensile strength or elasticity.

In vivo fluorescence lifetime optical projection tomography

PubMed Central

McGinty, James; Taylor, Harriet B.; Chen, Lingling; Bugeon, Laurence; Lamb, Jonathan R.; Dallman, Margaret J.; French, Paul M. W.

2011-01-01

We demonstrate the application of fluorescence lifetime optical projection tomography (FLIM-OPT) to in vivo imaging of lysC:GFP transgenic zebrafish embryos (Danio rerio). This method has been applied to unambiguously distinguish between the fluorescent protein (GFP) signal in myeloid cells from background autofluorescence based on the fluorescence lifetime. The combination of FLIM, an inherently ratiometric method, in conjunction with OPT results in a quantitative 3-D tomographic technique that could be used as a robust method for in vivo biological and pharmaceutical research, for example as a readout of Förster resonance energy transfer based interactions. PMID:21559145

Scanning force microscopy at the air-water interface of an air bubble coated with pulmonary surfactant.

PubMed Central

Knebel, D; Sieber, M; Reichelt, R; Galla, H-J; Amrein, M

2002-01-01

To study the structure-function relationship of pulmonary surfactant under conditions close to nature, molecular films of a model system consisting of dipalmitoylphosphatidylcholine, dipalmitoylphosphatidylglycerol, and surfactant-associated protein C were prepared at the air-water interface of air bubbles about the size of human alveoli (diameter of 100 microm). The high mechanical stability as well as the absence of substantial film flow, inherent to small air bubbles, allowed for scanning force microscopy (SFM) directly at the air-water interface. The SFM topographical structure was correlated to the local distribution of fluorescent-labeled dipalmitoylphosphatidylcholine, as revealed from fluorescence light microscopy of the same bubbles. Although SFM has proven before to be exceptionally well suited to probe the structure of molecular films of pulmonary surfactant, the films so far had to be transferred onto a solid support from the air-water interface of a film balance, where they had been formed. This made them prone to artifacts imposed by the transfer. Moreover, the supported monolayers disallowed the direct observation of the structural dynamics associated with expansion and compression of the films as upon breathing. The current findings are compared in this respect to our earlier findings from films, transferred onto a solid support. PMID:11751334

Emerging applications of fluorescence spectroscopy in medical microbiology field.

PubMed

Shahzad, Aamir; Köhler, Gottfried; Knapp, Martin; Gaubitzer, Erwin; Puchinger, Martin; Edetsberger, Michael

2009-11-26

There are many diagnostic techniques and methods available for diagnosis of medically important microorganisms like bacteria, viruses, fungi and parasites. But, almost all these techniques and methods have some limitations or inconvenience. Most of these techniques are laborious, time consuming and with chances of false positive or false negative results. It warrants the need of a diagnostic technique which can overcome these limitations and problems. At present, there is emerging trend to use Fluorescence spectroscopy as a diagnostic as well as research tool in many fields of medical sciences. Here, we will critically discuss research studies which propose that Fluorescence spectroscopy may be an excellent diagnostic as well as excellent research tool in medical microbiology field with high sensitivity and specificity.

First upper limits on the radar cross section of cosmic-ray induced extensive air showers

DOE Office of Scientific and Technical Information (OSTI.GOV)

Abbasi, R. U.; Abe, M.; Abou Bakr Othman, M.

TARA (Telescope Array Radar) is a cosmic ray radar detection experiment colocated with Telescope Array, the conventional surface scintillation detector (SD) and fluorescence telescope detector (FD) near Delta, Utah, U.S.A. Furthermore, the TARA detector combines a 40 kW, 54.1 MHz VHF transmitter and high-gain transmitting antenna which broadcasts the radar carrier over the SD array and within the FD field of view, towards a 250 MS/s DAQ receiver. TARA has been collecting data since 2013 with the primary goal of observing the radar signatures of extensive air showers (EAS). Simulations indicate that echoes are expected to be short in durationmore » (~10 µs) and exhibit rapidly changing frequency, with rates on the order 1 MHz/µs. The EAS radar cross-section (RCS) is currently unknown although it is the subject of over 70 years of speculation. One novel signal search technique is described in which the expected radar echo of a particular air shower is used as a matched filter template and compared to waveforms obtained by triggering the radar DAQ using the Telescope Array fluorescence detector. No evidence for the scattering of radio frequency radiation by EAS is obtained to date. Finally, we report the first quantitative RCS upper limits using EAS that triggered the Telescope Array Fluorescence Detector.« less

First upper limits on the radar cross section of cosmic-ray induced extensive air showers

DOE PAGES

Abbasi, R. U.; Abe, M.; Abou Bakr Othman, M.; ...

2016-11-19

TARA (Telescope Array Radar) is a cosmic ray radar detection experiment colocated with Telescope Array, the conventional surface scintillation detector (SD) and fluorescence telescope detector (FD) near Delta, Utah, U.S.A. Furthermore, the TARA detector combines a 40 kW, 54.1 MHz VHF transmitter and high-gain transmitting antenna which broadcasts the radar carrier over the SD array and within the FD field of view, towards a 250 MS/s DAQ receiver. TARA has been collecting data since 2013 with the primary goal of observing the radar signatures of extensive air showers (EAS). Simulations indicate that echoes are expected to be short in durationmore » (~10 µs) and exhibit rapidly changing frequency, with rates on the order 1 MHz/µs. The EAS radar cross-section (RCS) is currently unknown although it is the subject of over 70 years of speculation. One novel signal search technique is described in which the expected radar echo of a particular air shower is used as a matched filter template and compared to waveforms obtained by triggering the radar DAQ using the Telescope Array fluorescence detector. No evidence for the scattering of radio frequency radiation by EAS is obtained to date. Finally, we report the first quantitative RCS upper limits using EAS that triggered the Telescope Array Fluorescence Detector.« less

Fluorescence lifetime based bioassays

NASA Astrophysics Data System (ADS)

Meyer-Almes, Franz-Josef

2017-12-01

Fluorescence lifetime (FLT) is a robust intrinsic property and material constant of fluorescent matter. Measuring this important physical indicator has evolved from a laboratory curiosity to a powerful and established technique for a variety of applications in drug discovery, medical diagnostics and basic biological research. This distinct trend was mainly driven by improved and meanwhile affordable laser and detection instrumentation on the one hand, and the development of suitable FLT probes and biological assays on the other. In this process two essential working approaches emerged. The first one is primarily focused on high throughput applications employing biochemical in vitro assays with no requirement for high spatial resolution. The second even more dynamic trend is the significant expansion of assay methods combining highly time and spatially resolved fluorescence data by fluorescence lifetime imaging. The latter approach is currently pursued to enable not only the investigation of immortal tumor cell lines, but also specific tissues or even organs in living animals. This review tries to give an actual overview about the current status of FLT based bioassays and the wide range of application opportunities in biomedical and life science areas. In addition, future trends of FLT technologies will be discussed.

Intraoperative real-time localization of parathyroid gland with near infrared fluorescence imaging

PubMed Central

Kim, Sung Won; Lee, Hyoung Shin

2017-01-01

Surgeons have cited difficulties in identifying the parathyroid glands (PG) during thyroidectomy. To overcome the limitation of naked eye, many studies on near-infrared fluorescence imaging of PGs have been introduced and suggested that fluorescence imaging is useful for both localizing PGs and evaluating their function. This imaging technique has been reported in two ways: (I) imaging using a fluorescent material called indocyanine green (ICG); and (II) autofluorescence using intrinsic fluorophores. These innovative and novel techniques are expected to have a significant impact on performing thyroid or parathyroid surgery. In this article, current papers that describe ICG fluorescence and autofluorescence imaging of PG during thyroid and parathyroid surgery are reviewed. PMID:29142843

Study on excitation and fluorescence spectrums of Japanese citruses to construct machine vision systems for acquiring fluorescent images

NASA Astrophysics Data System (ADS)

Momin, Md. Abdul; Kondo, Naoshi; Kuramoto, Makoto; Ogawa, Yuichi; Shigi, Tomoo

2011-06-01

Research was conducted to acquire knowledge of the ultraviolet and visible spectrums from 300 -800 nm of some common varieties of Japanese citrus, to investigate the best wave-lengths for fluorescence excitation and the resulting fluorescence wave-lengths and to provide a scientific background for the best quality fluorescent imaging technique for detecting surface defects of citrus. A Hitachi U-4000 PC-based microprocessor controlled spectrophotometer was used to measure the absorption spectrum and a Hitachi F-4500 spectrophotometer was used for the fluorescence and excitation spectrums. We analyzed the spectrums and the selected varieties of citrus were categorized into four groups of known fluorescence level, namely strong, medium, weak and no fluorescence.The level of fluorescence of each variety was also examined by using machine vision system. We found that around 340-380 nm LEDs or UV lamps are appropriate as lighting devices for acquiring the best quality fluorescent image of the citrus varieties to examine their fluorescence intensity. Therefore an image acquisition device was constructed with three different lighting panels with UV LED at peak 365 nm, Blacklight blue lamps (BLB) peak at 350 nm and UV-B lamps at peak 306 nm. The results from fluorescent images also revealed that the findings of the measured spectrums worked properly and can be used for practical applications such as for detecting rotten, injured or damaged parts of a wide variety of citrus.

High speed fluorescence imaging with compressed ultrafast photography

NASA Astrophysics Data System (ADS)

Thompson, J. V.; Mason, J. D.; Beier, H. T.; Bixler, J. N.

2017-02-01

Fluorescent lifetime imaging is an optical technique that facilitates imaging molecular interactions and cellular functions. Because the excited lifetime of a fluorophore is sensitive to its local microenvironment,1, 2 measurement of fluorescent lifetimes can be used to accurately detect regional changes in temperature, pH, and ion concentration. However, typical state of the art fluorescent lifetime methods are severely limited when it comes to acquisition time (on the order of seconds to minutes) and video rate imaging. Here we show that compressed ultrafast photography (CUP) can be used in conjunction with fluorescent lifetime imaging to overcome these acquisition rate limitations. Frame rates up to one hundred billion frames per second have been demonstrated with compressed ultrafast photography using a streak camera.3 These rates are achieved by encoding time in the spatial direction with a pseudo-random binary pattern. The time domain information is then reconstructed using a compressed sensing algorithm, resulting in a cube of data (x,y,t) for each readout image. Thus, application of compressed ultrafast photography will allow us to acquire an entire fluorescent lifetime image with a single laser pulse. Using a streak camera with a high-speed CMOS camera, acquisition rates of 100 frames per second can be achieved, which will significantly enhance our ability to quantitatively measure complex biological events with high spatial and temporal resolution. In particular, we will demonstrate the ability of this technique to do single-shot fluorescent lifetime imaging of cells and microspheres.

Fluorescent nanodiamonds and their use in biomedical research

NASA Astrophysics Data System (ADS)

Suarez-Kelly, Lorena P.; Rampersaud, Isaac V.; Moritz, Charles E.; Campbell, Amanda R.; Hu, Zhiwei; Alkahtani, Masfer H.; Alghannam, Fahad S.; Hemmer, Phillip; Carson, William E.; Rampersaud, Arfaan A.

2016-03-01

Nanodiamonds containing color-centers produce non-quenching fluorescence that is easily detected. This makes them useful for cellular, proteomic and genomic applications. However, fluorescent nanodiamonds have yet to become popular in the biomedical research community as labeling reagents. We discuss production of nanodiamonds with distinct color-centers and assess their biocompatibility and techniques for bioconjugation. Fluorescent diamonds were fabricated by electron irradiation of high-pressure, high-temperature micron-sized diamonds which generated diamonds with vacancy-related defects (V). These diamonds were annealed to create nitrogen vacancy (NV)-centers then following a milling step were fractionated into nanoparticle sizes of 30, 60, and 95 nm. Optical characterization of Vand NV-center diamonds demonstrated fluorescence in two distinct green and red channels, respectively. In vitro studies demonstrated that these nanodiamonds are biocompatible and readily taken up by murine macrophage cells. Quantification of NV-center nanodiamond uptake by flow cytometry, showed that uptake was independent of nanodiamond size. Confocal microscopy demonstrated that NV-center nanodiamonds accumulate within the cytoplasm of these cells. NV-center nanodiamonds were then conjugated with streptavidin using a short polyethylene chain as linker. Conjugation was confirmed via a catalytic assay employing biotinylated-horseradish peroxidase. We present a technique for large-scale production of biocompatible conjugated V- or NV-center nanodiamonds. Functional testing is essential for standardization of fluorescent nanodiamond bioconjugates and quality control. Large-scale production of bioconjugated fluorescent nanodiamonds is crucial to their development as novel tools for biological and medical applications.

Air pollution measurements from satellites

NASA Technical Reports Server (NTRS)

Ludwig, C. B.; Griggs, M.; Malkmus, W.; Bartle, E. R.

1973-01-01

A study is presented on the remote sensing of gaseous and particulate air pollutants which is an extension of a previous report. Pollutants can be observed by either active or passive remote sensing systems. Calculations discussed herein indicate that tropospheric CO, CO2, SO2, NO2, NH3, HCHO, and CH4 can be measured by means of nadir looking passive systems. Additional species such as NO, HNO3, O3, and H2O may be measured in the stratosphere through a horizon experiment. A brief theoretical overview of resonance Raman scattering and resonance fluorescence is given. It is found that radiance measurements are most promising for general global applications, and that stratospheric aerosols may be measured using a sun occultation technique. The instrumentation requirements for both active and passive systems are examined and various instruments now under development are described.

Snapshot imaging Fraunhofer line discriminator for detection of plant fluorescence

NASA Astrophysics Data System (ADS)

Gupta Roy, S.; Kudenov, M. W.

2015-05-01

Non-invasive quantification of plant health is traditionally accomplished using reflectance based metrics, such as the normalized difference vegetative index (NDVI). However, measuring plant fluorescence (both active and passive) to determine photochemistry of plants has gained importance. Due to better cost efficiency, lower power requirements, and simpler scanning synchronization, detecting passive fluorescence is preferred over active fluorescence. In this paper, we propose a high speed imaging approach for measuring passive plant fluorescence, within the hydrogen alpha Fraunhofer line at ~656 nm, using a Snapshot Imaging Fraunhofer Line Discriminator (SIFOLD). For the first time, the advantage of snapshot imaging for high throughput Fraunhofer Line Discrimination (FLD) is cultivated by our system, which is based on a multiple-image Fourier transform spectrometer and a spatial heterodyne interferometer (SHI). The SHI is a Sagnac interferometer, which is dispersion compensated using blazed diffraction gratings. We present data and techniques for calibrating the SIFOLD to any particular wavelength. This technique can be applied to quantify plant fluorescence at low cost and reduced complexity of data collection.

Fluorescence of fungi in superficial and deep fungal infections

PubMed Central

Elston, Dirk M

2001-01-01

Background Fluorescence of many fungi is noted when H&E stained sections are examined under a fluorescent microscope. In theory, this phenomenon could aid in the diagnosis of cutaneous and disseminated fungal infections without the delay associated with special stains. Seventy-six cases of superficial and deep fungal infections and 3 cases of protothecosis were studied to determine the clinical usefulness of this technique. Results In most cases, fluorescence was noted, but was not intense. Fluorescence of fungi did not correlate with the age of the specimen. In most cases, organisms in H&E stained sections were more easily identified with routine light microscopy than with fluorescent microscopy. Conclusion This report suggests that in H&E stained skin specimens, fluorescent microscopy is of little benefit in the identification of fungal organisms. PMID:11602016

Experimental recovery of intrinsic fluorescence and fluorophore concentration in the presence of hemoglobin: spectral effect of scattering and absorption on fluorescence

NASA Astrophysics Data System (ADS)

Du Le, Vinh Nguyen; Patterson, Michael S.; Farrell, Thomas J.; Hayward, Joseph E.; Fang, Qiyin

2015-12-01

The ability to recover the intrinsic fluorescence of biological fluorophores is crucial to accurately identify the fluorophores and quantify their concentrations in the media. Although some studies have successfully retrieved the fluorescence spectral shape of known fluorophores, the techniques usually came with heavy computation costs and did not apply for strongly absorptive media, and the intrinsic fluorescence intensity and fluorophore concentration were not recovered. In this communication, an experimental approach was presented to recover intrinsic fluorescence and concentration of fluorescein in the presence of hemoglobin (Hb). The results indicated that the method was efficient in recovering the intrinsic fluorescence peak and fluorophore concentration with an error of 3% and 10%, respectively. The results also suggested that chromophores with irregular absorption spectra (e.g., Hb) have more profound effects on fluorescence spectral shape than chromophores with monotonic absorption and scattering spectra (e.g., black India ink and polystyrene microspheres).

A Fresh Cadaver Study on Indocyanine Green Fluorescence Lymphography: A New Whole-Body Imaging Technique for Investigating the Superficial Lymphatics.

PubMed

Shinaoka, Akira; Koshimune, Seijiro; Yamada, Kiyoshi; Kumagishi, Kanae; Suami, Hiroo; Kimata, Yoshihiro; Ohtsuka, Aiji

2018-05-01

Identification of the lymphatic system in cadavers is painstaking because lymphatic vessels have very thin walls and are transparent. Selection of appropriate contrast agents is a key factor for successfully visualizing the lymphatics. In this study, the authors introduce a new imaging technique of lymphatic mapping in the whole bodies of fresh cadavers. Ten fresh human cadavers were used for this study. The authors injected 0.1 ml of indocyanine green fluorescence solution subcutaneously at multiple spots along the watershed lines between lymphatic territories and hand and foot regions. After the body was scanned by the near-infrared camera system, fluorescent tissues were harvested and histologic examination was performed under the microscope equipped with the infrared camera system to confirm that they were the lymphatics. Subcutaneously injected indocyanine green was immediately transported into the lymphatic vessels after gentle massage on the injection points. Sweeping massage along the lymphatic vessels facilitated indocyanine green transport inside the lymphatic vessel to move toward the lymph nodes. The lymphatic system was visualized well in the whole body. Histologic examinations confirmed that indocyanine green was detected in the lymphatic lumens specifically, even when located far from the injected points. The lymphatic system could be visualized in whole-body fresh cadavers, as in living bodies, using indocyanine green fluorescence lymphography. Compatibility of indocyanine green lymphography would facilitate the use of cadaveric specimens for macroscopic and microscopic analyses.

Tumor-stem cells interactions by fluorescence imaging

NASA Astrophysics Data System (ADS)

Meleshina, Aleksandra V.; Cherkasova, Elena I.; Sergeeva, Ekaterina; Turchin, Ilya V.; Kiseleva, Ekaterina V.; Dashinimaev, Erdem B.; Shirmanova, Marina V.; Zagaynova, Elena V.

2013-02-01

Recently, great deal of interest is investigation the function of the stem cells (SC) in tumors. In this study, we studied «recipient-tumor- fluorescent stem cells » system using the methods of in vivo imaging and laser scanning microscopy (LSM). We used adipose-derived adult stem (ADAS) cells of human lentiviral transfected with the gene of fluorescent protein Turbo FP635. ADAS cells were administrated into nude mice with transplanted tumor HeLa Kyoto (human cervical carcinoma) at different stages of tumor growth (0-8 days) intravenously or into tumor. In vivo imaging was performed on the experimental setup for epi - luminescence bioimaging (IAP RAS, Nizhny Novgorod). The results of the imaging showed localization of fluorophore tagged stem cells in the spleen on day 5-9 after injection. The sensitivity of the technique may be improved by spectral separation autofluorescence and fluorescence of stem cells. We compared the results of in vivo imaging and confocal laser scanning microscopy (LSM 510 META, Carl Zeiss, Germany). Internal organs of the animals and tumor tissue were investigated. It was shown that with i.v. injection of ADAS, bright fluorescent structures with spectral characteristics corresponding to TurboFP635 protein are locally accumulated in the marrow, lungs and tumors of animals. These findings indicate that ADAS cells integrate in the animal body with transplanted tumor and can be identified by fluorescence bioimaging techniques in vivo and ex vivo.

Nanoscale characterization of vesicle adhesion by normalized total internal reflection fluorescence microscopy.

PubMed

Cardoso Dos Santos, Marcelina; Vézy, Cyrille; Jaffiol, Rodolphe

2016-06-01

We recently proposed a straightforward fluorescence microscopy technique to study adhesion of Giant Unilamellar Vesicles. This technique is based on dual observations which combine epi-fluorescence microscopy and total internal reflection fluorescence (TIRF) microscopy: TIRF images are normalized by epi-fluorescence ones. By this way, it is possible to map the membrane/substrate separation distance with a nanometric resolution, typically ~20 nm, with a maximal working range of 300-400 nm. The purpose of this paper is to demonstrate that this technique is useful to quantify vesicle adhesion from ultra-weak to strong membrane-surface interactions. Thus, we have examined unspecific and specific adhesion conditions. Concerning unspecific adhesion, we have controlled the strength of electrostatic forces between negatively charged vesicles and various functionalized surfaces which exhibit a positive or a negative effective charge. Specific adhesion was highlighted with lock-and-key forces mediated by the well defined biotin/streptavidin recognition. Copyright © 2016 Elsevier B.V. All rights reserved.

Flextime: A Modified Work Force Scheduling Technique for Selected Headquarters Air Force Logistics Command Organizations.

ERIC Educational Resources Information Center

Kimzey, Reed T.; Prince, Samuel M. O.

The thesis discusses the advantages and disadvantages of one work force scheduling technique--flextime. The authors were interested in determining if a flextime schedule could be put into effect in a governmental organization such as Headquarters Air Force Logistics Command (AFLC). The study objectives were to determine the feasibility,…

A-TEEMTM, a new molecular fingerprinting technique: simultaneous absorbance-transmission and fluorescence excitation-emission matrix method

NASA Astrophysics Data System (ADS)

Quatela, Alessia; Gilmore, Adam M.; Steege Gall, Karen E.; Sandros, Marinella; Csatorday, Karoly; Siemiarczuk, Alex; (Ben Yang, Boqian; Camenen, Loïc

2018-04-01

We investigate the new simultaneous absorbance-transmission and fluorescence excitation-emission matrix method for rapid and effective characterization of the varying components from a mixture. The absorbance-transmission and fluorescence excitation-emission matrix method uniquely facilitates correction of fluorescence inner-filter effects to yield quantitative fluorescence spectral information that is largely independent of component concentration. This is significant because it allows one to effectively monitor quantitative component changes using multivariate methods and to generate and evaluate spectral libraries. We present the use of this novel instrument in different fields: i.e. tracking changes in complex mixtures including natural water, wine as well as monitoring stability and aggregation of hormones for biotherapeutics.

Hyper-filter-fluorescer spectrometer for x-rays above 120 keV

DOEpatents

Wang, Ching L.

1983-01-01

An apparatus utilizing filter-fluorescer combinations is provided to measure short bursts of high fluence x-rays above 120 keV energy, where there are no practical absorption edges available for conventional filter-fluorescer techniques. The absorption edge of the prefilter is chosen to be less than that of the fluorescer, i.e., E.sub.PRF technique E.sub.PRF >E.sub.F. In this way, the response function is virtually zero between E.sub.PRF and E.sub.F and well defined and enhanced in an energy band of less than 1000 keV above the 120 keV energy.

Modelling and analysis of ozone concentration by artificial intelligent techniques for estimating air quality

NASA Astrophysics Data System (ADS)

Taylan, Osman

2017-02-01

High ozone concentration is an important cause of air pollution mainly due to its role in the greenhouse gas emission. Ozone is produced by photochemical processes which contain nitrogen oxides and volatile organic compounds in the lower atmospheric level. Therefore, monitoring and controlling the quality of air in the urban environment is very important due to the public health care. However, air quality prediction is a highly complex and non-linear process; usually several attributes have to be considered. Artificial intelligent (AI) techniques can be employed to monitor and evaluate the ozone concentration level. The aim of this study is to develop an Adaptive Neuro-Fuzzy inference approach (ANFIS) to determine the influence of peripheral factors on air quality and pollution which is an arising problem due to ozone level in Jeddah city. The concentration of ozone level was considered as a factor to predict the Air Quality (AQ) under the atmospheric conditions. Using Air Quality Standards of Saudi Arabia, ozone concentration level was modelled by employing certain factors such as; nitrogen oxide (NOx), atmospheric pressure, temperature, and relative humidity. Hence, an ANFIS model was developed to observe the ozone concentration level and the model performance was assessed by testing data obtained from the monitoring stations established by the General Authority of Meteorology and Environment Protection of Kingdom of Saudi Arabia. The outcomes of ANFIS model were re-assessed by fuzzy quality charts using quality specification and control limits based on US-EPA air quality standards. The results of present study show that the ANFIS model is a comprehensive approach for the estimation and assessment of ozone level and is a reliable approach to produce more genuine outcomes.

Spatial and temporal air quality pattern recognition using environmetric techniques: a case study in Malaysia.

PubMed

Syed Abdul Mutalib, Sharifah Norsukhairin; Juahir, Hafizan; Azid, Azman; Mohd Sharif, Sharifah; Latif, Mohd Talib; Aris, Ahmad Zaharin; Zain, Sharifuddin M; Dominick, Doreena

2013-09-01

The objective of this study is to identify spatial and temporal patterns in the air quality at three selected Malaysian air monitoring stations based on an eleven-year database (January 2000-December 2010). Four statistical methods, Discriminant Analysis (DA), Hierarchical Agglomerative Cluster Analysis (HACA), Principal Component Analysis (PCA) and Artificial Neural Networks (ANNs), were selected to analyze the datasets of five air quality parameters, namely: SO2, NO2, O3, CO and particulate matter with a diameter size of below 10 μm (PM10). The three selected air monitoring stations share the characteristic of being located in highly urbanized areas and are surrounded by a number of industries. The DA results show that spatial characterizations allow successful discrimination between the three stations, while HACA shows the temporal pattern from the monthly and yearly factor analysis which correlates with severe haze episodes that have happened in this country at certain periods of time. The PCA results show that the major source of air pollution is mostly due to the combustion of fossil fuel in motor vehicles and industrial activities. The spatial pattern recognition (S-ANN) results show a better prediction performance in discriminating between the regions, with an excellent percentage of correct classification compared to DA. This study presents the necessity and usefulness of environmetric techniques for the interpretation of large datasets aiming to obtain better information about air quality patterns based on spatial and temporal characterizations at the selected air monitoring stations.

Assessment of air quality in Haora River basin using fuzzy multiple-attribute decision making techniques.

PubMed

Singh, Ajit Pratap; Chakrabarti, Sumanta; Kumar, Sumit; Singh, Anjaney

2017-08-01

This paper deals with assessment of air quality in Haora River basin using two techniques. Initially, air quality indices were evaluated using a modified EPA method. The indices were also evaluated using a fuzzy comprehensive assessment (FCA) method. The results obtained from the fuzzy comprehensive assessment method were compared to that obtained from the modified EPA method. To illustrate the applicability of the methodology proposed herein, a case study has been presented. Air samples have been collected at 10 sampling sites located along Haora River. Six important air pollutants, namely, carbon monoxide, sulfur dioxide, nitrogen dioxide, suspended particulate matter (SPM), PM 10 , and lead, were monitored continuously, and air quality maps were generated on the GIS platform. Comparison of the methodologies has clearly highlighted superiority and robustness of the fuzzy comprehensive assessment method in determining air quality indices under study. It has effectively addressed the inherent uncertainties involved in the evaluation, modeling, and interpretation of sampling data, which was beyond the scope of the traditional weighted approaches employed otherwise. The FCA method is robust and prepares a credible platform of air quality evaluation and identification, in face of the uncertainties that remain eclipsed in the traditional approaches like the modified EPA method. The insights gained through the present study are believed to be of pivotal significance in guiding the development and implementation of effective environmental remedial action plans in the study area.

Intravital Fluorescence Excitation in Whole-Animal Optical Imaging.

PubMed

Nooshabadi, Fatemeh; Yang, Hee-Jeong; Bixler, Joel N; Kong, Ying; Cirillo, Jeffrey D; Maitland, Kristen C

2016-01-01

Whole-animal fluorescence imaging with recombinant or fluorescently-tagged pathogens or cells enables real-time analysis of disease progression and treatment response in live animals. Tissue absorption limits penetration of fluorescence excitation light, particularly in the visible wavelength range, resulting in reduced sensitivity to deep targets. Here, we demonstrate the use of an optical fiber bundle to deliver light into the mouse lung to excite fluorescent bacteria, circumventing tissue absorption of excitation light in whole-animal imaging. We present the use of this technology to improve detection of recombinant reporter strains of tdTomato-expressing Mycobacterium bovis BCG (Bacillus Calmette Guerin) bacteria in the mouse lung. A microendoscope was integrated into a whole-animal fluorescence imager to enable intravital excitation in the mouse lung with whole-animal detection. Using this technique, the threshold of detection was measured as 103 colony forming units (CFU) during pulmonary infection. In comparison, the threshold of detection for whole-animal fluorescence imaging using standard epi-illumination was greater than 106 CFU.

Air flow measurement techniques applied to noise reduction of a centrifugal blower

NASA Astrophysics Data System (ADS)

Laage, John W.; Armstrong, Ashli J.; Eilers, Daniel J.; Olsen, Michael G.; Mann, J. Adin

2005-09-01

The air flow in a centrifugal blower was studied using a variety of flow and sound measurement techniques. The flow measurement techniques employed included Particle Image Velocimetry (PIV), pitot tubes, and a five hole spherical probe. PIV was used to measure instantaneous and ensemble-averaged velocity fields over large area of the outlet duct as a function of fan position, allowing for the visualization of the flow as it leave the fan blades and progressed downstream. The results from the flow measurements were reviewed along side the results of the sound measurements with the goal of identifying sources of noise and inefficiencies in flow performance. The radiated sound power was divided into broadband and tone noise and measures of the flow. The changes in the tone and broadband sound were compared to changes in flow quantities such as the turbulent kinetic energy and Reynolds stress. Results for each method will be presented to demonstrate the strengths of each flow measurement technique as well as their limitations. Finally, the role that each played in identifying noise sources is described.

Designing of fluorescent and magnetic imprinted polymer for rapid, selective and sensitive detection of imidacloprid via activators regenerated by the electron transfer-atom transfer radical polymerization (ARGET-ATRP) technique

NASA Astrophysics Data System (ADS)

Kumar, Sunil; Karfa, Paramita; Madhuri, Rashmi; Sharma, Prashant K.

2018-05-01

In this work, we report on a dual-behavior electrochemical/optical sensor for sensitive determination of Imidacloprid by fluorescent dye (fluorescein, FL) and imprinted polymer modified europium doped superparamagnetic iron oxide nanoparticles (FL@SPIONs@MIP). The imidacloprid (IMD)-imprinted polymer was directly synthesized on the Eu-SPIONs surface via Activators regenerated by the electron transfer-atom transfer radical polymerization (ARGET-ATRP) technique. Preparation, characterization and application of the prepared FL@SPIONs@MIP were systematically investigated using scanning electron microscopy (SEM), X-ray diffraction (XRD), vibrating sample magnetometer (VSM), fluorescence spectroscopy and electrochemical techniques. The electrochemical experiments exhibited a remarkable selectivity of the prepared sensor towards IMD. Determination of IMD by the square wave stripping voltammetry method represented a wide linear range of 0.059-0.791 μg L-1 with a detection limit of 0.0125 μg L-1. In addition, the fluorescence method shows a linear range of 0.039-0.942 μg L-1 and LOD of 0.0108 μg L-1. The fluorescence property of prepared FL@SPIONs@MIP was used for rapid, on-spot but selective detection of IMD in real samples. The proposed electrode displayed excellent repeatability and long-term stability and was successfully applied for quantitative and trace level determination of IMD in several real samples.

Microanalysis of dental caries using laser-scanned fluorescence

NASA Astrophysics Data System (ADS)

Barron, Joseph R.; Paton, Barry E.; Zakariasen, Kenneth L.

1992-06-01

It is well known that enamel and dentin fluoresce when illuminated by short-wavelength optical radiation. Fluorescence emission from carious and non-carious regions of teeth have been studied using a new experimental scanning technique for fluorescence analysis of dental sections. Scanning in 2 dimensions will allow surface maps of dental caries to be created. These surface images are then enhanced using the conventional and newer image processing techniques. Carious regions can be readily identified and contour maps can be used to graphically display the degree of damage on both surfaces and transverse sections. Numerous studies have shown that carious fluorescence is significantly different than non-carious regions. The scanning laser fluorescence spectrometer focuses light from a 25 mW He-Cd laser at 442 nm through an objective lens onto a cross-section area as small as 3 micrometers in diameter. Microtome prepared dental samples 100 micrometers thick are laid flat onto an optical bench perpendicular to the incident beam. The sample is moved under computer control in X & Y with an absolute precision of 0.1 micrometers . The backscattered light is both spatial and wavelength filtered before being measured on a long wavelength sensitized photomultiplier tube. High precision analysis of dental samples allow detailed maps of carious regions to be determined. Successive images allow time studies of caries growth and even the potential for remineralization studies of decalcified regions.

Room temperature preparation of fluorescent starch nanoparticles from starch-dopamine conjugates and their biological applications.

PubMed

Shi, Yingge; Xu, Dazhuang; Liu, Meiying; Fu, Lihua; Wan, Qing; Mao, Liucheng; Dai, Yanfeng; Wen, Yuanqing; Zhang, Xiaoyong; Wei, Yen

2018-01-01

Fluorescent organic nanoparticles (FONs) have been regarded as the promising candidates for biomedical applications owing to their well adjustment of chemical structure and optical properties and good biological properties. However, the preparation of FONs from the natural derived polymers has been rarely reported thus far. In current work, we reported a novel strategy for preparation of FONs based on the self-polymerization of starch-dopamine conjugates and polyethyleneimine in rather mild experimental conditions, including air atmosphere, aqueous solution, absent catalysts and at room temperature. The morphology, chemical structure and optical properties of the resultant starch-based FONs were investigated by different characterization techniques. Biological evaluation results demonstrated that these starch-based FONs possess good biocompatibility and fluorescent imaging performance. More importantly, the novel strategy might also be extended for the preparation of many other carbohydrate polymers based FONs with different structure and functions. Therefore, this work opens a new avenue for the preparation and biomedical applications of luminescent carbohydrate polymers. Copyright © 2017 Elsevier B.V. All rights reserved.

Recent trends in atomic fluorescence spectrometry towards miniaturized instrumentation-A review.

PubMed

Zou, Zhirong; Deng, Yujia; Hu, Jing; Jiang, Xiaoming; Hou, Xiandeng

2018-08-17

Atomic fluorescence spectrometry (AFS), as one of the common atomic spectrometric techniques with high sensitivity, simple instrumentation, and low acquisition and running cost, has been widely used in various fields for trace elemental analysis, notably the determination of hydride-forming elements by hydride generation atomic fluorescence spectrometry (HG-AFS). In recent years, the soaring demand of field analysis has significantly promoted the miniaturization of analytical atomic spectrometers or at least instrumental components. Various techniques have also been developed to approach the goal of portable/miniaturized AFS instrumentation for field analysis. In this review, potentially portable/miniaturized AFS techniques, primarily involving advanced instrumental components and whole instrumentation with references since 2000, are summarized and discussed. The discussion mainly includes five aspects: radiation source, atomizer, detector, sample introduction, and miniaturized atomic fluorescence spectrometer/system. Copyright © 2018 Elsevier B.V. All rights reserved.

Trace Chemical Detection through Vegetation Sentinels and Fluorescence Spectroscopy

Treesearch

John E. Anderson; Robert L. Fischer; Jean D. Nelson

2006-01-01

Detection of environmental contaminants through vegetation sentinels has long been a goal of remote sensing scientists. A promising technique that should be scalable to wide-area applications is the combined use of genetically modified vascular plants and fluorescence imaging. The ultimate goal of our research is to produce a bioreporter that will express fluorescence...

Antibody-mediated fluorescence enhancement based on shifting the intramolecular dimer<-->monomer equilibrium of fluorescent dyes.

PubMed

Wei, A P; Blumenthal, D K; Herron, J N

1994-05-01

A novel concept is described for directly coupling fluorescence emission to protein-ligand binding. It is based on shifting the intramolecular monomer<-->dimer equilibrium of two fluorescent dyes linked by a short spacer. A 13-residue peptide, recognized by a monoclonal antibody against human chorionic gonadotrophin (hCG), was labeled with fluorescein (F) and tetramethylrhodamine (T) at its N- and C-terminus, respectively. Spectral evidence suggests that when the conjugate is free in solution, F and T exist as an intramolecular dimer. Fluorescence quenching of fluorescein and rhodamine is approximately 98% and approximately 90%, respectively, due to dimerization. When the double-labeled peptide is bound to anti-hCG, however, the rhodamine fluorescence increases by up to 7.8-fold, depending upon the excitation wavelength. This is attributed to the dissociation of intramolecular dimers brought about by conformational changes of the conjugate upon binding. Fluorescein fluorescence, on the other hand, was still quenched because of excited-state energy transfer and residual ground-state interactions. Antibody binding also resulted in a approximately 3.4-fold increase in fluorescence anisotropy of the peptide. These changes in intensity and anisotropy allow direct measurement of antigen-antibody binding with a fluorescence plate reader or a polarization analyzer, without the need for separation steps and labeling antibodies. Because recent advances in peptide technology have allowed rapid and economical identification of antigen-mimicking peptides, the double-labeled peptide approach offers many opportunities for developing new diagnostic assays and screening new therapeutic drugs. It also has many potential applications to techniques involving recombinant antibodies, biosensors, cell sorting, and DNA probes.

Fluorescence Lifetime Imaging Microscopy Using Near-Infrared Contrast Agents

PubMed Central

Nothdurft, Ralph; Sarder, Pinaki; Bloch, Sharon; Culver, Joseph; Achilefu, Samuel

2013-01-01

Although single-photon fluorescence lifetime imaging microscopy (FLIM) is widely used to image molecular processes using a wide range of excitation wavelengths, the captured emission of this technique is confined to the visible spectrum. Here, we explore the feasibility of utilizing near-infrared (NIR) fluorescent molecular probes with emission >700 nm for FLIM of live cells. The confocal microscope is equipped with a 785 nm laser diode, a red-enhanced photomultiplier tube, and a time-correlated single photon counting card. We demonstrate that our system reports the lifetime distributions of NIR fluorescent dyes, cypate and DTTCI, in cells. In cells labeled separately or jointly with these dyes, NIR FLIM successfully distinguishes their lifetimes, providing a method to sort different cell populations. In addition, lifetime distributions of cells co-incubated with these dyes allow estimate of the dyes’ relative concentrations in complex cellular microenvironments. With the heightened interest in fluorescence lifetime-based small animal imaging using NIR fluorophores, this technique further serves as a bridge between in vitro spectroscopic characterization of new fluorophore lifetimes and in vivo tissue imaging. PMID:22788550

Applications of two-photon fluorescence microscopy in deep-tissue imaging

NASA Astrophysics Data System (ADS)

Dong, Chen-Yuan; Yu, Betty; Hsu, Lily L.; Kaplan, Peter D.; Blankschstein, D.; Langer, Robert; So, Peter T. C.

2000-07-01

Based on the non-linear excitation of fluorescence molecules, two-photon fluorescence microscopy has become a significant new tool for biological imaging. The point-like excitation characteristic of this technique enhances image quality by the virtual elimination of off-focal fluorescence. Furthermore, sample photodamage is greatly reduced because fluorescence excitation is limited to the focal region. For deep tissue imaging, two-photon microscopy has the additional benefit in the greatly improved imaging depth penetration. Since the near- infrared laser sources used in two-photon microscopy scatter less than their UV/glue-green counterparts, in-depth imaging of highly scattering specimen can be greatly improved. In this work, we will present data characterizing both the imaging characteristics (point-spread-functions) and tissue samples (skin) images using this novel technology. In particular, we will demonstrate how blind deconvolution can be used further improve two-photon image quality and how this technique can be used to study mechanisms of chemically-enhanced, transdermal drug delivery.

Flame imaging using planar laser induced fluorescence of sulfur dioxide

NASA Astrophysics Data System (ADS)

Honza, Rene; Ding, Carl-Philipp; Dreizler, Andreas; Böhm, Benjamin

2017-09-01

Laser induced fluorescence of sulfur dioxide (SO2-PLIF) has been demonstrated as a useful tool for flame imaging. Advantage was taken from the strong temperature dependence of the SO2 fluorescence signal. SO2 fluorescence intensity increases by more than one order of magnitude if the temperature changes from ambient conditions to adiabatic flame temperatures of stoichiometric methane-air flames. This results in a steep gradient of SO2-PLIF intensities at the reaction zone and therefore can be used as a reliable flame marker. SO2 can be excited electronically using the fourth-harmonic of an Nd:YAG laser at 266 nm. This is an attractive alternative to OH-LIF, a well-recognized flame front marker, because no frequency-doubled dye lasers are needed. This simplifies the experimental setup and is advantageous for measurements at high repetition rates where dye bleaching can become an issue. To prove the performance of this approach, SO2-PLIF measurements were performed simultaneously with OH-PLIF on laminar premixed methane-air Bunsen flames for equivalence ratios between 0.9 and 1.25. These measurements were compared to 1D laminar flamelet simulations. The SO2 fluorescence signal was found to follow the temperature rise of the flame and is located closer to the steep temperature gradient than OH. Finally, the combined SO2- and OH-PLIF setup was applied to a spark ignition IC-engine to visualize the development of the early flame kernel.

Time-resolved laser fluorescence spectroscopy of organic ligands by europium: Fluorescence quenching and lifetime properties

NASA Astrophysics Data System (ADS)

Nouhi, A.; Hajjoul, H.; Redon, R.; Gagné, J. P.; Mounier, S.

2018-03-01

Time-resolved Laser Fluorescence Spectroscopy (TRLFS) has proved its usefulness in the fields of biophysics, life science and geochemistry to characterize the fluorescence probe molecule with its chemical environment. The purpose of this study is to demonstrate the applicability of this powerful technique combined with Steady-State (S-S) measurements. A multi-mode factor analysis, in particular CP/PARAFAC, was used to analyze the interaction between Europium (Eu) and Humic substances (HSs) extracted from Saint Lawrence Estuary in Canada. The Saint Lawrence system is a semi-enclosed water stream with connections to the Atlantic Ocean and is an excellent natural laboratory. CP/PARAFAC applied to fluorescence S-S data allows introspecting ligands-metal interactions and the one-site 1:1 modeling gives information about the stability constants. From the spectral signatures and decay lifetimes data given by TRLFS, one can deduce the fluorescence quenching which modifies the fluorescence and discuss its mechanisms. Results indicated a relatively strong binding ability between europium and humic substances samples (Log K value varies from 3.38 to 5.08 at pH 7.00). Using the Stern-Volmer plot, it has been concluded that static and dynamic quenching takes places in the case of salicylic acid and europium interaction while for HSs interaction only a static quenching is observed.

Intestine pH measurements using fluorescence imaging: an in-vivo preliminary study

NASA Astrophysics Data System (ADS)

Marechal, Xavier-Marie; Mordon, Serge R.; Devoisselle, Jean-Marie; Begu, Sylvie; Mathieu, D.; Buys, Bruno; Dhelin, Guy; Lesage, Jean C.; Neviere, Remi; Chopin, Claude

1999-02-01

Measurement of gastrointestinal intramucosal pH has been recognized as an important factor in the detection of hypoxia-induced dysfunctions. However, current pH measurement techniques are limited in terms of time and spatial resolution. A major advance in accurate pH measurement was the development of the ratiometric fluorescent indicator dye, 2',7'-bis(carboxyethyl)-4,5- carboxyfluorescein (BCECF). This study aimed to demonstrate the feasibility of fluorescence imaging technique to measure in vivo the pH of intestine. The intestine was inserted in an optical chamber placed under a microscope. Animals were injected i.v. with the pH-sensitive fluorescent dye BCECF. Fluorescence was visualized by illuminating the intestine alternately at 490 and 470 nm. The emitted fluorescence was directed to an intensified camera. The ratio of emitted fluorescence at excitation wavelengths of 490 and 470 nm was measured, corrected and converted to pH by constructing a calibration curve. The pH controls were performed with a pH microelectrode correlated with venous blood gas sampling. We concluded that accurate pH measurements of rat intestine can be obtained by fluorescence imaging using BCECF. This technology could be easily adapted for endoscopic pH measurement.

Autonomous unmanned air vehicles (UAV) techniques

NASA Astrophysics Data System (ADS)

Hsu, Ming-Kai; Lee, Ting N.

2007-04-01

The UAVs (Unmanned Air Vehicles) have great potentials in different civilian applications, such as oil pipeline surveillance, precision farming, forest fire fighting (yearly), search and rescue, boarder patrol, etc. The related industries of UAVs can create billions of dollars for each year. However, the road block of adopting UAVs is that it is against FAA (Federal Aviation Administration) and ATC (Air Traffic Control) regulations. In this paper, we have reviewed the latest technologies and researches on UAV navigation and obstacle avoidance. We have purposed a system design of Jittering Mosaic Image Processing (JMIP) with stereo vision and optical flow to fulfill the functionalities of autonomous UAVs.

KrF laser-induced OH fluorescence imaging in a supersonic combustion tunnel

NASA Technical Reports Server (NTRS)

Quagliaroli, T. M.; Laufer, G.; Hollo, S. D.; Krauss, R. H.; Whitehurst, R. B., III; Mcdaniel, J. C., Jr.

1992-01-01

Planar fluorescence images of OH in a continuous-flow, electrical-resistively heated, high enthalpy, hydrogen-air combustion tunnel, induced by a tunable KrF laser, were recorded. These images were compared to previously recorded fluorescence images induced by a doubled-dye laser under similar conditions. Images induced by the doubled-dye laser system demonstrated a severe distortion caused by absorption and fluorescence trapping. By contrast, images of the fluorescence induced by the tunable KrF laser retained the symmetry properties of the flow. Based on signal-to-noise ratio measurements the yield of the fluorescence induced by the doubled-dye laser is larger than the fluorescence yield induced by the KrF laser. The measurements in the present facility of OH fluorescence induced by the KrF laser were limited by the photon-statistical noise. Based 2 on this result, doubled-dye laser systems are recommended for OH imaging in small and OH lean (less than 10 exp 15/cu cm) facilities. KrF lasers should be selected otherwise.

Measurement of the fluorescence of crop residues: A tool for controlling soil erosion

NASA Technical Reports Server (NTRS)

Daughtry, C. S. T.; Mcmurtrey, J. E., III; Chappelle, E. W.; Hunter, W. J.

1994-01-01

Management of crop residues, the portion of a crop left in the field after harvest, is an important conservation practice for minimizing soil erosion and for improving water quality. Quantification of crop residue cover is required to evaluate the effectiveness of conservation tillage practices. Methods are needed to quantify residue cover that are rapid, accurate, and objective. The fluorescence of crop residue was found to be a broadband phenomenon with emission maxima at 420 to 495 nm for excitations of 350 to 420 nm. Soils had low intensity broadband emissions over the 400 to 690 nm region for excitations of 300 to 600 nm. The range of relative fluorescence intensities for the crop residues was much greater than the fluorescence observed of the soils. As the crop residues decompose their blue fluorescence values approach the fluorescence of the soil. Fluorescence techniques are concluded to be less ambiguous and better suited for discriminating crop residues and soils than reflectance methods. If properly implemented, fluorescence techniques can be used to quantify, not only crop residue cover, but also photosynthetic efficiency in the field.

Heat transfer assembly for a fluorescent lamp and fixture

DOEpatents

Siminovitch, M.J.; Rubenstein, F.M.; Whitman, R.E.

1992-12-29

In a lighting fixture including a lamp and a housing, a heat transfer structure is disclosed for reducing the minimum lamp wall temperature of a fluorescent light bulb. The heat transfer structure, constructed of thermally conductive material, extends from inside the housing to outside the housing, transferring heat energy generated from a fluorescent light bulb to outside the housing where the heat energy is dissipated to the ambient air outside the housing. Also disclosed is a method for reducing minimum lamp wall temperatures. Further disclosed is an improved lighting fixture including a lamp, a housing and the aforementioned heat transfer structure. 11 figs.

Heat transfer assembly for a fluorescent lamp and fixture

DOEpatents

Siminovitch, Michael J.; Rubenstein, Francis M.; Whitman, Richard E.

1992-01-01

In a lighting fixture including a lamp and a housing, a heat transfer structure is disclosed for reducing the minimum lamp wall temperature of a fluorescent light bulb. The heat transfer structure, constructed of thermally conductive material, extends from inside the housing to outside the housing, transferring heat energy generated from a fluorescent light bulb to outside the housing where the heat energy is dissipated to the ambient air outside the housing. Also disclosed is a method for reducing minimum lamp wall temperatures. Further disclosed is an improved lighting fixture including a lamp, a housing and the aforementioned heat transfer structure.

Rapid purification of fluorescent enzymes by ultrafiltration

NASA Technical Reports Server (NTRS)

Benjaminson, M. A.; Satyanarayana, T.

1983-01-01

In order to expedite the preparation of fluorescently tagged enzymes for histo-cyctochemistry, a previously developed method employing gel column purification was compared with a more rapid modern technique using the Millipore Immersible CX-ultrafilter. Microscopic evaluation of the resulting conjugates showed comparable products. Much time and effort is saved using the new technique.

Rapid purification of fluorescent enzymes by ultrafiltration

NASA Technical Reports Server (NTRS)

Benjaminson, M. A.; Satyanarayana, T.

1983-01-01

In order to expedite the preparation of fluorescently tagged enzymes for histo/cytochemistry, a previously developed method employing gel column purification was compared with a more rapid modern technique using the Millipore Immersible CX-ultrafilter. Microscopic evaluation of the resulting conjugates showed comparable products. Much time and effort is saved using the new technique.

Free volume dependent fluorescence property of PMMA composite: Positron annihilation studies

NASA Astrophysics Data System (ADS)

Ravindrachary, V.; Praveena, S. D.; Bhajantri, R. F.; Ismayil, Crasta, Vincent

2013-02-01

The free volume related fluorescence properties of chalcone chromophore [1-(4-methylphenyl)-3-(4-N, N, dimethylaminophenyl)-2-propen-1-one doped Poly(methyl methacrylate) have been studied using fluorescence spectroscopy and Positron Annihilation lifetime spectroscopy techniques. The fluorescence spectra show that the fluorescence behavior depends on the free volume dependent polymer microstructure and varies with dopant concentration with in the composite. The origin and variation of fluorescence is understood by twisted internal charge transfer state as well as free volume. The Positron annihilation study shows that the free volume related microstructure of the composite is vary with doping level.

Fluorescence detection of esophageal neoplasia

NASA Astrophysics Data System (ADS)

Borisova, E.; Vladimirov, B.; Avramov, L.

2008-06-01

White-light endoscopy is well-established and wide used modality. However, despite the many technological advances that have been occurred, conventional endoscopy is suboptimal and usually detects advanced stage lesions. The limitations of standard endoscopy initiate development of spectroscopic techniques, additional to standard endoscopic equipment. One of the most sensitive approaches is fluorescence spectroscopy of gastrointestinal mucosa for neoplasia detection. In the recent study delta-aminolevulinic acid/Protoporphyrin IX (5-ALA/PpIX) is used as fluorescent marker for dysplasia and tumor detection in esophagus. The 5-ALA is administered per os six hours before measurements at dose 20 mg/kg weight. Excitation source has max of emission at 405 nm and light is delivered by the standard light guide of the endoscopic equipment. Through endoscopic instrumental channel a fiber is applied to return information about fluorescence to microspectrometer. Spectral features observed during endoscopic investigations could be distinct as the next regions: 450-630 nm region, where tissue autofluorescence is observed; 630-710 nm region, where fluorescence of PpIX is clearly pronounced; 530-580 nm region, where minima in the autofluorescence signal are observed, related to reabsorption of blood. The lack of fluorescence peaks in the red spectral area for normal mucosa is an indication for selective accumulation of 5-ALA/PpIX only in abnormal sites Very good correlation between fluorescence signals and histology examination of the lesions investigated is achieved.

Correlative Fluorescence and Electron Microscopy

PubMed Central

Schirra, Randall T.; Zhang, Peijun

2014-01-01

Correlative fluorescence and electron microscopy (CFEM) is a multimodal technique that combines dynamic and localization information from fluorescence methods with ultrastructural data from electron microscopy, to give new information about how cellular components change relative to the spatiotemporal dynamics within their environment. In this review, we will discuss some of the basic techniques and tools of the trade for utilizing this attractive research method, which is becoming a very powerful tool for biology labs. The information obtained from correlative methods has proven to be invaluable in creating consensus between the two types of microscopy, extending the capability of each, and cutting the time and expense associate with using each method separately for comparative analysis. The realization of the advantages of these methods in cell biology have led to rapid improvement in the protocols and have ushered in a new generation of instruments to reach the next level of correlation – integration. PMID:25271959

Toward quantitative fluorescence microscopy with DNA origami nanorulers.

PubMed

Beater, Susanne; Raab, Mario; Tinnefeld, Philip

2014-01-01

The dynamic development of fluorescence microscopy has created a large number of new techniques, many of which are able to overcome the diffraction limit. This chapter describes the use of DNA origami nanostructures as scaffold for quantifying microscope properties such as sensitivity and resolution. The DNA origami technique enables placing of a defined number of fluorescent dyes in programmed geometries. We present a variety of DNA origami nanorulers that include nanorulers with defined labeling density and defined distances between marks. The chapter summarizes the advantages such as practically free choice of dyes and labeling density and presents examples of nanorulers in use. New triangular DNA origami nanorulers that do not require photoinduced switching by imaging transient binding to DNA nanostructures are also reported. Finally, we simulate fluorescence images of DNA origami nanorulers and reveal that the optimal DNA nanoruler for a specific application has an intermark distance that is roughly 1.3-fold the expected optical resolution. © 2014 Elsevier Inc. All rights reserved.

Radio morphing - towards a full parametrisation of the radio signal from air showers

NASA Astrophysics Data System (ADS)

Zilles, A.; Charrier, D.; Kotera, K.; Le Coz, S.; Martineau-Huynh, O.; Medina, C.; Niess, V.; Tueros, M.; de Vries, K.

2017-12-01

Over the last decades, radio detection of air showers has been established as a detection technique for ultra-high-energy cosmic-rays impinging on the Earth's atmosphere with energies far beyond LHC energies. Today’s second-generation of digital radio-detection experiments, as e.g. AERA or LOFAR, are becoming competitive in comparison to already standard techniques e.g. fluorescence light detection. Thanks to a detailed understanding of the physics of the radio emission in extensive air showers, simulations of the radio signal are already successfully tested and applied in the reconstruction of cosmic rays. However the limits of the computational power resources are easily reached when it comes to computing electric fields at the numerous positions requested by large or dense antenna arrays. In the case of mountainous areas as e.g. for the GRAND array, where 3D shower simulations are necessary, the problem arises with even stronger acuity. Therefore we developed a full parametrisation of the emitted radio signal on the basis of generic shower simulations which will reduce the simulation time by orders of magnitudes. In this talk we will present this concept after a short introduction to the concept of the radio detection of air-shower induced by cosmic rays.

X-ray Fluorescence Holography: Principles, Apparatus, and Applications

NASA Astrophysics Data System (ADS)

Hayashi, Kouichi; Korecki, Pawel

2018-06-01

X-ray fluorescence holography (XFH) is an atomic structure determination technique that combines the capabilities of X-ray diffraction and X-ray fluorescence spectroscopy. It provides a unique means of gaining fully three-dimensional information about the local atomic structure and lattice site positions of selected elements inside compound samples. In this work, we discuss experimental and theoretical aspects that are essential for the efficient recording and analysis of X-ray fluorescence holograms and review the most recent advances in XFH. We describe experiments performed with brilliant synchrotron radiation as well as with tabletop setups that employ conventional X-ray tubes.

Étude comparative des techniques d'analyse par fluorescence X à dispersion d'énergie (ED-XRF) et à dispersion de longueur d'onde (WD-XRF), et par spectrométrie d'émission atomique à source plasma couplée par induction (ICP-AES)

NASA Astrophysics Data System (ADS)

Rahmani, A.; Benyaïch, F.; Bounakhla, M.; Bilal, E.; Moutte, J.; Gruffat, J. J.; Zahry, F.

2004-11-01

Dans ce travail, nous présentons une étude comparative des techniques d'analyse par fluorescence X à dispersion d'énergie (ED-XRF) et à dispersion de longueur d'onde (WD-XRF), et par spectrométrie d'émission atomique à source plasma couplée par induction (ICP-AES). Les résultats de la calibration des spectromètres à dispersion d'énergie, à excitation par sources radioactives (55Fe, 109Cd et 241Am) et à excitation secondaire (cible secondaire Mo et Cu) du Centre National pour l'Energie, les Sciences et les Techniques Nucléaires (CNESTEN, Rabat, Maroc) sur des échantillons étalons de références de l'Agence International de l'Energie Atomique (AIEA) et du Community Bureau of Référence (BCR) ont été comparés aux résultats d'analyse des mêmes échantillons étalons par la spectrométrie X à dispersion de longueur d'onde (WD-XRF) et par spectrométrie d'émission atomique à source plasma couplé par induction (ICP-AES) au département GENERIC du centre SPIN à l'Ecole des Mines de Saint-Etienne (France). Les trois techniques d'analyse utilisées donnent des résultats comparables pour le dosage des éléments majeurs, alors que pour les traces on note des déviations importantes à cause des effets de matrice qui sont difficiles à corriger dans le cas de la fluorescence X.

Detection of hepatocarcinoma in rats by integration of the fluorescence spectrum: Experimental model

NASA Astrophysics Data System (ADS)

Marcassa, J. C.; Ferreira, J.; Zucoloto, S.; Castro E Silva, O., Jr.; Marcassa, L. G.; Bagnato, V. S.

2006-05-01

The incorporation of spectroscopic techniques into diagnostic procedures may greatly improve the chances for precise diagnostics. One promising technique is fluorescence spectroscopy, which has recently been used to detect many different types of diseases. In this work, we use laser-induced tissue fluorescence to detect hepatocarcinoma in rats using excitation light at wavelengths of 443 and 532 nm. Hepatocarcinoma was induced chemically in Wistar rats. The collected fluorescence spectrum ranges from the excitation wavelength up to 850 nm. A mathematical procedure carried out on the spectrum determines a figure of merit value, which allows the detection of hepatocarcinoma. The figure of merit involves a procedure which evaluates the ratio between the backscattered excitation wavelength and the broad emission fluorescence band. We demonstrate that a normalization allowed by integration of the fluorescence spectra is a simple operation that may allow the detection of hepatocarcinoma.

Let's Exploit Available Knowledge on Vegetation Fluorescence

NASA Technical Reports Server (NTRS)

Magnani, Federico; Raddi, Sabrina; Mohammed, Gina; Middleton, Elizabeth M.

2014-01-01

The potential to measure vegetation fluorescence from space (1) and to derive from it direct information on the gross primary productivity (GPP) of terrestrial ecosystems is probably the most thrilling development in remote sensing and global ecology of recent years, as it moves Earth observation techniques from the detection of canopy biophysics (e.g., fraction of absorbed radiation) and biochemistry (chlorophyll and nitrogen content) to the realm of ecosystem function. The existence of a functional relationship between fluorescence and photosynthesis has been elucidated over the last decade by several laboratories, notably as part of the preliminary studies of the European Space Agency Fluorescence Explorer (FLEX) Earth Explorer Mission. The empirical observation presented by Guanter et al. (2) of a linear relationship between fluorescence radiance and GPP, however, provides the first experimental confirmation of the feasibility of the approach— already thoroughly tested at leaf level—at the desired scale, despite the confounding effects associated with the satellite detection of such a faint signal. A word of clarification is needed here. The use of fluorescence as a probe of leaf photochemistry has been a staple of plant ecophysiology for decades, rooted in a sound understanding of photosynthetic energy dissipation. However, most past studies had to rely for the interpretation of results on active (pulse-saturated) techniques, making them unsuitable for remote-sensing applications. Over recent years, however, novel process based models have been developed for the interpretation of steady-state, solar-induced fluorescence at the leaf to canopy scale (3). We are therefore in a position to move beyond the mere empirical observation of an association between GPP and fluorescence radiance. In particular, Guanter et al. (2) base their analysis on the assumption of a constant ratio between photosynthetic and fluorescence light use efficiencies (equation 3 in ref

Complex formation between chlorophyll a and cytochrome c: surface properties at the air-water interface. Absorbance, fluorescence and fluorescence-lifetime in Langmuir-Blodgett films.

PubMed

Lamarche, F; Picard, G; Téchy, F; Aghion, J; Leblanc, R M

1991-04-23

The binding of cytochrome c to an insoluble monolayer of chlorophyll a was studied. Surface pressure (II), surface potential (delta V) and [14C]cytochrome c surface-concentration (gamma) isotherms were measured versus molecular area (sigma) in mixed films. Compared to the successive-addition method, this procedure allows the formation of homogeneous mixed films. The cytochrome c is incorporated into a chlorophyll a monolayer, compressed at a surface pressure of 20 mN.m-1. On expansion, the quantity of protein incorporated into the monolayer gradually increases. Subsequent compression-expansion cycles result in similar isotherms, distinct from that measured during the first expansion. All surface properties measured, but more specifically the surface radioactivity of [14C]cytochrome c, indicate the irreversibility of protein incorporation into the chlorophyll a monolayer. In fact, surface properties of the binary film are completely different from the properties of either of the pure components. As a result, calculated values of surface potentials for mixed films using the additivity law deviate from experimentally measured potentials. The absorption and fluorescence spectra of mixed films transferred onto a solid substrate by the Langmuir-Blodgett technique, indicate a dilution effect of chlorophyll a by cytochrome c. However, the dilution effect cannot be detected by the fluorescence lifetimes of pure chlorophyll a and mixed chlorophyll a-cytochrome c films, both shorter than 0.2 ns. This provides support for the existence of an energy-transfer mechanism between chlorophyll a monomer and chlorophyll a aggregates which could serve as an energy trap. The role of the protein could be related to that of the matrix.

Laboratory and airborne techniques for measuring fluoresence of natural surfaces

NASA Technical Reports Server (NTRS)

Stoertz, G. E.; Hemphill, W. R.

1972-01-01

Techniques are described for obtaining fluorescence spectra from samples of natural surfaces that can be used to predict spectral regions in which these surfaces would emit solar-stimulated or laser-stimulated fluorescence detectable by remote sensor. Scattered or reflected stray light caused large errors in spectrofluorometer analysis or natural sample surfaces. Most spurious light components can be eliminated by recording successive fluorescence spectra for each sample, using identical instrument settings, first with an appropriate glass or gelatin filter on the excitation side of the sample, and subsequently with the same filter on the emission side of the sample. This technique appears more accurate than any alternative technique for testing the fluorescence of natural surfaces.

Engineering fluorescent poly(dopamine) capsules.

PubMed

Chen, Xi; Yan, Yan; Müllner, Markus; van Koeverden, Martin P; Noi, Ka Fung; Zhu, Wei; Caruso, Frank

2014-03-18

The recent development of poly(dopamine) (PDA) capsules provides new opportunities for their application in biology and medicine. To advance the biomedical application of PDA capsules, strategies that enable the preparation of fluorescently labeled PDA (F-PDA) capsules are required, as this will allow evaluation of their cellular interactions using a range of fluorescence-based techniques. Herein, we report a facile approach for the fabrication of F-PDA capsules via the polymerization of dopamine (DA) on sacrificial templates in the presence of hydrogen peroxide (H2O2). F-PDA capsules with well-defined sizes are prepared by templating different organic and inorganic particles. The resulting F-PDA capsules show negligible cytotoxicity in HeLa cells after incubation for 48 h. We also demonstrate visualization of the F-PDA capsules following internalization by HeLa cells using conventional fluorescence microscopy, en route toward detailed investigations on their biological interactions.

In vivo near-infrared fluorescence imaging of Leishmania amazonensis expressing infrared fluorescence protein (iRFP) for real-time monitoring of cutaneous leishmaniasis in mice.

PubMed

Oliveira, Janaina Correia; da Silva, Aline Caroline; Oliveira, Renato Antonio Dos Santos; Pereira, Valéria Rêgo Alves; Gil, Laura Helena Vega Gonzales

2016-11-01

The use of Leishmania amazonensis-infected BALB/c mice is an important model for the study of experimental cutaneous leishmaniasis. Here we report the development of a non-invasive method to directly evaluate and measure parasite burden during the course of the infection, based on the near-infrared fluorescence detection of a recombinant L. amazonensis strain. So, we generated a L. amazonensis strain that stably expresses the near-infrared protein (iRFP) gene and compared the maintenance of its vitro and in vivo characteristics, such as fitness, pathogenicity and fluorescence emission. After that, we followed the disease development, as well as the parasite burden in BALB/c mice footpads infected with L. amazonensis-iRFP, by using an in vivo near-infrared fluorescence scanner. In vitro results showed a linear correlation between the fluorescence emission and the number of parasites. The in vivo study showed that the use of iRFP-transfected L. amazonensis enables the monitoring of parasite burden by measuring fluorescence signals. Therefore, this technique can be confidently used to directly monitor parasitic load and infection overtime and could be an excellent tool for in vitro and in vivo screening of anti-leishmanial drugs and vaccine efficiency. This is the first report of the use of the near-infrared fluorescence imaging technique for monitoring in vivo cutaneous leishmaniasis. Copyright © 2016 Elsevier B.V. All rights reserved.

Ozone Air Quality over North America: Part II-An Analysis of Trend Detection and Attribution Techniques.

PubMed

Porter, P Steven; Rao, S Trivikrama; Zurbenko, Igor G; Dunker, Alan M; Wolff, George T

2001-02-01

Assessment of regulatory programs aimed at improving ambient O 3 air quality is of considerable interest to the scientific community and to policymakers. Trend detection, the identification of statistically significant long-term changes, and attribution, linking change to specific clima-tological and anthropogenic forcings, are instrumental to this assessment. Detection and attribution are difficult because changes in pollutant concentrations of interest to policymakers may be much smaller than natural variations due to weather and climate. In addition, there are considerable differences in reported trends seemingly based on similar statistical methods and databases. Differences arise from the variety of techniques used to reduce nontrend variation in time series, including mitigating the effects of meteorology and the variety of metrics used to track changes. In this paper, we review the trend assessment techniques being used in the air pollution field and discuss their strengths and limitations in discerning and attributing changes in O 3 to emission control policies.

Fluorescent probes for lipid rafts: from model membranes to living cells.

PubMed

Klymchenko, Andrey S; Kreder, Rémy

2014-01-16

Membrane microdomains (rafts) remain one of the controversial issues in biophysics. Fluorescent molecular probes, which make these lipid nanostructures visible through optical techniques, are one of the tools currently used to study lipid rafts. The most common are lipophilic fluorescent probes that partition specifically into liquid ordered or liquid disordered phase. Their partition depends on the lipid composition of a given phase, which complicates their use in cellular membranes. A second class of probes is based on environment-sensitive dyes, which partition into both phases, but stain them by different fluorescence color, intensity, or lifetime. These probes can directly address the properties of each separate phase, but their cellular applications are still limited. The present review focuses on summarizing the current state in the field of developing and applying fluorescent molecular probes to study lipid rafts. We highlight an urgent need to develop new probes, specifically adapted for cell plasma membranes and compatible with modern fluorescence microscopy techniques to push the understanding of membrane microdomains forward. Copyright © 2014 Elsevier Ltd. All rights reserved.

Kinetics of bacterial fluorescence staining with 3,3'-diethylthiacyanine.

PubMed

Thomas, Marlon S; Nuñez, Vicente; Upadhyayula, Srigokul; Zielins, Elizabeth R; Bao, Duoduo; Vasquez, Jacob M; Bahmani, Baharak; Vullev, Valentine I

2010-06-15

For more than a century, colorimetric and fluorescence staining have been the foundation of a broad range of key bioanalytical techniques. The dynamics of such staining processes, however, still remains largely unexplored. We investigated the kinetics of fluorescence staining of two gram-negative and two gram-positive species with 3,3'-diethylthiacyanine (THIA) iodide. An increase in the THIA fluorescence quantum yield, induced by the bacterial dye uptake, was the principal reason for the observed emission enhancement. The fluorescence quantum yield of THIA depended on the media viscosity and not on the media polarity, which suggested that the microenvironment of the dye molecules taken up by the cells was restrictive. The kinetics of fluorescence staining did not manifest a statistically significant dependence neither on the dye concentration, nor on the cell count. In the presence of surfactant additives, however, the fluorescence-enhancement kinetic patterns manifested species specificity with statistically significant discernibility.

Two-photon excitation fluorescence bioassays.

PubMed

Hänninen, Pekka; Soukka, Jori; Soini, Juhani T

2008-01-01

Application of two-photon excitation of fluorescence in microscopy is one of the major discoveries of the "renaissance" of light microscopy that started in the 1980s. The technique derives its advantages from the biologically "smooth" wavelength of the excitation light and the confinement of the excitation. Difficult, and seemingly nontransparent, samples may be imaged with the technique with good resolution. Although the bioresearch has been concentrating mostly on the positive properties of the technique for imaging, the same properties may be applied successfully to nonimaging bioassays. This article focuses on the development path of two-photon excitation-based assay system.

Characterization of humic acids by two-dimensional correlation fluorescence spectroscopy

NASA Astrophysics Data System (ADS)

Nakashima, K.; Xing, Shaoyong; Gong, Yongkuan; Miyajima, Toru

2008-07-01

We have investigated interaction between humic acids and heavy metal ions by fluorescence spectroscopy. The humic acids examined are Aldrich humic acid (AHA) and Dando humic acid (DHA), and heavy metal ions are Cu 2+ and Pb 2+. The binding constants between the humic acids and the heavy metal ions are obtained by a conventional fluorescence quenching technique. The two prominent bands in the fluorescence spectra of the humic acids give different binding constants, implying that the two bands are originated from different fluorescent species in the matrices of the humic acids. This was confirmed by two-dimensional correlation analysis based on the quenching perturbation on the fluorescence spectra. Two prominent cross peaks corresponding to the two fluorescence bands are obtained in the asynchronous maps, indicating that the two fluorescence bands belong to different species. The order of the response of the two fluorescence bands to the quenching perturbation is also elucidated based on Noda's rule.

Automatic enhancement of skin fluorescence localization due to refractive index matching

NASA Astrophysics Data System (ADS)

Churmakov, Dmitry Y.; Meglinski, Igor V.; Piletsky, Sergey A.; Greenhalgh, Douglas A.

2004-07-01

Fluorescence diagnostic techniques are notable amongst many other optical methods, as they offer high sensitivity and non-invasive measurements of tissue properties. However, a combination of multiple scattering and physical heterogeneity of biological tissues hampers the interpretation of the fluorescence measurements. The analyses of the spatial distribution of endogenous and exogenous fluorophores excitations within tissues and their contribution to the detected signal localization are essential for many applications. We have developed a novel Monte Carlo technique that gives a graphical perception of how the excitation and fluorescence detected signal are localized in tissues. Our model takes into account spatial distribution of fluorophores and their quantum yields. We demonstrate that matching of the refractive indices of ambient medium and topical skin layer improves spatial localization of the detected fluorescence signal within the tissue. This result is consistent with the recent conclusion that administering biocompatible agents results in higher image contrast.

Simultaneous measurements of temperature and density in air flows using UV laser spectroscopy

NASA Technical Reports Server (NTRS)

Fletcher, D. G.; Mckenzie, R. L.

1991-01-01

The simultaneous measurement of temperature and density using laser-induced fluorescence of oxygen in combination with Q-branch Raman scattering of nitrogen and oxygen is demonstrated in a low-speed air flow. The lowest density and temperature measured in the experiment correspond to the freestream values at Mach 5 in the Ames 3.5-Foot Hypersonic Wind Tunnel for stagnation conditions of 100 atm and 1000 K. The experimental results demonstrate the viability of the optical technique for measurements that support the study of compressible turbulence and the validation of numerical codes in supersonic and hypersonic wind tunnel flows.

Selective detection of Fe2+ by combination of CePO4:Tb3+ nanocrystal-H2O2 hybrid system with synchronous fluorescence scan technique.

PubMed

Chen, Hongqi; Ren, Jicun

2012-04-21

A new method for quenching kinetic discrimination of Fe(2+) and Fe(3+), and sensitive detection of trace amount of Fe(2+) was developed by using synchronous fluorescence scan technique. The principle of this assay is based on the quenching kinetic discrimination of Fe(2+) and Fe(3+) in CePO(4):Tb(3+) nanocrytals-H(2)O(2) hybrid system and the Fenton reaction between Fe(2+) and H(2)O(2). Stable, water-soluble and well-dispersible CePO(4):Tb(3+) nanocrystals were synthesized in aqueous solutions, and characterized by transmission electron microscopy (TEM) and electron diffraction spectroscopy (EDS). We found that both Fe(2+) and Fe(3+) could quench the synchronous fluorescence of CePO(4):Tb(3+) nanocrytals-H(2)O(2) system, but their quenching kinetics velocities were quite different. In the presence of Fe(3+), the synchronous fluorescent intensity was unchanged after only one minute, but in the presence of Fe(2+), the synchronous fluorescent intensity decreased slowly until 28 min later. The Fenton reaction between Fe(2+) and H(2)O(2) resulted in hydroxyl radicals which effectively quenched the synchronous fluorescence of the CePO(4):Tb(3+) nanocrystals due to the oxidation of Ce(3+) into Ce(4+) by hydroxyl radicals. Under optimum conditions, the linear range for Fe(2+) is 3 nM-2 μM, and the limit of detection is 2.0 nM. The method was used to analyze water samples.

The air afterglow revisited

NASA Technical Reports Server (NTRS)

Kaufman, F.

1972-01-01

The air afterglow, 0 + NO2 chemiluminescence, is discussed in terms of fluorescence, photodissociation, and quantum theoretical calculations of NO2. The experimental results presented include pressure dependence, M-dependence, spectral dependence of P and M, temperature dependence, and infrared measurements. The NO2 energy transfer model is also discussed.

Automated detection of fluorescent cells in in-resin fluorescence sections for integrated light and electron microscopy.

PubMed

Delpiano, J; Pizarro, L; Peddie, C J; Jones, M L; Griffin, L D; Collinson, L M

2018-04-26

Integrated array tomography combines fluorescence and electron imaging of ultrathin sections in one microscope, and enables accurate high-resolution correlation of fluorescent proteins to cell organelles and membranes. Large numbers of serial sections can be imaged sequentially to produce aligned volumes from both imaging modalities, thus producing enormous amounts of data that must be handled and processed using novel techniques. Here, we present a scheme for automated detection of fluorescent cells within thin resin sections, which could then be used to drive automated electron image acquisition from target regions via 'smart tracking'. The aim of this work is to aid in optimization of the data acquisition process through automation, freeing the operator to work on other tasks and speeding up the process, while reducing data rates by only acquiring images from regions of interest. This new method is shown to be robust against noise and able to deal with regions of low fluorescence. © 2018 The Authors. Journal of Microscopy published by JohnWiley & Sons Ltd on behalf of Royal Microscopical Society.

Simulating a Measurement of the 2nd Knee in the Cosmic Ray Spectrum with an Atmospheric Fluorescence Telescope Tower Array

PubMed Central

Liu, Jiali; Yang, Qunyu; Bai, Yunxiang; Cao, Zhen

2014-01-01

A fluorescence telescope tower array has been designed to measure cosmic rays in the energy range of 1017–1018 eV. A full Monte Carlo simulation, including air shower production, light generation and propagation, detector response, electronics, and trigger system, has been developed for that purpose. Using such a simulation tool, the detector configuration, which includes one main tower array and two side-trigger arrays, 24 telescopes in total, has been optimized. The aperture and the event rate have been estimated. Furthermore, the performance of the X max⁡ technique in measuring composition has also been studied. PMID:24737964

Review of fluorescence guided surgery visualization and overlay techniques

PubMed Central

Elliott, Jonathan T.; Dsouza, Alisha V.; Davis, Scott C.; Olson, Jonathan D.; Paulsen, Keith D.; Roberts, David W.; Pogue, Brian W.

2015-01-01

In fluorescence guided surgery, data visualization represents a critical step between signal capture and display needed for clinical decisions informed by that signal. The diversity of methods for displaying surgical images are reviewed, and a particular focus is placed on electronically detected and visualized signals, as required for near-infrared or low concentration tracers. Factors driving the choices such as human perception, the need for rapid decision making in a surgical environment, and biases induced by display choices are outlined. Five practical suggestions are outlined for optimal display orientation, color map, transparency/alpha function, dynamic range compression, and color perception check. PMID:26504628

Genetically encoded sensors and fluorescence microscopy for anticancer research

NASA Astrophysics Data System (ADS)

Zagaynova, Elena V.; Shirmanova, Marina V.; Sergeeva, Tatiana F.; Klementieva, Natalia V.; Mishin, Alexander S.; Gavrina, Alena I.; Zlobovskay, Olga A.; Furman, Olga E.; Dudenkova, Varvara V.; Perelman, Gregory S.; Lukina, Maria M.; Lukyanov, Konstantin A.

2017-02-01

Early response of cancer cells to chemical compounds and chemotherapeutic drugs were studied using novel fluorescence tools and microscopy techniques. We applied confocal microscopy, two-photon fluorescence lifetime imaging microscopy and super-resolution localization-based microscopy to assess structural and functional changes in cancer cells in vitro. The dynamics of energy metabolism, intracellular pH, caspase-3 activation during staurosporine-induced apoptosis as well as actin cytoskeleton rearrangements under chemotherapy were evaluated. We have showed that new genetically encoded sensors and advanced fluorescence microscopy methods provide an efficient way for multiparameter analysis of cell activities

Laser induced fluorescence spectrum analysis of OH from photo-dissociation of water in gas phase

NASA Astrophysics Data System (ADS)

Li, Guohua; Ye, Jingfeng; Zhang, Zhengrong; Wang, Sheng; Hu, Zhiyun; Zhao, Xinyan

2017-05-01

The OH can be generated from photo-dissociation of water in the gas phase, and the generated OH has served in tagging velocimetry using the time-flight method. The hydroxyl tagging mechanism has the advantages of non-seeding, kindly flow following character, but its application in the reaction region is limited for the fluorescence interference from nascent OH. In this paper, we explored the laser induced fluorescence spectrum of OH both from burning and photodissociation. A photo-dissociation laser induced fluorescence (PD-LIF) system with optical multichannel analysis instrument (OMA) for spectrum analysis was developed. Based on multichannel mechanism, the LIF spectrum of OH from photo-dissociation and burning were acquired simultaneously. The temporal spectrum profiles of dissociation OH both in flame and air were taken by varying the pump-probe delay. The normalized emission spectrum in flame showed a process of rotational relaxation while in air the spectrum was almost not changed. The fluorescence intensity was precisely proportional to the base states population, so we can get certain states that the OH from dissociation was predominant from the fluorescence intensity ratio of OH. This result can be further utilized for hydroxyl tagging velocimetry technology (HTV) which was less affected by burning OH.

Redox-Responsive Fluorescent Probes with Different Design Strategies.

PubMed

Lou, Zhangrong; Li, Peng; Han, Keli

2015-05-19

In an aerobic organism, reactive oxygen species (ROS) are an inevitable metabolic byproduct. Endogenously produced ROS have a significant role in physiological processes, but excess ROS can cause oxidative stress and can damage tissue. Cells possess elaborate mechanisms to regulate their internal redox status. The intracellular redox homeostasis plays an essential role in maintaining cellular function. However, moderate alterations in redox balance can accompany major transitions in a cell's life cycle. Because of the role of ROS in physiology and in pathology, researchers need new tools to study redox chemistry in biological systems.In recent years, researchers have made remarkable progress in developing new, highly sensitive and selective fluorescent probes that respond to redox changes, and in this Account we highlight related research, primarily from our own group. We present an overview of the design, photophysical properties, and fluorescence transduction mechanisms of reported molecules that probe redox changes. We have designed and synthesized a series of fluorescent probes for redox cycles in biological systems relying on the active center of glutathione peroxidase (GPx). We have also constructed probes based on the oxidation and reduction of hydroquinone and of 2,2,6,6-tetramethylpiperidinooxy (TEMPO). Most of these probes exhibit high sensitivity and good selectivity, absorb in the near-infrared, and respond rapidly. Such probes are useful for confocal fluorescence microscopy, a dynamic imaging technique that could allow researchers to observe biologically important ROS and antioxidants in real time. This technique and these probes provide potentially useful tools for exploring the generation, transport, physiological function, and pathogenic mechanisms of ROS and antioxidants.We also describe features that could improve the properties of redox-responsive fluorescent probes: greater photostability; rapid, dynamic, cyclic and ratiometric responses; and

Intracellular applications of fluorescence correlation spectroscopy: prospects for neuroscience.

PubMed

Kim, Sally A; Schwille, Petra

2003-10-01

Based on time-averaging fluctuation analysis of small fluorescent molecular ensembles in equilibrium, fluorescence correlation spectroscopy has recently been applied to investigate processes in the intracellular milieu. The exquisite sensitivity of fluorescence correlation spectroscopy provides access to a multitude of measurement parameters (rates of diffusion, local concentration, states of aggregation and molecular interactions) in real time with fast temporal and high spatial resolution. The introduction of dual-color cross-correlation, imaging, two-photon excitation, and coincidence analysis coupled with fluorescence correlation spectroscopy has expanded the utility of the technique to encompass a wide range of promising applications in living cells that may provide unprecedented insight into understanding the molecular mechanisms of intracellular neurobiological processes.

Tomography of epidermal growth factor receptor binding to fluorescent Affibody in vivo studied with magnetic resonance guided fluorescence recovery in varying orthotopic glioma sizes

NASA Astrophysics Data System (ADS)

Holt, Robert W.; Demers, Jennifer-Lynn H.; Sexton, Kristian J.; Gunn, Jason R.; Davis, Scott C.; Samkoe, Kimberley S.; Pogue, Brian W.

2015-02-01

The ability to image targeted tracer binding to epidermal growth factor receptor (EGFR) was studied in vivo in orthotopically grown glioma tumors of different sizes. The binding potential was quantified using a dual-tracer approach, which employs a fluorescently labeled peptide targeted to EGFR and a reference tracer with similar pharmacokinetic properties but no specific binding, to estimate the relative bound fraction from kinetic compartment modeling. The recovered values of binding potential did not vary significantly as a function of tumor size (1 to 33 mm3), suggesting that binding potential may be consistent in the U251 tumors regardless of size or stage after implantation. However, the fluorescence yield of the targeted fluorescent tracers in the tumor was affected significantly by tumor size, suggesting that dual-tracer imaging helps account for variations in absolute uptake, which plague single-tracer imaging techniques. Ex vivo analysis showed relatively high spatial heterogeneity in each tumor that cannot be resolved by tomographic techniques. Nonetheless, the dual-tracer tomographic technique is a powerful tool for longitudinal bulk estimation of receptor binding.

Effect of surfactant and budesonide on the pulmonary distribution of fluorescent dye in mice.

PubMed

Huang, Liang-Ti; Yeh, Tsu-Fu; Kuo, Yu-Lin; Chen, Pin-Chuan; Chen, Chung-Ming

2015-02-01

Surfactant is a useful vehicle for the intratracheal delivery of medicine to the distal lung. The aim of this study was to analyze the effect of intratracheal surfactant and budesonide instillation on the pulmonary distribution of fluorescent dye in mice. Male athymic nude mice were assigned randomly as controls, fluorescent dye, fluorescent dye + surfactant (50 mg/kg), fluorescent dye + budesonide (0.25 mg/kg), and fluorescent dye + surfactant + budesonide groups. A total volume of 60 μL fluorescent solutions was intratracheally injected and followed by 60 μL of air. We photographed and measured fluorescence in the lungs, from the back, 15 minutes after intratracheal administration using an IVIS Xenogen imaging instrument. The fluorescent dye (1,1'-dioctadecyltetramethyl indotricarbocyanine iodide) was most strongly detected near the trachea and weakly detected in the lungs in mice administered with fluorescent solutions. Almost no fluorescence was seen in the lung region of control mice. Intratracheal administration of surfactant or budesonide increased fluorescent intensity compared with control mice. Combined administration of surfactant and budesonide further increased fluorescent intensity compared with mice given surfactant or budesonide alone. Surfactant and budesonide enhance the pulmonary distribution of fluorescent dye in mice. Copyright © 2014. Published by Elsevier B.V.

Plasmonics Enhanced Smartphone Fluorescence Microscopy.

PubMed

Wei, Qingshan; Acuna, Guillermo; Kim, Seungkyeum; Vietz, Carolin; Tseng, Derek; Chae, Jongjae; Shir, Daniel; Luo, Wei; Tinnefeld, Philip; Ozcan, Aydogan

2017-05-18

Smartphone fluorescence microscopy has various applications in point-of-care (POC) testing and diagnostics, ranging from e.g., quantification of immunoassays, detection of microorganisms, to sensing of viruses. An important need in smartphone-based microscopy and sensing techniques is to improve the detection sensitivity to enable quantification of extremely low concentrations of target molecules. Here, we demonstrate a general strategy to enhance the detection sensitivity of a smartphone-based fluorescence microscope by using surface-enhanced fluorescence (SEF) created by a thin metal-film. In this plasmonic design, the samples are placed on a silver-coated glass slide with a thin spacer, and excited by a laser-diode from the backside through a glass hemisphere, generating surface plasmon polaritons. We optimized this mobile SEF system by tuning the metal-film thickness, spacer distance, excitation angle and polarization, and achieved ~10-fold enhancement in fluorescence intensity compared to a bare glass substrate, which enabled us to image single fluorescent particles as small as 50 nm in diameter and single quantum-dots. Furthermore, we quantified the detection limit of this platform by using DNA origami-based brightness standards, demonstrating that ~80 fluorophores per diffraction-limited spot can be readily detected by our mobile microscope, which opens up new opportunities for POC diagnostics and sensing applications in resource-limited-settings.

Motor Oil Classification Based on Time-Resolved Fluorescence

PubMed Central

Mu, Taotao; Chen, Siying; Zhang, Yinchao; Guo, Pan; Chen, He; Meng, Fandong

2014-01-01

A time-resolved fluorescence (TRF) technique is presented for classifying motor oils. The system is constructed with a third harmonic Nd:YAG laser, a spectrometer, and an intensified charge coupled device (ICCD) camera. Steady-state and time-resolved fluorescence (TRF) measurements are reported for several motor oils. It is found that steady-state fluorescence is insufficient to distinguish the motor oil samples. Then contour diagrams of TRF intensities (CDTRFIs) are acquired to serve as unique fingerprints to identify motor oils by using the distinct TRF of motor oils. CDTRFIs are preferable to steady-state fluorescence spectra for classifying different motor oils, making CDTRFIs a particularly choice for the development of fluorescence-based methods for the discrimination and characterization of motor oils. The two-dimensional fluorescence contour diagrams contain more information, not only the changing shapes of the LIF spectra but also the relative intensity. The results indicate that motor oils can be differentiated based on the new proposed method, which provides reliable methods for analyzing and classifying motor oils. PMID:24988439

Intravital Fluorescence Excitation in Whole-Animal Optical Imaging

PubMed Central

Bixler, Joel N.; Kong, Ying; Cirillo, Jeffrey D.; Maitland, Kristen C.

2016-01-01

Whole-animal fluorescence imaging with recombinant or fluorescently-tagged pathogens or cells enables real-time analysis of disease progression and treatment response in live animals. Tissue absorption limits penetration of fluorescence excitation light, particularly in the visible wavelength range, resulting in reduced sensitivity to deep targets. Here, we demonstrate the use of an optical fiber bundle to deliver light into the mouse lung to excite fluorescent bacteria, circumventing tissue absorption of excitation light in whole-animal imaging. We present the use of this technology to improve detection of recombinant reporter strains of tdTomato-expressing Mycobacterium bovis BCG (Bacillus Calmette Guerin) bacteria in the mouse lung. A microendoscope was integrated into a whole-animal fluorescence imager to enable intravital excitation in the mouse lung with whole-animal detection. Using this technique, the threshold of detection was measured as 103 colony forming units (CFU) during pulmonary infection. In comparison, the threshold of detection for whole-animal fluorescence imaging using standard epi-illumination was greater than 106 CFU. PMID:26901051

Two-Photon/Laser-Induced Fluorescence (TP/LIF) sensor

NASA Technical Reports Server (NTRS)

Bradshaw, John D.

1994-01-01

The Two-Photon/Laser-Induced Fluorescence (TP/LIF) technique is based on the stepwise excitation of the OH transitions, X(exp 2)II, v(exp '') = 0 yields X(exp 2)II, v(exp '') = 1 (lambda = 2.9 microns) and X(exp 2)II v(exp '') = 1 yields A(exp2)Sigma, v' = 0 (lambda = 345 nm) with background free fluorescence monitoring of the A(exp 2)Sigma, v' = 0 yields X(exp 2)II, v(exp '') = 0 transition near 309 nm. This technique has awaited the advent of a suitable mid-infrared (2.9 microns) laser source. Turnable mid-IR lasers now exist that are capable of meeting the specifications required of a high sensitivity TP/LIF OH sensor.

Airborne particle characterization by spatial scattering and fluorescence

NASA Astrophysics Data System (ADS)

Barton, John; Hirst, Edwin; Kaye, Paul; Saunders, Spencer; Clark, Don

1999-11-01

Several workers have reported the development of systems which allow the measurement of intrinsic fluorescence from particles irradiated with ultra-violet radiation. The fluorescence data are frequently recorded in conjunction with other parameters such as particle size, measured either as a function of optical scatter or as an aerodynamic size. The motivation for this work has been principally the detection of bioaerosols within an ambient environment. Previous work by the authors has shown that an analysis of the scattering profile of a particle, i.e.: the spatial distribution of light scattered by the particle carried in a sample air-stream, can provide an effective means of particle characterization and classification in terms of both size and shape parameters. Current work is aimed at the simultaneous recording of both spatial scattering and fluorescence data from individual particles with a view to substantially enhanced discrimination of biological aerosols. A prototype instrument has recently been completed which employs a cw 266 nm laser source to produce both elastic (spatial scattering) and inelastic (fluorescence) signals from individual airborne particles. The instrument incorporates a custom designed high-gain multi- pixel hybrid photodiode (HPD) to record the spatial scattering data and a single photomultiplier to record total fluorescence from the illuminated particle. Recorded data are processed to allow the classification of airborne particles on the basis of size, shape, and fluorescence for both biological and non- biological aerosols.

Dual-detector X-ray fluorescence imaging of ancient artifacts with surface relief

PubMed Central

Smilgies, Detlef-M.; Powers, Judson A.; Bilderback, Donald H.; Thorne, Robert E.

2012-01-01

Interpretation of X-ray fluorescence images of archeological artifacts is complicated by the presence of surface relief and roughness. Using two symmetrically arranged fluorescence detectors in a back-reflection geometry, the proper X-ray fluorescence yield can be distinguished from intensity variations caused by surface topography. This technique has been applied to the study of Roman inscriptions on marble. PMID:22713888

Rapid fluorescence detection of pathogenic bacteria using magnetic enrichment technique combined with magnetophoretic chromatography.

PubMed

Che, Yulan; Xu, Yi; Wang, Renjie; Chen, Li

2017-08-01

A rapid and sensitive analytical method was developed to detect pathogenic bacteria which combined magnetic enrichment, fluorescence labeling with polyethylene glycol (PEG) magnetophoretic chromatography. As pathogenic bacteria usually exist in complex matrixes at low concentration, an efficient enrichment is essential for diagnosis. In order to capture series types of pathogenic bacteria in samples, amino-modified magnetic nanoparticles (Fe 3 O 4 @SiO 2 -NH 2 ) were prepared for efficient enrichment by the electrostatic interaction with pathogenic bacteria. It was shown that the capture efficiency reached up to 95.4% for Escherichia coli (E. coli). Furthermore, quantitative analysis of the bacteria was achieved by using acridine orange (AO) as a fluorescence probe for the captured E. coli due to its ability of staining series types of bacteria and rapid labeling. In order to remove the free magnetic nanoparticles and redundant fluorescent reagent, the labeled suspension was poured into a PEG separation column and was separated by applying an external magnetic field. The presence of 100 cfu mL -1 E. coli could be detected for semi-quantitative analysis by observing the separation column with the naked eye, and the concentration could be further evaluated by fluorescence detection. All the above processes were finished within 80 min. It was demonstrated that a good linear relationship existed between the fluorescence intensity and the concentration of E. coli ranging from 10 2 to 10 6  cfu mL -1 , with a detection limit of 100 cfu mL -1 when E. coli acted as target bacteria. The recovery rate of E. coli was 93.6∼102.0% in tap water and cooked meat samples, and the RSD was lower than 7% (n = 6); the result coincided with the conventional plate count method. Graphical abstract ᅟ.

The effect of flow reduction on microphytobenthos development in an alpine river stretch using novel fluorescence techniques

NASA Astrophysics Data System (ADS)

Ganglbauer, A.; Bondar-Kunze, E.; Hein, T.; Zeiringer, B.

2009-04-01

. The measurements included field surveys and two experimental settings. During May 2008 we conducted an in-situ experiment with artificial substrata to investigate the effect of flow velocity changes. We exposed glass slides in baskets along two transects in the River Ybbs at two sampling sites and eight different positions. After a period of about four weeks with weekly recurrent measurements including flow velocity, water depth, chlorophyll a content and electron transport rate (ETR) we started our experiment. Glass slides were taken from each position and were exposed in a flow reduced impounded area in the river Ybbs near Göstling. There low flow velocity was used to test the effect on microphytobenthos development. The next ten days daily measurements of flow velocity, water depth, chlorophyll-a content and electron transport rate (ETR) with the pulse amplitude modulated fluorescence method and microscopic analysis were undertaken. Based on these daily measurements under almost stable environmental conditions we could ascertain a shift in the benthic algae community. To assess the distribution along a river stretch we measured 70 sampling points at each sampling side. To characterize the biomass and activity of the microphytobenthos we used Pulse Amplitude Modulated Fluorescence (PAM-Fluorescence). Using this technique allow to measure the biomass (Chlorophyll a) and the ETR (electron transport rate) simultaneously without destroying the structure. With this technique it is possible to The PAM technique measure directly the fluorescence of chlorophyll a in the photosystem two. The quantum yield you get is the probability that a photon can be used photochemically. The quantum yield offers the possibility to illustrate the fitness of algae. Based on these measurements short term responses can be measured and combined with the results of field surveys. These analytical results were used for a habitat modelling approach to describe the microphytobenthos development

Micro-scale temperature measurement method using fluorescence polarization

NASA Astrophysics Data System (ADS)

Tatsumi, K.; Hsu, C.-H.; Suzuki, A.; Nakabe, K.

2016-09-01

A novel method that can measure the fluid temperature in microscopic scale by measuring the fluorescence polarization is described in this paper. The measurement technique is not influenced by the quenching effects which appears in conventional LIF methods and is believed to show a higher reliability in temperature measurements. Experiment was performed using a microchannel flow and fluorescent molecule probes, and the effects of the fluid temperature, fluid viscosity, measurement time, and pH of the solution on the measured fluorescence polarization degree are discussed to understand the basic characteristics of the present method. The results showed that fluorescence polarization is considerably less sensible to these quenching factors. A good correlation with the fluid temperature, on the other hand, was obtained and agreed well with the theoretical values confirming the feasibility of the method.

Estimating background-subtracted fluorescence transients in calcium imaging experiments: a quantitative approach.

PubMed

Joucla, Sébastien; Franconville, Romain; Pippow, Andreas; Kloppenburg, Peter; Pouzat, Christophe

2013-08-01

Calcium imaging has become a routine technique in neuroscience for subcellular to network level investigations. The fast progresses in the development of new indicators and imaging techniques call for dedicated reliable analysis methods. In particular, efficient and quantitative background fluorescence subtraction routines would be beneficial to most of the calcium imaging research field. A background-subtracted fluorescence transients estimation method that does not require any independent background measurement is therefore developed. This method is based on a fluorescence model fitted to single-trial data using a classical nonlinear regression approach. The model includes an appropriate probabilistic description of the acquisition system's noise leading to accurate confidence intervals on all quantities of interest (background fluorescence, normalized background-subtracted fluorescence time course) when background fluorescence is homogeneous. An automatic procedure detecting background inhomogeneities inside the region of interest is also developed and is shown to be efficient on simulated data. The implementation and performances of the proposed method on experimental recordings from the mouse hypothalamus are presented in details. This method, which applies to both single-cell and bulk-stained tissues recordings, should help improving the statistical comparison of fluorescence calcium signals between experiments and studies. Copyright © 2013 Elsevier Ltd. All rights reserved.

Maximizing the Biochemical Resolving Power of Fluorescence Microscopy

PubMed Central

Esposito, Alessandro; Popleteeva, Marina; Venkitaraman, Ashok R.

2013-01-01

Most recent advances in fluorescence microscopy have focused on achieving spatial resolutions below the diffraction limit. However, the inherent capability of fluorescence microscopy to non-invasively resolve different biochemical or physical environments in biological samples has not yet been formally described, because an adequate and general theoretical framework is lacking. Here, we develop a mathematical characterization of the biochemical resolution in fluorescence detection with Fisher information analysis. To improve the precision and the resolution of quantitative imaging methods, we demonstrate strategies for the optimization of fluorescence lifetime, fluorescence anisotropy and hyperspectral detection, as well as different multi-dimensional techniques. We describe optimized imaging protocols, provide optimization algorithms and describe precision and resolving power in biochemical imaging thanks to the analysis of the general properties of Fisher information in fluorescence detection. These strategies enable the optimal use of the information content available within the limited photon-budget typically available in fluorescence microscopy. This theoretical foundation leads to a generalized strategy for the optimization of multi-dimensional optical detection, and demonstrates how the parallel detection of all properties of fluorescence can maximize the biochemical resolving power of fluorescence microscopy, an approach we term Hyper Dimensional Imaging Microscopy (HDIM). Our work provides a theoretical framework for the description of the biochemical resolution in fluorescence microscopy, irrespective of spatial resolution, and for the development of a new class of microscopes that exploit multi-parametric detection systems. PMID:24204821

Nuclear Resonance Fluorescence of U-235

DOE Office of Scientific and Technical Information (OSTI.GOV)

Warren, Glen A.; Caggiano, Joseph A.; Hensley, Walter K.

Nuclear resonance fluorescence is a physical process that provides an isotopic-specific signature that could be used for the identification and characterization of materials. The technique involves the detection of prompt discrete-energy photons emitted from a sample which is exposed to photons in the MeV energy range. Potential applications of the technique range from detection of high explosives to characterization of special nuclear materials. One isotope of significant interest is 235U. Pacific Northwest National Laboratory and Passport Systems have collaborated to conduct measurements to search for a nuclear resonance fluorescence response of 235U below 3 MeV using a 200 g samplemore » of highly enriched uranium. Nine 235U resonances between 1650 and 2010 keV were identified in the preliminary analysis. Analysis of the measurement data to determine the integrated cross sections of the resonances is in progress.« less

Fate of the CHBrsub2O radical in air

NASA Technical Reports Server (NTRS)

Bayes, K. D.; Friedl, R. F.

2003-01-01

Trace amounts of bromoform in air have been photolyzed at 266 and 303 nm to form Br atoms and CHBr2 radicals. The Br concentration as a funtion of time is followed by resonance fluorescence. The CHBr2 radicals react with O2 in the air to form peroxy radicals.

Laser-saturated fluorescence measurements in laminar sooting diffusion flames

NASA Technical Reports Server (NTRS)

Wey, Changlie

1993-01-01

The hydroxyl radical is known to be one of the most important intermediate species in the combustion processes. The hydroxyl radical has also been considered a dominant oxidizer of soot particles in flames. In this investigation the hydroxyl concentration profiles in sooting diffusion flames were measured by the laser-saturated fluorescence (LSF) method. The temperature distributions in the flames were measured by the two-line LSF technique and by thermocouple. In the sooting region the OH fluorescence was too weak to make accurate temperature measurements. The hydroxyl fluorescence profiles for all four flames presented herein show that the OH fluorescence intensities peaked near the flame front. The OH fluorescence intensity dropped sharply toward the dark region of the flame and continued declining to the sooting region. The OH fluorescence profiles also indicate that the OH fluorescence decreased with increasing height in the flames for all flames investigated. Varying the oxidizer composition resulted in a corresponding variation in the maximum OH concentration and the flame temperature. Furthermore, it appears that the maximum OH concentration for each flame increased with increasing flame temperature.

Fluorescence lifetime imaging microscopy using near-infrared contrast agents.

PubMed

Nothdurft, R; Sarder, P; Bloch, S; Culver, J; Achilefu, S

2012-08-01

Although single-photon fluorescence lifetime imaging microscopy (FLIM) is widely used to image molecular processes using a wide range of excitation wavelengths, the captured emission of this technique is confined to the visible spectrum. Here, we explore the feasibility of utilizing near-infrared (NIR) fluorescent molecular probes with emission >700 nm for FLIM of live cells. The confocal microscope is equipped with a 785 nm laser diode, a red-enhanced photomultiplier tube, and a time-correlated single photon counting card. We demonstrate that our system reports the lifetime distributions of NIR fluorescent dyes, cypate and DTTCI, in cells. In cells labelled separately or jointly with these dyes, NIR FLIM successfully distinguishes their lifetimes, providing a method to sort different cell populations. In addition, lifetime distributions of cells co-incubated with these dyes allow estimate of the dyes' relative concentrations in complex cellular microenvironments. With the heightened interest in fluorescence lifetime-based small animal imaging using NIR fluorophores, this technique further serves as a bridge between in vitro spectroscopic characterization of new fluorophore lifetimes and in vivo tissue imaging. © 2012 The Author Journal of Microscopy © 2012 Royal Microscopical Society.

Success of segmentation in a sequence of images tracking the growth of endogenously fluorescent kidneys

NASA Astrophysics Data System (ADS)

Goldberg, Robert R.; Goldberg, Michael R.

1999-05-01

A previous paper by the authors presented an algorithm that successfully segmented organs grown in vitro from their surroundings. It was noticed that one difficulty in standard dyeing techniques for the analysis of contours in organs was due to the fact that the antigen necessary to bind with the fluorescent dye was not uniform throughout the cell borders. To address these concerns, a new fluorescent technique was utilized. A transgenic mouse line was genetically engineered utilizing the hoxb7/gfp (green fluorescent protein). Whereas the original technique (fixed and blocking) required a numerous number of noise removal filtering and sophisticated segmentation techniques, segmentation on the GFP kidney required only an adaptive binary threshold technique which yielded excellent results without the need for specific noise reduction. This is important for tracking the growth of kidney development through time.

Diolistics: incorporating fluorescent dyes into biological samples using a gene gun

PubMed Central

O’Brien, John A.; Lummis, Sarah C.R.

2007-01-01

The hand-held gene gun provides a rapid and efficient method of incorporating fluorescent dyes into cells, a technique that is becoming known as diolistics. Transporting fluorescent dyes into cells has, in the past, used predominantly injection or chemical methods. The use of the gene gun, combined with the new generation of fluorescent dyes, circumvents some of the problems of using these methods and also enables the study of cells that have proved difficult traditionally to transfect (e.g. those deep in tissues and/or terminally differentiated); in addition, the use of ion- or metabolite-sensitive dyes provides a route to study cellular mechanisms. Diolistics is also ideal for loading cells with optical nanosensors – nanometre-sized sensors linked to fluorescent probes. Here, we discuss the theoretical considerations of using diolistics, the advantages compared with other methods of inserting dyes into cells and the current uses of the technique, with particular consideration of nanosensors. PMID:17945370

Observed OH and HO2 concentrations in the upper troposphere inside and outside of Asian monsoon influenced air.

NASA Astrophysics Data System (ADS)

Marno, D. R.; Künstler, C.; Hens, K.; Tatum Ernest, C.; Broch, S.; Fuchs, H.; Martinez, M.; Bourtsoukidis, E.; Williams, J.; Holland, F.; Hofzumahaus, A.; Tomsche, L.; Fischer, H.; Klausner, T.; Schlager, H.; Eirenschmalz, L.; Stratmann, G.; Stock, P.; Ziereis, H.; Roiger, A.; Bohn, B.; Zahn, A.; Wahner, A.; Lelieveld, J.; Harder, H.

2016-12-01

The Asian monsoon convectively transports pollutants like volatile organic compounds (VOCs), NOx, and SO2 from the boundary layer over South Asia into the upper troposphere where they can potentially enter the stratosphere, or be dispersed globally. Therefore, it is crucial to understand the oxidizing capacity of this system regarding the rate of aerosol formation, and conversion of pollutants into compounds that have much shorter atmospheric lifetimes. OH plays a central role in this oxidation process. During the OMO-ASIA campaign in the summer of 2015, OH and HO2 were measured onboard the High Altitude Long-Range (HALO) Research Aircraft. Two laser-induced fluorescence instruments based on the fluorescence assay by gas expansion technique (LIF-FAGE) had been deployed, the AIR-LIF instrument from Forschungszentrum Jülich GmbH and the HORUS instrument from the Max Planck Institute for Chemistry, Mainz. To measure the chemical background of OH potentially produced inside the HORUS instrument from highly oxidized VOCs, atmospheric OH is scavenged by an Inlet Pre-injector (IPI) system. This was the first time an IPI system was implemented within an airborne LIF-FAGE instrument measuring OH and HO2. Throughout this campaign OH and HO2 were measured at 12 to 15km within the Asian monsoon anticyclone. These measurements have been contrasted by probing air outside the anticyclone in air masses influenced by North American emissions, and in very clean air masses originated from the southern hemisphere.

Detection of peroxyl radicals from polluted air by free radical reaction combined with liquid chromatography signal amplification technique.

PubMed

Wang, Guoying; Jia, Shiming; Niu, Xiuli; Liu, Yanrong; Tian, Haoqi; Chen, Xuefu; Shi, Gaofeng

2018-01-22

Free radicals play an important role in the oxidizing power of polluted air, the development of aging-related diseases, the formation of ozone, and the production of secondary particulate matter. The high variability of peroxyl radical concentration has prevented the detection of possible trends or distributions in the concentration of free radicals. We present a new method, free radical reaction combined with liquid chromatography photodiode array detection, for identifying and quantifying peroxyl radicals in polluted air. Functionalized graphene was used for loading peroxyl radicals and reactive molecules in air sampling system, which can facilitate reaction kinetics (charge transfers) between peroxyl radicals and reaction molecules. Separation was performed with and without a preliminary exposure of the polluted air sample to reactive molecule(s) system. The integral chromatographic peak areas before and after air sampling are used to quantify the atmospheric peroxyl radicals in polluted air. The utility of the new technique was tested with measurements carried out in the field. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Pulp tissue in sex determination: A fluorescent microscopic study

PubMed Central

Nayar, Amit; Singh, Harkanwal Preet; Leekha, Swati

2014-01-01

Aims: To determine and compare the reliability of pulp tissue in determination of sex and to analyze whether caries have any effect on fluorescent body test. Materials and Methods: This study was carried on 50 maxillary and mandibular teeth (25 male teeth and 25 female teeth), which were indicated for extraction. The teeth are categorized into 5 groups, 10 each (5 from males and 5 from females) on the basis of caries progression. The pulp cells are stained with quinacrine hydrochloride and observed with fluorescent microscope for fluorescent body. Gender is determined by identification of Y chromosome fluorescence in dental pulp. Results: Fluorescent bodies were found to be more in sound teeth in males as the caries increase the mean percentage of fluorescent bodies observed decreases in males. We also observed the fluorescent spots in females, and the value of the spot increases in female as the caries progresses, thereby giving false positive results in females. Conclusion: Sex determination by fluorescent staining of the Y chromosome is a reliable technique in teeth with healthy pulps or caries with enamel or up to half way of dentin. Teeth with caries involving pulp cannot be used for sex determination. PMID:25125912

Dual-Mode Combined Infra Red and Air-Coupled Ultrasonic Technique for Real-Time Industrial Process Control with Special Reference to the Food Industry

NASA Astrophysics Data System (ADS)

Pallav, P.; Hutchins, D. A.; Diamond, G. G.; Gan, T. H.; Hellyer, J. E.

2008-02-01

This paper describes the use of air-coupled ultrasound and Near Infra red (NIR) as complimentary techniques for food quality assessment. A major study has been performed, in collaboration with four industrial food companies, to investigate the use of air-coupled ultrasound and NIR to both detect foreign bodies, and to measure certain parameters of interest, such as the amount of a certain additive. The research has demonstrated that air-coupled ultrasound can be used in on-line situations, measuring food materials such as chocolate and cheese. It is also capable of performing measurements on moving sealed metal cans containing food, and is able to detect foreign bodies with the top removed, as encountered just before sealing. NIR has been used as a complimentary technique to test food materials where propagation of air-coupled ultrasound was found to be difficult. This could be due to the presence of air pockets within the food material, as in the case of bread dough.

Laser line illumination scheme allowing the reduction of background signal and the correction of absorption heterogeneities effects for fluorescence reflectance imaging.

PubMed

Fantoni, Frédéric; Hervé, Lionel; Poher, Vincent; Gioux, Sylvain; Mars, Jérôme I; Dinten, Jean-Marc

2015-10-01

Intraoperative fluorescence imaging in reflectance geometry is an attractive imaging modality as it allows to noninvasively monitor the fluorescence targeted tumors located below the tissue surface. Some drawbacks of this technique are the background fluorescence decreasing the contrast and absorption heterogeneities leading to misinterpretations concerning fluorescence concentrations. We propose a correction technique based on a laser line scanning illumination scheme. We scan the medium with the laser line and acquire, at each position of the line, both fluorescence and excitation images. We then use the finding that there is a relationship between the excitation intensity profile and the background fluorescence one to predict the amount of signal to subtract from the fluorescence images to get a better contrast. As the light absorption information is contained both in fluorescence and excitation images, this method also permits us to correct the effects of absorption heterogeneities. This technique has been validated on simulations and experimentally. Fluorescent inclusions are observed in several configurations at depths ranging from 1 mm to 1 cm. Results obtained with this technique are compared with those obtained with a classical wide-field detection scheme for contrast enhancement and with the fluorescence by an excitation ratio approach for absorption correction.

Chlorophyll fluorescence emission as a reporter on cold tolerance in Arabidopsis thaliana accessions

PubMed Central

Mishra, Anamika; Höermiller, Imke I; Heyer, Arnd G; Nedbal, Ladislav

2011-01-01

Non-invasive, high-throughput screening methods are valuable tools in breeding for abiotic stress tolerance in plants. Optical signals such as chlorophyll fluorescence emission can be instrumental in developing new screening techniques. In order to examine the potential of chlorophyll fluorescence to reveal plant tolerance to low temperatures, we used a collection of nine Arabidopsis thaliana accessions and compared their fluorescence features with cold tolerance quantified by the well established electrolyte leakage method on detached leaves. We found that, during progressive cooling, the minimal chlorophyll fluorescence emission rose strongly and that this rise was highly dependent on the cold tolerance of the accessions. Maximum quantum yield of PSII photochemistry and steady state fluorescence normalized to minimal fluorescence were also highly correlated to the cold tolerance measured by the electrolyte leakage method. In order to further increase the capacity of the fluorescence detection to reveal the low temperature tolerance, we applied combinatorial imaging that employs plant classification based on multiple fluorescence features. We found that this method, by including the resolving power of several fluorescence features, can be well employed to detect cold tolerance already at mild sub-zero temperatures. Therefore, there is no need to freeze the screened plants to the largely damaging temperatures of around −15°C. This, together with the method's easy applicability, represents a major advantage of the fluorescence technique over the conventional electrolyte leakage method. PMID:21427532

Cherenkov flashes and fluorescence flares on telescopes: New lights on UHECR spectroscopy while unveiling neutrinos astronomy

NASA Astrophysics Data System (ADS)

Fargion, D.; Oliva, P.; Massa, F.; Moreno, G.

2008-04-01

Multi-GeV and TeVs gamma sources are currently observed by their Cherenkov flashes on telescopes (as Magic, Hess and Veritas), looking vertically up into the sky. These detectors while pointing horizontally should also reveal the fluorescence flare tails of nearby down-going air-showers. Such air-showers, born at higher (tens of km) altitudes, are growing and extending up to lowest atmospheres (EeVs) or up to higher (few km) quotas (PeVs). These fluorescence signals extend the Cherenkov telescopes to a much higher cosmic ray spectroscopy. Vice versa, as it has been foreseen and only recently observed, the opposite takes place. Fluorescence telescopes made for UHECR detection (as AUGER ones) may be blazed by inclined Cherenkov lights: less energetic but frequent (PeVs) CR are expected to be often detected. Nearly dozens of blazing Cherenkov at EeV should be already found each year in AUGER, possibly in hybrid mode (FD SD, fluorescence and/or surface detector). Many more CR events (tens of thousands or hundreds of thousands) at PeVs energies should blaze Cherenkov lights each year on the AUGER fluorescence telescopes. Their UV filter may partially hide their signals and they cannot, unfortunately, be seen in any hybrid mode. At these comparable energies, the rarest UHE resonant anti-neutrino ν+e interactions in air at MW2/2me=6.3PeV energy offer enhanced W neutrino astronomy showering at air horizon, at ˜90, while crossing deep atmosphere column depth or Earth (Ande) boundaries. However, AUGER fluorescence detector (FD) are facing opposite way. An additional decay channel also rises (after resonant neutrino skimming Earth) via their secondary τ exit in air, by decay in flight via amplified showering: ν+e→W→ν+τ. Moreover, expected horizontal UHE GZK neutrinos ντν at EeVs energy, powered by guaranteed cosmogenic GZK [K.Greisen, Phys. Rev. Lett. 16 (1966) 748; G.T. Zatsepin, V.A. Kuz’min, Zh. Eks. Teor. Fiz., Pis’ma Red. 4 (1966) 144], νμν flavor

Enhanced speed in fluorescence imaging using beat frequency multiplexing

NASA Astrophysics Data System (ADS)

Mikami, Hideharu; Kobayashi, Hirofumi; Wang, Yisen; Hamad, Syed; Ozeki, Yasuyuki; Goda, Keisuke

2016-03-01

Fluorescence imaging using radiofrequency-tagged emission (FIRE) is an emerging technique that enables higher imaging speed (namely, temporal resolution) in fluorescence microscopy compared to conventional fluorescence imaging techniques such as confocal microscopy and wide-field microscopy. It works based on the principle that it uses multiple intensity-modulated fields in an interferometric setup as excitation fields and applies frequency-division multiplexing to fluorescence signals. Unfortunately, despite its high potential, FIRE has limited imaging speed due to two practical limitations: signal bandwidth and signal detection efficiency. The signal bandwidth is limited by that of an acousto-optic deflector (AOD) employed in the setup, which is typically 100-200 MHz for the spectral range of fluorescence excitation (400-600 nm). The signal detection efficiency is limited by poor spatial mode-matching between two interfering fields to produce a modulated excitation field. Here we present a method to overcome these limitations and thus to achieve higher imaging speed than the prior version of FIRE. Our method achieves an increase in signal bandwidth by a factor of two and nearly optimal mode matching, which enables the imaging speed limited by the lifetime of the target fluorophore rather than the imaging system itself. The higher bandwidth and better signal detection efficiency work synergistically because higher bandwidth requires higher signal levels to avoid the contribution of shot noise and amplifier noise to the fluorescence signal. Due to its unprecedentedly high-speed performance, our method has a wide variety of applications in cancer detection, drug discovery, and regenerative medicine.

Fluorescence Molecular Tomography: Principles and Potential for Pharmaceutical Research

PubMed Central

Stuker, Florian; Ripoll, Jorge; Rudin, Markus

2011-01-01

Fluorescence microscopic imaging is widely used in biomedical research to study molecular and cellular processes in cell culture or tissue samples. This is motivated by the high inherent sensitivity of fluorescence techniques, the spatial resolution that compares favorably with cellular dimensions, the stability of the fluorescent labels used and the sophisticated labeling strategies that have been developed for selectively labeling target molecules. More recently, two and three-dimensional optical imaging methods have also been applied to monitor biological processes in intact biological organisms such as animals or even humans. These whole body optical imaging approaches have to cope with the fact that biological tissue is a highly scattering and absorbing medium. As a consequence, light propagation in tissue is well described by a diffusion approximation and accurate reconstruction of spatial information is demanding. While in vivo optical imaging is a highly sensitive method, the signal is strongly surface weighted, i.e., the signal detected from the same light source will become weaker the deeper it is embedded in tissue, and strongly depends on the optical properties of the surrounding tissue. Derivation of quantitative information, therefore, requires tomographic techniques such as fluorescence molecular tomography (FMT), which maps the three-dimensional distribution of a fluorescent probe or protein concentration. The combination of FMT with a structural imaging method such as X-ray computed tomography (CT) or Magnetic Resonance Imaging (MRI) will allow mapping molecular information on a high definition anatomical reference and enable the use of prior information on tissue's optical properties to enhance both resolution and sensitivity. Today many of the fluorescent assays originally developed for studies in cellular systems have been successfully translated for experimental studies in animals. The opportunity of monitoring molecular processes non

Broadband Fluorescence Enhancement with Self-Assembled Silver Nanoparticle Optical Antennas.

PubMed

Vietz, Carolin; Kaminska, Izabela; Sanz Paz, Maria; Tinnefeld, Philip; Acuna, Guillermo P

2017-05-23

Plasmonic structures are known to affect the fluorescence properties of dyes placed in close proximity. This effect has been exploited in combination with single-molecule techniques for several applications in the field of biosensing. Among these plasmonic structures, top-down zero-mode waveguides stand out due to their broadband capabilities. In contrast, optical antennas based on gold nanostructures exhibit fluorescence enhancement on a narrow fraction of the visible spectrum typically restricted to the red to near-infrared region. In this contribution, we exploit the DNA origami technique to self-assemble optical antennas based on large (80 nm) silver nanoparticles. We have studied the performance of these antennas with far- and near-field simulations and characterized them experimentally with single-molecule fluorescence measurements. We demonstrate that silver-based optical antennas can yield a fluorescence enhancement of more than 2 orders of magnitude throughout the visible spectral range for high intrinsic quantum yield dyes. Additionally, a comparison between the performance of gold and silver-based antennas is included. The results indicate that silver-based antennas strongly outperform their gold counterparts in the blue and green ranges and exhibit marginal differences in the red range. These characteristics render silver-based optical antennas ready for applications involving several fluorescently labeled species across the visible spectrum.

Fluorescence lifetime evaluation of whole soils from the Amazon rainforest.

PubMed

Nicolodelli, Gustavo; Tadini, Amanda Maria; Nogueira, Marcelo Saito; Pratavieira, Sebastião; Mounier, Stephane; Huaman, Jose Luis Clabel; Dos Santos, Cléber Hilário; Montes, Célia Regina; Milori, Débora Marcondes Bastos Pereira

2017-08-20

Time-resolved fluorescence spectroscopy (TRFS) is a new tool that can be used to investigate processes of interaction between metal ions and organic matter (OM) in soils, providing a specific analysis of the structure and dynamics of macromolecules. To the best of our knowledge, there are no studies in the literature reporting the use of this technique applied to whole/non-fractionated soil samples, making it a potential method for use in future studies. This work describes the use of TRFS to evaluate the fluorescence lifetimes of OM of whole soils from the Amazon region. Analysis was made of pellets of soils from an oxisol-spodosol system, collected in São Gabriel da Cachoeira (Amazonas, Brazil). The fluorescence lifetimes in the oxisol-spodosol system were attributed to two different fluorophores. One was related to complexation of an OM fraction with metals, resulting in a shorter fluorophore lifetime. A short fluorescence lifetime (2-12 ns) could be associated with simpler structures of the OM, while a long lifetime (19-66 ns) was associated with more complex OM structures. This new TRFS technique for analysis of the fluorescence lifetime in whole soil samples complies with the principles of green chemistry.

Photonic crystal enhanced fluorescence immunoassay on diatom biosilica.

PubMed

Squire, Kenneth; Kong, Xianming; LeDuff, Paul; Rorrer, Gregory L; Wang, Alan X

2018-05-16

Fluorescence biosensing is one of the most established biosensing methods, particularly fluorescence spectroscopy and microscopy. These are two highly sensitive techniques but require high grade electronics and optics to achieve the desired sensitivity. Efforts have been made to implement these methods using consumer grade electronics and simple optical setups for applications such as point-of-care diagnostics, but the sensitivity inherently suffers. Sensing substrates, capable of enhancing fluorescence are thus needed to achieve high sensitivity for such applications. In this paper, we demonstrate a photonic crystal-enhanced fluorescence immunoassay biosensor using diatom biosilica, which consists of silica frustules with sub-100 nm periodic pores. Utilizing the enhanced local optical field, the Purcell effect and increased surface area from the diatom photonic crystals, we create ultrasensitive immunoassay biosensors that can significantly enhance fluorescence spectroscopy as well as fluorescence imaging. Using standard antibody-antigen-labeled antibody immunoassay protocol, we experimentally achieved 100× and 10× better detection limit with fluorescence spectroscopy and fluorescence imaging respectively. The limit of detection of the mouse IgG goes down to 10 -16 M (14 fg/mL) and 10 -15 M (140 fg/mL) for the two respective detection modalities, virtually sensing a single mouse IgG molecule on each diatom frustule. The effectively enhanced fluorescence imaging in conjunction with the simple hot-spot counting analysis method used in this paper proves the great potential of diatom fluorescence immunoassay for point-of-care biosensing. Scanning electron microscope image of biosilica diatom frustule that enables significant enhancement of fluorescence spectroscopy and fluorescence image. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Experimental design and quality assurance: in situ fluorescence instrumentation

USGS Publications Warehouse

Conmy, Robyn N.; Del Castillo, Carlos E.; Downing, Bryan D.; Chen, Robert F.

2014-01-01

Both instrument design and capabilities of fluorescence spectroscopy have greatly advanced over the last several decades. Advancements include solid-state excitation sources, integration of fiber optic technology, highly sensitive multichannel detectors, rapid-scan monochromators, sensitive spectral correction techniques, and improve data manipulation software (Christian et al., 1981, Lochmuller and Saavedra, 1986; Cabniss and Shuman, 1987; Lakowicz, 2006; Hudson et al., 2007). The cumulative effect of these improvements have pushed the limits and expanded the application of fluorescence techniques to numerous scientific research fields. One of the more powerful advancements is the ability to obtain in situ fluorescence measurements of natural waters (Moore, 1994). The development of submersible fluorescence instruments has been made possible by component miniaturization and power reduction including advances in light sources technologies (light-emitting diodes, xenon lamps, ultraviolet [UV] lasers) and the compatible integration of new optical instruments with various sampling platforms (Twardowski et at., 2005 and references therein). The development of robust field sensors skirt the need for cumbersome and or time-consuming filtration techniques, the potential artifacts associated with sample storage, and coarse sampling designs by increasing spatiotemporal resolution (Chen, 1999; Robinson and Glenn, 1999). The ability to obtain rapid, high-quality, highly sensitive measurements over steep gradients has revolutionized investigations of dissolved organic matter (DOM) optical properties, thereby enabling researchers to address novel biogeochemical questions regarding colored or chromophoric DOM (CDOM). This chapter is dedicated to the origin, design, calibration, and use of in situ field fluorometers. It will serve as a review of considerations to be accounted for during the operation of fluorescence field sensors and call attention to areas of concern when making

Characterizing non-photochemical quenching in leaves through fluorescence lifetime snapshots

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sylak-Glassman, Emily J.; Zaks, Julia; Amarnath, Kapil

2015-03-12

A technique is described to measure the fluorescence decay profiles of intact leaves during adaptation to high light and subsequent relaxation to dark conditions. We illustrate how to ensure that photosystem II reaction centers are closed and compare data for wild type Arabidopsis thaliana with conventional pulse-amplitude modulated (PAM) fluorescence measurements. Unlike PAM measurements, the lifetime measurements are not sensitive to photobleaching or chloroplast shielding, and the form of the fluorescence decay provides additional information to test quantitative models of excitation dynamics in intact leaves.

Imaging the environment of green fluorescent protein.

PubMed Central

Suhling, Klaus; Siegel, Jan; Phillips, David; French, Paul M W; Lévêque-Fort, Sandrine; Webb, Stephen E D; Davis, Daniel M

2002-01-01

An emerging theme in cell biology is that cell surface receptors need to be considered as part of supramolecular complexes of proteins and lipids facilitating specific receptor conformations and distinct distributions, e.g., at the immunological synapse. Thus, a new goal is to develop bioimaging that not only locates proteins in live cells but can also probe their environment. Such a technique is demonstrated here using fluorescence lifetime imaging of green fluorescent protein (GFP). We first show, by time-correlated single-photon counting, that the fluorescence decay of GFP depends on the local refractive index. This is in agreement with the Strickler Berg formula, relating the Einstein A and B coefficients for absorption and spontaneous emission in molecules. We then quantitatively image, by wide-field time-gated fluorescence lifetime imaging, the refractive index of the environment of GFP. This novel approach paves the way for imaging the biophysical environment of specific GFP-tagged proteins in live cells. PMID:12496126

Development of Functional Fluorescent Molecular Probes for the Detection of Biological Substances

PubMed Central

Suzuki, Yoshio; Yokoyama, Kenji

2015-01-01

This review is confined to sensors that use fluorescence to transmit biochemical information. Fluorescence is, by far, the most frequently exploited phenomenon for chemical sensors and biosensors. Parameters that define the application of such sensors include intensity, decay time, anisotropy, quenching efficiency, and luminescence energy transfer. To achieve selective (bio)molecular recognition based on these fluorescence phenomena, various fluorescent elements such as small organic molecules, enzymes, antibodies, and oligonucleotides have been designed and synthesized over the past decades. This review describes the immense variety of fluorescent probes that have been designed for the recognitions of ions, small and large molecules, and their biological applications in terms of intracellular fluorescent imaging techniques. PMID:26095660

Applications of fluorescence spectroscopy for predicting percent wastewater in an urban stream

USGS Publications Warehouse

Goldman, Jami H.; Rounds, Stewart A.; Needoba, Joseph A.

2012-01-01

Dissolved organic carbon (DOC) is a significant organic carbon reservoir in many ecosystems, and its characteristics and sources determine many aspects of ecosystem health and water quality. Fluorescence spectroscopy methods can quantify and characterize the subset of the DOC pool that can absorb and re-emit electromagnetic energy as fluorescence and thus provide a rapid technique for environmental monitoring of DOC in lakes and rivers. Using high resolution fluorescence techniques, we characterized DOC in the Tualatin River watershed near Portland, Oregon, and identified fluorescence parameters associated with effluent from two wastewater treatment plants and samples from sites within and outside the urban region. Using a variety of statistical approaches, we developed and validated a multivariate linear regression model to predict the amount of wastewater in the river as a function of the relative abundance of specific fluorescence excitation/emission pairs. The model was tested with independent data and predicts the percentage of wastewater in a sample within 80% confidence. Model results can be used to develop in situ instrumentation, inform monitoring programs, and develop additional water quality indicators for aquatic systems.

Laser-Induced Fluorescence Measurements and Modeling of Nitric Oxide in Counterflow Diffusion Flames

NASA Technical Reports Server (NTRS)

Ravikrishna, Rayavarapu V.

2000-01-01

The feasibility of making quantitative nonintrusive NO concentration ([NO]) measurements in nonpremixed flames has been assessed by obtaining laser-induced fluorescence (LIF) measurements of [NO] in counterflow diffusion flames at atmospheric and higher pressures. Comparisons at atmospheric pressure between laser-saturated fluorescence (LSF) and linear LIF measurements in four diluted ethane-air counterflow diffusion flames with strain rates from 5 to 48/s yielded excellent agreement from fuel-lean to moderately fuel-rich conditions, thus indicating the utility of a model-based quenching correction technique, which was then extended to higher pressures. Quantitative LIF measurements of [NO] in three diluted methane-air counterflow diffusion flames with strain rates from 5 to 35/s were compared with OPPDIF model predictions using the GRI (version 2.11) chemical kinetic mechanism. The comparisons revealed that the GRI mechanism underpredicts prompt-NO by 30-50% at atmospheric pressure. Based on these measurements, a modified reaction rate coefficient for the prompt-NO initiation reaction was proposed which causes the predictions to match experimental data. Temperature measurements using thin filament pyrometry (TFP) in conjunction with a new calibration method utilizing a near-adiabatic H2-air Hencken burner gave very good comparisons with model predictions in these counterflow diffusion flames. Quantitative LIF measurements of [NO] were also obtained in four methane-air counterflow partially-premixed flames with fuel-side equivalence ratios (phi(sub B)) of 1.45, 1.6, 1.8 and 2.0. The measurements were in excellent agreement with model predictions when accounting for radiative heat loss. Spatial separation between regions dominated by the prompt and thermal NO mechanisms was observed in the phi(sub B) = 1.45 flame. The modified rate coefficient proposed earlier for the prompt-NO initiation reaction improved agreement between code predictions and measurements in the

Magnetic field dependent measurement techniques of surface tension of magnetic fluid at an air interface

NASA Astrophysics Data System (ADS)

Nair, Nishant; Virpura, Hiral; Patel, Rajesh

2015-06-01

We describe here two measurement techniques to determine surface tension of magnetic fluid. (i) magneti c field dependent capillary rise method and (ii) Taylor wavelength method in which the distance between the consecutive stable spikes was measured and then surface tension was calculated. The surface tension measurements from both the methods are compared. It is observed that surface tension of magnetic fluid increases with increase in magnetic field due to field dependent structure formation in magnetic fluid at an air interface. We have also measured magnetic susceptibility and surface tension for different volume fractions. The measurement of magnetic susceptibility is carried out using Quincke's experimental techniques.

Optical molecular fluorescence determination of ultra-trace beryllium in occupational and environmental samples using highly alkaline conditions.

PubMed

Adams, Lori; Agrawal, Anoop; Cronin, John P; Ashley, Kevin

2017-01-01

Exposures to beryllium (Be), even at extremely low levels, can cause severe health effects in a percentage of those exposed; consequently, occupational exposure limits (OELs) promulgated for this element are the lowest established for any element. This work describes the advantages of using highly alkaline dye solutions for determination of Be in occupational hygiene and environmental samples by means of an optical molecular fluorescence technique after sample extraction in 1-3% (w˖w -1 ) aqueous ammonium bifluoride (NH 4 HF 2 ). Improved attributes include the ability to further enhance the detection limits of Be in extraction solutions of high acidity with minimal dilution, which is particularly beneficial when NH 4 HF 2 solutions of higher concentration are used for extraction of Be from soil samples. Significant improvements in Be method detection limits (MDLs) are obtained at levels many-fold below those reported previously for this methodology. Notably, MDLs for Be of <0.01 ng l -1 / 0.1 ng per sample have been attained, which are superior to MDLs routinely reported for this element by means of the most widely used ultra-trace elemental measurement technique, inductively coupled plasma mass spectrometry (ICP-MS). Very low MDLs for Be are essential in consideration of reductions in OELs for this element in workplace air by health organizations and regulatory agencies in the USA and internationally. Applications of enhanced Be measurements to air filter samples, surface wipe samples, soils and newly-designed occupational air sampler inserts are illustrated.

Calibration of fluorescence resonance energy transfer in microscopy

DOEpatents

Youvan, Dougalas C.; Silva, Christopher M.; Bylina, Edward J.; Coleman, William J.; Dilworth, Michael R.; Yang, Mary M.

2003-12-09

Imaging hardware, software, calibrants, and methods are provided to visualize and quantitate the amount of Fluorescence Resonance Energy Transfer (FRET) occurring between donor and acceptor molecules in epifluorescence microscopy. The MicroFRET system compensates for overlap among donor, acceptor, and FRET spectra using well characterized fluorescent beads as standards in conjunction with radiometrically calibrated image processing techniques. The MicroFRET system also provides precisely machined epifluorescence cubes to maintain proper image registration as the sample is illuminated at the donor and acceptor excitation wavelengths. Algorithms are described that pseudocolor the image to display pixels exhibiting radiometrically-corrected fluorescence emission from the donor (blue), the acceptor (green) and FRET (red). The method is demonstrated on samples exhibiting FRET between genetically engineered derivatives of the Green Fluorescent Protein (GFP) bound to the surface of Ni chelating beads by histidine-tags.

Calibration of fluorescence resonance energy transfer in microscopy

DOEpatents

Youvan, Douglas C.; Silva, Christopher M.; Bylina, Edward J.; Coleman, William J.; Dilworth, Michael R.; Yang, Mary M.

2002-09-24

Imaging hardware, software, calibrants, and methods are provided to visualize and quantitate the amount of Fluorescence Resonance Energy Transfer (FRET) occurring between donor and acceptor molecules in epifluorescence microscopy. The MicroFRET system compensates for overlap among donor, acceptor, and FRET spectra using well characterized fluorescent beads as standards in conjunction with radiometrically calibrated image processing techniques. The MicroFRET system also provides precisely machined epifluorescence cubes to maintain proper image registration as the sample is illuminated at the donor and acceptor excitation wavelengths. Algorithms are described that pseudocolor the image to display pixels exhibiting radiometrically-corrected fluorescence emission from the donor (blue), the acceptor (green) and FRET (red). The method is demonstrated on samples exhibiting FRET between genetically engineered derivatives of the Green Fluorescent Protein (GFP) bound to the surface of Ni chelating beads by histidine-tags.

Protein subcellular localization assays using split fluorescent proteins

DOEpatents

Waldo, Geoffrey S [Santa Fe, NM; Cabantous, Stephanie [Los Alamos, NM

2009-09-08

The invention provides protein subcellular localization assays using split fluorescent protein systems. The assays are conducted in living cells, do not require fixation and washing steps inherent in existing immunostaining and related techniques, and permit rapid, non-invasive, direct visualization of protein localization in living cells. The split fluorescent protein systems used in the practice of the invention generally comprise two or more self-complementing fragments of a fluorescent protein, such as GFP, wherein one or more of the fragments correspond to one or more beta-strand microdomains and are used to "tag" proteins of interest, and a complementary "assay" fragment of the fluorescent protein. Either or both of the fragments may be functionalized with a subcellular targeting sequence enabling it to be expressed in or directed to a particular subcellular compartment (i.e., the nucleus).

Chlorophyll fluorescence analysis: a guide to good practice and understanding some new applications.

PubMed

Murchie, E H; Lawson, T

2013-10-01

Chlorophyll fluorescence is a non-invasive measurement of photosystem II (PSII) activity and is a commonly used technique in plant physiology. The sensitivity of PSII activity to abiotic and biotic factors has made this a key technique not only for understanding the photosynthetic mechanisms but also as a broader indicator of how plants respond to environmental change. This, along with low cost and ease of collecting data, has resulted in the appearance of a large array of instrument types for measurement and calculated parameters which can be bewildering for the new user. Moreover, its accessibility can lead to misuse and misinterpretation when the underlying photosynthetic processes are not fully appreciated. This review is timely because it sits at a point of renewed interest in chlorophyll fluorescence where fast measurements of photosynthetic performance are now required for crop improvement purposes. Here we help the researcher make choices in terms of protocols using the equipment and expertise available, especially for field measurements. We start with a basic overview of the principles of fluorescence analysis and provide advice on best practice for taking pulse amplitude-modulated measurements. We also discuss a number of emerging techniques for contemporary crop and ecology research, where we see continual development and application of analytical techniques to meet the new challenges that have arisen in recent years. We end the review by briefly discussing the emerging area of monitoring fluorescence, chlorophyll fluorescence imaging, field phenotyping, and remote sensing of crops for yield and biomass enhancement.

Convection venting lensed reflector-type compact fluorescent lamp system

DOEpatents

Pelton, Bruce A.; Siminovitch, Michael

1997-01-01

Disclosed herein is a fluorescent lamp housing assembly capable of providing convection cooling to the lamp and the ballast. The lens of the present invention includes two distinct portions, a central portion and an apertured portion. The housing assembly further includes apertures so that air mass is able to freely move up through the assembly and out ventilation apertures.

Application of cytoplasmic Ca2+ fluorescence imaging techniques to study the molecular mechanisms of exercise-induced fatigue eliminated by Chinese medicine ginseng extract

NASA Astrophysics Data System (ADS)

Liu, Yi; Zhao, Yanping; Zhang, Heming; Liu, Songhao