Correlated cryo-fluorescence and cryo-electron microscopy with high spatial precision and improved sensitivity

Energy Technology Data Exchange (ETDEWEB)

Schorb, Martin [Structural and Computational Biology Unit, European Molecular Biology Laboratory, D-69117 Heidelberg (Germany); Briggs, John A.G., E-mail: john.briggs@embl.de [Structural and Computational Biology Unit, European Molecular Biology Laboratory, D-69117 Heidelberg (Germany); Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, D-69117 Heidelberg (Germany)

2014-08-01

Performing fluorescence microscopy and electron microscopy on the same sample allows fluorescent signals to be used to identify and locate features of interest for subsequent imaging by electron microscopy. To carry out such correlative microscopy on vitrified samples appropriate for structural cryo-electron microscopy it is necessary to perform fluorescence microscopy at liquid-nitrogen temperatures. Here we describe an adaptation of a cryo-light microscopy stage to permit use of high-numerical aperture objectives. This allows high-sensitivity and high-resolution fluorescence microscopy of vitrified samples. We describe and apply a correlative cryo-fluorescence and cryo-electron microscopy workflow together with a fiducial bead-based image correlation procedure. This procedure allows us to locate fluorescent bacteriophages in cryo-electron microscopy images with an accuracy on the order of 50 nm, based on their fluorescent signal. It will allow the user to precisely and unambiguously identify and locate objects and events for subsequent high-resolution structural study, based on fluorescent signals. - Highlights: • Workflow for correlated cryo-fluorescence and cryo-electron microscopy. • Cryo-fluorescence microscopy setup incorporating a high numerical aperture objective. • Fluorescent signals located in cryo-electron micrographs with 50 nm spatial precision.

Combination of panoramic and fluorescence endoscopic images to obtain tumor spatial distribution information useful for bladder cancer detection

Science.gov (United States)

Olijnyk, S.; Hernández Mier, Y.; Blondel, W. C. P. M.; Daul, C.; Wolf, D.; Bourg-Heckly, G.

2007-07-01

Bladder cancer is widely spread. Moreover, carcinoma in situ can be difficult to diagnose as it may be difficult to see, and become invasive in 50 % of case. Non invasive diagnosis methods like photodynamic or autofluorescence endoscopy allow enhancing sensitivity and specificity. Besides, bladder tumors can be multifocal. Multifocality increases the probability of recurrence and infiltration into bladder muscle. Analysis of spatial distribution of tumors could be used to improve diagnosis. We explore the feasibility to combine fluorescence and spatial information on phantoms. We developed a system allowing the acquisition of consecutive images under white light or UV excitation alternatively and automatically along the video sequence. We also developed an automatic image processing algorithm to build a partial panoramic image from a cystoscopic sequence of images. Fluorescence information is extracted from wavelength bandpass filtered images and superimposed over the cartography. Then, spatial distribution measures of fluorescent spots can be computed. This cartography can be positioned on a 3D generic shape of bladder by selecting some reference points. Our first results on phantoms show that it is possible to obtain cartography with fluorescent spots and extract quantitative information of their spatial distribution on a "wide" field of view basis.

Cardiac Calcium ATPase Dimerization Measured by Cross-Linking and Fluorescence Energy Transfer.

Science.gov (United States)

Blackwell, Daniel J; Zak, Taylor J; Robia, Seth L

2016-09-20

The cardiac sarco/endoplasmic reticulum calcium ATPase (SERCA) establishes the intracellular calcium gradient across the sarcoplasmic reticulum membrane. It has been proposed that SERCA forms homooligomers that increase the catalytic rate of calcium transport. We investigated SERCA dimerization in rabbit left ventricular myocytes using a photoactivatable cross-linker. Western blotting of cross-linked SERCA revealed higher-molecular-weight species consistent with SERCA oligomerization. Fluorescence resonance energy transfer measurements in cells transiently transfected with fluorescently labeled SERCA2a revealed that SERCA readily forms homodimers. These dimers formed in the absence or presence of the SERCA regulatory partner, phospholamban (PLB) and were unaltered by PLB phosphorylation or changes in calcium or ATP. Fluorescence lifetime data are compatible with a model in which PLB interacts with a SERCA homodimer in a stoichiometry of 1:2. Together, these results suggest that SERCA forms constitutive homodimers in live cells and that dimer formation is not modulated by SERCA conformational poise, PLB binding, or PLB phosphorylation. Copyright © 2016 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Laser-induced fluorescence imaging of subsurface tissue structures with a volume holographic spatial-spectral imaging system.

Science.gov (United States)

Luo, Yuan; Gelsinger-Austin, Paul J; Watson, Jonathan M; Barbastathis, George; Barton, Jennifer K; Kostuk, Raymond K

2008-09-15

A three-dimensional imaging system incorporating multiplexed holographic gratings to visualize fluorescence tissue structures is presented. Holographic gratings formed in volume recording materials such as a phenanthrenquinone poly(methyl methacrylate) photopolymer have narrowband angular and spectral transmittance filtering properties that enable obtaining spatial-spectral information within an object. We demonstrate this imaging system's ability to obtain multiple depth-resolved fluorescence images simultaneously.

Study of the Conformational State of Non-Cross-Linked and Cross-Linked Poly(alkylmethyldiallylammonium chlorides) in Aqueous Solution by Fluorescence Probing

NARCIS (Netherlands)

Wang, Guang-Jia; Engberts, Jan B.F.N.

The aggregation behaviour of novel non-cross-linked and cross-linked poly(alkylmethyldiallylammonium chlorides) in aqueous solutions has been investigated by fluorescence spectroscopy using pyrene as a probe. These copolymers were found to exhibit similar aggregate properties as the corresponding

X-ray fluorescence in Member States (Italy): Full field X-ray fluorescence imaging with high-energy and high-spatial resolution

Energy Technology Data Exchange (ETDEWEB)

Romano, F. P.; Masini, N.; Pappalardo, L., E-mail: romanop@lns.infn.it [IBAM, CNR, Via Biblioteca 4, 95124 Catania (Italy); Cosentino, L.; Gammino, S.; Mascali, D.; Rizzo, F. [INFN-LNS, Via S. Sofia 62, 95123 Catania (Italy)

2014-02-15

A full field X-ray camera for the X-Ray Fluorescence imaging of materials with high-energy and high-spatial resolution was designed and developed. The system was realized by coupling a pinhole collimator with a positionsensitive CCD detector. X-Ray fluorescence is induced on the samples by irradiation with an external X-ray tube. The characteristic X-ray spectra of the investigated materials are obtained by using a multi-frames acquisition in single-photon counting. The energy resolution measured at the Fe-Kα line was 157 eV. The spatial resolution of the system was determined by the analysis of a sharp-edge at different magnification values; it was estimated to be 90 μm at a magnification value of 3.2x and 190 μm at 0.8x. The present set-up of the system is suited to analyze samples with dimensions up to 5x4 cm{sup 2}. Typical measurement time is in the range between 1h to 4 h. (author)

Reorientational motion of a cross-link junction in a poly(dimethylsiloxane) network measured by time-resolved fluorescence depolarization

International Nuclear Information System (INIS)

Stein, A.D.; Hoffman, D.A.; Frank, C.W.; Fayer, M.D.

1992-01-01

The reorientational dynamics of a cross-link junction in poly(dimethylsiloxane) networks, measured by the fluorescence anisotropy decay of a chromophore tagged to the cross-link, have been investigated over a range of temperatures from T g +75 to T g +150. The probe chromophore, 1-dimethylamino-5-sulfonylnaphthalene amide (dansyl amide), is pendant to a trifunctional silane that acts as a cross-linking molecule. In cyclohexanol, the fluorescence anisotropy decay is in agreement with Debye--Stokes--Einstein hydrodynamic theory (rotational diffusion) demonstrating that the cross-linker can be used as a probe of orientational relaxation. The fluorescence anisotropy decays at a rapid rate in an end-linked poly(dimethyl siloxane) network reflecting fast reorientational motion of the cross-link junction. This reorientation appears diffusive and has a temperature dependence in accord with the Williams--Landel--Ferry equation. A model is proposed that suggests that reorientation and translational motion of the cross-link occur simultaneously and are both coupled to fluctuations of the polymer chain ends

Reorientational motion of a cross-link junction in a poly(dimethylsiloxane) network measured by time-resolved fluorescence depolarization

Energy Technology Data Exchange (ETDEWEB)

Stein, A.D. (Department of Chemistry, Stanford University, Stanford, California 94305 (United States)); Hoffman, D.A. (Department of Materials Science and Engineering, Stanford University, Stanford, California 94305 (United States)); Frank, C.W. (Department of Chemical Engineering, Stanford University, Stanford, California 94305 (United States)); Fayer, M.D. (Department of Chemistry, Stanford University, Stanford, California 94305 (United States))

1992-02-15

The reorientational dynamics of a cross-link junction in poly(dimethylsiloxane) networks, measured by the fluorescence anisotropy decay of a chromophore tagged to the cross-link, have been investigated over a range of temperatures from {ital T}{sub {ital g}}+75 to {ital T}{sub {ital g}}+150. The probe chromophore, 1-dimethylamino-5-sulfonylnaphthalene amide (dansyl amide), is pendant to a trifunctional silane that acts as a cross-linking molecule. In cyclohexanol, the fluorescence anisotropy decay is in agreement with Debye--Stokes--Einstein hydrodynamic theory (rotational diffusion) demonstrating that the cross-linker can be used as a probe of orientational relaxation. The fluorescence anisotropy decays at a rapid rate in an end-linked poly(dimethyl siloxane) network reflecting fast reorientational motion of the cross-link junction. This reorientation appears diffusive and has a temperature dependence in accord with the Williams--Landel--Ferry equation. A model is proposed that suggests that reorientation and translational motion of the cross-link occur simultaneously and are both coupled to fluctuations of the polymer chain ends.

Measurements of excited-state-to-excited-state transition probabilities and photoionization cross-sections using laser-induced fluorescence and photoionization signals

International Nuclear Information System (INIS)

Shah, M.L.; Sahoo, A.C.; Pulhani, A.K.; Gupta, G.P.; Dikshit, B.; Bhatia, M.S.; Suri, B.M.

2014-01-01

Laser-induced photoionization and fluorescence signals were simultaneously observed in atomic samarium using Nd:YAG-pumped dye lasers. Two-color, three-photon photoionization and two-color fluorescence signals were recorded simultaneously as a function of the second-step laser power for two photoionization pathways. The density matrix formalism has been employed to analyze these signals. Two-color laser-induced fluorescence signal depends on the laser powers used for the first and second-step transitions as well as the first and second-step transition probability whereas two-color, three-photon photoionization signal depends on the third-step transition cross-section at the second-step laser wavelength along with the laser powers and transition probability for the first and second-step transitions. Two-color laser-induced fluorescence was used to measure the second-step transition probability. The second-step transition probability obtained was used to infer the photoionization cross-section. Thus, the methodology combining two-color, three-photon photoionization and two-color fluorescence signals in a single experiment has been established for the first time to measure the second-step transition probability as well as the photoionization cross-section. - Highlights: • Laser-induced photoionization and fluorescence signals have been simultaneously observed. • The density matrix formalism has been employed to analyze these signals. • Two-color laser-induced fluorescence was used to measure the second-step transition probability. • The second-step transition probability obtained was used to infer the photoionization cross-section. • Transition probability and photoionization cross-section have been measured in a single experiment

Alloying effect on K shell X-ray fluorescence cross-sections and yields in Ti-Ni based shape memory alloys

Directory of Open Access Journals (Sweden)

Bünyamin Alım

2018-04-01

Full Text Available K shell X-ray fluorescence cross-sections (σKα, σKβ and σK, and K shell fluorescence yields (ωK of Ti, Ni both in pure metals and in different alloy compositions (TixNi1-x; x = 0.3, 0.4, 0.5, 0.6, 0.7 were measured by using energy dispersive X-ray fluorescence (EDXRF technique. The samples were excited by 22.69 keV X-rays from a 10 mCi Cd-109 radioactive point source and K X rays emitted by samples were counted by a high resolution Si(Li solid-state detector coupled to a 4 K multichannel analyzer (MCA. The alloying effects on the X-ray fluorescence (XRF parameters of Ti-Ni shape memory alloys (SMAs were investigated. It is clearly observed that alloying effect causes to change in K shell XRF parameter values in Ti-Ni based SMAs for different compositions of x. Also, the present investigation makes it possible to perform reliable interpretation of experimental σKα, σKβ and ωK values for Ti and Ni in SMAs and can also provide quantitative information about the changes of K shell X-ray fluorescence cross sections and fluorescence yields of these metals with alloy composition. Keywords: Alloying effect, XRF, K X-ray fluorescence cross-section, K shell fluorescence yield, Shape memory alloy

Circumvention of fluorophore photobleaching in fluorescence fluctuation experiments: a beam scanning approach.

Science.gov (United States)

Satsoura, Dmitri; Leber, Brian; Andrews, David W; Fradin, Cécile

2007-04-23

Photobleaching is a fluorophore-damaging process that commonly afflicts single-molecule fluorescence studies. It becomes an especially severe problem in fluorescence fluctuation experiments when studying slowly diffusing particles. One way to circumvent this problem is to use beam scanning to decrease the residence time of the fluorophores in the excitation volume. We report a systematic study of the effects of circular beam scanning on the photobleaching of fluorescent particles as observed in single-photon excitation fluorescence fluctuation experiments. We start by deriving a simple expression relating the average detected fluorescence to the photobleaching cross section of the fluorophores. We then perform numerical calculations of the spatial distribution of fluorescent particles in order to understand under which conditions beam scanning can prevent the formation of a photobleaching hole. To support these predictions, we show experimental results obtained for large unilamellar vesicles containing a small amount of the fluorescent lipophilic tracer DiD. We establish the required scanning radius and frequency range in order to obtain sufficient reduction of the photobleaching effect for that system. From the detected increase in fluorescence upon increase in scanning speed, we estimate the photobleaching cross section of DiD.

Near infrared spatial frequency domain fluorescence imaging of tumor phantoms containing erythrocyte-derived optical nanoplatforms

Science.gov (United States)

Burns, Joshua M.; Schaefer, Elise; Anvari, Bahman

2018-02-01

Light-activated theranostic constructs provide a multi-functional platform for optical imaging and phototherapeutic applications. Our group has engineered nano-sized vesicles derived from erythrocytes that encapsulate the FDAapproved near infrared (NIR) absorber indocyanine green (ICG). We refer to these constructs as NIR erythrocytemimicking transducers (NETs). Once photo-excited by NIR light these constructs can transduce the photons energy to emit fluorescence, generate heat, or induce chemical reactions. In this study, we investigated fluorescence imaging of NETs embedded within tumor phantoms using spatial frequency domain imaging (SFDI). Using SFDI, we were able to fluorescently image simulated tumors doped with different concentration of NETs. These preliminary results suggest that NETs can be used in conjunction with SFDI for potential tumor imaging applications.

Collimated dual species oven source and its characterisation via spatially resolved fluorescence spectroscopy

Science.gov (United States)

Cooper, N.; Da Ros, E.; Nute, J.; Baldolini, D.; Jouve, P.; Hackermüller, L.; Langer, M.

2018-03-01

We describe the design, construction and characterisation of a collimated, dual-species oven source for generating intense beams of lithium and caesium in UHV environments. Our design produces full beam overlap for the two species. Using an aligned microtube array the FWHM of the output beam is restricted to  ˜75 milliradians, with an estimated axial brightness of 3.6× 1014 atoms s-1 sr-1 for Li and 7.4× 1015 atoms s-1 sr-1 for Cs. We measure the properties of the output beam using a spatially-resolved fluorescence technique, which allows for the extraction of additional information not accessible without spatial resolution.

An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy

International Nuclear Information System (INIS)

Dunsby, C; Lanigan, P M P; McGinty, J; Elson, D S; Requejo-Isidro, J; Munro, I; Galletly, N; McCann, F; Treanor, B; Oenfelt, B; Davis, D M; Neil, M A A; French, P M W

2004-01-01

Fluorescence imaging is used widely in microscopy and macroscopic imaging applications for fields ranging from biomedicine to materials science. A critical component for any fluorescence imaging system is the excitation source. Traditionally, wide-field systems use filtered thermal or arc-generated white light sources, while point scanning confocal microscope systems require spatially coherent (point-like) laser sources. Unfortunately, the limited range of visible wavelengths available from conventional laser sources constrains the design and usefulness of fluorescent probes in confocal microscopy. A 'hands-off' laser-like source, electronically tunable across the visible spectrum, would be invaluable for fluorescence imaging and provide new opportunities, e.g. automated excitation fingerprinting and in situ measurement of excitation cross-sections. Yet more information can be obtained using fluorescence lifetime imaging (FLIM), which requires that the light source be pulsed or rapidly modulated. We show how a white light continuum, generated by injecting femtosecond optical radiation into a micro-structured optical fibre, coupled with a simple prism-based tunable filter arrangement, can fulfil all these roles as a continuously electronically tunable (435-1150 nm) visible ultrafast light source in confocal, wide-field and FLIM systems

Spatial Variability Analysis of Within-Field Winter Wheat Nitrogen and Grain Quality Using Canopy Fluorescence Sensor Measurements

Directory of Open Access Journals (Sweden)

Xiaoyu Song

2017-03-01

Full Text Available Wheat grain protein content (GPC is a key component when evaluating wheat nutrition. It is also important to determine wheat GPC before harvest for agricultural and food process enterprises in order to optimize the wheat grading process. Wheat GPC across a field is spatially variable due to the inherent variability of soil properties and position in the landscape. The objectives of this field study were: (i to assess the spatial and temporal variability of wheat nitrogen (N attributes related to the grain quality of winter wheat production through canopy fluorescence sensor measurements; and (ii to examine the influence of spatial variability of soil N and moisture across different growth stages on the wheat grain quality. A geostatistical approach was used to analyze data collected from 110 georeferenced locations. In particular, Ordinary Kriging Analysis (OKA was used to produce maps of wheat GPC, GPC yield, and wheat canopy fluorescence parameters, including simple florescence ratio and Nitrogen Balance Indices (NBI. Soil Nitrate-Nitrogen (NO3-N content and soil Time Domain Reflectometry (TDR value in the study field were also interpolated through the OKA method. The fluorescence parameter maps, soil NO3-N and soil TDR maps obtained from the OKA output were compared with the wheat GPC and GPC yield maps in order to assess their relationships. The results of this study indicate that the NBI spatial variability map in the late stage of wheat growth can be used to distinguish areas that produce higher GPC.

Spatially resolved analyses of uranium species using a coupled system made up of confocal laser-scanning microscopy (CLSM) and laser induced fluorescence spectroscopy (LIFS)

International Nuclear Information System (INIS)

Brockmann, S.; Grossmann, K.; Arnold, T.

2014-01-01

The fluorescent properties of uranium when excited by UV light are used increasingly for spectroscope analyses of uranium species within watery samples. Here, alongside the fluorescent properties of the hexavalent oxidation phases, the tetra and pentavalent oxidation phases also play an increasingly important role. The detection of fluorescent emission spectrums on solid and biological samples using (time-resolved) laser induced fluorescence spectroscopy (TRLFS or LIFS respectively) has, however, the disadvantage that no statements regarding the spatial localisation of the uranium can be made. However, particularly in complex, biological samples, such statements on the localisation of the uranium enrichment in the sample are desired, in order to e.g. be able to distinguish between intra and extra-cellular uranium bonds. The fluorescent properties of uranium (VI) compounds and minerals can also be used to detect their localisation within complex samples. So the application of fluorescent microscopic methods represents one possibility to localise and visualise uranium precipitates and enrichments in biological samples, such as biofilms or cells. The confocal laser-scanning microscopy (CLSM) is especially well suited to this purpose. Coupling confocal laser-scanning microscopy (CLSM) with laser induced fluorescence spectroscopy (LIFS) makes it possible to localise and visualise fluorescent signals spatially and three-dimensionally, while at the same time being able to detect spatially resolved, fluorescent-spectroscopic data. This technology is characterised by relatively low detection limits from up to 1.10 -6 M for uranium (VI) compounds within the confocal volume. (orig.)

Spatial discrimination against background with different optical systems for collection of fluorescence in laser-excited atomic fluorescence spectrometry with a graphite tube electrothermal atomizer.

Science.gov (United States)

Yuzefovsky, A I; Lonardo, R F; Michel, R G

1995-07-01

A single 90 degrees off-axis ellipsoidal mirror fragment was used in a dispersive detection system for electrothermal atomization laser-excited atomic fluorescence spectrometry. The performance of the new optical arrangement was compared with those of optical arrangements that employed a plane mirror in combination with biconvex or plano-convex lenses. All the optical arrangements collected fluorescence in a scheme called front surface illustration. BEAM-4, an optical ray tracing program, was used for calculations of spatial ray distributions and optical collection efficiency for the various optical configurations. Experimentally, the best collection efficiency was obtained by use of the ellipsoidal mirror, in qualitative agreement with simulations done by use of the BEAM-4 software. The best detection limit for cobalt with the new optical arrangement was 20 fg, which was a factor of 5 better than that obtained with conventional optical arrangements with otherwise the same instrumentation. The signal-to-background ratio and the fluorescence collection efficiency were also studied as a function of position of the optical components for the various optical arrangements. For both cobalt and phosphorus, the signal-to-background ratio with the new optical arrangement remained stable within 10-20% during +/- 8 mm shifts in the position of the detection system from the focal plane of the optics. Overall, the new optical arrangement offered high collection efficiency, excellent sensitivity, and facile optical alignment due to efficient spatial separation between the fluorescence signal and the background radiation. The advantages of the new optical arrangement were particularly important during measurements in the presence of high levels of blackbody radiation.

Multiphoton-Excited Fluorescence of Silicon-Vacancy Color Centers in Diamond

Science.gov (United States)

Higbie, J. M.; Perreault, J. D.; Acosta, V. M.; Belthangady, C.; Lebel, P.; Kim, M. H.; Nguyen, K.; Demas, V.; Bajaj, V.; Santori, C.

2017-05-01

Silicon-vacancy color centers in nanodiamonds are promising as fluorescent labels for biological applications, with a narrow, nonbleaching emission line at 738 nm. Two-photon excitation of this fluorescence offers the possibility of low-background detection at significant tissue depth with high three-dimensional spatial resolution. We measure the two-photon fluorescence cross section of a negatively charged silicon vacancy (Si -V- ) in ion-implanted bulk diamond to be 0.74 (19 )×10-50 cm4 s /photon at an excitation wavelength of 1040 nm. Compared to the diamond nitrogen-vacancy center, the expected detection threshold of a two-photon excited Si -V center is more than an order of magnitude lower, largely due to its much narrower linewidth. We also present measurements of two- and three-photon excitation spectra, finding an increase in the two-photon cross section with decreasing wavelength, and we discuss the physical interpretation of the spectra in the context of existing models of the Si -V energy-level structure.

The Pierre Auger fluorescence detector. Cross-checking the absolute calibration using a drone

Energy Technology Data Exchange (ETDEWEB)

Tomankova, Lenka [Institute for Nuclear Physics (IKP), Karlsruhe Institute of Technology (KIT), 76021 Karlsruhe (Germany); Collaboration: Pierre-Auger-Collaboration

2016-07-01

The Pierre Auger Observatory combines the air shower fluorescence and surface array methods to study ultra-high energy cosmic rays. As the energy scale of the experiment is derived from calorimetric measurements by the fluorescence telescopes, their accurate calibration is of primary importance to all Auger data. We discuss a novel calibration method based on a remotely flown drone equipped with a specially designed light source that mimics a snapshot of an air shower traversing the atmosphere. Several drone measurement campaigns have been performed to study the properties of the Auger fluorescence telescopes and to derive an end-to-end calibration. We give an overview of the measurements and present the basic analysis chain as well as the first results of an independent cross-check of the Auger energy scale.

A library for X-ray-matter interaction cross sections for X-ray fluorescence applications

Energy Technology Data Exchange (ETDEWEB)

Brunetti, A. [Istituto di Matematica e Fisica, Universita di Sassari, via Vienna 2, 07100 Sassari (Italy) and INFN, Sezione di Cagliari (Italy)]. E-mail: brunetti@uniss.it; Sanchez del Rio, M. [European Synchrotron Radiation Facility, 6 rue Jules Horowitz, 38043 Grenoble Cedex (France); Golosio, B. [INFN, Sezione di Cagliari (Italy); European Synchrotron Radiation Facility, 6 rue Jules Horowitz, 38043 Grenoble Cedex (France); Simionovici, A. [European Synchrotron Radiation Facility, 6 rue Jules Horowitz, 38043 Grenoble Cedex (France); Laboratoire de Sciences de la Terre, Ecole Normale Superieure, Lyon, F-69364 (France); Somogyi, A. [European Synchrotron Radiation Facility, 6 rue Jules Horowitz, 38043 Grenoble Cedex (France)

2004-10-08

Quantitative estimate of elemental composition by spectroscopic and imaging techniques using X-ray fluorescence requires the availability of accurate data of X-ray interaction with matter. Although a wide number of computer codes and data sets are reported in literature, none of them is presented in the form of freely available library functions which can be easily included in software applications for X-ray fluorescence. This work presents a compilation of data sets from different published works and an xraylib interface in the form of callable functions. Although the target applications are on X-ray fluorescence, cross sections of interactions like photoionization, coherent scattering and Compton scattering, as well as form factors and anomalous scattering functions, are also available.

Cross-Linked Fluorescent Supramolecular Nanoparticles as Finite Tattoo Pigments with Controllable Intradermal Retention Times.

Science.gov (United States)

Choi, Jin-Sil; Zhu, Yazhen; Li, Hongsheng; Peyda, Parham; Nguyen, Thuy Tien; Shen, Mo Yuan; Yang, Yang Michael; Zhu, Jingyi; Liu, Mei; Lee, Mandy M; Sun, Shih-Sheng; Yang, Yang; Yu, Hsiao-Hua; Chen, Kai; Chuang, Gary S; Tseng, Hsian-Rong

2017-01-24

Tattooing has been utilized by the medical community for precisely demarcating anatomic landmarks. This practice is especially important for identifying biopsy sites of nonmelanoma skin cancer (NMSC) due to the long interval (i.e., up to 3 months) between the initial diagnostic biopsy and surgical treatment. Commercially available tattoo pigments possess several issues, which include causing poor cosmesis, being mistaken for a melanocytic lesion, requiring additional removal procedures when no longer desired, and potentially inducing inflammatory responses. The ideal tattoo pigment for labeling of skin biopsy sites for NMSC requires (i) invisibility under ambient light, (ii) fluorescence under a selective light source, (iii) a finite intradermal retention time (ca. 3 months), and (iv) biocompatibility. Herein, we introduce cross-linked fluorescent supramolecular nanoparticles (c-FSNPs) as a "finite tattoo" pigment, with optimized photophysical properties and intradermal retention time to achieve successful in vivo finite tattooing. Fluorescent supramolecular nanoparticles encapsulate a fluorescent conjugated polymer, poly[5-methoxy-2-(3-sulfopropoxy)-1,4-phenylenevinylene] (MPS-PPV), into a core via a supramolecular synthetic approach. FSNPs which possess fluorescent properties superior to those of the free MPS-PPV are obtained through a combinatorial screening process. Covalent cross-linking of FSNPs results in micrometer-sized c-FSNPs, which exhibit a size-dependent intradermal retention. The 1456 nm sized c-FSNPs display an ideal intradermal retention time (ca. 3 months) for NMSC lesion labeling, as observed in an in vivo tattoo study. In addition, the c-FSNPs induce undetectable inflammatory responses after tattooing. We believe that the c-FSNPs can serve as a "finite tattoo" pigment to label potential malignant NMSC lesions.

Red fluorescence of dental plaque in children -A cross-sectional study.

Science.gov (United States)

Volgenant, Catherine M C; Zaura, Egija; Brandt, Bernd W; Buijs, Mark J; Tellez, Marisol; Malik, Gayatri; Ismail, Amid I; Ten Cate, Jacob M; van der Veen, Monique H

2017-03-01

The relation between the presence of red fluorescent plaque and the caries status in children was studied. In addition, the microbial composition of dental plaque from sites with red fluorescent plaque (RFP) and from sites with no red fluorescent plaque (NFP) was assessed. Fluorescence photographs were taken from fifty children (6-14 years old) with overnight plaque. Full-mouth caries scores (ICDAS II) were obtained. The composition of a saliva sample and two plaque samples (RFP and NFP) was assessed using 16S rDNA sequencing. At the site level, no clinically relevant correlations were found between the presence of RFP and the caries status. At the subject level, a weak correlation was found between RFP and the caries status when non-cavitated lesions were included (r s =0.37, p=0.007). The microbial composition of RFP differed significantly from NFP. RFP had more anaerobes and more Gram-negative bacterial taxa. The most discriminative operational taxonomic units (OTUs) for RFP were Corynebacterium, Leptotrichia, Porphyromonas and Selenomonas, while the most discriminative OTUs for NFP were Neisseria, Actinomyces, Streptococcus and Rothia. There were no clinical relevant correlations in this cross-sectional study between the presence of RFP and (early) caries lesions. There were differences in the composition of these phenotypically different plaque samples: RFP contained more Gram-negative, anaerobic taxa and was more diverse than NFP. The study outcomes provide more insight in the possibilities to use plaque fluorescence in oral health risk assessments. Copyright © 2017 Elsevier Ltd. All rights reserved.

Pedestrian temporal and spatial gap acceptance at mid-block street crossing in developing world.

Science.gov (United States)

Pawar, Digvijay S; Patil, Gopal R

2015-02-01

Most of the midblock pedestrian crossings on urban roads in India are uncontrolled; wherein the high degree of discretion in pedestrians' behavior while crossing the traffic stream, has made the situation complex to analyze. Vehicles do not yield to pedestrians, even though the traffic laws give priority to pedestrians over motorized vehicles at unsignalized pedestrian crossings. Therefore, a pedestrian has to decide if an available gap is safe or not for crossing. This paper aims to investigate pedestrian temporal and spatial gap acceptance for midblock street crossings. Field data were collected using video camera at two midblock pedestrian crossings. The data extraction in laboratory resulted in 1107 pedestrian gaps. Available gaps, pedestrians' decision, traffic volume, etc. were extracted from the videos. While crossing a road with multiple lanes, rolling gap acceptance behavior was observed. Using binary logit analysis, six utility models were developed, three each for temporal and spatial gaps. The 50th percentile temporal and spatial gaps ranged from 4.1 to 4.8s and 67 to 79 m respectively, whereas the 85th percentile temporal and spatial gaps ranged from 5 to 5.8s and 82 to 95 m respectively. These gap values were smaller than that reported in the studies in developed countries. The speed of conflicting vehicle was found to be significant in spatial gap but not in temporal gap acceptance. The gap acceptance decision was also found to be affected by the type of conflicting vehicles. The insights from this study can be used for the safety and performance evaluation of uncontrolled midblock street crossings in developing countries. Copyright © 2014 Elsevier Ltd and National Safety Council. All rights reserved.

Single molecule fluorescence image patterns linked to dipole orientation and axial position: application to myosin cross-bridges in muscle fibers.

Directory of Open Access Journals (Sweden)

Thomas P Burghardt

2011-02-01

Full Text Available Photoactivatable fluorescent probes developed specifically for single molecule detection extend advantages of single molecule imaging to high probe density regions of cells and tissues. They perform in the native biomolecule environment and have been used to detect both probe position and orientation.Fluorescence emission from a single photoactivated probe captured in an oil immersion, high numerical aperture objective, produces a spatial pattern on the detector that is a linear combination of 6 independent and distinct spatial basis patterns with weighting coefficients specifying emission dipole orientation. Basis patterns are tabulated for single photoactivated probes labeling myosin cross-bridges in a permeabilized muscle fiber undergoing total internal reflection illumination. Emitter proximity to the glass/aqueous interface at the coverslip implies the dipole near-field and dipole power normalization are significant affecters of the basis patterns. Other characteristics of the basis patterns are contributed by field polarization rotation with transmission through the microscope optics and refraction by the filter set. Pattern recognition utilized the generalized linear model, maximum likelihood fitting, for Poisson distributed uncertainties. This fitting method is more appropriate for treating low signal level photon counting data than χ(2 minimization.Results indicate that emission dipole orientation is measurable from the intensity image except for the ambiguity under dipole inversion. The advantage over an alternative method comparing two measured polarized emission intensities using an analyzing polarizer is that information in the intensity spatial distribution provides more constraints on fitted parameters and a single image provides all the information needed. Axial distance dependence in the emission pattern is also exploited to measure relative probe position near focus. Single molecule images from axial scanning fitted

Alexa Fluor-labeled Fluorescent Cellulose Nanocrystals for Bioimaging Solid Cellulose in Spatially Structured Microenvironments

Energy Technology Data Exchange (ETDEWEB)

Grate, Jay W.; Mo, Kai-For; Shin, Yongsoon; Vasdekis, Andreas; Warner, Marvin G.; Kelly, Ryan T.; Orr, Galya; Hu, Dehong; Dehoff, Karl J.; Brockman, Fred J.; Wilkins, Michael J.

2015-03-18

Cellulose nanocrystal materials have been labeled with modern Alexa Fluor dyes in a process that first links the dye to a cyanuric chloride molecule. Subsequent reaction with cellulose nanocrystals provides dyed solid microcrystalline cellulose material that can be used for bioimaging and suitable for deposition in films and spatially structured microenvironments. It is demonstrated with single molecular fluorescence microscopy that these films are subject to hydrolysis by cellulose enzymes.

The impact of path crossing on visuo-spatial serial memory: encoding or rehearsal effect?

Science.gov (United States)

Parmentier, Fabrice B R; Andrés, Pilar

2006-11-01

The determinants of visuo-spatial serial memory have been the object of little research, despite early evidence that not all sequences are equally remembered. Recently, empirical evidence was reported indicating that the complexity of the path formed by the to-be-remembered locations impacted on recall performance, defined for example by the presence of crossings in the path formed by successive locations (Parmentier, Elford, & Maybery, 2005). In this study, we examined whether this effect reflects rehearsal or encoding processes. We examined the effect of a retention interval and spatial interference on the ordered recall of spatial sequences with and without path crossings. Path crossings decreased recall performance, as did a retention interval. In line with the encoding hypothesis, but in contrast with the rehearsal hypothesis, the effect of crossing was not affected by the retention interval nor by tapping. The possible nature of the impact of path crossing on encoding mechanisms is discussed.

The nanosizing of fluorescent objects by 458 nm spatially modulated illumination microscopy using a simplified size evaluation algorithm

Energy Technology Data Exchange (ETDEWEB)

Schweitzer, Andreas; Wagner, Christian; Cremer, Christoph [Kirchhoff-Institute for Physics of the University, Im Neuenheimer Feld 227, 69120 Heidelberg (Germany)

2004-07-07

In fluorescent light microscopy, structured illumination approaches have emerged as a novel tool to analyse subwavelength sized objects in thick transparent specimens. In this report, new size measurements ('nanosizing') of small subwavelength sized fluorescent objects applying spatially modulated illumination (SMI) microscopy with an excitation wavelength of {lambda}{sub ex} 458 nm are presented. These measurements were made using fluorescent particles with a given diameter. From the SMI data achieved, the size (diameter) was determined using special calibration curves derived from analytical considerations assuming a Gaussian dye distribution within the object. The results showed that with SMI microscopy combined with suitable calibration, size measurements of objects considerably smaller than the epifluorescent optical resolution at {lambda}{sub ex} = 458 nm are feasible.

Superresolution size determination in fluorescence microscopy: A comparison between spatially modulated illumination and confocal laser scanning microscopy

International Nuclear Information System (INIS)

Spoeri, Udo; Failla, Antonio Virgilio; Cremer, Christoph

2004-01-01

Recently developed far field light optical methods are a powerful tool to analyze biological nanostructures and their dynamics, in particular including the interior of three-dimensionally conserved cells. In this article, the recently described method of spatially modulated illumination (SMI) microscopy has been further extended to the online determination of the extension of small, subwavelength sized, fluorescent objects (nanosizing). Using fluorescence excitation with 488 nm, the determination of fluorescent labeled object diameters down to 40 nm corresponding to about 1/12th of the wavelength used for one-photon excitation could be shown. The results of the SMI nanosizing procedure for a detailed, systematic variation of the object diameter are presented together with a fast algorithm for online size evaluation. In addition, we show a direct comparison of the diameter of 'colocalization volumes' between SMI nanosizing and conventional confocal laser scanning microscopy

Fluorescence spectroscopic study of the aggregation behavior of non-cross-linked and cross-linked poly(alkylmethyldiallylammonium bromides) having decyl, octyl, and hexyl side chains in aqueous solution

NARCIS (Netherlands)

Wang, G.J; Engberts, J.B.F.N.

1996-01-01

The conformational state of a series of non-cross-linked and cross-linked poly(alkylmethyldiallylammonium bromides) bearing decyl, octyl, and hexyl side chains ((CL)-CopolC1-10, (CL)-CopolC1-8, and (CL)-CopolC1-6, respectively) in aqueous solutions were investigated by fluorescence spectroscopy

Spatially varying cross-correlation coefficients in the presence of nugget effects

KAUST Repository

Kleiber, William; Genton, Marc G.

2012-01-01

We derive sufficient conditions for the cross-correlation coefficient of a multivariate spatial process to vary with location when the spatial model is augmented with nugget effects. The derived class is valid for any choice of covariance functions, and yields substantial flexibility between multiple processes. The key is to identify the cross-correlation coefficient matrix with a contraction matrix, which can be either diagonal, implying a parsimonious formulation, or a fully general contraction matrix, yielding greater flexibility but added model complexity. We illustrate the approach with a bivariate minimum and maximum temperature dataset in Colorado, allowing the two variables to be positively correlated at low elevations and nearly independent at high elevations, while still yielding a positive definite covariance matrix. © 2012 Biometrika Trust.

Spatially varying cross-correlation coefficients in the presence of nugget effects

KAUST Repository

Kleiber, William

2012-11-29

We derive sufficient conditions for the cross-correlation coefficient of a multivariate spatial process to vary with location when the spatial model is augmented with nugget effects. The derived class is valid for any choice of covariance functions, and yields substantial flexibility between multiple processes. The key is to identify the cross-correlation coefficient matrix with a contraction matrix, which can be either diagonal, implying a parsimonious formulation, or a fully general contraction matrix, yielding greater flexibility but added model complexity. We illustrate the approach with a bivariate minimum and maximum temperature dataset in Colorado, allowing the two variables to be positively correlated at low elevations and nearly independent at high elevations, while still yielding a positive definite covariance matrix. © 2012 Biometrika Trust.

Enhanced spatial resolution in fluorescence molecular tomography using restarted L1-regularized nonlinear conjugate gradient algorithm.

Science.gov (United States)

Shi, Junwei; Liu, Fei; Zhang, Guanglei; Luo, Jianwen; Bai, Jing

2014-04-01

Owing to the high degree of scattering of light through tissues, the ill-posedness of fluorescence molecular tomography (FMT) inverse problem causes relatively low spatial resolution in the reconstruction results. Unlike L2 regularization, L1 regularization can preserve the details and reduce the noise effectively. Reconstruction is obtained through a restarted L1 regularization-based nonlinear conjugate gradient (re-L1-NCG) algorithm, which has been proven to be able to increase the computational speed with low memory consumption. The algorithm consists of inner and outer iterations. In the inner iteration, L1-NCG is used to obtain the L1-regularized results. In the outer iteration, the restarted strategy is used to increase the convergence speed of L1-NCG. To demonstrate the performance of re-L1-NCG in terms of spatial resolution, simulation and physical phantom studies with fluorescent targets located with different edge-to-edge distances were carried out. The reconstruction results show that the re-L1-NCG algorithm has the ability to resolve targets with an edge-to-edge distance of 0.1 cm at a depth of 1.5 cm, which is a significant improvement for FMT.

Spatially resolved density and ionization measurements of shocked foams using x-ray fluorescence

Energy Technology Data Exchange (ETDEWEB)

MacDonald, M. J.; Keiter, P. A.; Montgomery, D. S.; Scott, H. A.; Biener, M. M.; Fein, J. R.; Fournier, K. B.; Gamboa, E. J.; Kemp, G. E.; Klein, S. R.; Kuranz, C. C.; LeFevre, H. J.; Manuel, M. J. -E.; Wan, W. C.; Drake, R. P.

2016-09-28

We present experiments at the Trident laser facility demonstrating the use of x-ray fluorescence (XRF) to simultaneously measure density, ionization state populations, and electron temperature in shocked foams. An imaging x-ray spectrometer obtained spatially resolved measurements of Ti K-α emission. Density profiles were measured from K-α intensity. Ti ionization state distributions and electron temperatures were inferred by fitting K-α spectra to spectra from CRETIN simulations. This work shows that XRF provides a powerful tool to complement other diagnostics to make equation of state measurements of shocked materials containing a suitable tracer element.

Spatial interpolation of point velocities in stream cross-section

Directory of Open Access Journals (Sweden)

Hasníková Eliška

2015-03-01

Full Text Available The most frequently used instrument for measuring velocity distribution in the cross-section of small rivers is the propeller-type current meter. Output of measuring using this instrument is point data of a tiny bulk. Spatial interpolation of measured data should produce a dense velocity profile, which is not available from the measuring itself. This paper describes the preparation of interpolation models.

Spatially Controlled Fabrication of Brightly Fluorescent Nanodiamond-Array with Enhanced Far-Red Si-V Luminescence

Science.gov (United States)

Singh, Sonal; Thomas, Vinoy; Martyshkin, Dmitry; Kozlovskaya, Veronika; Kharlampieva, Eugenia

2014-01-01

We demonstrate a novel approach to precise pattern fluorescent nanodiamond-arrays with enhanced far-red intense photostable luminescence from silicon-vacancy (Si-V) defect centers. The precision-patterned pre-growth seeding of nanodiamonds is achieved by scanning probe “Dip-Pen” nanolithography technique using electrostatically-driven transfer of nanodiamonds from “inked” cantilevers to a UV-treated hydrophilic SiO2 substrate. The enhanced emission from nanodiamond-dots in the far-red is achieved by incorporating Si-V defect centers in subsequent chemical vapor deposition treatment. The development of a suitable nanodiamond ink, mechanism of ink transport, and effect of humidity, dwell time on nanodiamond patterning are investigated. The precision-patterning of as-printed (pre-CVD) arrays with dot diameter and dot height as small as 735 nm ± 27 nm, 61 nm ± 3 nm, respectively and CVD-treated fluorescent ND-arrays with consistently patterned dots having diameter and height as small as 820 nm ± 20 nm, 245 nm ± 23 nm, respectively using 1 s dwell time and 30% RH is successfully achieved. We anticipate that the far-red intense photostable luminescence (~738 nm) observed from Si-V defect centers integrated in spatially arranged nanodiamonds could be beneficial for the development of the next generation fluorescent based devices and applications. PMID:24394286

Zero-crossing detection algorithm for arrays of optical spatial filtering velocimetry sensors

DEFF Research Database (Denmark)

Jakobsen, Michael Linde; Pedersen, Finn; Hanson, Steen Grüner

2008-01-01

This paper presents a zero-crossing detection algorithm for arrays of compact low-cost optical sensors based on spatial filtering for measuring fluctuations in angular velocity of rotating solid structures. The algorithm is applicable for signals with moderate signal-to-noise ratios, and delivers...... repeating the same measurement error for each revolution of the target, and to gain high performance measurement of angular velocity. The traditional zero-crossing detection is extended by 1) inserting an appropriate band-pass filter before the zero-crossing detection, 2) measuring time periods between zero...

Balanced cross-rate model for saturated molecular fluorescence in flames using a nanosecond pulse length laser

International Nuclear Information System (INIS)

Lucht, R.P.; Sweeney, D.W.; Laurendeau, N.M.

1980-01-01

The balanced cross-rate model is proposed to analyze laser-induced molecular fluorescence signals when the laser pulse length is of the order of nanoseconds. Nanosecond pulse length lasers. specifically Q-switched Nd:YAG-pumped dye lasers, are attractive for saturated molecular fluorescence spectroscopy because of their high peak power and because of their short pulse length minimizes the risk of laser-induced chemistry. In the balanced cross-rate model, single upper and lower rotational levels are assumed to be directly coupled by the laser radiation. Because the laser-induced processes which couple these levels are so fast at saturation intensities, a steady state is established between the two levels within picoseconds. Provided that the total population of the two laser-coupled rotational levels is constant during the laser pulse, the total molecular population can be calculated from the observed upper rotational level population using a two-level saturation model and Boltzmann statistics. Numerical simulation of the laser excitation dynamics of OH in an atmospheric pressure H 2 /O 2 /N 2 flame indicates that the balanced cross-rate model will give accurate results provided that the rotational relaxation rates in the upper and lower sets of rotational levels are approximately equal

Cross-coherent vector sensor processing for spatially distributed glider networks.

Science.gov (United States)

Nichols, Brendan; Sabra, Karim G

2015-09-01

Autonomous underwater gliders fitted with vector sensors can be used as a spatially distributed sensor array to passively locate underwater sources. However, to date, the positional accuracy required for robust array processing (especially coherent processing) is not achievable using dead-reckoning while the gliders remain submerged. To obtain such accuracy, the gliders can be temporarily surfaced to allow for global positioning system contact, but the acoustically active sea surface introduces locally additional sensor noise. This letter demonstrates that cross-coherent array processing, which inherently mitigates the effects of local noise, outperforms traditional incoherent processing source localization methods for this spatially distributed vector sensor network.

FLUORESCENCE PROBING OF THE FORMATION OF HYDROPHOBIC MICRODOMAINS BY CROSS-LINKED POLY(ALKYLMETHYLDIALLYLAMMONIUM BROMIDES) IN AQUEOUS-SOLUTION

NARCIS (Netherlands)

WANG, GJ; ENGBERTS, J B F N

Pyrene has been used as a fluorescence probe to investigate the conformational behavior of cross-linked poly(alkylmethyldiallylammonium bromides) in aqueous solutions. Binding of pyrene to hydrophobic microdomains, formed by the polysoaps, is reflected by a change in the ratio I-1/I-3 of the

A 32-channel photon counting module with embedded auto/cross-correlators for real-time parallel fluorescence correlation spectroscopy

Energy Technology Data Exchange (ETDEWEB)

Gong, S.; Labanca, I.; Rech, I.; Ghioni, M. [Dipartimento di Elettronica, Informazione e Bioingegneria, Politecnico di Milano, Piazza Leonardo da Vinci 32, 20133 Milano (Italy)

2014-10-15

Fluorescence correlation spectroscopy (FCS) is a well-established technique to study binding interactions or the diffusion of fluorescently labeled biomolecules in vitro and in vivo. Fast FCS experiments require parallel data acquisition and analysis which can be achieved by exploiting a multi-channel Single Photon Avalanche Diode (SPAD) array and a corresponding multi-input correlator. This paper reports a 32-channel FPGA based correlator able to perform 32 auto/cross-correlations simultaneously over a lag-time ranging from 10 ns up to 150 ms. The correlator is included in a 32 × 1 SPAD array module, providing a compact and flexible instrument for high throughput FCS experiments. However, some inherent features of SPAD arrays, namely afterpulsing and optical crosstalk effects, may introduce distortions in the measurement of auto- and cross-correlation functions. We investigated these limitations to assess their impact on the module and evaluate possible workarounds.

A 32-channel photon counting module with embedded auto/cross-correlators for real-time parallel fluorescence correlation spectroscopy

International Nuclear Information System (INIS)

Gong, S.; Labanca, I.; Rech, I.; Ghioni, M.

2014-01-01

Fluorescence correlation spectroscopy (FCS) is a well-established technique to study binding interactions or the diffusion of fluorescently labeled biomolecules in vitro and in vivo. Fast FCS experiments require parallel data acquisition and analysis which can be achieved by exploiting a multi-channel Single Photon Avalanche Diode (SPAD) array and a corresponding multi-input correlator. This paper reports a 32-channel FPGA based correlator able to perform 32 auto/cross-correlations simultaneously over a lag-time ranging from 10 ns up to 150 ms. The correlator is included in a 32 × 1 SPAD array module, providing a compact and flexible instrument for high throughput FCS experiments. However, some inherent features of SPAD arrays, namely afterpulsing and optical crosstalk effects, may introduce distortions in the measurement of auto- and cross-correlation functions. We investigated these limitations to assess their impact on the module and evaluate possible workarounds

Skin fluorescence model based on the Monte Carlo technique

Science.gov (United States)

Churmakov, Dmitry Y.; Meglinski, Igor V.; Piletsky, Sergey A.; Greenhalgh, Douglas A.

2003-10-01

The novel Monte Carlo technique of simulation of spatial fluorescence distribution within the human skin is presented. The computational model of skin takes into account spatial distribution of fluorophores following the collagen fibers packing, whereas in epidermis and stratum corneum the distribution of fluorophores assumed to be homogeneous. The results of simulation suggest that distribution of auto-fluorescence is significantly suppressed in the NIR spectral region, while fluorescence of sensor layer embedded in epidermis is localized at the adjusted depth. The model is also able to simulate the skin fluorescence spectra.

Assessing Photosynthesis by Fluorescence Imaging

Science.gov (United States)

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…

Spatial and temporal superresolution concepts to study plasma membrane organization by single molecule fluorescence techniques

International Nuclear Information System (INIS)

Ruprecht, V.

2010-01-01

Fluorescence microscopy techniques are currently among the most important experimental tools to study cellular processes. Ultra-sensitive detection devices nowadays allow for measuring even individual farnesylacetate labeled target molecules with nanometer spatial accuracy and millisecond time resolution. The emergence of single molecule fluorescence techniques especially contributed to the field of membrane biology and provided basic knowledge on structural and dynamic features of the cellular plasma membrane. However, we are still confronted with a rather fragmentary understanding of the complex architecture and functional interrelations of membrane constituents. In this thesis new concepts in one- and dual-color single molecule fluorescence techniques are presented that allow for addressing organization principles and interaction dynamics in the live cell plasma membrane. Two complementary experimental strategies are described which differ in their detection principle: single molecule fluorescence imaging and fluorescence correlation spectroscopy. The presented methods are discussed in terms of their implementation, accuracy, quantitative and statistical data analysis, as well as live cell applications. State-of-the-art dual color single molecule imaging is introduced as the most direct experimental approach to study interaction dynamics between differently labeled target molecules. New analytical estimates for robust data analysis are presented that facilitate quantitative recording and identification of co localizations in dual color single molecule images. A novel dual color illumination scheme is further described that profoundly extends the current range and sensitivity of conventional dual color single molecule experiments. The method enables working at high surface densities of fluorescent molecules - a feature typically incommensurable with single molecule imaging - and is especially suited for the detection of rare interactions by tracking co localized

L X-ray fluorescence cross sections experimentally determined for elements with 45

Energy Technology Data Exchange (ETDEWEB)

Bonzi, Edgardo V., E-mail: bonzie@famaf.unc.edu.ar [Facultad de Matematica, Astronomia y Fisica, Universidad Nacional de Cordoba, Ciudad Universitaria-5010, Cordoba (Argentina); Badiger, Nagappa M. [Departments of Physics, Karnatak University, Dharwad 580 003, Karnataka (India); Grad, Gabriela B. [Facultad de Matematica, Astronomia y Fisica, Universidad Nacional de Cordoba, Ciudad Universitaria-5010, Cordoba (Argentina); Barrea, Raul A. [The Biophysics Collaborative Access Team (BioCAT), Dept of Biological Chemical, and Physical Sciences, Illinois Institute of Technology, Chicago, IL 60616 (United States); Figueroa, Rodolo G. [Departamento de Cs. Fisicas, Universidad de La Frontera, Temuco (Chile)

2012-04-15

Experimental determination of L fluorescence cross-sections for elements with 45cross sections obtained in this work were compared with data calculated using coefficients from , , and Scofield and Puri et al. (1993, 1995) - Highlights: Black-Right-Pointing-Pointer Experimental data of L fluorescence cross-sections with 45cross sections Ll, L{alpha}, L{beta}{sub I}, L{beta}{sub II}, L{gamma}{sub {Iota}} and L{gamma}{sub {Iota}{Iota}} obtained, were compared with calculated data. Black-Right-Pointing-Pointer The Hypermet function was used to fit the data, because it considers a tail on the left side of the peak. Black-Right-Pointing-Pointer The tail is relevant when a small peak has another one on the right side with a big area.

Fluorescent microthermographic imaging

Energy Technology Data Exchange (ETDEWEB)

Barton, D.L.

1993-09-01

In the early days of microelectronics, design rules and feature sizes were large enough that sub-micron spatial resolution was not needed. Infrared or IR thermal techniques were available that calculated the object`s temperature from infrared emission. There is a fundamental spatial resolution limitation dependent on the wavelengths of light being used in the image formation process. As the integrated circuit feature sizes began to shrink toward the one micron level, the limitations imposed on IR thermal systems became more pronounced. Something else was needed to overcome this limitation. Liquid crystals have been used with great success, but they lack the temperature measurement capabilities of other techniques. The fluorescent microthermographic imaging technique (FMI) was developed to meet this need. This technique offers better than 0.01{degrees}C temperature resolution and is diffraction limited to 0.3 {mu}m spatial resolution. While the temperature resolution is comparable to that available on IR systems, the spatial resolution is much better. The FMI technique provides better spatial resolution by using a temperature dependent fluorescent film that emits light at 612 nm instead of the 1.5 {mu}m to 12 {mu}m range used by IR techniques. This tutorial starts with a review of blackbody radiation physics, the process by which all heated objects emit radiation to their surroundings, in order to understand the sources of information that are available to characterize an object`s surface temperature. The processes used in infrared thermal imaging are then detailed to point out the limitations of the technique but also to contrast it with the FMI process. The FMI technique is then described in detail, starting with the fluorescent film physics and ending with a series of examples of past applications of FMI.

Sheet Fluorescence and Annular Analysis of Ultracold Neutral Plasmas

International Nuclear Information System (INIS)

Castro, J.; Gao, H.; Killian, T. C.

2009-01-01

Annular analysis of fluorescence imaging measurements on Ultracold Neutral Plasmas (UNPs) is demonstrated. Spatially-resolved fluorescence imaging of the strontium ions produces a spectrum that is Doppler-broadened due to the thermal ion velocity and shifted due to the ion expansion velocity. The fluorescence excitation beam is spatially narrowed into a sheet, allowing for localized analysis of ion temperatures within a volume of the plasma with small density variation. Annular analysis of fluorescence images permits an enhanced signal-to-noise ratio compared to previous fluorescence measurements done in strontium UNPs. Using this technique and analysis, plasma ion temperatures are measured and shown to display characteristics of plasmas with strong coupling such as disorder induced heating and kinetic energy oscillations.

Fluorescence lifetime based bioassays

Science.gov (United States)

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.

Integrated image presentation of transmission and fluorescent X-ray CT using synchrotron radiation

Energy Technology Data Exchange (ETDEWEB)

Zeniya, T.; Takeda, T. E-mail: ttakeda@md.tsukuba.ac.jp; Yu, Q.; Hasegawa, Y.; Hyodo, K.; Yuasa, T.; Hiranaka, Y.; Itai, Y.; Akatsuka, T

2001-07-21

We have developed a computed tomography (CT) system with synchrotron radiation (SR) to detect fluorescent X-rays and transmitted X-rays simultaneously. Both SR transmission X-ray CT (SR-TXCT) and SR fluorescent X-ray CT (SR-FXCT) can describe cross-sectional images with high spatial and contrast resolutions as compared to conventional CT. TXCT gives morphological information and FXCT gives functional information of organs. So, superposed display system for SR-FXCT and SR-TXCT images has been developed for clinical diagnosis with higher reliability. Preliminary experiment with brain phantom was carried out and the superposition of both images was performed. The superposed SR-CT image gave us both functional and morphological information easily with high reliability, thus demonstrating the usefulness of this system.

Integrated image presentation of transmission and fluorescent X-ray CT using synchrotron radiation

Science.gov (United States)

Zeniya, T.; Takeda, T.; Yu, Q.; Hasegawa, Y.; Hyodo, K.; Yuasa, T.; Hiranaka, Y.; Itai, Y.; Akatsuka, T.

2001-07-01

We have developed a computed tomography (CT) system with synchrotron radiation (SR) to detect fluorescent X-rays and transmitted X-rays simultaneously. Both SR transmission X-ray CT (SR-TXCT) and SR fluorescent X-ray CT (SR-FXCT) can describe cross-sectional images with high spatial and contrast resolutions as compared to conventional CT. TXCT gives morphological information and FXCT gives functional information of organs. So, superposed display system for SR-FXCT and SR-TXCT images has been developed for clinical diagnosis with higher reliability. Preliminary experiment with brain phantom was carried out and the superposition of both images was performed. The superposed SR-CT image gave us both functional and morphological information easily with high reliability, thus demonstrating the usefulness of this system.

Vertical profiles of atmospheric fluorescent aerosols observed by a mutil-channel lidar spectrometer system

Science.gov (United States)

Huang, Z.; Huang, J.; Zhou, T.; Sugimoto, N.; Bi, J.

2015-12-01

Zhongwei Huang1*, Jianping Huang1, Tian Zhou1, Nobuo Sugimoto2, Jianrong Bi1 and Jinsen Shi11Key Laboratory for Semi-Arid Climate Change of the Ministry of Education, College of Atmospheric Sciences, Lanzhou University, Lanzhou, China. 2Atmospheric Environment Division, National Institutes for Environmental Studies, Tsukuba, Japan Email: huangzhongwei@lzu.edu.cn Abstract Atmospheric aerosols have a significant impact on regional and globe climate. The challenge in quantifying aerosol direct radiative forcing and aerosol-cloud interactions arises from large spatial and temporal heterogeneity of aerosol concentrations, compositions, sizes, shape and optical properties (IPCC, 2007). Lidar offers some remarkable advantages for determining the vertical structure of atmospheric aerosols and their related optical properties. To investigate the characterization of atmospheric aerosols (especially bioaerosols) with high spatial and temporal resolution, we developed a Raman/fluorescence/polarization lidar system employed a multi-channel spectrometer, with capabilities of providing measurements of Raman scattering and laser-induced fluorescence excitation at 355 nm from atmospheric aerosols. Meanwhile, the lidar system operated polarization measurements both at 355nm and 532nm wavelengths, aiming to obtain more information of aerosols. It employs a high power pulsed laser and a received telescope with 350mm diameter. The receiver could simultaneously detect a wide fluorescent spectrum about 178 nm with spectral resolution 5.7 nm, mainly including an F/3.7 Crossed Czerny-Turner spectrograph, a grating (1200 gr/mm) and a PMT array with 32 photocathode elements. Vertical structure of fluorescent aerosols in the atmosphere was observed by the developed lidar system at four sites across northwest China, during 2014 spring field observation that conducted by Lanzhou University. It has been proved that the developed lidar could detect the fluorescent aerosols with high temporal and

Spatially controlled fabrication of a bright fluorescent nanodiamond-array with enhanced far-red Si-V luminescence.

Science.gov (United States)

Singh, Sonal; Thomas, Vinoy; Martyshkin, Dmitry; Kozlovskaya, Veronika; Kharlampieva, Eugenia; Catledge, Shane A

2014-01-31

We demonstrate a novel approach to precisely pattern fluorescent nanodiamond-arrays with enhanced far-red intense photostable luminescence from silicon-vacancy (Si-V) defect centers. The precision-patterned pre-growth seeding of nanodiamonds is achieved by a scanning probe 'dip-pen' nanolithography technique using electrostatically driven transfer of nanodiamonds from 'inked' cantilevers to a UV-treated hydrophilic SiO2 substrate. The enhanced emission from nanodiamond dots in the far-red is achieved by incorporating Si-V defect centers in a subsequent chemical vapor deposition treatment. The development of a suitable nanodiamond ink and mechanism of ink transport, and the effect of humidity and dwell time on nanodiamond patterning are investigated. The precision patterning of as-printed (pre-CVD) arrays with dot diameter and dot height as small as 735 nm ± 27 nm and 61 nm ± 3 nm, respectively, and CVD-treated fluorescent ND-arrays with consistently patterned dots having diameter and height as small as 820 nm ± 20 nm and, 245 nm ± 23 nm, respectively, using 1 s dwell time and 30% RH is successfully achieved. We anticipate that the far-red intense photostable luminescence (~738 nm) observed from Si-V defect centers integrated in spatially arranged nanodiamonds could be beneficial for the development of next generation fluorescence-based devices and applications.

Spatial heterogeneity of biofouling under different cross-flow velocities in reverse osmosis membrane systems

KAUST Repository

Farhat, Nadia; Staal, M.; Bucs, Szilard; Van Loosdrecht, M.C.M.; Vrouwenvelder, Johannes S.

2016-01-01

the spatial heterogeneity of biofilm development over the membrane fouling simulator (MFS) length (inlet and outlet part) at three different cross-flow velocities (0.08, 0.12 and 0.16 m/s). The MFS contained sheets of membrane and feed spacer and simulatedComparison of the inlet and outlet position of the MFS showed a more (i) heterogeneous biofilm distribution and a (ii) higher biological activity at the inlet side (first 2.5 cm) for all cross-flow velocities. The lowest cross-flow velocity had

The detection of T-Nos, a genetic element present in GMOs, by cross-priming isothermal amplification with real-time fluorescence.

Science.gov (United States)

Zhang, Fang; Wang, Liu; Fan, Kai; Wu, Jian; Ying, Yibin

2014-05-01

An isothermal cross-priming amplification (CPA) assay for Agrobacterium tumefaciens nopaline synthase terminator (T-Nos) was established and investigated in this work. A set of six specific primers, recognizing eight distinct regions on the T-Nos sequence, was designed. The CPA assay was performed at a constant temperature, 63 °C, and detected by real-time fluorescence. The results indicated that real-time fluorescent CPA had high specificity, and the limit of detection was 1.06 × 10(3) copies of rice genomic DNA, which could be detected in 40 min. Comparison of real-time fluorescent CPA and conventional polymerase chain reaction (PCR) was also performed. Results revealed that real-time fluorescent CPA had a comparable sensitivity to conventional real-time PCR and had taken a shorter time. In addition, different contents of genetically modified (GM)-contaminated rice seed powder samples were detected for practical application. The result showed real-time fluorescent CPA could detect 0.5 % GM-contaminated samples at least, and the whole reaction could be finished in 35 min. Real-time fluorescent CPA is sensitive enough to monitor labeling systems and provides an attractive method for the detection of GMO.

Confocal microscopy evaluation of stromal fluorescence intensity after standard and accelerated iontophoresis-assisted corneal cross-linking.

Science.gov (United States)

Lanzini, Manuela; Curcio, Claudia; Spoerl, Eberhard; Calienno, Roberta; Mastropasqua, Alessandra; Colasante, Martina; Mastropasqua, Rodolfo; Nubile, Mario; Mastropasqua, Leonardo

2017-02-01

The aim of this study is to determine modifications in stromal fluorescence intensity after different corneal cross-linking (CXL) procedures and to correlate stromal fluorescence to corneal biomechanical resistance. For confocal microscopy study, 15 human cadaver corneas were examined. Three served as control (group 1), three were just soaked with iontophoresis procedure (group 2), three were treated with standard epi-off technique (group 3), and six underwent iontophoresis imbibition. Three of later six were irradiated for 30 min with 3 mW/cm 2 UVA (group 4) and three for 9 min at 10 mW/cm 2 UVA (group 5). Confocal microscopy was performed to quantify the fluorescence intensity in the cornea at different stromal depths. For biomechanical study, 30 human cadaver corneas were randomly divided into five groups and treated as previously described. Static stress-strain measurements of the corneas were performed. Iontophoresis imbibition followed by 10mW/cm 2 irradiation proved to increase stromal fluorescence into the corneal stroma and significant differences were revealed between group 3 and 5 both at 100 (p = 0.0171) and 250 µm (p = 0.0024), respectively. Biomechanical analysis showed an improvement of corneal resistance in group 5. Iontophoresis imbibition followed by accelerated irradiation increased the stromal fluorescence and is related to an improvement of biomechanical resistance. This approach may represent a new strategy to achieve greater concentrations of riboflavin without removing corneal epithelium and improve clinical results while reducing the side effects of CXL.

Fluorescence microscopy.

Science.gov (United States)

Sanderson, Michael J; Smith, Ian; Parker, Ian; Bootman, Martin D

2014-10-01

Fluorescence microscopy is a major tool with which to monitor cell physiology. Although the concepts of fluorescence and its optical separation using filters remain similar, microscope design varies with the aim of increasing image contrast and spatial resolution. The basics of wide-field microscopy are outlined to emphasize the selection, advantages, and correct use of laser scanning confocal microscopy, two-photon microscopy, scanning disk confocal microscopy, total internal reflection, and super-resolution microscopy. In addition, the principles of how these microscopes form images are reviewed to appreciate their capabilities, limitations, and constraints for operation. © 2014 Cold Spring Harbor Laboratory Press.

Spatial-temporal consistency between gross primary productivity and solar-induced chlorophyll fluorescence of vegetation in China during 2007-2014

Science.gov (United States)

Ma, J.; Xiao, X.; Zhang, Y.; Chen, B.; Zhao, B.

2017-12-01

Great significance exists in accurately estimating spatial-temporal patterns of gross primary production (GPP) because of its important role in global carbon cycle. Satellite-based light use efficiency (LUE) models are regarded as an efficient tool in simulating spatially time-sires GPP. However, the estimation of the accuracy of GPP simulations from LUE at both spatial and temporal scales is still a challenging work. In this study, we simulated GPP of vegetation in China during 2007-2014 using a LUE model (Vegetation Photosynthesis Model, VPM) based on MODIS (moderate-resolution imaging spectroradiometer) images of 8-day temporal and 500-m spatial resolutions and NCEP (National Center for Environmental Prediction) climate data. Global Ozone Monitoring Instrument 2 (GOME-2) solar-induced chlorophyll fluorescence (SIF) data were used to compare with VPM simulated GPP (GPPVPM) temporally and spatially using linear correlation analysis. Significant positive linear correlations exist between monthly GPPVPM and SIF data over both single year (2010) and multiple years (2007-2014) in China. Annual GPPVPM is significantly positive correlated with SIF (R2>0.43) spatially for all years during 2007-2014 and all seasons in 2010 (R2>0.37). GPP dynamic trends is high spatial-temporal heterogeneous in China during 2007-2014. The results of this study indicate that GPPVPM is temporally and spatially in line with SIF data, and space-borne SIF data have great potential in validating and parameterizing GPP estimation of LUE-based models.

Spatially controlled fabrication of a bright fluorescent nanodiamond-array with enhanced far-red Si-V luminescence

International Nuclear Information System (INIS)

Singh, Sonal; Thomas, Vinoy; Kharlampieva, Eugenia; Catledge, Shane A; Martyshkin, Dmitry; Kozlovskaya, Veronika

2014-01-01

We demonstrate a novel approach to precisely pattern fluorescent nanodiamond-arrays with enhanced far-red intense photostable luminescence from silicon-vacancy (Si-V) defect centers. The precision-patterned pre-growth seeding of nanodiamonds is achieved by a scanning probe ‘dip-pen’ nanolithography technique using electrostatically driven transfer of nanodiamonds from ‘inked’ cantilevers to a UV-treated hydrophilic SiO 2 substrate. The enhanced emission from nanodiamond dots in the far-red is achieved by incorporating Si-V defect centers in a subsequent chemical vapor deposition treatment. The development of a suitable nanodiamond ink and mechanism of ink transport, and the effect of humidity and dwell time on nanodiamond patterning are investigated. The precision patterning of as-printed (pre-CVD) arrays with dot diameter and dot height as small as 735 nm ± 27 nm and 61 nm ± 3 nm, respectively, and CVD-treated fluorescent ND-arrays with consistently patterned dots having diameter and height as small as 820 nm ± 20 nm and, 245 nm ± 23 nm, respectively, using 1 s dwell time and 30% RH is successfully achieved. We anticipate that the far-red intense photostable luminescence (∼738 nm) observed from Si-V defect centers integrated in spatially arranged nanodiamonds could be beneficial for the development of next generation fluorescence-based devices and applications. (paper)

Manipulating Bodily Presence Affects Cross-Modal Spatial Attention: A Virtual-Reality-Based ERP Study

OpenAIRE

Harjunen, Ville J.; Ahmed, Imtiaj; Jacucci, Giulio; Ravaja, Niklas; Spap?, Michiel M.

2017-01-01

Earlier studies have revealed cross-modal visuo-tactile interactions in endogenous spatial attention. The current research used event-related potentials (ERPs) and virtual reality (VR) to identify how the visual cues of the perceiver’s body affect visuo-tactile interaction in endogenous spatial attention and at what point in time the effect takes place. A bimodal oddball task with lateralized tactile and visual stimuli was presented in two VR conditions, one with and one without visible hands...

Disparity in formulations used for fluorescent X-ray intensity measurements

International Nuclear Information System (INIS)

Mittal, Raj; Gupta, Sheenu

2011-01-01

The paper presents a problem in computations of X-ray fluorescence cross-sections, shell/sub-shell fluorescence yields, Coster-Kronig yields, vacancy alignment, etc. from X-ray fluorescence (XRF) studies. While using barn/atom as a unit for cross-sections if the atomic masses are not considered it causes a discrepancy in the measured cross-section, yield and alignment values. Most of the earlier publications are being quoted where such an oversight has occurred and discrepancy is evident. - Highlights: → Manuscript gives basic formulation for measurements of fluorescent X-ray intensities. → Most publications ignored the fact that use of barn/atom units for cross-sections requires atomic masses. → Published experimental results higher by a factor ≥2 or less by factor 1/M K . → Inspection of published data on XRF parameters needed.

A new X-ray pinhole camera for energy dispersive X-ray fluorescence imaging with high-energy and high-spatial resolution

Energy Technology Data Exchange (ETDEWEB)

Romano, F.P., E-mail: romanop@lns.infn.it [IBAM, CNR, Via Biblioteca 4, 95124 Catania (Italy); INFN-LNS, Via S. Sofia 62, 95123 Catania (Italy); Altana, C. [INFN-LNS, Via S. Sofia 62, 95123 Catania (Italy); Dipartimento di Fisica e Astronomia, Università di Catania, Via S. Sofia 64, 95123 Catania (Italy); Cosentino, L.; Celona, L.; Gammino, S.; Mascali, D. [INFN-LNS, Via S. Sofia 62, 95123 Catania (Italy); Pappalardo, L. [IBAM, CNR, Via Biblioteca 4, 95124 Catania (Italy); INFN-LNS, Via S. Sofia 62, 95123 Catania (Italy); Rizzo, F. [INFN-LNS, Via S. Sofia 62, 95123 Catania (Italy); Dipartimento di Fisica e Astronomia, Università di Catania, Via S. Sofia 64, 95123 Catania (Italy)

2013-08-01

A new X-ray pinhole camera for the Energy Dispersive X-ray Fluorescence (ED-XRF) imaging of materials with high-energy and high-spatial resolution, was designed and developed. It consists of a back-illuminated and deep depleted CCD detector (composed of 1024 × 1024 pixels with a lateral size of 13 μm) coupled to a 70 μm laser-drilled pinhole-collimator, positioned between the sample under analysis and the CCD. The X-ray pinhole camera works in a coaxial geometry allowing a wide range of magnification values. The characteristic X-ray fluorescence is induced on the samples by irradiation with an external X-ray tube working at a maximum power of 100 W (50 kV and 2 mA operating conditions). The spectroscopic capabilities of the X-ray pinhole camera were accurately investigated. Energy response and energy calibration of the CCD detector were determined by irradiating pure target-materials emitting characteristic X-rays in the energy working-domain of the system (between 3 keV and 30 keV). Measurements were performed by using a multi-frame acquisition in single-photon counting. The characteristic X-ray spectra were obtained by an automated processing of the acquired images. The energy resolution measured at the Fe–Kα line is 157 eV. The use of the X-ray pinhole camera for the 2D resolved elemental analysis was investigated by using reference-patterns of different materials and geometries. The possibility of the elemental mapping of samples up to an area of 3 × 3 cm{sup 2} was demonstrated. Finally, the spatial resolution of the pinhole camera was measured by analyzing the profile function of a sharp-edge. The spatial resolution determined at the magnification values of 3.2 × and 0.8 × (used as testing values) is about 90 μm and 190 μm respectively. - Highlights: • We developed an X-ray pinhole camera for the 2D X-ray fluorescence imaging. • X-ray spectra are obtained by a multi-frame acquisition in single photon mode. • The energy resolution in the X

Improving visual spatial working memory in younger and older adults: effects of cross-modal cues.

Science.gov (United States)

Curtis, Ashley F; Turner, Gary R; Park, Norman W; Murtha, Susan J E

2017-11-06

Spatially informative auditory and vibrotactile (cross-modal) cues can facilitate attention but little is known about how similar cues influence visual spatial working memory (WM) across the adult lifespan. We investigated the effects of cues (spatially informative or alerting pre-cues vs. no cues), cue modality (auditory vs. vibrotactile vs. visual), memory array size (four vs. six items), and maintenance delay (900 vs. 1800 ms) on visual spatial location WM recognition accuracy in younger adults (YA) and older adults (OA). We observed a significant interaction between spatially informative pre-cue type, array size, and delay. OA and YA benefitted equally from spatially informative pre-cues, suggesting that attentional orienting prior to WM encoding, regardless of cue modality, is preserved with age.  Contrary to predictions, alerting pre-cues generally impaired performance in both age groups, suggesting that maintaining a vigilant state of arousal by facilitating the alerting attention system does not help visual spatial location WM.

Fluorescent Probes and Fluorescence (Microscopy Techniques — Illuminating Biological and Biomedical Research

Directory of Open Access Journals (Sweden)

Gregor P. C. Drummen

2012-11-01

Full Text Available Fluorescence, the absorption and re-emission of photons with longer wavelengths, is one of those amazing phenomena of Nature. Its discovery and utilization had, and still has, a major impact on biological and biomedical research, since it enables researchers not just to visualize normal physiological processes with high temporal and spatial resolution, to detect multiple signals concomitantly, to track single molecules in vivo, to replace radioactive assays when possible, but also to shed light on many pathobiological processes underpinning disease states, which would otherwise not be possible. Compounds that exhibit fluorescence are commonly called fluorochromes or fluorophores and one of these fluorescent molecules in particular has significantly enabled life science research to gain new insights in virtually all its sub-disciplines: Green Fluorescent Protein. Because fluorescent proteins are synthesized in vivo, integration of fluorescent detection methods into the biological system via genetic techniques now became feasible. Currently fluorescent proteins are available that virtually span the whole electromagnetic spectrum. Concomitantly, fluorescence imaging techniques were developed, and often progress in one field fueled innovation in the other. Impressively, the properties of fluorescence were utilized to develop new assays and imaging modalities, ranging from energy transfer to image molecular interactions to imaging beyond the diffraction limit with super-resolution microscopy. Here, an overview is provided of recent developments in both fluorescence imaging and fluorochrome engineering, which together constitute the “fluorescence toolbox” in life science research.

Spatial Exploration and Characterization of Endozoicomonas spp. Bacteria in Stylophora pistillata Using Fluorescence In Situ Hybridization

KAUST Repository

Alsheikh-­Hussain, Areej

2011-12-12

Studies of coral-­associated bacterial communities have repeatedly demonstrated that the microbial assemblages of the coral host are highly specific and complex. In particular, bacterial community surveys of scleractinian and soft corals from geographically diverse reefs continually uncover a high abundance of sequences affiliated with the Gammaproteobacteria genus Endozoicomonas. The role of these bacteria within the complex coral holobiont is currently unknown. In order to localize these cells and gain an understanding of their potential interactions within the coral, we developed a fluorescence in situ hybridization(FISH) approach for reef-­building coral tissues. Using a custom small-­subunit ribosomal RNA gene database, we developed two Endozoicomonas-­specific probes that cover almost all known coral-­associated Endozoicomonas sequences. Probe hybridization conditions were quantitatively evaluated against target and non-­target bacterial cultures using fluorescence microscopy. Using these experimentally tested conditions, probes were then hybridized to the branching coral Stylophora pistillata, obtained from the Red Sea, using whole mount and paraffin embedding techniques. This study allowed preliminary spatial exploration and characterization of Endozoicomonas in coral, which has provided insight into their functional role and interactions within the coral holobiont.

Mitigating fluorescence spectral overlap in wide-field endoscopic imaging

Science.gov (United States)

Hou, Vivian; Nelson, Leonard Y.; Seibel, Eric J.

2013-01-01

Abstract. The number of molecular species suitable for multispectral fluorescence imaging is limited due to the overlap of the emission spectra of indicator fluorophores, e.g., dyes and nanoparticles. To remove fluorophore emission cross-talk in wide-field multispectral fluorescence molecular imaging, we evaluate three different solutions: (1) image stitching, (2) concurrent imaging with cross-talk ratio subtraction algorithm, and (3) frame-sequential imaging. A phantom with fluorophore emission cross-talk is fabricated, and a 1.2-mm ultrathin scanning fiber endoscope (SFE) is used to test and compare these approaches. Results show that fluorophore emission cross-talk could be successfully avoided or significantly reduced. Near term, the concurrent imaging method of wide-field multispectral fluorescence SFE is viable for early stage cancer detection and localization in vivo. Furthermore, a means to enhance exogenous fluorescence target-to-background ratio by the reduction of tissue autofluorescence background is demonstrated. PMID:23966226

A Strategic Spatial Planning Approach to Cross-Border Place Branding with References to Galicia and Northern Portugal

NARCIS (Netherlands)

da Silva Oliveira, Eduardo; Zenker, Sebastian; Jacobsen, Björn

2015-01-01

This chapter adopts a strategic spatial planning approach to think strategically about potential joint place-branding initiatives between cross-border regions. The case study focuses on the extended cross-border European region composed of the NUTS III Alto Minho, Cávado, Ave, Área Metropolitina do

Spatial heterogeneity of biofouling under different cross-flow velocities in reverse osmosis membrane systems

KAUST Repository

Farhat, Nadia

2016-09-06

The spatially heterogeneous distribution of biofouling in spiral wound membrane systems restricts (i) the water distribution over the membrane surface and therefore (ii) the membrane-based water treatment. The objective of the study was to assess the spatial heterogeneity of biofilm development over the membrane fouling simulator (MFS) length (inlet and outlet part) at three different cross-flow velocities (0.08, 0.12 and 0.16 m/s). The MFS contained sheets of membrane and feed spacer and simulated the first 0.20 m of spiral-wound membrane modules where biofouling accumulates the most in practice. In-situ non-destructive oxygen imaging using planar optodes was applied to determine the biofilm spatially resolved activity and heterogeneity.

Breaking the deadlock: lessons from cross-border spatial projects in Flanders and the Netherlands

NARCIS (Netherlands)

de Vries, J.

2008-01-01

This paper explores the specific characteristics of decision-making in cross-border spatial projects and draws some lessons for European territorial cooperation. This is done by examining three major infrastructure projects between the Netherlands and Flanders: the construction of a high-speed rail

Quantification of Material Fluorescence and Light Scattering Cross Sections Using Ratiometric Bandwidth-Varied Polarized Resonance Synchronous Spectroscopy.

Science.gov (United States)

Xu, Joanna Xiuzhu; Hu, Juan; Zhang, Dongmao

2018-05-25

Presented herein is the ratiometric bandwidth-varied polarized resonance synchronous spectroscopy (BVPRS2) method for quantification of material optical activity spectra. These include the sample light absorption and scattering cross-section spectrum, the scattering depolarization spectrum, and the fluorescence emission cross-section and depolarization spectrum in the wavelength region where the sample both absorbs and emits. This ratiometric BVPRS2 spectroscopic method is a self-contained technique capable of quantitatively decoupling material fluorescence and light scattering signal contribution to its ratiometric BVPRS2 spectra through the linear curve-fitting of the ratiometric BVPRS2 signal as a function of the wavelength bandwidth used in the PRS2 measurements. Example applications of this new spectroscopic method are demonstrated with materials that can be approximated as pure scatterers, simultaneous photon absorbers/emitters, simultaneous photon absorbers/scatterers, and finally simultaneous photon absorbers/scatterers/emitters. Because the only instruments needed for this ratiometric BVPRS2 technique are the conventional UV-vis spectrophotometer and spectrofluorometer, this work should open doors for routine decomposition of material UV-vis extinction spectrum into its absorption and scattering component spectra. The methodology and insights provided in this work should be of broad significance to all chemical research that involves photon/matter interactions.

Integrating cross-scale analysis in the spatial and temporal domains for classification of behavioral movement

Directory of Open Access Journals (Sweden)

Ali Soleymani

2014-06-01

Full Text Available Since various behavioral movement patterns are likely to be valid within different, unique ranges of spatial and temporal scales (e.g., instantaneous, diurnal, or seasonal with the corresponding spatial extents, a cross-scale approach is needed for accurate classification of behaviors expressed in movement. Here, we introduce a methodology for the characterization and classification of behavioral movement data that relies on computing and analyzing movement features jointly in both the spatial and temporal domains. The proposed methodology consists of three stages. In the first stage, focusing on the spatial domain, the underlying movement space is partitioned into several zonings that correspond to different spatial scales, and features related to movement are computed for each partitioning level. In the second stage, concentrating on the temporal domain, several movement parameters are computed from trajectories across a series of temporal windows of increasing sizes, yielding another set of input features for the classification. For both the spatial and the temporal domains, the ``reliable scale'' is determined by an automated procedure. This is the scale at which the best classification accuracy is achieved, using only spatial or temporal input features, respectively. The third stage takes the measures from the spatial and temporal domains of movement, computed at the corresponding reliable scales, as input features for behavioral classification. With a feature selection procedure, the most relevant features contributing to known behavioral states are extracted and used to learn a classification model. The potential of the proposed approach is demonstrated on a dataset of adult zebrafish (Danio rerio swimming movements in testing tanks, following exposure to different drug treatments. Our results show that behavioral classification accuracy greatly increases when firstly cross-scale analysis is used to determine the best analysis scale, and

Nonmydriatic fluorescence-based quantitative imaging of human macular pigment distributions

Science.gov (United States)

Sharifzadeh, Mohsen; Bernstein, Paul S.; Gellermann, Werner

2006-10-01

We have developed a CCD-camera-based nonmydriatic instrument that detects fluorescence from retinal lipofuscin chromophores ("autofluorescence") as a means to indirectly quantify and spatially image the distribution of macular pigment (MP). The lipofuscin fluorescence intensity is reduced at all retinal locations containing MP, since MP has a competing absorption in the blue-green wavelength region. Projecting a large diameter, 488 nm excitation spot onto the retina, centered on the fovea, but extending into the macular periphery, and comparing lipofuscin fluorescence intensities outside and inside the foveal area, it is possible to spatially map out the distribution of MP. Spectrally selective detection of the lipofuscin fluorescence reveals an important wavelength dependence of the obtainable image contrast and deduced MP optical density levels, showing that it is important to block out interfering fluorescence contributions in the detection setup originating from ocular media such as the lens. Measuring 70 healthy human volunteer subjects with no ocular pathologies, we find widely varying spatial extent of MP, distinctly differing distribution patterns of MP, and strongly differing absolute MP levels among individuals. Our population study suggests that MP imaging based on lipofuscin fluorescence is useful as a relatively simple, objective, and quantitative noninvasive optical technique suitable to rapidly screen MP levels and distributions in healthy humans with undilated pupils.

Anatomy of Teaching Anatomy: Do Prosected Cross Sections Improve Students Understanding of Spatial and Radiological Anatomy?

Directory of Open Access Journals (Sweden)

L. B. Samarakoon

2016-01-01

Full Text Available Introduction. Cadaveric dissections and prosections have traditionally been part of undergraduate medical teaching. Materials and Methods. Hundred and fifty-nine first-year students in the Faculty of Medicine, University of Colombo, were invited to participate in the above study. Students were randomly allocated to two age and gender matched groups. Both groups were exposed to identical series of lectures regarding anatomy of the abdomen and conventional cadaveric prosections of the abdomen. The test group (n=77, 48.4% was also exposed to cadaveric cross-sectional slices of the abdomen to which the control group (n=82, 51.6% was blinded. At the end of the teaching session both groups were assessed by using their performance in a timed multiple choice question paper as well as ability to identify structures in abdominal CT films. Results. Scores for spatial and radiological anatomy were significantly higher among the test group when compared with the control group (P<0.05, CI 95%. Majority of the students in both control and test groups agreed that cadaveric cross section may be useful for them to understand spatial and radiological anatomy. Conclusion. Introduction of cadaveric cross-sectional prosections may help students to understand spatial and radiological anatomy better.

Mechanisms of crossing for an X-junction based on dark spatial solitons

International Nuclear Information System (INIS)

Torres-Cisneros, M; Aguilera-Cortes, L A; Meneses-Nava, M A; Sanchez-Mondragon, J J; Torres-Cisneros, G E

2004-01-01

We present a fundamental study on the capability of a crossing of two optical waveguides based on dark spatial solitons to act as a controllable optical beam splitter. Our study is based on the fact that the guided beam is diffracted at the waveguide crossing by an effective phase screen formed by the soliton collision profile. We find that when the two dark solitons are immersed into the same finite bright background, the energy of a guided beam can be split into the desired optical channel according to the collision angle. We also found that even the corresponding phase diffractive screen possesses a quite different structure in the bright and dark soliton cases; the physics involved is the same

Automatic enhancement of skin fluorescence localization due to refractive index matching

Science.gov (United States)

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.

Design and implementation of a dual-wavelength intrinsic fluorescence camera system

Science.gov (United States)

Ortega-Martinez, Antonio; Musacchia, Joseph J.; Gutierrez-Herrera, Enoch; Wang, Ying; Franco, Walfre

2017-03-01

Intrinsic UV fluorescence imaging is a technique that permits the observation of spatial differences in emitted fluorescence. It relies on the fluorescence produced by the innate fluorophores in the sample, and thus can be used for marker-less in-vivo assessment of tissue. It has been studied as a tool for the study of the skin, specifically for the classification of lesions, the delimitation of lesion borders and the study of wound healing, among others. In its most basic setup, a sample is excited with a narrow-band UV light source and the resulting fluorescence is imaged with a UV sensitive camera filtered to the emission wavelength of interest. By carefully selecting the excitation/emission pair, we can observe changes in fluorescence associated with physiological processes. One of the main drawbacks of this simple setup is the inability to observe more than a single excitation/emission pair at the same time, as some phenomena are better studied when two or more different pairs are studied simultaneously. In this work, we describe the design and the hardware and software implementation of a dual wavelength portable UV fluorescence imaging system. Its main components are an UV camera, a dual wavelength UV LED illuminator (295 and 345 nm) and two different emission filters (345 and 390 nm) that can be swapped by a mechanical filter wheel. The system is operated using a laptop computer and custom software that performs basic pre-processing to improve the image. The system was designed to allow us to image fluorescent peaks of tryptophan and collagen cross links in order to study wound healing progression.

Cross sections for energy transfer in collisions between two excited sodium atoms

International Nuclear Information System (INIS)

Huennekens, J.; Gallagher, A.

1983-01-01

We have measured cross sections, sigma/sub n/L, for the excitation transfer process Na(3P)+Na(3P)→Na(3S)+Na(nL), where nL is the 4D or 5S level. Our results are sigma/sub 4D/ = 23 A 2 +- 35% and sigma/sub 5S/ = 16 A 2 +- 35% at Tapprox.600 K. To obtain these cross sections we have used pulsed excitation and measured the intensities of 4D, 5S, and 3P fluorescence emissions, and the spatial distribution of excited atoms resulting from radiation diffusion, as well as the excited atom density as a function of time. Additionally, we have accounted for (time-dependent) radiation trapping of 3P and nL level radiation and for the resulting anisotropies of these fluorescence emissions. Comparisons of our results with theory have been made, and their relevance to other experiments is discussed

Spatial and temporal single-cell volume estimation by a fluorescence imaging technique with application to astrocytes in primary culture

Science.gov (United States)

Khatibi, Siamak; Allansson, Louise; Gustavsson, Tomas; Blomstrand, Fredrik; Hansson, Elisabeth; Olsson, Torsten

1999-05-01

Cell volume changes are often associated with important physiological and pathological processes in the cell. These changes may be the means by which the cell interacts with its surrounding. Astroglial cells change their volume and shape under several circumstances that affect the central nervous system. Following an incidence of brain damage, such as a stroke or a traumatic brain injury, one of the first events seen is swelling of the astroglial cells. In order to study this and other similar phenomena, it is desirable to develop technical instrumentation and analysis methods capable of detecting and characterizing dynamic cell shape changes in a quantitative and robust way. We have developed a technique to monitor and to quantify the spatial and temporal volume changes in a single cell in primary culture. The technique is based on two- and three-dimensional fluorescence imaging. The temporal information is obtained from a sequence of microscope images, which are analyzed in real time. The spatial data is collected in a sequence of images from the microscope, which is automatically focused up and down through the specimen. The analysis of spatial data is performed off-line and consists of photobleaching compensation, focus restoration, filtering, segmentation and spatial volume estimation.

Thermally activated delayed fluorescence organic dots for two-photon fluorescence lifetime imaging

Science.gov (United States)

He, Tingchao; Ren, Can; Li, Zhuohua; Xiao, Shuyu; Li, Junzi; Lin, Xiaodong; Ye, Chuanxiang; Zhang, Junmin; Guo, Lihong; Hu, Wenbo; Chen, Rui

2018-05-01

Autofluorescence is a major challenge in complex tissue imaging when molecules present in the biological tissue compete with the fluorophore. This issue may be resolved by designing organic molecules with long fluorescence lifetimes. The present work reports the two-photon absorption (TPA) properties of a thermally activated delayed fluorescence (TADF) molecule with carbazole as the electron donor and dicyanobenzene as the electron acceptor (i.e., 4CzIPN). The results indicate that 4CzIPN exhibits a moderate TPA cross-section (˜9 × 10-50 cm4 s photon-1), high fluorescence quantum yield, and a long fluorescence lifetime (˜1.47 μs). 4CzIPN was compactly encapsulated into an amphiphilic copolymer via nanoprecipitation to achieve water-soluble organic dots. Interestingly, 4CzIPN organic dots have been utilized in applications involving two-photon fluorescence lifetime imaging (FLIM). Our work aptly demonstrates that TADF molecules are promising candidates of nonlinear optical probes for developing next-generation multiphoton FLIM applications.

Standard Deviation of Spatially-Averaged Surface Cross Section Data from the TRMM Precipitation Radar

Science.gov (United States)

Meneghini, Robert; Jones, Jeffrey A.

2010-01-01

We investigate the spatial variability of the normalized radar cross section of the surface (NRCS or Sigma(sup 0)) derived from measurements of the TRMM Precipitation Radar (PR) for the period from 1998 to 2009. The purpose of the study is to understand the way in which the sample standard deviation of the Sigma(sup 0) data changes as a function of spatial resolution, incidence angle, and surface type (land/ocean). The results have implications regarding the accuracy by which the path integrated attenuation from precipitation can be inferred by the use of surface scattering properties.

Memory-induced sign reversals of the spatial cross-correlation for particles in viscoelastic shear flows

International Nuclear Information System (INIS)

Sauga, Ako; Laas, Katrin; Mankin, Romi

2015-01-01

Highlights: • Cross-correlation (CC) of coordinates of particles in viscoelastic shear flows is discussed. • Expressions for CC functions subjected to both internal and external noises are presented. • Impact of internal and external noises on CC functions are compared. • Memory-induced reentrant sign reversals of the spatial cross-moment are established. - Abstract: The behavior of shear-induced cross-correlation functions between particle fluctuations along orthogonal directions in the shear plane for harmonically trapped Brownian particles in a viscoelastic shear flow is studied. A generalized Langevin equation with a power-law-type memory kernel is used to model the complex structure of the viscoelastic media. Interaction with fluctuations of environmental parameters is modeled by a multiplicative white Gaussian noise, by an internal fractional Gaussian noise, and by an additive external white noise. It is shown that the presence of a memory has a profound effect on the behavior of the cross-correlation functions. Particularly, memory-induced reentrant sign reversals of the spatial cross-moment between orthogonal random displacements of a particle are established, i.e., an increase of the memory exponent can cause the sign reversal from positive to negative, but by a further increase of the memory exponent a reentrant transition from negative to positive values appears. Similarities and differences between the behavior of the models with additive internal and external noises are considered. It is shown that additive external and internal noises cause qualitatively different dependencies of the cross-correlation functions on the time lag. The occurrence of energetic instability due to the influence of multiplicative noise is also discussed.

Spatial-temporal consistency between gross primary productivity and solar-induced chlorophyll fluorescence of vegetation in China during 2007-2014.

Science.gov (United States)

Ma, Jun; Xiao, Xiangming; Zhang, Yao; Doughty, Russell; Chen, Bangqian; Zhao, Bin

2018-10-15

Accurately estimating spatial-temporal patterns of gross primary production (GPP) is important for the global carbon cycle. Satellite-based light use efficiency (LUE) models are regarded as an efficient tool in simulating spatial-temporal dynamics of GPP. However, the accuracy assessment of GPP simulations from LUE models at both spatial and temporal scales remains a challenge. In this study, we simulated GPP of vegetation in China during 2007-2014 using a LUE model (Vegetation Photosynthesis Model, VPM) based on MODIS (moderate-resolution imaging spectroradiometer) images with 8-day temporal and 500-m spatial resolutions and NCEP (National Center for Environmental Prediction) climate data. Global Ozone Monitoring Instrument 2 (GOME-2) solar-induced chlorophyll fluorescence (SIF) data were used to compare with VPM simulated GPP (GPP VPM ) temporally and spatially using linear correlation analysis. Significant positive linear correlations exist between monthly GPP VPM and SIF data over a single year (2010) and multiple years (2007-2014) in most areas of China. GPP VPM is also significantly positive correlated with GOME-2 SIF (R 2  > 0.43) spatially for seasonal scales. However, poor consistency was detected between GPP VPM and SIF data at yearly scale. GPP dynamic trends have high spatial-temporal variation in China during 2007-2014. Temperature, leaf area index (LAI), and precipitation are the most important factors influence GPP VPM in the regions of East Qinghai-Tibet Plateau, Loss Plateau, and Southwestern China, respectively. The results of this study indicate that GPP VPM is temporally and spatially in line with GOME-2 SIF data, and space-borne SIF data have great potential for evaluating LUE-based GPP models. Copyright © 2018 Elsevier B.V. All rights reserved.

Submicrometre geometrically encoded fluorescent barcodes self-assembled from DNA

Science.gov (United States)

Lin, Chenxiang; Jungmann, Ralf; Leifer, Andrew M.; Li, Chao; Levner, Daniel; Church, George M.; Shih, William M.; Yin, Peng

2012-10-01

The identification and differentiation of a large number of distinct molecular species with high temporal and spatial resolution is a major challenge in biomedical science. Fluorescence microscopy is a powerful tool, but its multiplexing ability is limited by the number of spectrally distinguishable fluorophores. Here, we used (deoxy)ribonucleic acid (DNA)-origami technology to construct submicrometre nanorods that act as fluorescent barcodes. We demonstrate that spatial control over the positioning of fluorophores on the surface of a stiff DNA nanorod can produce 216 distinct barcodes that can be decoded unambiguously using epifluorescence or total internal reflection fluorescence microscopy. Barcodes with higher spatial information density were demonstrated via the construction of super-resolution barcodes with features spaced by ˜40 nm. One species of the barcodes was used to tag yeast surface receptors, which suggests their potential applications as in situ imaging probes for diverse biomolecular and cellular entities in their native environments.

Spatially resolved analyses of uranium species using a coupled system made up of confocal laser-scanning microscopy (CLSM) and laser induced fluorescence spectroscopy (LIFS); Ortsaufgeloeste Analyse von Uranspezies mittels einem Gekoppelten System aus Konfokaler Laser-Scanning Mikroskopie (CLSM) und Laser Induzierter Fluoreszenzspektroskopie (LIFS)

Energy Technology Data Exchange (ETDEWEB)

Brockmann, S. [Verein fuer Kernverfahrenstechnik und Analytik Rossendorf e.V. (VKTA), Dresden (Germany); Grossmann, K.; Arnold, T. [Helmholtz-Zentrum Dresden-Rossendorf e.V. (Germany). Inst. fuer Ressourcenoekologie

2014-01-15

The fluorescent properties of uranium when excited by UV light are used increasingly for spectroscope analyses of uranium species within watery samples. Here, alongside the fluorescent properties of the hexavalent oxidation phases, the tetra and pentavalent oxidation phases also play an increasingly important role. The detection of fluorescent emission spectrums on solid and biological samples using (time-resolved) laser induced fluorescence spectroscopy (TRLFS or LIFS respectively) has, however, the disadvantage that no statements regarding the spatial localisation of the uranium can be made. However, particularly in complex, biological samples, such statements on the localisation of the uranium enrichment in the sample are desired, in order to e.g. be able to distinguish between intra and extra-cellular uranium bonds. The fluorescent properties of uranium (VI) compounds and minerals can also be used to detect their localisation within complex samples. So the application of fluorescent microscopic methods represents one possibility to localise and visualise uranium precipitates and enrichments in biological samples, such as biofilms or cells. The confocal laser-scanning microscopy (CLSM) is especially well suited to this purpose. Coupling confocal laser-scanning microscopy (CLSM) with laser induced fluorescence spectroscopy (LIFS) makes it possible to localise and visualise fluorescent signals spatially and three-dimensionally, while at the same time being able to detect spatially resolved, fluorescent-spectroscopic data. This technology is characterised by relatively low detection limits from up to 1.10{sup -6} M for uranium (VI) compounds within the confocal volume. (orig.)

Problems of fluorescent imaging and its solution using nanofluorophores. Part I: Advantages of fluorescent nanoparticles over conventional organic fluorophores

International Nuclear Information System (INIS)

Zhelev, Z.; Hadjidekov, G.; Zlateva, G.; Spasov, L.; Bakalova, R.

2011-01-01

The application of fluorescence in deep-tissue imaging is rapidly expanding in fast several years. The progress in fluorescent molecular probes and fluorescent imaging techniques gives an opportunity to detect single cells and even molecules in live organisms. The highly sensitive and high-speed fluorescent molecular sensors and detection devices allow the application of fluorescence in functional imaging. With development of novel bright fluorophores based on nano-technologies and 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. This review outlines the current status and future trends of fluorescent nanoparticles - quantum dots (QDs), as a new generation of fluorophores in experimental and pre-clinical fluorescent imaging diagnostic. Part 1 focuses on the advantages of quantum dots over conventional organic fluorophores and defines the major requirements to the 'perfect' fluorophore for fluorescent deep-tissue imaging diagnostic. The analysis is based on the limitations of fluorescent imaging in vivo and overcome by using quantum dots

Enhancing molecular logic through modulation of temporal and spatial constraints with quantum dot-based systems that use fluorescent (Förster) resonance energy transfer

Science.gov (United States)

Claussen, Jonathan C.; Algar, W. Russ; Hildebrandt, Niko; Susumu, Kimihiro; Ancona, Mario G.; Medintz, Igor L.

2013-10-01

Luminescent semiconductor nanocrystals or quantum dots (QDs) contain favorable photonic properties (e.g., resistance to photobleaching, size-tunable PL, and large effective Stokes shifts) that make them well-suited for fluorescence (Förster) resonance energy transfer (FRET) based applications including monitoring proteolytic activity, elucidating the effects of nanoparticles-mediated drug delivery, and analyzing the spatial and temporal dynamics of cellular biochemical processes. Herein, we demonstrate how unique considerations of temporal and spatial constraints can be used in conjunction with QD-FRET systems to open up new avenues of scientific discovery in information processing and molecular logic circuitry. For example, by conjugating both long lifetime luminescent terbium(III) complexes (Tb) and fluorescent dyes (A647) to a single QD, we can create multiple FRET lanes that change temporally as the QD acts as both an acceptor and donor at distinct time intervals. Such temporal FRET modulation creates multi-step FRET cascades that produce a wealth of unique photoluminescence (PL) spectra that are well-suited for the construction of a photonic alphabet and photonic logic circuits. These research advances in bio-based molecular logic open the door to future applications including multiplexed biosensing and drug delivery for disease diagnostics and treatment.

Up-Conversion Intersystem Crossing Rates in Organic Emitters for Thermally Activated Delayed Fluorescence: Impact of the Nature of Singlet vs Triplet Excited States

KAUST Repository

Samanta, Pralok Kumar; Kim, Dongwook; Coropceanu, Veaceslav; Bredas, Jean-Luc

2017-01-01

The rates for up-conversion intersystem crossing (UISC) from the T1 state to the S1 state are calculated for a series of organic emitters with an emphasis on thermally activated delayed fluorescence (TADF) materials. Both the spin-orbit coupling

X-ray fluorescence method for trace analysis and imaging

International Nuclear Information System (INIS)

Hayakawa, Shinjiro

2000-01-01

X-ray fluorescence analysis has a long history as conventional bulk elemental analysis with medium sensitivity. However, with the use of synchrotron radiation x-ray fluorescence method has become a unique analytical technique which can provide tace elemental information with the spatial resolution. To obtain quantitative information of trace elemental distribution by using the x-ray fluorescence method, theoretical description of x-ray fluorescence yield is described. Moreover, methods and instruments for trace characterization with a scanning x-ray microprobe are described. (author)

Manipulating Bodily Presence Affects Cross-Modal Spatial Attention: A Virtual-Reality-Based ERP Study

Science.gov (United States)

Harjunen, Ville J.; Ahmed, Imtiaj; Jacucci, Giulio; Ravaja, Niklas; Spapé, Michiel M.

2017-01-01

Earlier studies have revealed cross-modal visuo-tactile interactions in endogenous spatial attention. The current research used event-related potentials (ERPs) and virtual reality (VR) to identify how the visual cues of the perceiver’s body affect visuo-tactile interaction in endogenous spatial attention and at what point in time the effect takes place. A bimodal oddball task with lateralized tactile and visual stimuli was presented in two VR conditions, one with and one without visible hands, and one VR-free control with hands in view. Participants were required to silently count one type of stimulus and ignore all other stimuli presented in irrelevant modality or location. The presence of hands was found to modulate early and late components of somatosensory and visual evoked potentials. For sensory-perceptual stages, the presence of virtual or real hands was found to amplify attention-related negativity on the somatosensory N140 and cross-modal interaction in somatosensory and visual P200. For postperceptual stages, an amplified N200 component was obtained in somatosensory and visual evoked potentials, indicating increased response inhibition in response to non-target stimuli. The effect of somatosensory, but not visual, N200 enhanced when the virtual hands were present. The findings suggest that bodily presence affects sustained cross-modal spatial attention between vision and touch and that this effect is specifically present in ERPs related to early- and late-sensory processing, as well as response inhibition, but do not affect later attention and memory-related P3 activity. Finally, the experiments provide commeasurable scenarios for the estimation of the signal and noise ratio to quantify effects related to the use of a head mounted display (HMD). However, despite valid a-priori reasons for fearing signal interference due to a HMD, we observed no significant drop in the robustness of our ERP measurements. PMID:28275346

Manipulating Bodily Presence Affects Cross-Modal Spatial Attention: A Virtual-Reality-Based ERP Study.

Science.gov (United States)

Harjunen, Ville J; Ahmed, Imtiaj; Jacucci, Giulio; Ravaja, Niklas; Spapé, Michiel M

2017-01-01

Earlier studies have revealed cross-modal visuo-tactile interactions in endogenous spatial attention. The current research used event-related potentials (ERPs) and virtual reality (VR) to identify how the visual cues of the perceiver's body affect visuo-tactile interaction in endogenous spatial attention and at what point in time the effect takes place. A bimodal oddball task with lateralized tactile and visual stimuli was presented in two VR conditions, one with and one without visible hands, and one VR-free control with hands in view. Participants were required to silently count one type of stimulus and ignore all other stimuli presented in irrelevant modality or location. The presence of hands was found to modulate early and late components of somatosensory and visual evoked potentials. For sensory-perceptual stages, the presence of virtual or real hands was found to amplify attention-related negativity on the somatosensory N140 and cross-modal interaction in somatosensory and visual P200. For postperceptual stages, an amplified N200 component was obtained in somatosensory and visual evoked potentials, indicating increased response inhibition in response to non-target stimuli. The effect of somatosensory, but not visual, N200 enhanced when the virtual hands were present. The findings suggest that bodily presence affects sustained cross-modal spatial attention between vision and touch and that this effect is specifically present in ERPs related to early- and late-sensory processing, as well as response inhibition, but do not affect later attention and memory-related P3 activity. Finally, the experiments provide commeasurable scenarios for the estimation of the signal and noise ratio to quantify effects related to the use of a head mounted display (HMD). However, despite valid a-priori reasons for fearing signal interference due to a HMD, we observed no significant drop in the robustness of our ERP measurements.

Depth profile analysis of non-specific fluorescence and color of tooth tissues after peroxide bleaching.

Science.gov (United States)

Klukowska, Malgorzata; Götz, Hermann; White, Donald J; Zoladz, James; Schwarz, Björn-Olaf; Duschner, Heinz

2013-02-01

To examine laboratory changes of endogenous non-specific fluorescence and color throughout subsurface of tooth structures prior to and following peroxide bleaching. Extracted human teeth were cross sectioned and mounted on glass slides. Cross sections were examined for internal color (digital camera) and nonspecific fluorescence (microRaman spectroscopy) throughout the tooth structure at specified locations. Surfaces of sections were then saturation bleached for 70 hours with a gel containing 6% hydrogen peroxide. Cross sections were reexamined for color and non-specific fluorescence changes. Unbleached enamel, dentin-enamel junction and dentin exhibit different CIELab color and non-specific fluorescence properties. Bleaching of teeth produced significant changes in color of internal cross sections and substantial reductions of non-specific fluorescence levels within enamel dentin and DEJ. Enamel and dentin non-specific fluorescence were reduced to common values with bleaching with enamel and the DEJ showing larger reductions than dentin.

Spatially resolved data on sediment transport: 1) field application examining fluorescent soil particle movement from tillage

Science.gov (United States)

Quinton, John; Hardy, Robert; Pates, Jacqueline; James, Michael

2017-04-01

Understanding where sediment originates from and where it travels to, in what quantities and at which rate is at the heart of many questions surrounding sediment transport. Progress towards unravelling these questions and deepening our understanding has come from a wide range of approaches, including laboratory and field experiments conducted at a variety of scales. In seeking to understand the connectivity of sources and sinks of sediment scientists have spent considerable energy in developing tracing technologies. These have included numerous studies that have relied on the chemical properties of the soil and sediment to establish source-sink connectivity, and the use of 137Ceasium, from radioactive fall-out, to map sediment redistribution. More recently there has been an upsurge in interest in the use of artificially applied soil tracers, including rare earth element oxides and magnetic minerals. However all these tracing methods have a significant drawback: they rely on the collection of samples to assess their concentration. This means that their spatial distribution cannot easily be established in situ and that the environment that is being studied is damaged by the sampling process; nor can data be collected in real time which allows a dynamic understanding of erosion and transport processes to be developed. Here we report on the field application of a fluorescent sand sized tracer at the hillslope scale during a tillage erosion experiment. Here we trialled both intensity based and particle counting methodologies for tracer enumeration. After simulating seven years of tillage on a hillslope we were able to precisely determine the distribution of the fluorescent tracer and also its incorporation and distribution within the soil profile. Single grains of tracer could be found over 35 m from the insertion point. In a second abstract we report on an application that combines novel fluorescent videography techniques with custom image processing to trace the

Analysis of hyperspectral fluorescence images for poultry skin tumor inspection

Science.gov (United States)

Kong, Seong G.; Chen, Yud-Ren; Kim, Intaek; Kim, Moon S.

2004-02-01

We present a hyperspectral fluorescence imaging system with a fuzzy inference scheme for detecting skin tumors on poultry carcasses. Hyperspectral images reveal spatial and spectral information useful for finding pathological lesions or contaminants on agricultural products. Skin tumors are not obvious because the visual signature appears as a shape distortion rather than a discoloration. Fluorescence imaging allows the visualization of poultry skin tumors more easily than reflectance. The hyperspectral image samples obtained for this poultry tumor inspection contain 65 spectral bands of fluorescence in the visible region of the spectrum at wavelengths ranging from 425 to 711 nm. The large amount of hyperspectral image data is compressed by use of a discrete wavelet transform in the spatial domain. Principal-component analysis provides an effective compressed representation of the spectral signal of each pixel in the spectral domain. A small number of significant features are extracted from two major spectral peaks of relative fluorescence intensity that have been identified as meaningful spectral bands for detecting tumors. A fuzzy inference scheme that uses a small number of fuzzy rules and Gaussian membership functions successfully detects skin tumors on poultry carcasses. Spatial-filtering techniques are used to significantly reduce false positives.

Integrated cross-domain object storage in working memory: evidence from a verbal-spatial memory task.

Science.gov (United States)

Morey, Candice C

2009-11-01

Working-memory theories often include domain-specific verbal and visual stores (e.g., the phonological and visuospatial buffers of Baddeley, 1986), and some also posit more general stores thought to be capable of holding verbal or visuospatial materials (Baddeley, 2000; Cowan, 2005). However, it is currently unclear which type of store is primarily responsible for maintaining objects that include components from multiple domains. In these studies, a spatial array of letters was followed by a single probe identical to an item in the array or differing systematically in spatial location, letter identity, or their combination. Concurrent verbal rehearsal suppression impaired memory in each of these trial types in a task that required participants to remember verbal-spatial binding, but did not impair memory for spatial locations if the task did not require verbal-spatial binding for a correct response. Thus, spatial information might be stored differently when it must be bound to verbal information. This suggests that a cross-domain store such as the episodic buffer of Baddeley (2000) or the focus of attention of Cowan (2001) might be used for integrated object storage, rather than the maintenance of associations between features stored in separate domain-specific buffers.

Temporo-spatial gait parameters during street crossing conditions: a comparison between younger and older adults.

Science.gov (United States)

Vieira, Edgar R; Lim, Hyun-Hwa; Brunt, Denis; Hallal, Camilla Z; Kinsey, Laura; Errington, Lisa; Gonçalves, Mauro

2015-02-01

Most traffic accidents involving pedestrians happen during street crossing. Safe street crossing by older adults requires complex planning and imposes high cognitive demands. Understanding how street crossing situations affect younger and older adults' gait is important to create evidence-based policies, education and training. The objective of this study was to develop and test a method to evaluate temporo-spatial gait parameters of younger and older adults during simulated street crossing situations. Twenty-two younger (25±2 years old) and 22 older adults (73±6 years old) who lived independently in the community completed 3 walking trials at preferred gait speed and during simulated street crossing with regular and with reduced time. There were significant differences between groups (pstreet crossing walking speed was higher than their preferred speed (pstreet crossing resulted in significant and progressive gait changes. The methods developed and tested can be used to (1) evaluate if people are at risk of falls and accidents during street crossing situations, (2) to compare among different groups, and (3) to help establish appropriate times for older pedestrians to cross streets safely. The current time to cross streets is too short even for healthy older adults. Copyright © 2014 Elsevier B.V. All rights reserved.

Spatial graduation of fuel taxes; consequences for cross-border and domestic fuelling

Energy Technology Data Exchange (ETDEWEB)

Rietveld, P.; Bruinsma, F.R.; Vuuren, D.J. van [Vrije University, Amsterdam (Niger). Faculty of Economics, Business Administration and Econometrics, Department of Spatial Economics and Tinbergen Institute

2001-07-01

Substantial differences exist among fuel taxes between various countries. These differences represent a form of fiscal competition that has undesirable side effects because it leads to cross-border fuelling and hence to extra kilometres driven. One possible way of dealing with this problem of low fuel taxes in neighbouring countries is to introduce a spatial differentiation of taxes: low near the border and higher farther away. This paper contains an empirical analysis of the consequences of such a spatial graduation of fuel taxes for The Netherlands. Impacts on fuelling behaviour, vehicle kilometres driven, tax receipts, and sales by owners of gas stations are analysed. The appropriate slope of the graduation curve in order to prevent fuel-fetching trips is also discussed. Our conclusion is that in a small country such as The Netherlands, a spatial graduation of fuel taxes will lead to considerable problems, even when the graduation curve is not steep that fuel-fetching trips are prevented. The reason is that - given their activity patterns - car drivers will change the location of their fuelling activity leading to substantial problems for owners of gas stations in areas with high taxes. (author)

Mapping absolute tissue endogenous fluorophore concentrations with chemometric wide-field fluorescence microscopy

Science.gov (United States)

Xu, Zhang; Reilley, Michael; Li, Run; Xu, Min

2017-06-01

We report chemometric wide-field fluorescence microscopy for imaging the spatial distribution and concentration of endogenous fluorophores in thin tissue sections. Nonnegative factorization aided by spatial diversity is used to learn both the spectral signature and the spatial distribution of endogenous fluorophores from microscopic fluorescence color images obtained under broadband excitation and detection. The absolute concentration map of individual fluorophores is derived by comparing the fluorescence from "pure" fluorophores under the identical imaging condition following the identification of the fluorescence species by its spectral signature. This method is then demonstrated by characterizing the concentration map of endogenous fluorophores (including tryptophan, elastin, nicotinamide adenine dinucleotide, and flavin adenine dinucleotide) for lung tissue specimens. The absolute concentrations of these fluorophores are all found to decrease significantly from normal, perilesional, to cancerous (squamous cell carcinoma) tissue. Discriminating tissue types using the absolute fluorophore concentration is found to be significantly more accurate than that achievable with the relative fluorescence strength. Quantification of fluorophores in terms of the absolute concentration map is also advantageous in eliminating the uncertainties due to system responses or measurement details, yielding more biologically relevant data, and simplifying the assessment of competing imaging approaches.

RNA Imaging with Multiplexed Error Robust Fluorescence in situ Hybridization

Science.gov (United States)

Moffitt, Jeffrey R.; Zhuang, Xiaowei

2016-01-01

Quantitative measurements of both the copy number and spatial distribution of large fractions of the transcriptome in single-cells could revolutionize our understanding of a variety of cellular and tissue behaviors in both healthy and diseased states. Single-molecule Fluorescence In Situ Hybridization (smFISH)—an approach where individual RNAs are labeled with fluorescent probes and imaged in their native cellular and tissue context—provides both the copy number and spatial context of RNAs but has been limited in the number of RNA species that can be measured simultaneously. Here we describe Multiplexed Error Robust Fluorescence In Situ Hybridization (MERFISH), a massively parallelized form of smFISH that can image and identify hundreds to thousands of different RNA species simultaneously with high accuracy in individual cells in their native spatial context. We provide detailed protocols on all aspects of MERFISH, including probe design, data collection, and data analysis to allow interested laboratories to perform MERFISH measurements themselves. PMID:27241748

Homogeneous immunoassay for the cancer marker alpha-fetoprotein using single wavelength excitation fluorescence cross-correlation spectroscopy and CdSe/ZnS quantum dots and fluorescent dyes as labels

International Nuclear Information System (INIS)

Wang, Jinjie; Liu, Heng; Huang, Xiangyi; Ren, Jicun

2016-01-01

The article describes sensitive and selective homogeneous immunoassays for the liver cancer biomarker alpha-fetoprotein (AFP) in human serum by using single wavelength excitation fluorescence cross-correlation spectroscopy (SW-FCCS). Both competitive and sandwich immunoassay modes were applied, and AFP served as a model analyte. Fluorescent CdSe/ZnS quantum dots (with a 655 nm emission peak) and the fluorophore Alexa Fluor 488 (520 nm emission) were chosen to label the antibodies in the sandwich mode, and the antibody and the antigen in the competitive mode. Under optimized conditions, the sandwich assay has a linear dynamic range that covers the 20 pM to 5.0 nM concentration range. The competitive assay, in turn, extends from 180 pM to 15.0 nM. The respective detection limits are 20 pM and 180 pM. The method was successfully applied to directly determine AFP in (spiked) clinical samples, and results were in good agreement with data obtained via ELISAs. (author)

Spatially modulated imaging system

International Nuclear Information System (INIS)

Barrett, H.H.

1975-01-01

Noncoherent radiation, such as x-rays, is spatially coded, directed through an object and spatially detected to form a spatially coded pattern, from which an image of the object may be reconstructed. The x-ray source may be formed by x-ray fluorescence and substration of the holographic images formed by two sources having energy levels predominantly above and below the maximum absorption range of an agent in the object may be used to enhance contrast in the reproduced image. (Patent Office Record)

Detection of rheumatoid arthritis in humans by fluorescence imaging

Science.gov (United States)

Ebert, Bernd; Dziekan, Thomas; Weissbach, Carmen; Mahler, Marianne; Schirner, Michael; Berliner, Birgitt; Bauer, Daniel; Voigt, Jan; Berliner, Michael; Bahner, Malte L.; Macdonald, Rainer

2010-02-01

The blood pool agent indo-cyanine green (ICG) has been investigated in a prospective clinical study for detection of rheumatoid arthritis using fluorescence imaging. Temporal behavior as well as spatial distribution of fluorescence intensity are suited to differentiate healthy and inflamed finger joints after i.v. injection of an ICG bolus.

Connecting active to passive fluorescence with photosynthesis: a method for evaluating remote sensing measurements of Chl fluorescence.

Science.gov (United States)

Magney, Troy S; Frankenberg, Christian; Fisher, Joshua B; Sun, Ying; North, Gretchen B; Davis, Thomas S; Kornfeld, Ari; Siebke, Katharina

2017-09-01

Recent advances in the retrieval of Chl fluorescence from space using passive methods (solar-induced Chl fluorescence, SIF) promise improved mapping of plant photosynthesis globally. However, unresolved issues related to the spatial, spectral, and temporal dynamics of vegetation fluorescence complicate our ability to interpret SIF measurements. We developed an instrument to measure leaf-level gas exchange simultaneously with pulse-amplitude modulation (PAM) and spectrally resolved fluorescence over the same field of view - allowing us to investigate the relationships between active and passive fluorescence with photosynthesis. Strongly correlated, slope-dependent relationships were observed between measured spectra across all wavelengths (F λ , 670-850 nm) and PAM fluorescence parameters under a range of actinic light intensities (steady-state fluorescence yields, F t ) and saturation pulses (maximal fluorescence yields, F m ). Our results suggest that this method can accurately reproduce the full Chl emission spectra - capturing the spectral dynamics associated with changes in the yields of fluorescence, photochemical (ΦPSII), and nonphotochemical quenching (NPQ). We discuss how this method may establish a link between photosynthetic capacity and the mechanistic drivers of wavelength-specific fluorescence emission during changes in environmental conditions (light, temperature, humidity). Our emphasis is on future research directions linking spectral fluorescence to photosynthesis, ΦPSII, and NPQ. © 2017 The Authors. New Phytologist © 2017 New Phytologist Trust.

Electron detachment dissociation of fluorescently labeled sialylated oligosaccharides.

Science.gov (United States)

Zhou, Wen; Håkansson, Kristina

2011-12-01

We explored the application of electron detachment dissociation (EDD) and infrared multiphoton dissociation (IRMPD) tandem mass spectrometry to fluorescently labeled sialylated oligosaccharides. Standard sialylated oligosaccharides and a sialylated N-linked glycan released from human transferrin were investigated. EDD yielded extensive glycosidic cleavages and cross-ring cleavages in all cases studied, consistently providing complementary structural information compared with infrared multiphoton dissociation. Neutral losses and satellite ions such as C-2H ions were also observed following EDD. In addition, we examined the influence of different fluorescent labels. The acidic label 2-aminobenzoic acid (2-AA) enhanced signal abundance in negative-ion mode. However, few cross-ring fragments were observed for 2-AA-labeled oligosaccharides. The neutral label 2-aminobenzamide (2-AB) resulted in more cross-ring cleavages compared with 2-AA-labeled species, but not as extensive fragmentation as for native oligosaccharides, likely resulting from altered negative charge locations from introduction of the fluorescent tag. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Scanless multitarget-matching multiphoton excitation fluorescence microscopy

Directory of Open Access Journals (Sweden)

Junpeng Qiu

2018-03-01

Full Text Available Using the combination of a reflective blazed grating and a reflective phase-only diffractive spatial light modulator (SLM, scanless multitarget-matching multiphoton excitation fluorescence microscopy (SMTM-MPM was achieved. The SLM shaped an incoming mode-locked, near-infrared Ti:sapphire laser beam into an excitation pattern with addressable shapes and sizes that matched the samples of interest in the field of view. Temporal and spatial focusing were simultaneously realized by combining an objective lens and a blazed grating. The fluorescence signal from illuminated areas was recorded by a two-dimensional sCMOS camera. Compared with a conventional temporal focusing multiphoton microscope, our microscope achieved effective use of the laser power and decreased photodamage with higher axial resolution.

Wide-field fluorescent microscopy and fluorescent imaging flow cytometry on a cell-phone.

Science.gov (United States)

Zhu, Hongying; Ozcan, Aydogan

2013-04-11

Fluorescent microscopy and flow cytometry are widely used tools in biomedical research and clinical diagnosis. However these devices are in general relatively bulky and costly, making them less effective in the resource limited settings. To potentially address these limitations, we have recently demonstrated the integration of wide-field fluorescent microscopy and imaging flow cytometry tools on cell-phones using compact, light-weight, and cost-effective opto-fluidic attachments. In our flow cytometry design, fluorescently labeled cells are flushed through a microfluidic channel that is positioned above the existing cell-phone camera unit. Battery powered light-emitting diodes (LEDs) are butt-coupled to the side of this microfluidic chip, which effectively acts as a multi-mode slab waveguide, where the excitation light is guided to uniformly excite the fluorescent targets. The cell-phone camera records a time lapse movie of the fluorescent cells flowing through the microfluidic channel, where the digital frames of this movie are processed to count the number of the labeled cells within the target solution of interest. Using a similar opto-fluidic design, we can also image these fluorescently labeled cells in static mode by e.g. sandwiching the fluorescent particles between two glass slides and capturing their fluorescent images using the cell-phone camera, which can achieve a spatial resolution of e.g. - 10 μm over a very large field-of-view of - 81 mm(2). This cell-phone based fluorescent imaging flow cytometry and microscopy platform might be useful especially in resource limited settings, for e.g. counting of CD4+ T cells toward monitoring of HIV+ patients or for detection of water-borne parasites in drinking water.

High speed fluorescence imaging with compressed ultrafast photography

Science.gov (United States)

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.

Phenotyping of Arabidopsis Drought Stress Response Using Kinetic Chlorophyll Fluorescence and Multicolor Fluorescence Imaging

Directory of Open Access Journals (Sweden)

Jieni Yao

2018-05-01

Full Text Available Plant responses to drought stress are complex due to various mechanisms of drought avoidance and tolerance to maintain growth. Traditional plant phenotyping methods are labor-intensive, time-consuming, and subjective. Plant phenotyping by integrating kinetic chlorophyll fluorescence with multicolor fluorescence imaging can acquire plant morphological, physiological, and pathological traits related to photosynthesis as well as its secondary metabolites, which will provide a new means to promote the progress of breeding for drought tolerant accessions and gain economic benefit for global agriculture production. Combination of kinetic chlorophyll fluorescence and multicolor fluorescence imaging proved to be efficient for the early detection of drought stress responses in the Arabidopsis ecotype Col-0 and one of its most affected mutants called reduced hyperosmolality-induced [Ca2+]i increase 1. Kinetic chlorophyll fluorescence curves were useful for understanding the drought tolerance mechanism of Arabidopsis. Conventional fluorescence parameters provided qualitative information related to drought stress responses in different genotypes, and the corresponding images showed spatial heterogeneities of drought stress responses within the leaf and the canopy levels. Fluorescence parameters selected by sequential forward selection presented high correlations with physiological traits but not morphological traits. The optimal fluorescence traits combined with the support vector machine resulted in good classification accuracies of 93.3 and 99.1% for classifying the control plants from the drought-stressed ones with 3 and 7 days treatments, respectively. The results demonstrated that the combination of kinetic chlorophyll fluorescence and multicolor fluorescence imaging with the machine learning technique was capable of providing comprehensive information of drought stress effects on the photosynthesis and the secondary metabolisms. It is a promising

Fluorescence detection system for microfluidic droplets

Science.gov (United States)

Chen, Binyu; Han, Xiaoming; Su, Zhen; Liu, Quanjun

2018-05-01

In microfluidic detection technology, because of the universality of optical methods in laboratory, optical detection is an attractive solution for microfluidic chip laboratory equipment. In addition, the equipment with high stability and low cost can be realized by integrating appropriate optical detection technology on the chip. This paper reports a detection system for microfluidic droplets. Photomultiplier tubes (PMT) is used as a detection device to improve the sensitivity of detection. This system improves the signal to noise ratio by software filtering and spatial filter. The fluorescence intensity is proportional to the concentration of the fluorescence and intensity of the laser. The fluorescence micro droplets of different concentrations can be distinguished by this system.

On-chip transduction of nucleic acid hybridization using spatial profiles of immobilized quantum dots and fluorescence resonance energy transfer.

Science.gov (United States)

Tavares, Anthony J; Noor, M Omair; Vannoy, Charles H; Algar, W Russ; Krull, Ulrich J

2012-01-03

The glass surface of a glass-polydimethylsiloxane (PDMS) microfluidic channel was modified to develop a solid-phase assay for quantitative determination of nucleic acids. Electroosmotic flow (EOF) within channels was used to deliver and immobilize semiconductor quantum dots (QDs), and electrophoresis was used to decorate the QDs with oligonucleotide probe sequences. These processes took only minutes to complete. The QDs served as energy donors in fluorescence resonance energy transfer (FRET) for transduction of nucleic acid hybridization. Electrokinetic injection of fluorescent dye (Cy3) labeled oligonucleotide target into a microfluidic channel and subsequent hybridization (within minutes) provided the proximity for FRET, with emission from Cy3 being the analytical signal. The quantification of target concentration was achieved by measurement of the spatial length of coverage by target along a channel. Detection of femtomole quantities of target was possible with a dynamic range spanning an order of magnitude. The assay provided excellent resistance to nonspecific interactions of DNA. Further selectivity of the assay was achieved using 20% formamide, which allowed discrimination between a fully complementary target and a 3 base pair mismatch target at a contrast ratio of 4:1. © 2011 American Chemical Society

Assessment of an X-Ray Spectrometer for fluorescence cross sections measurements of elements with 22≤Ζ≤55 in a secondary target configuration

International Nuclear Information System (INIS)

Delabat Diaz, Y.

2015-01-01

A performance evaluation of an X-Ray Fluorescence Spectrometer for X-Ray Fluorescence (XRF) cross section measurements in a Secondary Target (ST) set-up has been carried out. Using Cd and Dy as STs, an annular 241 Am (∼1 Ci) radioactive source and an X-Ray Spectrometer with a Si(Li) semiconductor detector, the photon effective flux factors (Ι 0 Gε) were measured for some elements with 22≤Ζ≤55 as a function of the characteristics X-Rays energy for two different distances Source-St (0.5 cm and 1.0 cm). Thin high purity foils and a few pellets made out of composed materials were used as samples for the Ι 0 Gε calibrations. the contribution of 59.54 KeV scattered photons to the XRF was analysed and the Scattering Correction Factor (SCF) due to excitation by 59.54 keV scattered photons was estimated in the Cd configuration for further cross section measurements improvements. (Author)

Variability of sun-induced chlorophyll fluorescence according to stand age-related processes in a managed loblolly pine forest.

Science.gov (United States)

Colombo, Roberto; Celesti, Marco; Bianchi, Remo; Campbell, Petya K E; Cogliati, Sergio; Cook, Bruce D; Corp, Lawrence A; Damm, Alexander; Domec, Jean-Christophe; Guanter, Luis; Julitta, Tommaso; Middleton, Elizabeth M; Noormets, Asko; Panigada, Cinzia; Pinto, Francisco; Rascher, Uwe; Rossini, Micol; Schickling, Anke

2018-02-20

Leaf fluorescence can be used to track plant development and stress, and is considered the most direct measurement of photosynthetic activity available from remote sensing techniques. Red and far-red sun-induced chlorophyll fluorescence (SIF) maps were generated from high spatial resolution images collected with the HyPlant airborne spectrometer over even-aged loblolly pine plantations in North Carolina (United States). Canopy fluorescence yield (i.e., the fluorescence flux normalized by the light absorbed) in the red and far-red peaks was computed. This quantifies the fluorescence emission efficiencies that are more directly linked to canopy function compared to SIF radiances. Fluorescence fluxes and yields were investigated in relation to tree age to infer new insights on the potential of those measurements in better describing ecosystem processes. The results showed that red fluorescence yield varies with stand age. Young stands exhibited a nearly twofold higher red fluorescence yield than mature forest plantations, while the far-red fluorescence yield remained constant. We interpreted this finding in a context of photosynthetic stomatal limitation in aging loblolly pine stands. Current and future satellite missions provide global datasets of SIF at coarse spatial resolution, resulting in intrapixel mixture effects, which could be a confounding factor for fluorescence signal interpretation. To mitigate this effect, we propose a surrogate of the fluorescence yield, namely the Canopy Cover Fluorescence Index (CCFI) that accounts for the spatial variability in canopy structure by exploiting the vegetation fractional cover. It was found that spatial aggregation tended to mask the effective relationships, while the CCFI was still able to maintain this link. This study is a first attempt in interpreting the fluorescence variability in aging forest stands and it may open new perspectives in understanding long-term forest dynamics in response to future climatic

Complementary studies of lipid membrane dynamics using iSCAT and super-resolved fluorescence correlation spectroscopy

Science.gov (United States)

Reina, Francesco; Galiani, Silvia; Shrestha, Dilip; Sezgin, Erdinc; de Wit, Gabrielle; Cole, Daniel; Lagerholm, B. Christoffer; Kukura, Philipp; Eggeling, Christian

2018-06-01

Observation techniques with high spatial and temporal resolution, such as single-particle tracking based on interferometric scattering (iSCAT) microscopy, and fluorescence correlation spectroscopy applied on a super-resolution STED microscope (STED-FCS), have revealed new insights of the molecular organization of membranes. While delivering complementary information, there are still distinct differences between these techniques, most prominently the use of fluorescent dye tagged probes for STED-FCS and a need for larger scattering gold nanoparticle tags for iSCAT. In this work, we have used lipid analogues tagged with a hybrid fluorescent tag–gold nanoparticle construct, to directly compare the results from STED-FCS and iSCAT measurements of phospholipid diffusion on a homogeneous supported lipid bilayer (SLB). These comparative measurements showed that while the mode of diffusion remained free, at least at the spatial (>40 nm) and temporal (50  ⩽  t  ⩽  100 ms) scales probed, the diffussion coefficient was reduced by 20- to 60-fold when tagging with 20 and 40 nm large gold particles as compared to when using dye tagged lipid analogues. These FCS measurements of hybrid fluorescent tag–gold nanoparticle labeled lipids also revealed that commercially supplied streptavidin-coated gold nanoparticles contain large quantities of free streptavidin. Finally, the values of apparent diffusion coefficients obtained by STED-FCS and iSCAT differed by a factor of 2–3 across the techniques, while relative differences in mobility between different species of lipid analogues considered were identical in both approaches. In conclusion, our experiments reveal that large and potentially cross-linking scattering tags introduce a significant slow-down in diffusion on SLBs but no additional bias, and our labeling approach creates a new way of exploiting complementary information from STED-FCS and iSCAT measurements.

Single photon counting fluorescence lifetime detection of pericellular oxygen concentrations.

Science.gov (United States)

Hosny, Neveen A; Lee, David A; Knight, Martin M

2012-01-01

Fluorescence lifetime imaging microscopy offers a non-invasive method for quantifying local oxygen concentrations. However, existing methods are either invasive, require custom-made systems, or show limited spatial resolution. Therefore, these methods are unsuitable for investigation of pericellular oxygen concentrations. This study describes an adaptation of commercially available equipment which has been optimized for quantitative extracellular oxygen detection with high lifetime accuracy and spatial resolution while avoiding systematic photon pile-up. The oxygen sensitive fluorescent dye, tris(2,2'-bipyridyl)ruthenium(II) chloride hexahydrate [Ru(bipy)(3)](2+), was excited using a two-photon excitation laser. Lifetime was measured using a Becker & Hickl time-correlated single photon counting, which will be referred to as a TCSPC card. [Ru(bipy)(3)](2+) characterization studies quantified the influences of temperature, pH, cellular culture media and oxygen on the fluorescence lifetime measurements. This provided a precisely calibrated and accurate system for quantification of pericellular oxygen concentration based on measured lifetimes. Using this technique, quantification of oxygen concentrations around isolated viable chondrocytes, seeded in three-dimensional agarose gel, revealed a subpopulation of cells that exhibited significant spatial oxygen gradients such that oxygen concentration reduced with increasing proximity to the cell. This technique provides a powerful tool for quantifying spatial oxygen gradients within three-dimensional cellular models.

Silole-Based Red Fluorescent Organic Dots for Bright Two-Photon Fluorescence In vitro Cell and In vivo Blood Vessel Imaging.

Science.gov (United States)

Chen, Bin; Feng, Guangxue; He, Bairong; Goh, Chiching; Xu, Shidang; Ramos-Ortiz, Gabriel; Aparicio-Ixta, Laura; Zhou, Jian; Ng, Laiguan; Zhao, Zujin; Liu, Bin; Tang, Ben Zhong

2016-02-10

Robust luminescent dyes with efficient two-photon fluorescence are highly desirable for biological imaging applications, but those suitable for organic dots fabrication are still rare because of aggregation-caused quenching. In this work, a red fluorescent silole, 2,5-bis[5-(dimesitylboranyl)thiophen-2-yl]-1-methyl-1,3,4-triphenylsilole ((MesB)2 DTTPS), is synthesized and characterized. (MesB)2 DTTPS exhibits enhanced fluorescence efficiency in nanoaggregates, indicative of aggregation-enhanced emission (AEE). The organic dots fabricated by encapsulating (MesB)2 DTTPS within lipid-PEG show red fluorescence peaking at 598 nm and a high fluorescence quantum yield of 32%. Upon excitation at 820 nm, the dots show a large two-photon absorption cross section of 3.43 × 10(5) GM, which yields a two-photon action cross section of 1.09 × 10(5) GM. These (MesB)2 DTTPS dots show good biocompatibility and are successfully applied to one-photon and two-photon fluorescence imaging of MCF-7 cells and two-photon in vivo visualization of the blood vascular of mouse muscle in a high-contrast and noninvasive manner. Moreover, the 3D blood vasculature located at the mouse ear skin with a depth of over 100 μm can also be visualized clearly, providing the spatiotemporal information about the whole blood vascular network. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Electron Detachment Dissociation (EDD) of Fluorescently Labeled Sialylated Oligosaccharides

Science.gov (United States)

Zhou, Wen; Håkansson, Kristina

2012-01-01

We explored the application of electron detachment dissociation (EDD) and infrared multiphoton dissociation (IRMPD) tandem mass spectrometry to fluorescently labeled sialylated oligosaccharides. Standard sialylated oligosaccharides and a sialylated N-linked glycan released from human transferrin were investigated. EDD yielded extensive glycosidic cleavages and cross-ring cleavages in all cases studied, consistently providing complementary structural information compared to IRMPD. Neutral losses and satellite ions such as C – 2H ions were also observed following EDD. In addition, we examined the influence of different fluorescent labels. The acidic label 2-aminobenzoic acid (2-AA) enhanced signal abundance in negative-ion mode. However, few cross-ring fragments were observed for 2-AA labeled oligosaccharides. The neutral label 2-aminobenzamide (2-AB) resulted in more cross-ring cleavages compared to 2-AA labeled species, but not as extensive fragmentation as for native oligosaccharides, likely resulting from altered negative charge locations from introduction of the fluorescent tag. PMID:22120881

Crossed Leg Sign Is Associated With Severity of Unilateral Spatial Neglect After Stroke

Directory of Open Access Journals (Sweden)

Gustavo José Luvizutto

2018-04-01

Full Text Available BackgroundThe crossed leg sign in patients with right hemisphere stroke is thought to be associated with perceptual disorders, such as unilateral spatial neglect (USN. The aim of this study was to compare the crossed leg sign with the severity of USN during the acute phase of stroke.Experimental proceduresThis was an observational and prospective clinical study of individuals with a diagnosis of right parietal stroke, as confirmed by neuroimaging. The occurrence of the crossed leg sign, the time at which this occurred after the stroke, and a clinical diagnosis of USN were measured and recorded. The patients’ age, sex, and lesion severity, as determined by the National Institutes of Health Stroke Scale and Glasgow coma scale, were included in the analyses as confounding variables. The outcome of interest was the degree of USN, as measured by the cancellation and bisection tests. Binary logistic regression was used to analyze the effect of crossed leg syndrome on the severity of USN. In the adjusted multiple regression model, a p-value of <0.05 was considered statistically significant.ResultsOverall, 60 patients were included in this study. There were no associations between patient demographics and the presence of the crossed leg sign. There was, however, an association between the crossed leg sign and the absolute value of the deviation in the line bisection test (B = −0.234; p = 0.039. The crossed leg sign was not associated with other measures of USN.ConclusionBased on the results of our study, we can conclude that a crossed leg sign in the acute phase of stroke is associated with USN severity, specifically the misinterpretation of the midline.

Fluorescence imaging as a diagnostic of M-band x-ray drive condition in hohlraum with fluorescent Si targets

International Nuclear Information System (INIS)

Li, Qi; Hu, Zhimin; Yao, Li; Huang, Chengwu; Yuan, Zheng; Zhao, Yang; Xiong, Gang; Qing, Bo; Lv, Min; Zhu, Tuo; Deng, Bo; Li, Jin; Wei, Minxi; Zhan, Xiayu; Li, Jun; Yang, Yimeng; Su, Chunxiao; Yang, Guohong; Zhang, Jiyan; Li, Sanwei

2017-01-01

Fluorescence imaging of surrogate Si-doped CH targets has been used to provide a measurement for drive condition of high-energy x-ray (i.e. M-band x-ray) drive symmetry upon the capsule in hohlraum on Shenguang-II laser facility. A series of experiments dedicated to the study of photo-pumping and fluorescence effect in Si-plasma are presented. To investigate the feasibility of fluorescence imaging in Si-plasma, an silicon plasma in Si-foil target is pre-formed at ground state by the soft x-ray from a half-hohlraum, which is then photo-pumped by the K-shell lines from a spatially distinct laser-produced Si-plasma. The resonant Si photon pump is used to improve the fluorescence signal and cause visible image in the Si-foil. Preliminary fluorescence imaging of Si-ball target is performed in both Si-doped and pure Au hohlraum. The usual capsule at the center of the hohlraum is replaced with a solid Si-doped CH-ball (Si-ball). Since the fluorescence is proportional to the photon pump upon the Si-plasma, high-energy x-ray drive symmetry is equal to the fluorescence distribution of the Si-ball. (paper)

Shedding quantitative fluorescence light on novel regulatory mechanisms in skeletal biomedicine and biodentistry.

Science.gov (United States)

Lee, Ji-Won; Iimura, Tadahiro

2017-02-01

Digitalized fluorescence images contain numerical information such as color (wavelength), fluorescence intensity and spatial position. However, quantitative analyses of acquired data and their validation remained to be established. Our research group has applied quantitative fluorescence imaging on tissue sections and uncovered novel findings in skeletal biomedicine and biodentistry. This review paper includes a brief background of quantitative fluorescence imaging and discusses practical applications by introducing our previous research. Finally, the future perspectives of quantitative fluorescence imaging are discussed.

Fluorescent nanoparticles for intracellular sensing: A review

International Nuclear Information System (INIS)

Ruedas-Rama, Maria J.; Walters, Jamie D.; Orte, Angel; Hall, Elizabeth A.H.

2012-01-01

Highlights: ► Analytical applications of fluorescent nanoparticles (NPs) in intracellular sensing. ► Critical review on performance of QDots, metal NPs, silica NPs, and polymer NPs. ► Highlighted potential of fluorescence lifetime imaging microscopy (FLIM). - Abstract: Fluorescent nanoparticles (NPs), including semiconductor NPs (Quantum Dots), metal NPs, silica NPs, polymer NPs, etc., have been a major focus of research and development during the past decade. The fluorescent nanoparticles show unique chemical and optical properties, such as brighter fluorescence, higher photostability and higher biocompatibility, compared to classical fluorescent organic dyes. Moreover, the nanoparticles can also act as multivalent scaffolds for the realization of supramolecular assemblies, since their high surface to volume ratio allow distinct spatial domains to be functionalized, which can provide a versatile synthetic platform for the implementation of different sensing schemes. Their excellent properties make them one of the most useful tools that chemistry has supplied to biomedical research, enabling the intracellular monitoring of many different species for medical and biological purposes. In this review, we focus on the developments and analytical applications of fluorescent nanoparticles in chemical and biological sensing within the intracellular environment. The review also points out the great potential of fluorescent NPs for fluorescence lifetime imaging microscopy (FLIM). Finally, we also give an overview of the current methods for delivering of fluorescent NPs into cells, where critically examine the benefits and liabilities of each strategy.

Structural and dynamical aspects of skin studied by multiphoton excitation fluorescence microscopy-based methods

DEFF Research Database (Denmark)

Bloksgaard, Maria; Brewer, Jonathan R.; Bagatolli, Luis

2013-01-01

' parameters. Specifically, by applying these methods, spatially resolved maps of water dipolar relaxation (generalized polarization function using the 6-lauroyl-2-(N,N-dimethylamino)naphthale probe), activity of protons (fluorescence lifetime imaging using a proton sensitive fluorescence probe--2,7-bis-(2......-carboxyethyl)-5-(and-6)-carboxyfluorescein) and diffusion coefficients of distinct fluorescence probes (raster imaging correlation spectroscopy) can be obtained from different regions of the tissue. Comparative studies of different tissue strata, but also between equivalent regions of normal and abnormal......This mini-review reports on applications of particular multiphoton excitation microscopy-based methodologies employed in our laboratory to study skin. These approaches allow in-depth optical sectioning of the tissue, providing spatially resolved information on specific fluorescence probes...

Application of the X-ray fluorescence analysis and X-ray diffraction in geochemical studies of the Pleistocene tills from Holy Cross Mountains

International Nuclear Information System (INIS)

Kubala-Kukuś, A.; Ludwikowska-Kedzia, M.; Banaś, D.; Braziewicz, J.; Majewska, U.; Pajek, M.; Wudarczyk-Moćko, J.

2013-01-01

X-ray fluorescence analysis methods (wavelength dispersive X-ray fluorescence analysis (WDXRF) and total reflection X-ray fluorescence (TXRF)) and X-ray powder diffraction (XRPD) have been applied in complementary geochemical studies of the Pleistocene till samples. The XRPD technique gave information about the mineral composition of the analyzed samples while the WDXRF and TXRF studies allowed the fast elemental analysis. The till samples were collected from different regions of Holy Cross Mountains (located in central Poland) which are still not unambiguously described in the context of the geochemical studies of the Quaternary sediments. The analysis was concentrated on the geochemical composition of the till samples both for materials occurring on the surface (characterized by continuous weathering processes) and for samples taken from core borehole. The overriding purpose of these studies is determination of the local lithotype of the tills and its lithologic and petrographic diagnostic properties, including the chemical composition of clay and minerals found in the clay. In the presented work the experimental sets up, sample preparation procedure and measurements programme will be discussed in details. Finally, the elemental and mineral compositions will be presented for studied different groups of the samples. - Highlights: • XRF analysis and X-ray diffraction used in studies of the till samples. • The till samples were collected from different regions of Holy Cross Mountains. • The analysis concentrates both on the samples from surface and from core borehole. • The purpose is determination of the local lithotype of the tills. • The experimental setup, sample preparation, measurements and results are discussed

Cross-media color reproduction using the frequency-based spatial gamut mapping algorithm based on human color vision

Science.gov (United States)

Wu, Guangyuan; Niu, Shijun; Li, Xiaozhou; Hu, Guichun

2018-04-01

Due to the increasing globalization of printing industry, remoting proofing will become the inevitable development trend. Cross-media color reproduction will occur in different color gamuts using remote proofing technologies, which usually leads to the problem of incompatible color gamut. In this paper, to achieve equivalent color reproduction between a monitor and a printer, a frequency-based spatial gamut mapping algorithm is proposed for decreasing the loss of visual color information. The design of algorithm is based on the contrast sensitivity functions (CSF), which exploited CSF spatial filter to preserve luminance of the high spatial frequencies and chrominance of the low frequencies. First we show a general framework for how to apply CSF spatial filter in retention of relevant visual information. Then we compare the proposed framework with HPMINDE, CUSP, Bala's algorithm. The psychophysical experimental results indicated the good performance of the proposed algorithm.

Thermally activated delayed fluorescence of fluorescein derivative for time-resolved and confocal fluorescence imaging.

Science.gov (United States)

Xiong, Xiaoqing; Song, Fengling; Wang, Jingyun; Zhang, Yukang; Xue, Yingying; Sun, Liangliang; Jiang, Na; Gao, Pan; Tian, Lu; Peng, Xiaojun

2014-07-09

Compared with fluorescence imaging utilizing fluorophores whose lifetimes are in the order of nanoseconds, time-resolved fluorescence microscopy has more advantages in monitoring target fluorescence. In this work, compound DCF-MPYM, which is based on a fluorescein derivative, showed long-lived luminescence (22.11 μs in deaerated ethanol) and was used in time-resolved fluorescence imaging in living cells. Both nanosecond time-resolved transient difference absorption spectra and time-correlated single-photon counting (TCSPC) were employed to explain the long lifetime of the compound, which is rare in pure organic fluorophores without rare earth metals and heavy atoms. A mechanism of thermally activated delayed fluorescence (TADF) that considers the long wavelength fluorescence, large Stokes shift, and long-lived triplet state of DCF-MPYM was proposed. The energy gap (ΔEST) of DCF-MPYM between the singlet and triplet state was determined to be 28.36 meV by the decay rate of DF as a function of temperature. The ΔE(ST) was small enough to allow efficient intersystem crossing (ISC) and reverse ISC, leading to efficient TADF at room temperature. The straightforward synthesis of DCF-MPYM and wide availability of its starting materials contribute to the excellent potential of the compound to replace luminescent lanthanide complexes in future time-resolved imaging technologies.

An operational fluorescence system for crop assessment

Science.gov (United States)

Belzile, Charles; Belanger, Marie-Christine; Viau, Alain A.; Chamberland, Martin; Roy, Simon

2004-03-01

The development of precision farming requires new tools for plant nutritional stress monitoring. An operational fluorescence system has been designed for vegetation status mapping and stress detection at plant and field scale. The instrument gives relative values of fluorescence at different wavelengths induced by the two-excitation sources. Lightinduced fluorescence has demonstrated successful crop health monitoring and plant nutritional stress detection capabilities. The spectral response of the plants has first been measured with an hyperspectral imager using laser-induced fluorescence. A tabletop imaging fluorometer based on flash lamp technology has also been designed to study the spatial distribution of fluorescence on plant leaves. For field based non-imaging system, LED technology is used as light source to induce fluorescence of the plant. The operational fluorescence system is based on ultraviolet and blue LED to induce fluorescence. Four narrow fluorescence bands centered on 440, 520, 690 and 740nm are detected. The instrument design includes a modular approach for light source and detector. It can accommodate as many as four different light sources and six bands of fluorescence detection. As part of the design for field application, the instrument is compatible with a mobile platform equipped with a GPS and data acquisition system. The current system developed by Telops/GAAP is configured for potato crops fluorescence measurement but can easily be adapted for other crops. This new instrument offers an effective and affordable solution for precision farming.

Fluorescent nanoparticles for intracellular sensing: a review.

Science.gov (United States)

Ruedas-Rama, Maria J; Walters, Jamie D; Orte, Angel; Hall, Elizabeth A H

2012-11-02

Fluorescent nanoparticles (NPs), including semiconductor NPs (Quantum Dots), metal NPs, silica NPs, polymer NPs, etc., have been a major focus of research and development during the past decade. The fluorescent nanoparticles show unique chemical and optical properties, such as brighter fluorescence, higher photostability and higher biocompatibility, compared to classical fluorescent organic dyes. Moreover, the nanoparticles can also act as multivalent scaffolds for the realization of supramolecular assemblies, since their high surface to volume ratio allow distinct spatial domains to be functionalized, which can provide a versatile synthetic platform for the implementation of different sensing schemes. Their excellent properties make them one of the most useful tools that chemistry has supplied to biomedical research, enabling the intracellular monitoring of many different species for medical and biological purposes. In this review, we focus on the developments and analytical applications of fluorescent nanoparticles in chemical and biological sensing within the intracellular environment. The review also points out the great potential of fluorescent NPs for fluorescence lifetime imaging microscopy (FLIM). Finally, we also give an overview of the current methods for delivering of fluorescent NPs into cells, where critically examine the benefits and liabilities of each strategy. Copyright © 2012 Elsevier B.V. All rights reserved.

Endogenous synchronous fluorescence spectroscopy (SFS) of basal cell carcinoma-initial study

Science.gov (United States)

Borisova, E.; Zhelyazkova, Al.; Keremedchiev, M.; Penkov, N.; Semyachkina-Glushkovskaya, O.; Avramov, L.

2016-01-01

The human skin is a complex, multilayered and inhomogeneous organ with spatially varying optical properties. Analysis of cutaneous fluorescence spectra could be a very complicated task; therefore researchers apply complex mathematical tools for data evaluation, or try to find some specific approaches, that would simplify the spectral analysis. Synchronous fluorescence spectroscopy (SFS) allows improving the spectral resolution, which could be useful for the biological tissue fluorescence characterization and could increase the tumour detection diagnostic accuracy.

Spatially resolved synchrotron radiation induced X-ray fluorescence analyses of rare Rembrandt silverpoint drawings

International Nuclear Information System (INIS)

Reiche, I.; Radtke, M.; Berger, A.; Goerner, W.; Merchel, S.; Riesemeier, H.; Bevers, H.

2006-01-01

New analyses of a series of very rare silverpoint drawings that were executed by Rembrandt Harmensz. van Rijn (1606-1669) which are kept today in the Kupferstichkabinett (Museum of Prints and Drawings) of the State Museums of Berlin are reported here. Analysis of these drawings requires particular attention because the study has to be fully non-destructive and extremely sensitive. The metal alloy on the paper does not exceed some hundreds of μg/cm 2 . Therefore, synchrotron radiation induced X-ray fluorescence (SR-XRF) is - together with external micro-proton-induced X-ray emission - the only well-suited method for the analyses of metalpoint drawings. In some primary work, about 25 German and Flemish metalpoint drawings were investigated using spatially resolved SR-XRF analysis at the BAMline at BESSY. This study enlarges the existing French-German database of metalpoint drawings dating from the 15th and 16th centuries, as these Rembrandt drawings originate from the 17th century where this graphical technique was even rarer and already obsolete. It also illustrates how SR-XRF analysis can reinforce art historical assumptions on the dating of drawings and their connection. (orig.)

Cross-calibrating Spatial Positions of Light-viewing Diagnostics using Plasma Edge Sweeps in DIII-D

International Nuclear Information System (INIS)

Solomon, W.M.; Burrell, K.H.; Gohil, P.; Groebner, R.; Kaplan, D.

2003-01-01

An experimental technique is presented that permits diagnostics viewing light from the plasma edge to be spatially calibrated relative to one another. By sweeping the plasma edge, each chord of each diagnostic sweeps out a portion of the light emission profile. A nonlinear least-squares fit to such data provides superior cross-calibration of diagnostics located at different toroidal locations compared with simple surveying. Another advantage of the technique is that it can be used to monitor the position of viewing chords during an experimental campaign to ensure that alignment does not change over time. Moreover, should such a change occur, the data can still be cross-calibrated and its usefulness retained

Automated characterization of nerve fibers labeled fluorescently: determination of size, class and spatial distribution.

Science.gov (United States)

Prodanov, Dimiter; Feirabend, Hans K P

2008-10-03

Morphological classification of nerve fibers could help interpret the assessment of neural regeneration and the understanding of selectivity of nerve stimulation. Specific populations of myelinated nerve fibers can be investigated by retrograde tracing from a muscle followed by microscopic measurements of the labeled fibers at different anatomical levels. Gastrocnemius muscles of adult rats were injected with the retrograde tracer Fluoro-Gold. After a survival period of 3 days, cross-sections of spinal cords, ventral roots, sciatic, and tibial nerves were collected and imaged on a fluorescence microscope. Nerve fibers were classified using a variation-based criterion acting on the distribution of their equivalent diameters. The same criterion was used to classify the labeled axons using the size of the fluorescent marker. Measurements of the axons were paired to those of the entire fibers (axons+myelin sheaths) in order to establish the correspondence between so-established axonal and fiber classifications. It was found that nerve fibers in L6 ventral roots could be classified into four populations comprising two classes of Aalpha (denoted Aalpha1 and Aalpha2), Agamma, and an additional class of Agammaalpha fibers. Cut-off borders between Agamma and Agammaalpha fiber classes were estimated to be 5.00+/-0.09 microm (SEM); between Agammaalpha and Aalpha1 fiber classes to be 6.86+/-0.11 microm (SEM); and between Aalpha1 and Aalpha2 fiber classes to be 8.66+/-0.16 microm (SEM). Topographical maps of the nerve fibers that innervate the gastrocnemius muscles were constructed per fiber class for the spinal root L6. The major advantage of the presented approach consists of the combined indirect classification of nerve fiber types and the construction of topographical maps of so-identified fiber classes.

Determination of experimental K-shell fluorescence yield for ...

Indian Academy of Sciences (India)

X-ray; fluorescence yield; cross-section and chemical effects; potassium; cal- ... The X-ray production cross-section in Ti, V, Cr, Mn, Fe, Co, Ni and Cu .... where µinc (cm2 g−1) and µemt (cm2 g−1) are the mass attenuation coefficients at the.

Small-Animal Imaging Using Diffuse Fluorescence Tomography.

Science.gov (United States)

Davis, Scott C; Tichauer, Kenneth M

2016-01-01

Diffuse fluorescence tomography (DFT) has been developed to image the spatial distribution of fluorescence-tagged tracers in living tissue. This capability facilitates the recovery of any number of functional parameters, including enzymatic activity, receptor density, blood flow, and gene expression. However, deploying DFT effectively is complex and often requires years of know-how, especially for newer mutlimodal systems that combine DFT with conventional imaging systems. In this chapter, we step through the process of using MRI-DFT imaging of a receptor-targeted tracer in small animals.

Fluorescent nanoparticles for intracellular sensing: A review

Energy Technology Data Exchange (ETDEWEB)

Ruedas-Rama, Maria J., E-mail: mjruedas@ugr.esmailto [Department of Physical Chemistry, Faculty of Pharmacy, University of Granada, Campus Cartuja, 18071, Granada (Spain); Walters, Jamie D. [Department of Chemical Engineering and Biotechnology, University of Cambridge, Tennis Court Road, Cambridge, UK CB2 1QT (United Kingdom); Orte, Angel [Department of Physical Chemistry, Faculty of Pharmacy, University of Granada, Campus Cartuja, 18071, Granada (Spain); Hall, Elizabeth A.H., E-mail: lisa.hall@biotech.cam.ac.uk [Department of Chemical Engineering and Biotechnology, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QT (United Kingdom)

2012-11-02

Highlights: Black-Right-Pointing-Pointer Analytical applications of fluorescent nanoparticles (NPs) in intracellular sensing. Black-Right-Pointing-Pointer Critical review on performance of QDots, metal NPs, silica NPs, and polymer NPs. Black-Right-Pointing-Pointer Highlighted potential of fluorescence lifetime imaging microscopy (FLIM). - Abstract: Fluorescent nanoparticles (NPs), including semiconductor NPs (Quantum Dots), metal NPs, silica NPs, polymer NPs, etc., have been a major focus of research and development during the past decade. The fluorescent nanoparticles show unique chemical and optical properties, such as brighter fluorescence, higher photostability and higher biocompatibility, compared to classical fluorescent organic dyes. Moreover, the nanoparticles can also act as multivalent scaffolds for the realization of supramolecular assemblies, since their high surface to volume ratio allow distinct spatial domains to be functionalized, which can provide a versatile synthetic platform for the implementation of different sensing schemes. Their excellent properties make them one of the most useful tools that chemistry has supplied to biomedical research, enabling the intracellular monitoring of many different species for medical and biological purposes. In this review, we focus on the developments and analytical applications of fluorescent nanoparticles in chemical and biological sensing within the intracellular environment. The review also points out the great potential of fluorescent NPs for fluorescence lifetime imaging microscopy (FLIM). Finally, we also give an overview of the current methods for delivering of fluorescent NPs into cells, where critically examine the benefits and liabilities of each strategy.

A 3D imaging system integrating photoacoustic and fluorescence orthogonal projections for anatomical, functional and molecular assessment of rodent models

Science.gov (United States)

Brecht, Hans P.; Ivanov, Vassili; Dumani, Diego S.; Emelianov, Stanislav Y.; Anastasio, Mark A.; Ermilov, Sergey A.

2018-03-01

We have developed a preclinical 3D imaging instrument integrating photoacoustic tomography and fluorescence (PAFT) addressing known deficiencies in sensitivity and spatial resolution of the individual imaging components. PAFT is designed for simultaneous acquisition of photoacoustic and fluorescence orthogonal projections at each rotational position of a biological object, enabling direct registration of the two imaging modalities. Orthogonal photoacoustic projections are utilized to reconstruct large (21 cm3 ) volumes showing vascularized anatomical structures and regions of induced optical contrast with spatial resolution exceeding 100 µm. The major advantage of orthogonal fluorescence projections is significant reduction of background noise associated with transmitted or backscattered photons. The fluorescence imaging component of PAFT is used to boost detection sensitivity by providing low-resolution spatial constraint for the fluorescent biomarkers. PAFT performance characteristics were assessed by imaging optical and fluorescent contrast agents in tissue mimicking phantoms and in vivo. The proposed PAFT technology will enable functional and molecular volumetric imaging using fluorescent biomarkers, nanoparticles, and other photosensitive constructs mapped with high fidelity over robust anatomical structures, such as skin, central and peripheral vasculature, and internal organs.

Fluorescence correlation spectroscopy and fluorescence cross-correlation spectroscopy reveal the cytoplasmic origination of loaded nuclear RISC in vivo in human cells.

Science.gov (United States)

Ohrt, Thomas; Mütze, Jörg; Staroske, Wolfgang; Weinmann, Lasse; Höck, Julia; Crell, Karin; Meister, Gunter; Schwille, Petra

2008-11-01

Studies of RNA interference (RNAi) provide evidence that in addition to the well-characterized cytoplasmic mechanisms, nuclear mechanisms also exist. The mechanism by which the nuclear RNA-induced silencing complex (RISC) is formed in mammalian cells, as well as the relationship between the RNA silencing pathways in nuclear and cytoplasmic compartments is still unknown. Here we show by applying fluorescence correlation and cross-correlation spectroscopy (FCS/FCCS) in vivo that two distinct RISC exist: a large approximately 3 MDa complex in the cytoplasm and a 20-fold smaller complex of approximately 158 kDa in the nucleus. We further show that nuclear RISC, consisting only of Ago2 and a short RNA, is loaded in the cytoplasm and imported into the nucleus. The loaded RISC accumulates in the nucleus depending on the presence of a target, based on an miRNA-like interaction with impaired cleavage of the cognate RNA. Together, these results suggest a new RISC shuttling mechanism between nucleus and cytoplasm ensuring concomitant gene regulation by small RNAs in both compartments.

Hybrid system for in vivo real-time planar fluorescence and volumetric optoacoustic imaging

Science.gov (United States)

Chen, Zhenyue; Deán-Ben, Xosé Luís.; Gottschalk, Sven; Razansky, Daniel

2018-02-01

Fluorescence imaging is widely employed in all fields of cell and molecular biology due to its high sensitivity, high contrast and ease of implementation. However, the low spatial resolution and lack of depth information, especially in strongly-scattering samples, restrict its applicability for deep-tissue imaging applications. On the other hand, optoacoustic imaging is known to deliver a unique set of capabilities such as high spatial and temporal resolution in three dimensions, deep penetration and spectrally-enriched imaging contrast. Since fluorescent substances can generate contrast in both modalities, simultaneous fluorescence and optoacoustic readings can provide new capabilities for functional and molecular imaging of living organisms. Optoacoustic images can further serve as valuable anatomical references based on endogenous hemoglobin contrast. Herein, we propose a hybrid system for in vivo real-time planar fluorescence and volumetric optoacoustic tomography, both operating in reflection mode, which synergistically combines the advantages of stand-alone systems. Validation of the spatial resolution and sensitivity of the system were first carried out in tissue mimicking phantoms while in vivo imaging was further demonstrated by tracking perfusion of an optical contrast agent in a mouse brain in the hybrid imaging mode. Experimental results show that the proposed system effectively exploits the contrast mechanisms of both imaging modalities, making it especially useful for accurate monitoring of fluorescence-based signal dynamics in highly scattering samples.

Laser scanning endoscope via an imaging fiber bundle for fluorescence imaging

Science.gov (United States)

Yeboah, Lorenz D.; Nestler, Dirk; Steiner, Rudolf W.

1994-12-01

Based on a laser scanning endoscope via an imaging fiber bundle, a new approach for a tumor diagnostic system has been developed to assist physicians in the diagnosis before the actual PDT is carried out. Laser induced, spatially resolved fluorescence images of diseased tissue can be compared with images received by video endoscopy using a white light source. The set- up is required to produce a better contrast between infected and healthy tissue and might serve as a constructive diagnostic help for surgeons. The fundamental idea is to scan a low-power laser beam on an imaging fiber bundle and to achieve a spatially resolved projection on the tissue surface. A sufficiently high laser intensity from the diode laser is concentrated on each single spot of the tissue exciting fluorescence when a dye has previously been accumulated. Subsequently, video image of the tissue is recorded and stored. With an image processing unit, video and fluorescence images are overlaid producing a picture of the fluorescence intensity in the environment of the observed tissue.

X-ray fluorescence investigation of gilded and enamelled silver: The case study of four medieval processional crosses from central Italy

Science.gov (United States)

Ferretti, Marco; Polese, Claudia; Roldán García, Clodoaldo

2013-05-01

The presence of multilayered structures is common in such cultural artefacts as paintings, corroded metals, objects that underwent a whatever form of surface qualification. One of the most usual and complete ways to investigate such structures is observing a cross section, which requires sampling. There are however situations where at least part of the stratigraphic information can be derived non-destructively: the literature shows that X-ray fluorescence (XRF) has frequently been used, in recent years, for this purpose, with special regard to paintings and gilded metals. Aim of this paper is to further explore the suitability of XRF-based techniques to characterise multilayered structures. This is achieved by introducing improvements, with respect to previous works, in both equipment and data processing. The method, that has been developed for gilded and enamelled silver artefacts, relies on optimum excitation conditions provided for silver and on the relationship existing between the ratio AgKα/AgKβ of its fluorescence lines and the gilding thickness itself. The coating (gilding or enamel) thickness is derived by verifying the condition CKα,Ag = CKβ,Ag, where CKα,Ag and CKβ,Ag are the mass fractions of silver calculated on the lines AgKα and AgKβ, respectively. The calculations are carried out by PyMCA, a Fundamental Parameters code that implements the analysis of multilayered samples. As a case study we investigated in situ the four processional crosses of Borbona, Sant'Elpidio, Rosciolo and Forcella, made of a wood core with attached gilt and embossed silver sheets and enamelled silver plates. The analyses allowed to distinguish ancient restorations from original parts, to characterise the enamels and find their composition consistent with the dates of manufacturing and, as regards the cross of Rosciolo, to hypothesize the contribution of different "hands" in its manufacturing.

Fluorescent silica colloids for study and visualization of skin care products.

Science.gov (United States)

Iyer, Swaminathan; Kievsky, Yaroslav; Sokolov, Igor

2007-08-01

The efficacy of skin care products depends on the time and dynamics of their absorbance by the skin, and its spatial distribution on the skin. Regular scrape-based methods may depend on the operator and are destructive and invasive in nature. Here, we describe a novel method based on non-contact optical measurements to trace the location and dynamics of skin care products on the skin. We use fluorescent silica colloidal particles of micron sizes at a rather small concentration as non-invasive tracers. As an example of skin care products, we use two base materials: either glycerin or vaseline. A mixture of each product with fluorescent particles is applied on human skin. The amount of fluorescence is monitored by means of a fluorescent spectrometer. The scraping method is used to compare with the spectroscopic measurements. Fluorescent tracers make the skin care product visible under UV light. This allows obtaining an optical image of the spatial distribution of the product on the skin. The quantitative data of fluorescence are well correlated with the scrape data. Comparison of the difference in the spectral and scraped mass data reveals the details of accumulation of the skin products in skin cracks and crevices. We described an efficient non-invasive benign method to quantify dynamics and to perform mapping of emollients and humectants on the skin.

Spatial reliability analysis of a wind turbine blade cross section subjected to multi-axial extreme loading

DEFF Research Database (Denmark)

Dimitrov, Nikolay Krasimirov; Bitsche, Robert; Blasques, José Pedro Albergaria Amaral

2017-01-01

This paper presents a methodology for structural reliability analysis of wind turbine blades. The study introduces several novel elements by taking into account loading direction using a multiaxial probabilistic load model, considering random material strength, spatial correlation between material...... properties, progressive material failure, and system reliability effects. An example analysis of reliability against material failure is demonstrated for a blade cross section. Based on the study we discuss the implications of using a system reliability approach, the effect of spatial correlation length......, type of material degradation algorithm, and reliability methods on the system failure probability, as well as the main factors that have an influence on the reliability. (C) 2017 Elsevier Ltd. All rights reserved....

Lenses and effective spatial resolution in macroscopic optical mapping

International Nuclear Information System (INIS)

Bien, Harold; Parikh, Puja; Entcheva, Emilia

2007-01-01

Optical mapping of excitation dynamically tracks electrical waves travelling through cardiac or brain tissue by the use of fluorescent dyes. There are several characteristics that set optical mapping apart from other imaging modalities: dynamically changing signals requiring short exposure times, dim fluorescence demanding sensitive sensors and wide fields of view (low magnification) resulting in poor optical performance. These conditions necessitate the use of optics with good light gathering ability, i.e. lenses having high numerical aperture. Previous optical mapping studies often used sensor resolution to estimate the minimum spatial feature resolvable, assuming perfect optics and infinite contrast. We examine here the influence of finite contrast and real optics on the effective spatial resolution in optical mapping under broad-field illumination for both lateral (in-plane) resolution and axial (depth) resolution of collected fluorescence signals

Synthesis and Sensing Applications of Fluorescent 3-Cinnamoyl Coumarins

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Preeti Yadav

2015-12-01

Full Text Available We have synthesized two novel fluorescent 3-(4-diethylaminocinnamoyl coumarins that exhibit fluorescence quenching upon exposure to a nerve agent simulant, diethylchlorophosphate (DCP, providing a basis for rapid and sensitive DCP chemosensing. Furthermore, these coumarin derivatives display two-photon fluorescence upon illumination with near-infrared laser pulses and their two-photon (TP absorption cross-section was evaluated. The potential for TP bio-imaging of these compounds was investigated by their cellular uptake in HeLa cells by TP confocal microscopy.

Characterization of the photoreaction between DNA and aminomethyl-trimethylpsoralen using absorption and fluorescence spectroscopy

International Nuclear Information System (INIS)

Johnston, B.H.; Hearst, J.E.

1981-01-01

The use of absorption and fluorescence spectroscopy for following the progress of the photoreaction between DNA and 4'-aminomethyl-4,5',8-trimethylpsoralen (AMT) has been investigated. Absorption at long wavelengths and fluorescence both decline upon intercalation of AMT into the DNA helix. The loss of fluorescence from AMT and the accompanying appearance of monoadduct fluorescence upon irradiation by UV light can be easily followed by using the excitation beam of a spectrofluorometer as the source of irradiation and monitoring the changing emission spectrum. Where cross-link formation is possible, the subsequent decline of monoadduct fluorescence is seen as well. This suggests that the 4',5'-monoadduct is a precursor of cross-links. Both monoaddition and cross-linking are more rapid with poly d(A-T) than with calf thymus DNA or poly d(A.T). Excitation spectra can be helpful in resolving the levels of AMT and 4',5'-monoadduct when both are contributing to the emission spectrum. Some changes are observed in the emission spectrum of AMT-poly d(A.T) monoadducts after prolonged irradiation which indicate further photoreaction. (author)

Efficacy of NGR peptide-modified PEGylated quantum dots for crossing the blood-brain barrier and targeted fluorescence imaging of glioma and tumor vasculature.

Science.gov (United States)

Huang, Ning; Cheng, Si; Zhang, Xiang; Tian, Qi; Pi, Jiangli; Tang, Jun; Huang, Qing; Wang, Feng; Chen, Jin; Xie, Zongyi; Xu, Zhongye; Chen, Weifu; Zheng, Huzhi; Cheng, Yuan

2017-01-01

Delivery of imaging agents to brain glioma is challenging because the blood-brain barrier (BBB) functions as a physiological checkpoint guarding the central nervous system from circulating large molecules. Moreover, the ability of existing probes to target glioma has been insufficient and needs to be improved. In present study, PEG-based long circulation, CdSe/ZnS quantum dots (QDs)-based nanoscale and fluorescence, asparagines-glycine-arginine peptides (NGR)-based specific CD13 recognition were integrated to design and synthesize a novel nanoprobe by conjugating biotinylated NGR peptides to avidin-PEG-coated QDs. Our data showed that the NGR-PEG-QDs were nanoscale with less than 100 nm and were stable in various pH (4.0~8.0). These nanomaterials with non-toxic concentrations could cross the BBB and target CD13-overexpressing glioma and tumor vasculature in vitro and in vivo, contributing to fluorescence imaging of this brain malignancy. These achievements allowed groundbreaking technological advances in targeted fluorescence imaging for the diagnosis and surgical removal of glioma, facilitating potential transformation toward clinical nanomedicine. Copyright © 2016 Elsevier Inc. All rights reserved.

Cross-modal activation of auditory regions during visuo-spatial working memory in early deafness.

Science.gov (United States)

Ding, Hao; Qin, Wen; Liang, Meng; Ming, Dong; Wan, Baikun; Li, Qiang; Yu, Chunshui

2015-09-01

Early deafness can reshape deprived auditory regions to enable the processing of signals from the remaining intact sensory modalities. Cross-modal activation has been observed in auditory regions during non-auditory tasks in early deaf subjects. In hearing subjects, visual working memory can evoke activation of the visual cortex, which further contributes to behavioural performance. In early deaf subjects, however, whether and how auditory regions participate in visual working memory remains unclear. We hypothesized that auditory regions may be involved in visual working memory processing and activation of auditory regions may contribute to the superior behavioural performance of early deaf subjects. In this study, 41 early deaf subjects (22 females and 19 males, age range: 20-26 years, age of onset of deafness memory task than did the hearing controls. Compared with hearing controls, deaf subjects exhibited increased activation in the superior temporal gyrus bilaterally during the recognition stage. This increased activation amplitude predicted faster and more accurate working memory performance in deaf subjects. Deaf subjects also had increased activation in the superior temporal gyrus bilaterally during the maintenance stage and in the right superior temporal gyrus during the encoding stage. These increased activation amplitude also predicted faster reaction times on the spatial working memory task in deaf subjects. These findings suggest that cross-modal plasticity occurs in auditory association areas in early deaf subjects. These areas are involved in visuo-spatial working memory. Furthermore, amplitudes of cross-modal activation during the maintenance stage were positively correlated with the age of onset of hearing aid use and were negatively correlated with the percentage of lifetime hearing aid use in deaf subjects. These findings suggest that earlier and longer hearing aid use may inhibit cross-modal reorganization in early deaf subjects. Granger

Historical Evolution of Spatial Abilities

Directory of Open Access Journals (Sweden)

A. Ardila

1993-01-01

Full Text Available Historical evolution and cross-cultural differences in spatial abilities are analyzed. Spatial abilities have been found to be significantly associated with the complexity of geographical conditions and survival demands. Although impaired spatial cognition is found in cases of, exclusively or predominantly, right hemisphere pathology, it is proposed that this asymmetry may depend on the degree of training in spatial abilities. It is further proposed that spatial cognition might have evolved in a parallel way with cultural evolution and environmental demands. Contemporary city humans might be using spatial abilities in some new, conceptual tasks that did not exist in prehistoric times: mathematics, reading, writing, mechanics, music, etc. Cross-cultural analysis of spatial abilities in different human groups, normalization of neuropsychological testing instruments, and clinical observations of spatial ability disturbances in people with different cultural backgrounds and various spatial requirements, are required to construct a neuropsychological theory of brain organization of spatial cognition.

Fluorescence Lifetime Correlation Spectroscopy (FLCS): Concepts, Applications and Outlook

Czech Academy of Sciences Publication Activity Database

Kapusta, Peter; Macháň, Radek; Benda, A.; Hof, Martin

2012-01-01

Roč. 13, č. 10 (2012), s. 12890-12910 E-ISSN 1422-0067 R&D Projects: GA ČR GBP208/12/G016 Institutional support: RVO:61388955 Keywords : fluorescence correlation spectroscopy (FCS) * time correlated single photon counting (TCSPC) * fluorescence cross-correlation spectroscopy (FCCS) Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 2.464, year: 2012

Multi-color imaging of fluorescent nanodiamonds in living HeLa cells using direct electron-beam excitation.

Science.gov (United States)

Nawa, Yasunori; Inami, Wataru; Lin, Sheng; Kawata, Yoshimasa; Terakawa, Susumu; Fang, Chia-Yi; Chang, Huan-Cheng

2014-03-17

Multi-color, high spatial resolution imaging of fluorescent nanodiamonds (FNDs) in living HeLa cells has been performed with a direct electron-beam excitation-assisted fluorescence (D-EXA) microscope. In this technique, fluorescent materials are directly excited with a focused electron beam and the resulting cathodoluminescence (CL) is detected with nanoscale resolution. Green- and red-light-emitting FNDs were employed for two-color imaging, which were observed simultaneously in the cells with high spatial resolution. This technique could be applied generally for multi-color immunostaining to reveal various cell functions. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Plasticity of Human Spatial Cognition: Spatial Language and Cognition Covary across Cultures

Science.gov (United States)

Haun, Daniel B. M.; Rapold, Christian J.; Janzen, Gabriele; Levinson, Stephen C.

2011-01-01

The present paper explores cross-cultural variation in spatial cognition by comparing spatial reconstruction tasks by Dutch and Namibian elementary school children. These two communities differ in the way they predominantly express spatial relations in language. Four experiments investigate cognitive strategy preferences across different levels of…

Fluorescence imaging of soybean flavonol isolines

Science.gov (United States)

Kim, Moon S.; Lee, Edward H.; Mulchi, Charles L.; McMurtrey, James E., III; Chappelle, Emmett W.; Rowland, Randy A.

1998-07-01

Experiments were conducted to characterize the fluorescence emission of leaves from four soybean ('Harosoy') plants containing different concentrations of flavonols (kaempferol glycosides). The investigation utilized genetically mutated soybean flavonol isolines grown in a constant environment, thus limiting factors known to affect fluorescence emission characteristics other than different kaempferol glycosides concentrations. Flavonol isolines included OX922, OX941, OX942, OX944. The first two isolines contain kaempferol (K) glycosides; K3, K6, and K9, and the latter two did not have K3, K6, and K9. A fluorescence imaging system (FIS) was used to characterize steady state florescence images of the sample leaves measured at wavelengths centered at 450, 550, 680, and 740 nm with an excitation at 360 nm. Images taken with FIS greatly complement non-imaging fluorescence measurements by characterizing the spatial variation of fluorescence within leaves. We also acquired fluorescence emission spectra to characterize spectral features of the soybean flavonol isolines. The emission spectral shape of the fluorescence emission characteristics were not significantly different between the soybeans that contain kaempferol glycosides and the ones that do not contain kaempferol glycosides. Typical emission maxima of green vegetation in the blue, green, red, and far-red bands were noticed in all four soybean isolines. However, plants containing kaempferol glycosides, OX922 and OX941 had significantly lower intensities throughout the wavelength regions. These results imply that fluorescence emission intensities in the fluorescence emission bands studied are significantly affected by the presence and absence of kaempferol glycosides concentrations (UV radiation screening compounds). Pure kaempferol glycoside dissolved in solution show minimal fluorescence emission when excited with the absorption maximum radiation at 365 nm. However, a broad band emission can be seen in the green

Er3+ infrared fluorescence affected by spatial distribution synchronicity of Ba2+ and Er3+ in Er3+-doped BaO–SiO2 glasses

Directory of Open Access Journals (Sweden)

Atsunobu Masuno

2016-02-01

Full Text Available Glasses with the composition xBaO–(99.9 − xSiO2–0.1ErO3/2 (0 ≤x ≤ 34.9 were fabricated by a levitation technique. The glasses in the immiscibility region were opaque due to chemical inhomogeneity, while the other glasses were colorless and transparent. The scanning electron microscope observations and electron probe microanalysis scan profiles revealed that more Er3+ ions were preferentially distributed in the regions where more Ba2+ ions existed in the chemically inhomogeneous glasses. The synchronicity of the spatial distributions of the two ions initially increased with increasing x and then decreased when the Ba2+ concentration exceeded a certain value. The peak shape and lifetime of the fluorescence at 1.55 μm depended on x as well as the spatial distribution of both ions. These results indicate that although ErOn polyhedra are preferentially coordinated with Ba2+ ions and their local structure is affected by the coordination of Ba2+, there is a maximum in the amount of Ba2+ ions that can coordinate ErOn polyhedra since the available space for Ba2+ ions is limited. These findings provide us with efficient ways to design the chemical composition of glasses with superior Er3+ fluorescence properties for optical communication network systems.

Influence of auditory spatial attention on cross-modal semantic priming effect: evidence from N400 effect.

Science.gov (United States)

Wang, Hongyan; Zhang, Gaoyan; Liu, Baolin

2017-01-01

Semantic priming is an important research topic in the field of cognitive neuroscience. Previous studies have shown that the uni-modal semantic priming effect can be modulated by attention. However, the influence of attention on cross-modal semantic priming is unclear. To investigate this issue, the present study combined a cross-modal semantic priming paradigm with an auditory spatial attention paradigm, presenting the visual pictures as the prime stimuli and the semantically related or unrelated sounds as the target stimuli. Event-related potentials results showed that when the target sound was attended to, the N400 effect was evoked. The N400 effect was also observed when the target sound was not attended to, demonstrating that the cross-modal semantic priming effect persists even though the target stimulus is not focused on. Further analyses revealed that the N400 effect evoked by the unattended sound was significantly lower than the effect evoked by the attended sound. This contrast provides new evidence that the cross-modal semantic priming effect can be modulated by attention.

Spatial distribution and contamination assessment of heavy metals in street dust from Camagüey city (Cuba) using X-ray fluorescence

International Nuclear Information System (INIS)

Rizo, Oscar Díaz; Rivero Palma, Orestes; D’Alessandro Rodríguez, Katia; García Trápaga, César

2015-01-01

Concentrations of Cr, Co, Ni, Cu, Zn, Pb and Fe in the street dust from Camagüey city were studied by X-ray fluorescence analysis. The mean Cr, Co, Ni, Cu, Zn and Pb contents in the urban dust samples (97 ± 30, 14 ± 2, 35 ± 36, 94 ± 26, 199 ± 87 and 42 ± 29 mg.kg-1 dry weight, respectively) were compared with mean concentrations for other cities around the world. Spatial distribution maps indicated the same behaviour for Cr–Ni and Pb–Zn–Cu, respectively, whereas the spatial distribution of Co differs from other heavy metals. The metal-to-iron normalization, using Cuban average metal soil contents as background, showed that street dusts from Camagüey city are moderately or significantly Zn-Pb enriched in those areas associated with heavy traffic density and metallurgic plant location. However, the calculation of the potential ecological risk index shows that metal content in Camagüey street dust go not represent any risk for city population. (author)

Fluorescence-type Monochromatic X-ray Beam-position Monitor with High-spatial Resolution for the NSLS-II Beamlines

International Nuclear Information System (INIS)

Yoon, Phil S.; Siddons, D. Peter

2010-01-01

We developed a fluorescence-type monochromatic X-ray beam-position monitor (X-BPM) with high-spatial resolution for end-station experiments at the initial project beamlines of the NSLS-II. We designed a ring array of multi-segmented Si PIN-junction photodiodes to use as a position sensor. Further, we integrated a low-noise charge-preamplification HERMES4 ASIC chip into an electronic readout system for photon-counting application. A series of precision measurements to characterize electronically the Si-photodiode sensor and the ASIC chip demonstrated that the inherent noise from the detector system is sufficiently low to meet our stringent requirements. Using a Gaussian beam, we parametrically modeled the optimum working distance to ensure the detector's best performance. Based upon the results from the parametric modeling, prototypes of the next versions of the X-BPM are being developed. In this paper, we describe the methodology for developing the new compact monochromatic X-ray BPM, including its instrumentation, detector modeling, and future plan.

Quantitative in vivo fluorescence cross-correlation analyses highlight the importance of competitive effects in the regulation of protein-protein interactions.

Science.gov (United States)

Sadaie, Wakako; Harada, Yoshie; Matsuda, Michiyuki; Aoki, Kazuhiro

2014-09-01

Computer-assisted simulation is a promising approach for clarifying complicated signaling networks. However, this approach is currently limited by a deficiency of kinetic parameters determined in living cells. To overcome this problem, we applied fluorescence cross-correlation spectrometry (FCCS) to measure dissociation constant (Kd) values of signaling molecule complexes in living cells (in vivo Kd). Among the pairs of fluorescent molecules tested, that of monomerized enhanced green fluorescent protein (mEGFP) and HaloTag-tetramethylrhodamine was most suitable for the measurement of in vivo Kd by FCCS. Using this pair, we determined 22 in vivo Kd values of signaling molecule complexes comprising the epidermal growth factor receptor (EGFR)-Ras-extracellular signal-regulated kinase (ERK) mitogen-activated protein (MAP) kinase pathway. With these parameters, we developed a kinetic simulation model of the EGFR-Ras-ERK MAP kinase pathway and uncovered a potential role played by stoichiometry in Shc binding to EGFR during the peak activations of Ras, MEK, and ERK. Intriguingly, most of the in vivo Kd values determined in this study were higher than the in vitro Kd values reported previously, suggesting the significance of competitive bindings inside cells. These in vivo Kd values will provide a sound basis for the quantitative understanding of signal transduction. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

Once for All: A Novel Robust System for Co-expression of Multiple Chimeric Fluorescent Fusion Proteins in Plants

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Guitao Zhong

2017-06-01

Full Text Available Chimeric fluorescent fusion proteins have been employed as a powerful tool to reveal the subcellular localizations and dynamics of proteins in living cells. Co-expression of a fluorescent fusion protein with well-known organelle markers in the same cell is especially useful in revealing its spatial and temporal functions of the protein in question. However, the conventional methods for co-expressing multiple fluorescent tagged proteins in plants have the drawbacks of low expression efficiency, variations in the expression level and time-consuming genetic crossing. Here, we have developed a novel robust system that allows for high-efficient co-expression of multiple chimeric fluorescent fusion proteins in plants in a time-saving fashion. This system takes advantage of employing a single expression vector which consists of multiple semi-independent expressing cassettes for the protein co-expression thereby overcoming the limitations of using multiple independent expressing plasmids. In addition, it is a highly manipulable DNA assembly system, in which modification and recombination of DNA molecules are easily achieved through an optimized one-step assembly reaction. By employing this effective system, we demonstrated that co-expression of two chimeric fluorescent fusion reporter proteins of vacuolar sorting receptor and secretory carrier membrane protein gave rise to their perspective subcellular localizations in plants via both transient expression and stable transformation. Thus, we believed that this technical advance represents a promising approach for multi-color-protein co-expression in plant cells.

Example-Based Super-Resolution Fluorescence Microscopy.

Science.gov (United States)

Jia, Shu; Han, Boran; Kutz, J Nathan

2018-04-23

Capturing biological dynamics with high spatiotemporal resolution demands the advancement in imaging technologies. Super-resolution fluorescence microscopy offers spatial resolution surpassing the diffraction limit to resolve near-molecular-level details. While various strategies have been reported to improve the temporal resolution of super-resolution imaging, all super-resolution techniques are still fundamentally limited by the trade-off associated with the longer image acquisition time that is needed to achieve higher spatial information. Here, we demonstrated an example-based, computational method that aims to obtain super-resolution images using conventional imaging without increasing the imaging time. With a low-resolution image input, the method provides an estimate of its super-resolution image based on an example database that contains super- and low-resolution image pairs of biological structures of interest. The computational imaging of cellular microtubules agrees approximately with the experimental super-resolution STORM results. This new approach may offer potential improvements in temporal resolution for experimental super-resolution fluorescence microscopy and provide a new path for large-data aided biomedical imaging.

Time-resolved fluorescence monitoring of cholesterol in peripheral blood mononuclear cells

Science.gov (United States)

Martinakova, Z.; Horilova, J.; Lajdova, I.; Marcek Chorvatova, A.

2014-12-01

Precise evaluation of intracellular cholesterol distribution is crucial for improving diagnostics of diseased states associated with cholesterol alteration. Time-resolved fluorescence techniques are tested for non-invasive investigation of cholesterol in living cells. Fluorescent probe NBD attached to cholesterol was employed to evaluate cholesterol distribution in peripheral blood mononuclear cells (PBMC) isolated from the human blood. Fluorescence Lifetime Imaging Microscopy (FLIM) was successfully applied to simultaneously monitor the spatial distribution and the timeresolved characteristics of the NBD-cholesterol fluorescence in PBMC. Gathered data are the first step in the development of a new perspective non-invasive diagnostic method for evaluation of cholesterol modifications in diseases associated with disorders of lipid metabolism.

Fluorescence X-ray microscopy on hydrated tributyltin-clay mineral suspensions

Science.gov (United States)

Neuhäusler, U.; Schmidt, C.; Hoch, M.; Susini, J.

2003-03-01

Using the scanning transmission X-ray microscope at ID21 beamline of the ESRF in fluorescence mode, we mapped tin at a bulk concentration of 1000 μg(Sn)/ml within hydrated tributyltin (TBT)-clay mineral (Kaolinite) dispersion with sub-300 nm spatial resolution. Using the L absorption edges of tin at 3929, 4156 and 4465 eV fluorescence radiation was excited in tin atoms with incident photon energies of 4 and 4.5 keV. When using 4 keV radiation, only tin fluorescence is excited. For 4.5 keV X rays, both the fluorescence of tin and calcium (which is present in the solid phase) can be measured. Methodologically, we were interested in assessing and proving the possibilities and limitations of fluorescence mapping using the L absorption edges of tin, where the fluorescence yield is significantly lower compared to other elements with their K edges in the same energy range. Scientifically, organotin-clay mineral interactions are of environmental concern because this factor influences significantly the distribution of toxic TBT in the aquatic System. On one hand, the half-life of TBT deposited to the sediment phase increases, and consequently the time of its bioavailability. On the other hand, the adsorption process is reversible, which means that contaminated sediments can act as a source of pollution. The adsorption and desorption effects can be studied directly with high spatial resolution and brought into connection to the surface properties of the clay mineral under study as well as to other experimental parameters, like pH or salinity.

Detection and identification of Cu2+ and Hg2+ based on the cross-reactive fluorescence responses of a dansyl-functionalized film in different solvents.

Science.gov (United States)

Cao, Yuan; Ding, Liping; Wang, Shihuai; Liu, Yuan; Fan, Junmei; Hu, Wenting; Liu, Ping; Fang, Yu

2014-01-08

A dansyl-functionalized fluorescent film sensor was specially designed and prepared by assembling dansyl on a glass plate surface via a long flexible spacer containing oligo(oxyethylene) and amine units. The chemical attachment of dansyl moieties on the surface was verified by contact angle, XPS, and fluorescence measurements. Solvent effect examination revealed that the polarity-sensitivity was retained for the surface-confined dansyl moieties. Fluorescence quenching studies in water declared that the dansyl-functionalized SAM possesses a higher sensitivity towards Hg(2+) and Cu(2+) than the other tested divalent metal ions including Zn(2+), Cd(2+), Co(2+), and Pb(2+). Further measurements of the fluorescence responses of the film towards Cu(2+) and Hg(2+) in three solvents including water, acetonitrile, and THF evidenced that the present film exhibits cross-reactive responses to these two metal ions. The combined signals from the three solvents provide a recognition pattern for both metal ions at a certain concentration and realize the identification between Hg(2+) and Cu(2+). Moreover, using principle component analysis, this method can be extended to identify metal ions that are hard to detect by the film sensor in water such as Co(2+) and Ni(2+).

Towards real time spatially resolved data on sediment transport: 1) tracing the motion of the fluorescent soil particles under rainfall

Science.gov (United States)

Quinton, John; Hardy, Rob; Pates, Jackie; James, Mike

2017-04-01

Understanding where sediment originates from and where it travels to, in what quantities and at which rate is at the heart of many questions surrounding sediment transport, including the connectivity problem. Progress towards unravelling these questions and deepening our understanding has come from a wide range of approaches, including laboratory and field experiments conducted at a variety of scales. In seeking to understand the connectivity of sources and sinks of sediment scientists have spent considerable energy in developing tracing technologies. These have included numerous studies that have relied on the chemical properties of the soil and sediment to establish source-sink connectivity, and the use of 137Ceasium, from radioactive fall-out, to map sediment redistribution. More recently there has been an upsurge in interest in the use of artificially applied soil tracers, including rare earth element oxides and magnetic minerals. However all these tracing methods have a significant drawback: they rely on the collection of samples to assess their concentration. This means that their spatial distribution cannot easily be established in situ and that the environment that is being studied is damaged by the sampling process; nor can data be collected in real time which allows a dynamic understanding of erosion and transport processes to be developed. In this paper we present a methodology for use with a commercially available fluorescent tracer. The tracer is produced in a range of sizes and fluorescent signatures and can be applied to the soil surface. Here we report on an application that combines novel fluorescent videography techniques with custom image processing to trace the motion of the fluorescent soil particles under rainfall. Here we demonstrate the tracking of multiple sub-millimetre particles simultaneously, establishing their position 50 times a second with submillimetre precision. From this we are able to visualise and quantify parameters such as

Fluorescence imaging spectrometer optical design

Science.gov (United States)

Taiti, A.; Coppo, P.; Battistelli, E.

2015-09-01

The optical design of the FLuORescence Imaging Spectrometer (FLORIS) studied for the Fluorescence Explorer (FLEX) mission is discussed. FLEX is a candidate for the ESA's 8th Earth Explorer opportunity mission. FLORIS is a pushbroom hyperspectral imager foreseen to be embarked on board of a medium size satellite, flying in tandem with Sentinel-3 in a Sun synchronous orbit at a height of about 815 km. FLORIS will observe the vegetation fluorescence and reflectance within a spectral range between 500 and 780 nm. Multi-frames acquisitions on matrix detectors during the satellite movement will allow the production of 2D Earth scene images in two different spectral channels, called HR and LR with spectral resolution of 0.3 and 2 nm respectively. A common fore optics is foreseen to enhance by design the spatial co-registration between the two spectral channels, which have the same ground spatial sampling (300 m) and swath (150 km). An overlapped spectral range between the two channels is also introduced to simplify the spectral coregistration. A compact opto-mechanical solution with all spherical and plane optical elements is proposed, and the most significant design rationales are described. The instrument optical architecture foresees a dual Babinet scrambler, a dioptric telescope and two grating spectrometers (HR and LR), each consisting of a modified Offner configuration. The developed design is robust, stable vs temperature, easy to align, showing very high optical quality along the whole field of view. The system gives also excellent correction for transverse chromatic aberration and distortions (keystone and smile).

An instrument for the simultaneous acquisition of size, shape, and spectral fluorescence data from single aerosol particles

Science.gov (United States)

Hirst, Edwin; Kaye, Paul H.; Foot, Virginia E.; Clark, James M.; Withers, Philip B.

2004-12-01

We describe the construction of a bio-aerosol monitor designed to capture and record intrinsic fluorescence spectra from individual aerosol particles carried in a sample airflow and to simultaneously capture data relating to the spatial distribution of elastically scattered light from each particle. The spectral fluorescence data recorded by this PFAS (Particle Fluorescence and Shape) monitor contains information relating to the particle material content and specifically to possible biological fluorophores. The spatial scattering data from PFAS yields information relating to particle size and shape. The combination of these data can provide a means of aiding the discrimination of bio-aerosols from background or interferent aerosol particles which may have similar fluorescence properties but exhibit shapes and/or sizes not normally associated with biological particles. The radiation used both to excite particle fluorescence and generate the necessary spatially scattered light flux is provided by a novel compact UV fiber laser operating at 266nm wavelength. Particles drawn from the ambient environment traverse the laser beam in single file. Intrinsic particle fluorescence in the range 300-570nm is collected via an ellipsoidal concentrator into a concave grating spectrometer, the spectral data being recorded using a 16-anode linear array photomultiplier detector. Simultaneously, the spatial radiation pattern scattered by the particle over 5°-30° scattering angle and 360° of azimuth is recorded using a custom designed 31-pixel radial hybrid photodiode array. Data from up to ~5,000 particles per second may be acquired for analysis, usually performed by artificial neural network classification.

An orange fluorescent protein tagging system for real-time pollen tracking.

Science.gov (United States)

Rice, J Hollis; Millwood, Reginald J; Mundell, Richard E; Chambers, Orlando D; Abercrombie, Laura L; Davies, H Maelor; Stewart, C Neal

2013-09-27

Monitoring gene flow could be important for future transgenic crops, such as those producing plant-made-pharmaceuticals (PMPs) in open field production. A Nicotiana hybrid (Nicotiana. tabacum × Nicotiana glauca) shows limited male fertility and could be used as a bioconfined PMP platform. Effective assessment of gene flow from these plants is augmented with methods that utilize fluorescent proteins for transgenic pollen identification. We report the generation of a pollen tagging system utilizing an orange fluorescent protein to monitor pollen flow and as a visual assessment of transgene zygosity of the parent plant. This system was created to generate a tagged Nicotiana hybrid that could be used for the incidence of gene flow. Nicotiana tabacum 'TN 90' and Nicotiana glauca were successfully transformed via Agrobacterium tumefaciens to express the orange fluorescent protein gene, tdTomato-ER, in pollen and a green fluorescent protein gene, mgfp5-er, was expressed in vegetative structures of the plant. Hybrids were created that utilized the fluorescent proteins as a research tool for monitoring pollen movement and gene flow. Manual greenhouse crosses were used to assess hybrid sexual compatibility with N. tabacum, resulting in seed formation from hybrid pollination in 2% of crosses, which yielded non-viable seed. Pollen transfer to the hybrid formed seed in 19% of crosses and 10 out of 12 viable progeny showed GFP expression. The orange fluorescent protein is visible when expressed in the pollen of N. glauca, N. tabacum, and the Nicotiana hybrid, although hybrid pollen did not appear as bright as the parent lines. The hybrid plants, which show limited ability to outcross, could provide bioconfinement with the benefit of detectable pollen using this system. Fluorescent protein-tagging could be a valuable tool for breeding and in vivo ecological monitoring.

Tracking Lithium Ions via Widefield Fluorescence Microscopy for Battery Diagnostics.

Science.gov (United States)

Padilla, Nicolas A; Rea, Morgan T; Foy, Michael; Upadhyay, Sunil P; Desrochers, Kyle A; Derus, Tyler; Knapper, Kassandra A; Hunter, Nathanael H; Wood, Sharla; Hinton, Daniel A; Cavell, Andrew C; Masias, Alvaro G; Goldsmith, Randall H

2017-07-28

Direct tracking of lithium ions with time and spatial resolution can provide an important diagnostic tool for understanding mechanisms in lithium ion batteries. A fluorescent indicator of lithium ions, 2-(2-hydroxyphenyl)naphthoxazole, was synthesized and used for real-time tracking of lithium ions via widefield fluorescence microscopy. The fluorophore can be excited with visible light and was shown to enable quantitative determination of the lithium ion diffusion constant in a microfluidic model system for a plasticized polymer electrolyte lithium battery. The use of widefield fluorescence microscopy for in situ tracking of lithium ions in batteries is discussed.

Improved fluorescent X-ray image intensifying screen

International Nuclear Information System (INIS)

Landeghem, W.K. van; Suys, A.R.

1981-01-01

An X-ray image intensifying screen is described, which includes at least one fluorescent layer comprising phosphor particles dispersed in a binder and on top of such layer a protective layer containing a crosslinked polymer mass obtained by an acid-catalyzed reaction of a polymer or mixture of polymers containing reactive hydrogen atoms and a cross-linking agent, the cross-linking agent being an organic compound containing a plurality of etherified N-methylol groups. Examples are given of appropriate polymers and cross-linking agents. (author)

X-ray fluorescence investigation of gilded and enamelled silver: The case study of four medieval processional crosses from central Italy

International Nuclear Information System (INIS)

Ferretti, Marco; Polese, Claudia; Roldán García, Clodoaldo

2013-01-01

The presence of multilayered structures is common in such cultural artefacts as paintings, corroded metals, objects that underwent a whatever form of surface qualification. One of the most usual and complete ways to investigate such structures is observing a cross section, which requires sampling. There are however situations where at least part of the stratigraphic information can be derived non-destructively: the literature shows that X-ray fluorescence (XRF) has frequently been used, in recent years, for this purpose, with special regard to paintings and gilded metals. Aim of this paper is to further explore the suitability of XRF-based techniques to characterise multilayered structures. This is achieved by introducing improvements, with respect to previous works, in both equipment and data processing. The method, that has been developed for gilded and enamelled silver artefacts, relies on optimum excitation conditions provided for silver and on the relationship existing between the ratio AgKα/AgKβ of its fluorescence lines and the gilding thickness itself. The coating (gilding or enamel) thickness is derived by verifying the condition C Kα,Ag = C Kβ,Ag , where C Kα,Ag and C Kβ,Ag are the mass fractions of silver calculated on the lines AgKα and AgKβ, respectively. The calculations are carried out by PyMCA, a Fundamental Parameters code that implements the analysis of multilayered samples. As a case study we investigated in situ the four processional crosses of Borbona, Sant'Elpidio, Rosciolo and Forcella, made of a wood core with attached gilt and embossed silver sheets and enamelled silver plates. The analyses allowed to distinguish ancient restorations from original parts, to characterise the enamels and find their composition consistent with the dates of manufacturing and, as regards the cross of Rosciolo, to hypothesize the contribution of different “hands” in its manufacturing. - Highlights: ► We discuss a non-destructive XRF method for the

X-ray fluorescence investigation of gilded and enamelled silver: The case study of four medieval processional crosses from central Italy

Energy Technology Data Exchange (ETDEWEB)

Ferretti, Marco, E-mail: marco.ferretti@itabc.cnr.it [CNR, Istituto per le Tecnologie Applicate ai Beni Culturali, AdR RM1, Via Salaria km 29.300, 00016 Montelibretti (Roma) (Italy); Polese, Claudia, E-mail: claudia.polese87@hotmail.it [Dip. di Chimica, Università di Roma “Sapienza,” Piazzale Aldo Moro 5, 00185 Roma (Italy); Roldán García, Clodoaldo, E-mail: Clodoaldo.Roldan@uv.es [Instituto de Ciencia de Materiales, Parc Cientific Universidad de Valencia, C/Catedrático José Beltrán 2, 46980 Paterna (Valencia) (Spain)

2013-05-01

The presence of multilayered structures is common in such cultural artefacts as paintings, corroded metals, objects that underwent a whatever form of surface qualification. One of the most usual and complete ways to investigate such structures is observing a cross section, which requires sampling. There are however situations where at least part of the stratigraphic information can be derived non-destructively: the literature shows that X-ray fluorescence (XRF) has frequently been used, in recent years, for this purpose, with special regard to paintings and gilded metals. Aim of this paper is to further explore the suitability of XRF-based techniques to characterise multilayered structures. This is achieved by introducing improvements, with respect to previous works, in both equipment and data processing. The method, that has been developed for gilded and enamelled silver artefacts, relies on optimum excitation conditions provided for silver and on the relationship existing between the ratio AgKα/AgKβ of its fluorescence lines and the gilding thickness itself. The coating (gilding or enamel) thickness is derived by verifying the condition C{sub Kα,Ag} = C{sub Kβ,Ag}, where C{sub Kα,Ag} and C{sub Kβ,Ag} are the mass fractions of silver calculated on the lines AgKα and AgKβ, respectively. The calculations are carried out by PyMCA, a Fundamental Parameters code that implements the analysis of multilayered samples. As a case study we investigated in situ the four processional crosses of Borbona, Sant'Elpidio, Rosciolo and Forcella, made of a wood core with attached gilt and embossed silver sheets and enamelled silver plates. The analyses allowed to distinguish ancient restorations from original parts, to characterise the enamels and find their composition consistent with the dates of manufacturing and, as regards the cross of Rosciolo, to hypothesize the contribution of different “hands” in its manufacturing. - Highlights: ► We discuss a non

Fluorescent visualization of macromolecules in Drosophila whole mounts.

Science.gov (United States)

Ramos, Ricardo Guelerman Pinheiro; Machado, Luciana Claudia Herculano; Moda, Livia Maria Rosatto

2010-01-01

The ability to determine the expression dynamics of individual genes "in situ" by visualizing the precise spatial and temporal distribution of their products in whole mounts by histochemical and immunocytochemical reactions has revolutionized our understanding of cellular processes. Drosophila developmental genetics was one of the fields that benefited most from these technologies, and a variety of fluorescent methods were specifically designed for investigating the localization of developmentally important proteins and cell markers during embryonic and post embryonic stages of this model organism. In this chapter we present detailed protocols for fluorescence immunocytochemistry of whole mount embryos, imaginal discs, pupal retinas, and salivary glands of Drosophila melanogaster, as well as methods for fluorescent visualization of specific subcellular structures in these tissues.

Probing GFP-actin diffusion in living cells using fluorescence correlation spectroscopy

International Nuclear Information System (INIS)

Engelke, Hanna; Heinrich, Doris; Rädler, Joachim O.

2010-01-01

The cytoskeleton of eukaryotic cells is continuously remodeled by polymerization and depolymerization of actin. Consequently, the relative content of polymerized filamentous actin (F-actin) and monomeric globular actin (G-actin) is subject to temporal and spatial fluctuations. Since fluorescence correlation spectroscopy (FCS) can measure the diffusion of fluorescently labeled actin it seems likely that FCS allows us to determine the dynamics and hence indirectly the structural properties of the cytoskeleton components with high spatial resolution. To this end we investigate the FCS signal of GFP-actin in living Dictyostelium discoideum cells and explore the inherent spatial and temporal signatures of the actin cytoskeleton. Using the free green fluorescent protein (GFP) as a reference, we find that actin diffusion inside cells is dominated by G-actin and slower than diffusion in diluted cell extract. The FCS signal in the dense cortical F-actin network near the cell membrane is probed using the cytoskeleton protein LIM and is found to be slower than cytosolic G-actin diffusion. Furthermore, we show that polymerization of the cytoskeleton induced by Jasplakinolide leads to a substantial decrease of G-actin diffusion. Pronounced fluctuations in the distribution of the FCS correlation curves can be induced by latrunculin, which is known to induce actin waves. Our work suggests that the FCS signal of GFP-actin in combination with scanning or spatial correlation techniques yield valuable information about the local dynamics and concomitant cytoskeletal properties

Vivre la frontière : les pratiques spatiales transfrontalières dans l’Arc jurassien franco-suisse Living the border : cross-border spatial practices in the French-Swiss Jura region

Directory of Open Access Journals (Sweden)

Yann Dubois

2012-12-01

Full Text Available La construction européenne a redéfini la signification et les fonctions des frontières nationales. Cet article s’intéresse à cette mutation sous l’angle des pratiques des habitants de l’Arc jurassien franco-suisse. Une enquête par questionnaire a permis de mesurer l’intensité de certaines pratiques spatiales transfrontalières réalisées pendant le temps libre (achats, loisirs, etc. et d’en déterminer les logiques sous-jacentes. L’effet frontière se manifeste sous la forme d’un différentiel de prix (coût de la vie, taux de change, taxation de certains produits et d’un différentiel de connaissances (manque d’informations sur le pays voisin, habitude, etc. qui freinent ou incitent le franchissement de la frontière. L’effet frontière est toutefois atténué dans le contexte territorial étudié par un effet de centralité impliquant un différentiel d’offre (attraction des communes françaises à vocation résidentielle par les centres urbains helvétiques. La combinaison de ces effets explique l’intensité et l’orientation des pratiques spatiales transfrontalières.The European integration has been redefining the meaning and functions of national borders. This paper addresses this mutation from the perspective of the inhabitants’ spatial practices in the French-Swiss Jura region. Through a questionnaire survey we have measured the intensity of cross-border spatial practices done in the free time (purchasing, leisure, etc. and determined the underlying logics. The border effect appears in the form of a price differential (cost of living, exchange rate, taxation of some goods and a knowledge differential (lack of information on the neighbouring country, habits, etc. that curb or stimulate border crossing. The border effect is however mitigated in the spatial context under study by a centrality effect involving a supply differential (attraction of residential French municipalities by Swiss urban centres. The

Protein recognition by a pattern-generating fluorescent molecular probe

Science.gov (United States)

Pode, Zohar; Peri-Naor, Ronny; Georgeson, Joseph M.; Ilani, Tal; Kiss, Vladimir; Unger, Tamar; Markus, Barak; Barr, Haim M.; Motiei, Leila; Margulies, David

2017-12-01

Fluorescent molecular probes have become valuable tools in protein research; however, the current methods for using these probes are less suitable for analysing specific populations of proteins in their native environment. In this study, we address this gap by developing a unimolecular fluorescent probe that combines the properties of small-molecule-based probes and cross-reactive sensor arrays (the so-called chemical 'noses/tongues'). On the one hand, the probe can detect different proteins by generating unique identification (ID) patterns, akin to cross-reactive arrays. On the other hand, its unimolecular scaffold and selective binding enable this ID-generating probe to identify combinations of specific protein families within complex mixtures and to discriminate among isoforms in living cells, where macroscopic arrays cannot access. The ability to recycle the molecular device and use it to track several binding interactions simultaneously further demonstrates how this approach could expand the fluorescent toolbox currently used to detect and image proteins.

Measurement of OH free radical in magnetized sheet plasma crossed with vertical gas-flow by laser-induced fluorescence spectroscopy

International Nuclear Information System (INIS)

Tonegawa, Akira; Takatori, Masahiko; Kobayashi, Yukihiro; Kawamura, Kazutaka; Takayama, Kazuo

1997-01-01

We demonstrated the production of OH free radicals in an argon magnetized sheet plasma crossed with vertical gas-flow mixed with an oxygen gas O 2 and a hydrogen gas H 2 . The density and the rotational-vibrational temperature of the OH free radicals were measured by a laser-induced fluorescence (LIF). The density of OH free radicals increases with increasing O 2 gas-flow, while the high energy part of the electron-energy-distribution-function f e (E) above 8 eV decreases. These results suggest the high energy part of f e (E) is contributed to the dissociation of O 2 and the production of OH free radicals. (author)

Spatial Attention and Audiovisual Interactions in Apparent Motion

Science.gov (United States)

Sanabria, Daniel; Soto-Faraco, Salvador; Spence, Charles

2007-01-01

In this study, the authors combined the cross-modal dynamic capture task (involving the horizontal apparent movement of visual and auditory stimuli) with spatial cuing in the vertical dimension to investigate the role of spatial attention in cross-modal interactions during motion perception. Spatial attention was manipulated endogenously, either…

Automatic segmentation of lesions for the computer-assisted detection in fluorescence urology

Science.gov (United States)

Kage, Andreas; Legal, Wolfgang; Kelm, Peter; Simon, Jörg; Bergen, Tobias; Münzenmayer, Christian; Benz, Michaela

2012-03-01

Bladder cancer is one of the most common cancers in the western world. The diagnosis in Germany is based on the visual inspection of the bladder. This inspection performed with a cystoscope is a challenging task as some kinds of abnormal tissues do not differ much in their appearance from their surrounding healthy tissue. Fluorescence Cystoscopy has the potential to increase the detection rate. A liquid marker introduced into the bladder in advance of the inspection is concentrated in areas with high metabolism. Thus these areas appear as bright "glowing". Unfortunately, the fluorescence image contains besides the glowing of the suspicious lesions no more further visual information like for example the appearance of the blood vessels. A visual judgment of the lesion as well as a precise treatment has to be done using white light illumination. Thereby, the spatial information of the lesion provided by the fluorescence image has to be guessed by the clinical expert. This leads to a time consuming procedure due to many switches between the modalities and increases the risk of mistreatment. We introduce an automatic approach, which detects and segments any suspicious lesion in the fluorescence image automatically once the image was classified as a fluorescence image. The area of the contour of the detected lesion is transferred to the corresponding white light image and provide the clinical expert the spatial information of the lesion. The advantage of this approach is, that the clinical expert gets the spatial and the visual information of the lesion together in one image. This can save time and decrease the risk of an incomplete removal of a malign lesion.

Differentiation of Cariogenic Streptococci by Fluorescent Antibody1

Science.gov (United States)

Jablon, James M.; Zinner, Doran D.

1966-01-01

Jablon, J. M. (University of Miami, Miami, Fla.), and D. D. Zinner. Differentiation of cariogenic streptococci by fluorescent antibody. J. Bacteriol. 92:1590–1596. 1966.—Eight strains of streptococci were isolated from human carious lesions by the fluorescent-antibody (FA) technique. Seven of these strains produced experimental caries in hamsters or rats maintained on a high sucrose diet. The eighth strain was noncariogenic in animals but possessed some antigenic components in common with the cariogenic strains. On the basis of antigen-antibody reactions by microprecipitin and agar-gel diffusion patterns, the strains were divided into four groups; these groups differed with regard to their cariogenic activity in hamsters. Fluorescein-conjugated antisera, prepared against the human strains, showed some cross-reactions which interfered with the efficacy of the FA technique in differentiating between the related streptococcal groups. To eliminate these cross-reactions, a small amount of related-strain antisera was added to the fluorescein-conjugated antisera to the cariogenic strains. This technique is effective in blocking cross-reactions and should be tried wherever cross-reactions are encountered in the FA technique. Images PMID:5334765

Trends in environmental science using microscopic X-ray fluorescence

International Nuclear Information System (INIS)

Fittschen, Ursula Elisabeth Adriane; Falkenberg, Gerald

2011-01-01

Microscopic X-ray fluorescence (micro-XRF) is a versatile tool in environmental analysis. We review work done in this field from 2008 to 2010 and highlight new aspects. Overall, there is a strong trend to combine fluorescence data with other data like diffraction or absorption spectroscopy. Also, the use of laboratory based instrumentation has become wide spread as more commercial instruments are available. At laboratories and synchrotron sites the trend towards higher spatial resolution is still persistent hitting sub micrometer values in case of synchrotron set ups.

Trends in environmental science using microscopic X-ray fluorescence

Energy Technology Data Exchange (ETDEWEB)

Fittschen, Ursula Elisabeth Adriane, E-mail: ursula.fittschen@chemie.uni-hamburg.de [Department of Chemistry, University of Hamburg, Martin-Luther-King-Platz 6, 20146 Hamburg (Germany); Falkenberg, Gerald [Deutsches Elektronen-Synchrotron, Notkestr. 85, 22603 Hamburg (Germany)

2011-08-15

Microscopic X-ray fluorescence (micro-XRF) is a versatile tool in environmental analysis. We review work done in this field from 2008 to 2010 and highlight new aspects. Overall, there is a strong trend to combine fluorescence data with other data like diffraction or absorption spectroscopy. Also, the use of laboratory based instrumentation has become wide spread as more commercial instruments are available. At laboratories and synchrotron sites the trend towards higher spatial resolution is still persistent hitting sub micrometer values in case of synchrotron set ups.

Spatial distribution of carbon species in laser ablation of graphite target

International Nuclear Information System (INIS)

Ikegami, T.; Ishibashi, S.; Yamagata, Y.; Ebihara, K.; Thareja, R.K.; Narayan, J.

2001-01-01

We report on the temporal evolution and spatial distribution of C 2 and C 3 molecules produced by KrF laser ablation of a graphite target using laser induced fluorescence imaging and optical emission spectroscopy. Spatial density profiles of C 2 were measured using two-dimensional fluorescence in various pressures of different ambient (vacuum, nitrogen, oxygen, hydrogen, helium, and argon) gases at various ablation laser fluences and ablation area. A large yield of C 2 is observed in the central part of the plume and near the target surface and its density and distribution was affected by the laser fluence and ambient gas. Fluorescent C 3 was studied in Ar gas and the yield of C 3 is enhanced at higher gas pressure and longer delay times after ablation

Spatial and temporal dynamics of receptor for advanced glycation endproducts, integrins, and actin cytoskeleton as probed with fluorescence-based imaging techniques

Energy Technology Data Exchange (ETDEWEB)

Syed, Aleem [Iowa State Univ., Ames, IA (United States)

2016-01-01

Systematic spatial and temporal fluctuations are a fundamental part of any biological process. For example, lateral diffusion of membrane proteins is one of the key mechanisms in their cellular function. Lateral diffusion governs how membrane proteins interact with intracellular, transmembrane, and extracellular components to achieve their function. Herein, fluorescence-based techniques are used to elucidate the dynamics of receptor for advanced glycation end-products (RAGE) and integrin membrane proteins. RAGE is a transmembrane protein that is being used as a biomarker for various diseases. RAGE dependent signaling in numerous pathological conditions is well studied. However, RAGE lateral diffusion in the cell membrane is poorly understood. For this purpose, effect of cholesterol, cytoskeleton dynamics, and presence of ligand on RAGE lateral diffusion is investigated.

Global Monitoring of Terrestrial Chlorophyll Fluorescence from Moderate-spectral-resolution Near-infrared Satellite Measurements: Methodology, Simulations, and Application to GOME-2

Science.gov (United States)

Joiner, J.; Gaunter, L.; Lindstrot, R.; Voigt, M.; Vasilkov, A. P.; Middleton, E. M.; Huemmrich, K. F.; Yoshida, Y.; Frankenberg, C.

2013-01-01

Globally mapped terrestrial chlorophyll fluorescence retrievals are of high interest because they can provide information on the functional status of vegetation including light-use efficiency and global primary productivity that can be used for global carbon cycle modeling and agricultural applications. Previous satellite retrievals of fluorescence have relied solely upon the filling-in of solar Fraunhofer lines that are not significantly affected by atmospheric absorption. Although these measurements provide near-global coverage on a monthly basis, they suffer from relatively low precision and sparse spatial sampling. Here, we describe a new methodology to retrieve global far-red fluorescence information; we use hyperspectral data with a simplified radiative transfer model to disentangle the spectral signatures of three basic components: atmospheric absorption, surface reflectance, and fluorescence radiance. An empirically based principal component analysis approach is employed, primarily using cloudy data over ocean, to model and solve for the atmospheric absorption. Through detailed simulations, we demonstrate the feasibility of the approach and show that moderate-spectral-resolution measurements with a relatively high signal-to-noise ratio can be used to retrieve far-red fluorescence information with good precision and accuracy. The method is then applied to data from the Global Ozone Monitoring Instrument 2 (GOME-2). The GOME-2 fluorescence retrievals display similar spatial structure as compared with those from a simpler technique applied to the Greenhouse gases Observing SATellite (GOSAT). GOME-2 enables global mapping of far-red fluorescence with higher precision over smaller spatial and temporal scales than is possible with GOSAT. Near-global coverage is provided within a few days. We are able to show clearly for the first time physically plausible variations in fluorescence over the course of a single month at a spatial resolution of 0.5 deg × 0.5 deg

Photonic reagents for concentration measurement of flu-orescent proteins with overlapping spectra

Science.gov (United States)

Goun, Alexei; Bondar, Denys I.; Er, Ali O.; Quine, Zachary; Rabitz, Herschel A.

2016-05-01

By exploiting photonic reagents (i.e., coherent control by shaped laser pulses), we employ Optimal Dynamic Discrimination (ODD) as a novel means for quantitatively characterizing mixtures of fluorescent proteins with a large spectral overlap. To illustrate ODD, we simultaneously measured concentrations of in vitro mixtures of Enhanced Blue Fluorescent Protein (EBFP) and Enhanced Cyan Fluorescent Protein (ECFP). Building on this foundational study, the ultimate goal is to exploit the capabilities of ODD for parallel monitoring of genetic and protein circuits by suppressing the spectral cross-talk among multiple fluorescent reporters.

Collisional and radiative processes in fluorescent lamps

International Nuclear Information System (INIS)

Lister, Graeme G.

2003-01-01

Since electrode life is the major limiting factor in operating fluorescent lamps, many lighting companies have introduced 'electrodeless' fluorescent lamps, using inductively coupled discharges. These lamps often operate at much higher power loadings than standard lamps and numerical models have not been successful in reproducing experimental measurements in the parameter ranges of interest. A comprehensive research program was undertaken to study the fundamental physical processes of these discharges, co-funded by the Electric Power Research Institute (EPRI) and OSRAM SYLVANIA under the name of ALITE. The program included experiments and modeling of radiation transport, computations of electron-atom and atom-atom cross sections and the first comprehensive power balance studies of a highly loaded fluorescent lamp. Results from the program and their importance to the understanding of the physics of fluorescent lamps are discussed, with particular emphasis on the important collisional and radiative processes. Comparisons between results of experimental measurements and numerical models are presented

Sun-induced fluorescence - a new probe of photosynthesis: First maps from the imaging spectrometer HyPlant.

Science.gov (United States)

Rascher, U; Alonso, L; Burkart, A; Cilia, C; Cogliati, S; Colombo, R; Damm, A; Drusch, M; Guanter, L; Hanus, J; Hyvärinen, T; Julitta, T; Jussila, J; Kataja, K; Kokkalis, P; Kraft, S; Kraska, T; Matveeva, M; Moreno, J; Muller, O; Panigada, C; Pikl, M; Pinto, F; Prey, L; Pude, R; Rossini, M; Schickling, A; Schurr, U; Schüttemeyer, D; Verrelst, J; Zemek, F

2015-12-01

Variations in photosynthesis still cause substantial uncertainties in predicting photosynthetic CO2 uptake rates and monitoring plant stress. Changes in actual photosynthesis that are not related to greenness of vegetation are difficult to measure by reflectance based optical remote sensing techniques. Several activities are underway to evaluate the sun-induced fluorescence signal on the ground and on a coarse spatial scale using space-borne imaging spectrometers. Intermediate-scale observations using airborne-based imaging spectroscopy, which are critical to bridge the existing gap between small-scale field studies and global observations, are still insufficient. Here we present the first validated maps of sun-induced fluorescence in that critical, intermediate spatial resolution, employing the novel airborne imaging spectrometer HyPlant. HyPlant has an unprecedented spectral resolution, which allows for the first time quantifying sun-induced fluorescence fluxes in physical units according to the Fraunhofer Line Depth Principle that exploits solar and atmospheric absorption bands. Maps of sun-induced fluorescence show a large spatial variability between different vegetation types, which complement classical remote sensing approaches. Different crop types largely differ in emitting fluorescence that additionally changes within the seasonal cycle and thus may be related to the seasonal activation and deactivation of the photosynthetic machinery. We argue that sun-induced fluorescence emission is related to two processes: (i) the total absorbed radiation by photosynthetically active chlorophyll; and (ii) the functional status of actual photosynthesis and vegetation stress. © 2015 John Wiley & Sons Ltd.

Chlorophyll Fluorescence Imaging Uncovers Photosynthetic Fingerprint of Citrus Huanglongbing

Directory of Open Access Journals (Sweden)

Haiyan Cen

2017-08-01

Full Text Available Huanglongbing (HLB is one of the most destructive diseases of citrus, which has posed a serious threat to the global citrus production. This research was aimed to explore the use of chlorophyll fluorescence imaging combined with feature selection to characterize and detect the HLB disease. Chlorophyll fluorescence images of citrus leaf samples were measured by an in-house chlorophyll fluorescence imaging system. The commonly used chlorophyll fluorescence parameters provided the first screening of HLB disease. To further explore the photosynthetic fingerprint of HLB infected leaves, three feature selection methods combined with the supervised classifiers were employed to identify the unique fluorescence signature of HLB and perform the three-class classification (i.e., healthy, HLB infected, and nutrient deficient leaves. Unlike the commonly used fluorescence parameters, this novel data-driven approach by using the combination of the mean fluorescence parameters and image features gave the best classification performance with the accuracy of 97%, and presented a better interpretation for the spatial heterogeneity of photochemical and non-photochemical components in HLB infected citrus leaves. These results imply the potential of the proposed approach for the citrus HLB disease diagnosis, and also provide a valuable insight for the photosynthetic response to the HLB disease.

Molecular engineering of two-photon fluorescent probes for bioimaging applications

Science.gov (United States)

Liu, Hong-Wen; Liu, Yongchao; Wang, Peng; Zhang, Xiao-Bing

2017-03-01

During the past two decades, two-photon microscopy (TPM), which utilizes two near-infrared photons as the excitation source, has emerged as a novel, attractive imaging tool for biological research. Compared with one-photon microscopy, TPM offers several advantages, such as lowering background fluorescence in living cells and tissues, reducing photodamage to biosamples, and a photobleaching phenomenon, offering better 3D spatial localization, and increasing penetration depth. Small-molecule-based two-photon fluorescent probes have been well developed for the detection and imaging of various analytes in biological systems. In this review, we will give a general introduction of molecular engineering of two-photon fluorescent probes based on different fluorescence response mechanisms for bioimaging applications during the past decade. Inspired by the desired advantages of small-molecule two-photon fluorescent probes in biological imaging applications, we expect that more attention will be devoted to the development of new two-photon fluorophores and applications of TPM in areas of bioanalysis and disease diagnosis.

The use of yellow fluorescent hybrids to indicate mating in Trypanosoma brucei

Directory of Open Access Journals (Sweden)

Ferris Vanessa

2008-02-01

Full Text Available Abstract Background Trypanosoma brucei undergoes genetic exchange in its insect vector, the tsetse fly, by an unknown mechanism. The difficulties of working with this experimental system of genetic exchange have hampered investigation, particularly because the trypanosome life cycle stages involved cannot be cultured in vitro and therefore must be examined in the insect. Searching for small numbers of hybrid trypanosomes directly in the fly has become possible through the incorporation of fluorescent reporter genes, and we have previously carried out a successful cross using a reporter-repressor strategy. However, we could not be certain that all fluorescent trypanosomes observed in that cross were hybrids, due to mutations of the repressor leading to spontaneous fluorescence, and we have therefore developed an alternative strategy. Results To visualize the production of hybrids in the fly, parental trypanosome clones were transfected with a gene encoding Green Fluorescent Protein (GFP or Red Fluorescent Protein (RFP. Co-infection of flies with red and green fluorescent parental trypanosomes produced yellow fluorescent hybrids, which were easily visualized in the fly salivary glands. Yellow trypanosomes were not seen in midgut or proventricular samples and first appeared in the glands as epimastigotes as early as 13 days after fly infection. Cloned progeny originating from individual salivary glands had yellow, red, green or no fluorescence and were confirmed as hybrids by microsatellite, molecular karyotype and kinetoplast (mitochondrial DNA analyses. Hybrid clones showed biparental inheritance of both nuclear and kinetoplast genomes. While segregation and reassortment of the reporter genes and microsatellite alleles were consistent with Mendelian inheritance, flow cytometry measurement of DNA content revealed both diploid and polyploid trypanosomes among the hybrid progeny clones. Conclusion The strategy of using production of yellow hybrids

Fully time-resolved near-field scanning optical microscopy fluorescence imaging

International Nuclear Information System (INIS)

Kwak, Eun-Soo; Vanden Bout, David A.

2003-01-01

Time-correlated single photon counting has been coupled with near-field scanning optical microscopy (NSOM) to record complete fluorescence lifetime decays at each pixel in an NSOM image. The resulting three-dimensional data sets can be binned in the time dimension to create images of photons at particular time delays or images of the fluorescence lifetime. Alternatively, regions of interest identified in the topography and fluorescence images can be used to bin the data in the spatial dimensions resulting in high signal to noise fluorescence decays of particular regions of the sample. The technique has been demonstrated on films of poly(vinylalcohol), doped with the fluorescent dye, cascade blue (CB). The CB segregates into small circular regions of high concentration within the films during the drying process. The lifetime imaging shows that the spots have slightly faster excited state decays due to quenching of the luminescence as a result of the higher concentration. The technique is also used to image the fluorescence lifetime of an annealed film of poly(dihexylfluorene). The samples show high contrast in the total intensity fluorescence image, but the lifetime image reveals the sample to be extremely uniform

Studying flow close to an interface by total internal reflection fluorescence cross-correlation spectroscopy: Quantitative data analysis

Science.gov (United States)

Schmitz, R.; Yordanov, S.; Butt, H. J.; Koynov, K.; Dünweg, B.

2011-12-01

Total internal reflection fluorescence cross-correlation spectroscopy (TIR-FCCS) has recently [S. Yordanov , Optics ExpressOPEXFF1094-408710.1364/OE.17.021149 17, 21149 (2009)] been established as an experimental method to probe hydrodynamic flows near surfaces, on length scales of tens of nanometers. Its main advantage is that fluorescence occurs only for tracer particles close to the surface, thus resulting in high sensitivity. However, the measured correlation functions provide only rather indirect information about the flow parameters of interest, such as the shear rate and the slip length. In the present paper, we show how to combine detailed and fairly realistic theoretical modeling of the phenomena by Brownian dynamics simulations with accurate measurements of the correlation functions, in order to establish a quantitative method to retrieve the flow properties from the experiments. First, Brownian dynamics is used to sample highly accurate correlation functions for a fixed set of model parameters. Second, these parameters are varied systematically by means of an importance-sampling Monte Carlo procedure in order to fit the experiments. This provides the optimum parameter values together with their statistical error bars. The approach is well suited for massively parallel computers, which allows us to do the data analysis within moderate computing times. The method is applied to flow near a hydrophilic surface, where the slip length is observed to be smaller than 10nm, and, within the limitations of the experiments and the model, indistinguishable from zero.

Refractive Index Sensing of Green Fluorescent Proteins in Living Cells Using Fluorescence Lifetime Imaging Microscopy

Science.gov (United States)

van Manen, Henk-Jan; Verkuijlen, Paul; Wittendorp, Paul; Subramaniam, Vinod; van den Berg, Timo K.; Roos, Dirk; Otto, Cees

2008-01-01

We show that fluorescence lifetime imaging microscopy (FLIM) of green fluorescent protein (GFP) molecules in cells can be used to report on the local refractive index of intracellular GFP. We expressed GFP fusion constructs of Rac2 and gp91phox, which are both subunits of the phagocyte NADPH oxidase enzyme, in human myeloid PLB-985 cells and showed by high-resolution confocal fluorescence microscopy that GFP-Rac2 and GFP-gp91phox are targeted to the cytosol and to membranes, respectively. Frequency-domain FLIM experiments on these PLB-985 cells resulted in average fluorescence lifetimes of 2.70 ns for cytosolic GFP-Rac2 and 2.31 ns for membrane-bound GFP-gp91phox. By comparing these lifetimes with a calibration curve obtained by measuring GFP lifetimes in PBS/glycerol mixtures of known refractive index, we found that the local refractive indices of cytosolic GFP-Rac2 and membrane-targeted GFP-gp91phox are ∼1.38 and ∼1.46, respectively, which is in good correspondence with reported values for the cytosol and plasma membrane measured by other techniques. The ability to measure the local refractive index of proteins in living cells by FLIM may be important in revealing intracellular spatial heterogeneities within organelles such as the plasma and phagosomal membrane. PMID:18223002

Photo-triggered fluorescent theranostic prodrugs as DNA alkylating agents for mechlorethamine release and spatiotemporal monitoring.

Science.gov (United States)

Cao, Yanting; Pan, Rong; Xuan, Weimin; Wei, Yongyi; Liu, Kejian; Zhou, Jiahong; Wang, Wei

2015-06-28

We describe a new theranostic strategy for selective delivery and spatiotemporal monitoring of mechlorethamine, a DNA alkylating agent. A photo-responsive prodrug is designed and composed of a photolabile o-nitrophenylethyl group, a DNA alkylating mechlorethamine drug and a coumarin fluorophore. Masking of the "N" in mechlorethamine in a positively charged state in the prodrug renders it inactive, non-toxic, selective and non-fluorescent. Indeed, the stable prodrug shows negligible cytotoxicity towards normal cells with and without UV activation and is completely non-fluorescent. However, upon photo-irradiation, the active mechlorethamine is released and induces efficient DNA cross-links, accompanied by a strong fluorescence enhancement (152 fold). Furthermore, DNA cross-linking activity from the release can be transformed into anticancer activity observed in in vitro studies of tumor cells. Importantly, the drug release progress and the movement can be conveniently monitored by fluorescence spectroscopy. The mechanistic study proves that the DNA cross-linking activity is mainly due to the release of DNA alkylating mechlorethamine. Altogether, the studies show the power of the theranostic strategy for efficient therapy in cancer treatment.

Spatial Analysis Of Human Capital Structures

Directory of Open Access Journals (Sweden)

Gajdos Artur

2014-12-01

Full Text Available The main purpose of this paper is to analyse the interdependence between labour productivity and the occupational structure of human capital in a spatial cross-section. Research indicates (see Fischer 2009 the possibility to assess the impact of the quality of human capital (measured by means of the level of education on labour productivity in a spatial cross-section.

Measurement of L X-ray fluorescence cross-sections for elements with 45 ≤ Z ≤ 50 using synchrotron radiation at 8 keV

International Nuclear Information System (INIS)

Bonzi, Edgardo V.; Badiger, Nagappa M.; Grad, Gabriela B.; Barrea, Raul A.; Figueroa, Rodolfo G.

2011-01-01

The L shell fluorescence cross-sections of the elements in range 45 ≤ Z ≤ 50 have been determined at 8 keV using Synchrotron radiation. The individual L X-ray photons, Ll, Lα, Lβ I , Lβ II , Lγ I and Lγ II produced in the target were measured with high resolution Si(Li) detector. The experimental set-up provided a low background by using linearly polarized monoenergetic photon beam, improving the signal-to-noise ratio. The experimental cross-sections obtained in this work were compared with available experimental data from Scofield Krause and Scofield and Puri et al. . These experimental values closely agree with the theoretical values calculated using Scofield and Krause data, except for the case of Lγ, where values measured of this work are slighter higher.

Porphyrin involvement in redshift fluorescence in dentin decay

Science.gov (United States)

Slimani, A.; Panayotov, I.; Levallois, B.; Cloitre, T.; Gergely, C.; Bec, N.; Larroque, C.; Tassery, H.; Cuisinier, F.

2014-05-01

The aim of this study was to evaluate the porphyrin involvement in the red fluorescence observed in dental caries with Soprolife® light-induced fluorescence camera in treatments mode (SOPRO, ACTEON Group, La Ciotat, France) and Vistacam® camera (DÜRR DENTAL AG, Bietigheim-Bissingen, Germany). The International Caries Detection and Assessment System (ICDAS) was used to rand the samples. Human teeth cross-sections, ranked from ICDAS score 0 to 6, were examined by epi-fluorescence microscopy and Confocal Raman microscopy. Comparable studies were done with Protoporphyrin IX, Porphyrin I and Pentosidine solutions. An RGB analysis of Soprolife® images was performed using ImageJ Software (1.46r, National Institutes of Health, USA). Fluorescence spectroscopy and MicroRaman spectroscopy revealed the presence of Protoporphyrin IX, in carious enamel, dentin and dental plaque. However, the presence of porphyrin I and pentosidine cannot be excluded. The results indicated that not only porphyrin were implicated in the red fluorescence, Advanced Glygation Endproducts (AGEs) of the Maillard reaction also contributed to this phenomenon.

Optical imaging of non-fluorescent nanoparticle probes in live cells

Energy Technology Data Exchange (ETDEWEB)

Wang, Gufeng; Stender, Anthony S.; Sun, Wei; and Fang, Ning

2009-12-17

Precise imaging of cellular and subcellular structures and dynamic processes in live cells is crucial for fundamental research in life sciences and in medical applications. Non-fluorescent nanoparticles are an important type of optical probe used in live-cell imaging due to their photostability, large optical cross-sections, and low toxicity. Here, we provide an overview of recent developments in the optical imaging of non-fluorescent nanoparticle probes in live cells.

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

Science.gov (United States)

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.

Study of x-ray fluorescence : Development in Geant4 of new models of cross sections for simulation PIXE. Biological and archaeological applications

International Nuclear Information System (INIS)

Ben Abdelouahed, Haifa

2010-01-01

a great number of times, that makes it possible to reproduce the macroscopic effects and to calculate sizes like, in our case, x-ray fluorescence. Among several tools available for Monte Carlo simulation of the interactions particle-matter, the tool for simulation Monte Carlo Geant4 is characterized by a particularly flexible architecture based on directed technology object. This is why, to build our code of simulation, our choice was fixed on the use of this tool. We treat in the first chapter the guiding principle of x-ray fluorescence, which leads us to present the various phenomena which limit its sensitivity of analysis. We devote the second chapter to the presentation of the Geant4 tool, and our tests of validation of its electromagnetic processes. We describe the performances and the limits of this tool as for the simulation of the cross sections. We will expose in this same chapter our development, in the Geant4 tool, of new models of calculation of the cross sections of ionization of the atoms by the protons and the particles alpha. We discuss also our method of validation of the models in question. This work was object of publication. It was taken into account by the Geant4 project: our development currently replaces the data base EEDL of Geant4 in the process of determination of the cross sections of ionization by charged particles, and makes thus functional the Geant4 tool in the simulation of the x-ray fluorescence induced by charged particles. We describe in the third chapter our use of the code worked out for the simulation of the effectiveness of absorption of the standard Si(Li) detector which constitutes the essential piece of our device of analysis by x-ray fluorescence. We expose the adopted approach which enabled us to optimize the geometrical parameters of the studied detector. This work was presented at the second international conference of spectroscopy. We present, in the fourth chapter, some examples of concrete applications on the study of

Variations in cardiovascular disease under-diagnosis in England: national cross-sectional spatial analysis

Directory of Open Access Journals (Sweden)

Walford Hannah

2011-03-01

Full Text Available Abstract Background There is under-diagnosis of cardiovascular disease (CVD in the English population, despite financial incentives to encourage general practices to register new cases. We compared the modelled (expected and diagnosed (observed prevalence of three cardiovascular conditions- coronary heart disease (CHD, hypertension and stroke- at local level, their geographical variation, and population and healthcare predictors which might influence diagnosis. Methods Cross-sectional observational study in all English local authorities (351 and general practices (8,372 comparing model-based expected prevalence with diagnosed prevalence on practice disease registers. Spatial analyses were used to identify geographic clusters and variation in regression relationships. Results A total of 9,682,176 patients were on practice CHD, stroke and transient ischaemic attack, and hypertension registers. There was wide spatial variation in observed: expected prevalence ratios for all three diseases, with less than five per cent of expected cases diagnosed in some areas. London and the surrounding area showed statistically significant discrepancies in observed: expected prevalence ratios, with observed prevalence much lower than the epidemiological models predicted. The addition of general practitioner supply as a variable yielded stronger regression results for all three conditions. Conclusions Despite almost universal access to free primary healthcare, there may be significant and highly variable under-diagnosis of CVD across England, which can be partially explained by persistent inequity in GP supply. Disease management studies should consider the possible impact of under-diagnosis on population health outcomes. Compared to classical regression modelling, spatial analytic techniques can provide additional information on risk factors for under-diagnosis, and can suggest where healthcare resources may be most needed.

Exploration in vivo by X-ray fluorescence (thyroid-brain)

International Nuclear Information System (INIS)

Delcroix, V.; Allemand, R.; Laval, M.; Dipaola, M.; Tubiana, M.

1975-01-01

X-ray fluorescence methods of medical exploration avoid the use of radioactive tracers and hence reduce the total dose received by the patient. In addition the collimation to the excitation source and detector respectively produces a tomographic effect which improves the spatial resolution of the system and even allows organs to be charted. The physical principles involved in X-ray fluorescence are outlined, with emphasis on the fact that the only elements useful for such applications are those of high enough atomic number to emit a fluorescence radiation of energy sufficient to pass through the tissues. The apparatus used, the excitation sources (radioactive source or X-ray tube), the detector and the measurement equipment are described. The experimental results obtained are given in two fields: measurement of blood flow in the tissues; thyroid imagery [fr

Fluorescence Molecular Tomography: Principles and Potential for Pharmaceutical Research

Directory of Open Access Journals (Sweden)

Florian Stuker

2011-04-01

Full Text Available 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

Evaluation of CDOM sources and their links with water quality in the lakes of Northeast China using fluorescence spectroscopy

Science.gov (United States)

Zhao, Ying; Song, Kaishan; Wen, Zhidan; Fang, Chong; Shang, Yingxin; Lv, Lili

2017-07-01

The spatial distributions of the fluorescence intensities Fmax for chromophoric dissolved organic matter (CDOM) components, the fluorescence indices (FI370 and FI310) and their correlations with water quality of 19 lakes in the Songhua River Basin (SHRB) across semiarid regions of Northeast China were examined with the data collected in September 2012 and 2015. The 19 lakes were divided into two groups according to EC (threshold value = 800 μS cm-1): fresh water (N = 13) and brackish water lakes (N = 6). The fluorescent characteristics of CDOM in the 19 lakes were investigated using excitation-emission matrix fluorescence spectroscopy (EEM) coupled with parallel factor (PARAFAC) and multivariate analysis. Two humic-like components (C1 and C3), one tryptophan-like component (C2), and one tyrosine-like component (C4) were identified by PARAFAC. The component C4 was not included in subsequent analyses due to the strong scatter in some colloidal water samples from brackish water lakes. The correlations between Fmax for the three EEM-PARAFAC extracted CDOM components C1-C3, the fluorescence indices (FI370 and FI310) and the water quality parameters (i.e., TN, TP, Chl-a, pH, EC, turbidity (Turb) and dissolved organic carbon (DOC)) were determined by redundancy analysis (RDA). The results of RDA analysis showed that spatial variation in land cover, pollution sources, and salinity/EC gradients in water quality affected Fmax for the fluorescent components C1-C3 and the fluorescence indices (FI370 and FI310). Further examination indicated that the CDOM fluorescent components and the fluorescence indices (FI370 and FI310) did not significantly differ (t-test, p > 0.05) in fresh water (N = 13) and brackish water lakes (N = 6). There was a difference in the distribution of the average Fmax for the CDOM fluorescent components between C1 to C3 from agricultural sources and urban wastewater sources in hypereutrophic brackish water lakes. The Fmax for humic-like components C1 and

TU-G-207-03: High Spatial Resolution and High Sensitivity X-Ray Fluorescence Imaging

International Nuclear Information System (INIS)

Xing, L.

2015-01-01

Last few years has witnessed the development of novel of X-ray imaging modalities, such as spectral CT, phase contrast CT, and X-ray acoustic/fluorescence/luminescence imaging. This symposium will present the recent advances of these emerging X-ray imaging modalities and update the attendees with knowledge in various related topics, including X-ray photon-counting detectors, X-ray physics underlying the emerging applications beyond the traditional X-ray imaging, image reconstruction for the novel modalities, characterization and evaluation of the systems, and their practical implications. In addition, the concept and practical aspects of X-ray activatable targeted nanoparticles for molecular X-ray imaging will be discussed in the context of X-ray fluorescence and luminescence CT. Learning Objectives: Present background knowledge of various emerging X-ray imaging techniques, such as spectral CT, phase contrast CT and X-ray fluorescence/luminescence CT. Discuss the practical need, technical aspects and current status of the emerging X-ray imaging modalities. Describe utility and future impact of the new generation of X-ray imaging applications

Size effects in the quantum yield of Cd Te quantum dots for optimum fluorescence bioimaging

International Nuclear Information System (INIS)

Jacinto, C.; Rocha, U.S.; Maestro, L.M.; Garcia-Sole, J.; Jaque, D.

2011-01-01

Full text: Semiconductor nano-crystals, usually referred as Quantum Dots (QDs) are nowadays regarded as one of the building-blocks in modern photonics. They constitute bright and photostable fluorescence sources whose emission and absorption properties can be adequately tailored through their size. Recent advances on the controlled modification of their surface has made possible the development of water soluble QDs, without causing any deterioration in their fluorescence properties. This has made them excellent optical selective markers to be used in fluorescence bio-imaging experiments. The suitability of colloidal QDs for bio-imaging is pushed forward by their large two-photon absorption cross section so that their visible luminescence (associated to the recombination of electro-hole pairs) can be also efficiently excited under infrared excitation (two-photon excitation). This, in turns, allows for large penetration depths in tissues, minimization of auto-fluorescence and achievement of superior spatial imaging resolution. In addition, recent works have demonstrated the ability of QDs to act as nano-thermometers based on the thermal sensitivity of their fluorescence bands. Based on all these outstanding properties, QDs have been successfully used to mark individual receptors in cell membranes, to intracellular temperature measurements and to label living embryos at different stages. Most of the QD based bio-images reported up to now were obtained by using whether CdSe or CdTe QDs since both are currently commercial available with a high degree of quality. They show similar fluorescence properties and optical performance when used in bio-imaging. Nevertheless, CdTe-QDs have very recently attracted much attention since the hyper-thermal sensitivity of their fluorescence bands was discovered. Based on this, it has been postulated that intracellular thermal sensing with resolutions as large as 0.25 deg C can be achieved based on CdTe-QDs, three times better than

Experimental determination of L X-ray fluorescence cross sections for elements with 45 < Z < 50 at 10 keV

Directory of Open Access Journals (Sweden)

Edgardo Bonzi

2014-02-01

Full Text Available Synchrotron radiation at 10 keV was used to experimentally determine the Ll, L, LI, LII, LI and LII fluorescence cross sections for elements with 45 < Z < 50, as part of an ongoing investigation at low energies. The measured data were compared with calculated values obtained using coefficients from Scofield, Krause and Puri et al. Received: 19 December 2013, Accepted: 20 January 2014; Reviewed by: J. P. Marques, Departamento de Física, Centro de Física Atómica, Fac. de Ciências, Universidade de Lisboa, Portugal.; Edited by: P. Weck; DOI: http://dx.doi.org/10.4279/PIP.060001 Cite as: E V Bonzi, G B Grad, R A Barrea, Papers in Physics 6, 060001 (2014

Global monitoring of terrestrial chlorophyll fluorescence from moderate-spectral-resolution near-infrared satellite measurements: methodology, simulations, and application to GOME-2

Directory of Open Access Journals (Sweden)

J. Joiner

2013-10-01

Full Text Available Globally mapped terrestrial chlorophyll fluorescence retrievals are of high interest because they can provide information on the functional status of vegetation including light-use efficiency and global primary productivity that can be used for global carbon cycle modeling and agricultural applications. Previous satellite retrievals of fluorescence have relied solely upon the filling-in of solar Fraunhofer lines that are not significantly affected by atmospheric absorption. Although these measurements provide near-global coverage on a monthly basis, they suffer from relatively low precision and sparse spatial sampling. Here, we describe a new methodology to retrieve global far-red fluorescence information; we use hyperspectral data with a simplified radiative transfer model to disentangle the spectral signatures of three basic components: atmospheric absorption, surface reflectance, and fluorescence radiance. An empirically based principal component analysis approach is employed, primarily using cloudy data over ocean, to model and solve for the atmospheric absorption. Through detailed simulations, we demonstrate the feasibility of the approach and show that moderate-spectral-resolution measurements with a relatively high signal-to-noise ratio can be used to retrieve far-red fluorescence information with good precision and accuracy. The method is then applied to data from the Global Ozone Monitoring Instrument 2 (GOME-2. The GOME-2 fluorescence retrievals display similar spatial structure as compared with those from a simpler technique applied to the Greenhouse gases Observing SATellite (GOSAT. GOME-2 enables global mapping of far-red fluorescence with higher precision over smaller spatial and temporal scales than is possible with GOSAT. Near-global coverage is provided within a few days. We are able to show clearly for the first time physically plausible variations in fluorescence over the course of a single month at a spatial resolution of 0

Quantitative Fluorescence Sensing Through Highly Autofluorescent, Scattering, and Absorbing Media Using Mobile Microscopy

KAUST Repository

Gö rö cs, Zoltá n; Rivenson, Yair; Ceylan Koydemir, Hatice; Tseng, Derek; Troy, Tamara L.; Demas, Vasiliki; Ozcan, Aydogan

2016-01-01

Compact and cost-effective systems for in vivo fluorescence and near-infrared imaging in combination with activatable reporters embedded inside the skin to sample interstitial fluid or blood can enable a variety of biomedical applications. However, the strong autofluorescence of human skin creates an obstacle for fluorescence-based sensing. Here we introduce a method for quantitative fluorescence sensing through highly autofluorescent, scattering, and absorbing media. For this, we created a compact and cost-effective fluorescence microscope weighing <40 g and used it to measure various concentrations of a fluorescent dye embedded inside a tissue phantom, which was designed to mimic the optical characteristics of human skin. We used an elliptical Gaussian beam excitation to digitally separate tissue autofluorescence from target fluorescence, although they severely overlap in both space and optical spectrum. Using ∼10-fold less excitation intensity than the safety limit for skin radiation exposure, we successfully quantified the density of the embedded fluorophores by imaging the skin phantom surface and achieved a detection limit of ∼5 × 105 and ∼2.5 × 107 fluorophores within ∼0.01 μL sample volume that is positioned 0.5 and 2 mm below the phantom surface, corresponding to a concentration of 105.9 pg/mL and 5.3 ng/mL, respectively. We also confirmed that this approach can track the spatial misalignments of the mobile microscope with respect to the embedded target fluorescent volume. This wearable microscopy platform might be useful for designing implantable biochemical sensors with the capability of spatial multiplexing to continuously monitor a panel of biomarkers and chronic conditions even at patients’ home.

Quantitative Fluorescence Sensing Through Highly Autofluorescent, Scattering, and Absorbing Media Using Mobile Microscopy

KAUST Repository

Göröcs, Zoltán

2016-09-13

Compact and cost-effective systems for in vivo fluorescence and near-infrared imaging in combination with activatable reporters embedded inside the skin to sample interstitial fluid or blood can enable a variety of biomedical applications. However, the strong autofluorescence of human skin creates an obstacle for fluorescence-based sensing. Here we introduce a method for quantitative fluorescence sensing through highly autofluorescent, scattering, and absorbing media. For this, we created a compact and cost-effective fluorescence microscope weighing <40 g and used it to measure various concentrations of a fluorescent dye embedded inside a tissue phantom, which was designed to mimic the optical characteristics of human skin. We used an elliptical Gaussian beam excitation to digitally separate tissue autofluorescence from target fluorescence, although they severely overlap in both space and optical spectrum. Using ∼10-fold less excitation intensity than the safety limit for skin radiation exposure, we successfully quantified the density of the embedded fluorophores by imaging the skin phantom surface and achieved a detection limit of ∼5 × 105 and ∼2.5 × 107 fluorophores within ∼0.01 μL sample volume that is positioned 0.5 and 2 mm below the phantom surface, corresponding to a concentration of 105.9 pg/mL and 5.3 ng/mL, respectively. We also confirmed that this approach can track the spatial misalignments of the mobile microscope with respect to the embedded target fluorescent volume. This wearable microscopy platform might be useful for designing implantable biochemical sensors with the capability of spatial multiplexing to continuously monitor a panel of biomarkers and chronic conditions even at patients’ home.

Spatial heterogeneity in active chlorophyll fluorescence and PSII activity of coral tissues

DEFF Research Database (Denmark)

Ralph, P.J.; Gademann, R.; Larkum, A.W.D.

2002-01-01

Chlorophyll-a fluorescence was measured in six species of coral, using pulse-amplitude-modulated fluorometers employing fibre-optic probes with diameters of 8 mm, 1 mm and 140 µm. The 8-mm probe integrated responses over a large area, giving more weight to coenosarc than polyp tissue for Acropora...

Very High Spectral Resolution Imaging Spectroscopy: the Fluorescence Explorer (FLEX) Mission

Science.gov (United States)

Moreno, Jose F.; Goulas, Yves; Huth, Andreas; Middleton, Elizabeth; Miglietta, Franco; Mohammed, Gina; Nedbal, Ladislav; Rascher, Uwe; Verhoef, Wouter; Drusch, Matthias

2016-01-01

The Fluorescence Explorer (FLEX) mission has been recently selected as the 8th Earth Explorer by the European Space Agency (ESA). It will be the first mission specifically designed to measure from space vegetation fluorescence emission, by making use of very high spectral resolution imaging spectroscopy techniques. Vegetation fluorescence is the best proxy to actual vegetation photosynthesis which can be measurable from space, allowing an improved quantification of vegetation carbon assimilation and vegetation stress conditions, thus having key relevance for global mapping of ecosystems dynamics and aspects related with agricultural production and food security. The FLEX mission carries the FLORIS spectrometer, with a spectral resolution in the range of 0.3 nm, and is designed to fly in tandem with Copernicus Sentinel-3, in order to provide all the necessary spectral / angular information to disentangle emitted fluorescence from reflected radiance, and to allow proper interpretation of the observed fluorescence spatial and temporal dynamics.

Chlorophyll induced fluorescence retrieved from GOME2 for improving gross primary productivity estimates of vegetation

Science.gov (United States)

van Leth, Thomas C.; Verstraeten, Willem W.; Sanders, Abram F. J.

2014-05-01

Mapping terrestrial chlorophyll fluorescence is a crucial activity to obtain information on the functional status of vegetation and to improve estimates of light-use efficiency (LUE) and global primary productivity (GPP). GPP quantifies carbon fixation by plant ecosystems and is therefore an important parameter for budgeting terrestrial carbon cycles. Satellite remote sensing offers an excellent tool for investigating GPP in a spatially explicit fashion across different scales of observation. The GPP estimates, however, still remain largely uncertain due to biotic and abiotic factors that influence plant production. Sun-induced fluorescence has the ability to enhance our knowledge on how environmentally induced changes affect the LUE. This can be linked to optical derived remote sensing parameters thereby reducing the uncertainty in GPP estimates. Satellite measurements provide a relatively new perspective on global sun-induced fluorescence, enabling us to quantify spatial distributions and changes over time. Techniques have recently been developed to retrieve fluorescence emissions from hyperspectral satellite measurements. We use data from the Global Ozone Monitoring Instrument 2 (GOME2) to infer terrestrial fluorescence. The spectral signatures of three basic components atmospheric: absorption, surface reflectance, and fluorescence radiance are separated using reference measurements of non-fluorescent surfaces (desserts, deep oceans and ice) to solve for the atmospheric absorption. An empirically based principal component analysis (PCA) approach is applied similar to that of Joiner et al. (2013, ACP). Here we show our first global maps of the GOME2 retrievals of chlorophyll fluorescence. First results indicate fluorescence distributions that are similar with that obtained by GOSAT and GOME2 as reported by Joiner et al. (2013, ACP), although we find slightly higher values. In view of optimizing the fluorescence retrieval, we will show the effect of the references

Lead tolerance and cellular distribution in Elsholtzia splendens using synchrotron radiation micro-X-ray fluorescence

Energy Technology Data Exchange (ETDEWEB)

Zhang, Jie [MOE Key Laboratory of Environment Remediation and Ecosystem Health, College of Environmental and Resource Sciences, Zhejiang University, Hangzhou 310058 (China); Tian, Shengke [MOE Key Laboratory of Environment Remediation and Ecosystem Health, College of Environmental and Resource Sciences, Zhejiang University, Hangzhou 310058 (China); University of Florida, Institute of Food and Agricultural Science, Indian River Research and Education Center, Fort Pierce, FL 34945 (United States); Lu, Lingli; Shohag, M.J.I.; Liao, Haibing [MOE Key Laboratory of Environment Remediation and Ecosystem Health, College of Environmental and Resource Sciences, Zhejiang University, Hangzhou 310058 (China); Yang, Xiaoe, E-mail: xyang@zju.edu.cn [MOE Key Laboratory of Environment Remediation and Ecosystem Health, College of Environmental and Resource Sciences, Zhejiang University, Hangzhou 310058 (China)

2011-12-15

Highlights: Black-Right-Pointing-Pointer Elsholtzia splendens had a good ability of lead tolerance and accumulation. Black-Right-Pointing-Pointer Pb was mostly restricted to the vascular bundles and epidermis tissues. Black-Right-Pointing-Pointer Pb and Ca shared most similar distribution patterns in E. splendens. - Abstract: Hydroponic experiments were conducted to investigate the tolerance and spatial distribution of lead (Pb) in Elsholtzia splendens-a copper (Cu) accumulator plant using synchrotron-based micro-X-ray fluorescence. According to chlorophyll concentration and chlorophyll fluorescence parameters, E. splendens displayed certain tolerance at 100 {mu}M Pb treatment. Lead concentration in roots, stems and leaves of E. splendens reached 45,183.6, 1657.6, and 380.9 mg kg{sup -1}, respectively. Pb was mostly accumulated in the roots, and there were also high concentrations of Pb been transported into stems and leaves. Micro-XRF analysis of the stem and leaf cross section revealed that Pb was mostly restricted in the vascular bundles and epidermis tissues of both stem and leaf of E. splendens. The correlation between distribution of K, Ca, Zn and Pb were analyzed. There were significant positive correlations (P < 0.01) among Pb and Ca, K, Zn distribution both in stem and leaf of E. splendens. However, among the three elements, Ca shared the most similar distribution pattern and the highest correlation coefficients with Pb in both stem and leaf cross section of E. splendens. This suggests that Ca may play an important role in Pb accumulation in stem and leaf of E. splendens.

Snapshot imaging Fraunhofer line discriminator for detection of plant fluorescence

Science.gov (United States)

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.

Site-specific confocal fluorescence imaging of biological microstructures in a turbid medium

International Nuclear Information System (INIS)

Saloma, Caesar; Palmes-Saloma, Cynthia; Kondoh, Hisato

1998-01-01

Normally transparent biological structures in a turbid medium are imaged using a laser confocal microscope and multiwavelength site-specific fluorescence labelling. The spatial filtering capability of the detector pinhole in the confocal microscope limits the number of scattered fluorescent photons that reach the photodetector. Simultaneous application of different fluorescent markers on the same sample site minimizes photobleaching by reducing the excitation time for each marker. A high-contrast grey-level image is also produced by summing confocal images of the same site taken at different fluorescence wavelengths. Monte Carlo simulations are performed to obtain the quantitative behaviour of confocal fluorescence imaging in turbid media. Confocal images of the following samples were also obtained: (i) 15 μm diameter fluorescent spheres placed 1.16 mm deep beneath an aqueous suspension of 0.0823 μm diameter polystyrene latex spheres, and (ii) hindbrain of a whole-mount mouse embryo (age 10 days) that was stained to fluoresce at 515 nm and 580 nm peak wavelengths. Expression of RNA transcripts of a gene within the embryo hindbrain was detected by a fluorescence-based whole-mount in situ hybridization procedure that we recently tested. (author)

Fluorophore appended saccharide cyclophane: self-association, fluorescent properties, heterodimers with cyclodextrins, and cross-linking behavior with peanut agglutinin of dansyl-modified saccharide cyclophane.

Science.gov (United States)

Hayashida, Osamu; Hamachi, Itaru

2004-05-14

(PNA), galactoside-binding lectin, which was readily monitored by a visible turbidity of the solution due to a cross-linking agglutination of these components, as well as by fluorescence spectroscopy.

Heat shock-induced interactions among nuclear HSFs detected by fluorescence cross-correlation spectroscopy

Energy Technology Data Exchange (ETDEWEB)

Pack, Chan-Gi, E-mail: changipack@amc.seoul.kr [Asan Institute for Life Sciences, University of Ulsan, College of Medicine, Asan Medical Center, Seoul 138-736 (Korea, Republic of); Ahn, Sang-Gun [Dept. of Pathology, College of Dentistry, Chosun University, Seosuk-dong, Dong-gu, Gwangju 501-759 (Korea, Republic of)

2015-07-31

The cellular response to stress is primarily controlled in cells via transcriptional activation by heat shock factor 1 (HSF1). HSF1 is well-known to form homotrimers for activation upon heat shock and subsequently bind to target DNAs, such as heat-shock elements, by forming stress granules. A previous study demonstrated that nuclear HSF1 and HSF2 molecules in live cells interacted with target DNAs on the stress granules. However, the process underlying the binding interactions of HSF family in cells upon heat shock remains unclear. This study demonstrate for the first time that the interaction kinetics among nuclear HSF1, HSF2, and HSF4 upon heat shock can be detected directly in live cells using dual color fluorescence cross-correlation spectroscopy (FCCS). FCCS analyses indicated that the binding between HSFs was dramatically changed by heat shock. Interestingly, the recovery kinetics of interaction between HSF1 molecules after heat shock could be represented by changes in the relative interaction amplitude and mobility. - Highlights: • The binding interactions among nuclear HSFs were successfully detected. • The binding kinetics between HSF1s during recovery was quantified. • HSF2 and HSF4 strongly formed hetero-complex, even before heat shock. • Nuclear HSF2 and HSF4 bound to HSF1 only after heat shock.

Sexual Orientation-Related Differences in Virtual Spatial Navigation and Spatial Search Strategies.

Science.gov (United States)

Rahman, Qazi; Sharp, Jonathan; McVeigh, Meadhbh; Ho, Man-Ling

2017-07-01

Spatial abilities are generally hypothesized to differ between men and women, and people with different sexual orientations. According to the cross-sex shift hypothesis, gay men are hypothesized to perform in the direction of heterosexual women and lesbian women in the direction of heterosexual men on cognitive tests. This study investigated sexual orientation differences in spatial navigation and strategy during a virtual Morris water maze task (VMWM). Forty-four heterosexual men, 43 heterosexual women, 39 gay men, and 34 lesbian/bisexual women (aged 18-54 years) navigated a desktop VMWM and completed measures of intelligence, handedness, and childhood gender nonconformity (CGN). We quantified spatial learning (hidden platform trials), probe trial performance, and cued navigation (visible platform trials). Spatial strategies during hidden and probe trials were classified into visual scanning, landmark use, thigmotaxis/circling, and enfilading. In general, heterosexual men scored better than women and gay men on some spatial learning and probe trial measures and used more visual scan strategies. However, some differences disappeared after controlling for age and estimated IQ (e.g., in visual scanning heterosexual men differed from women but not gay men). Heterosexual women did not differ from lesbian/bisexual women. For both sexes, visual scanning predicted probe trial performance. More feminine CGN scores were associated with lower performance among men and greater performance among women on specific spatial learning or probe trial measures. These results provide mixed evidence for the cross-sex shift hypothesis of sexual orientation-related differences in spatial cognition.

Performance of a two-leaf light use efficiency model for mapping gross primary productivity against remotely sensed sun-induced chlorophyll fluorescence data.

Science.gov (United States)

Zan, Mei; Zhou, Yanlian; Ju, Weimin; Zhang, Yongguang; Zhang, Leiming; Liu, Yibo

2018-02-01

Estimating terrestrial gross primary production is an important task when studying the carbon cycle. In this study, the ability of a two-leaf light use efficiency model to simulate regional gross primary production in China was validated using satellite Global Ozone Monitoring Instrument - 2 sun-induced chlorophyll fluorescence data. The two-leaf light use efficiency model was used to estimate daily gross primary production in China's terrestrial ecosystems with 500-m resolution for the period from 2007 to 2014. Gross primary production simulated with the two-leaf light use efficiency model was resampled to a spatial resolution of 0.5° and then compared with sun-induced chlorophyll fluorescence. During the study period, sun-induced chlorophyll fluorescence and gross primary production simulated by the two-leaf light use efficiency model exhibited similar spatial and temporal patterns in China. The correlation coefficient between sun-induced chlorophyll fluorescence and monthly gross primary production simulated by the two-leaf light use efficiency model was significant (pproduction simulated by the two-leaf light use efficiency model were similar in spring and autumn in most vegetated regions, but dissimilar in winter and summer. The spatial variability of sun-induced chlorophyll fluorescence and gross primary production simulated by the two-leaf light use efficiency model was similar in spring, summer, and autumn. The proportion of spatial variations of sun-induced chlorophyll fluorescence and annual gross primary production simulated by the two-leaf light use efficiency model explained by ranged from 0.76 (2011) to 0.80 (2013) during the study period. Overall, the two-leaf light use efficiency model was capable of capturing spatial and temporal variations in gross primary production in China. However, the model needs further improvement to better simulate gross primary production in summer. Copyright © 2017 Elsevier B.V. All rights reserved.

A novel fluorescent in situ hybridization technique for detection of Rickettsia spp. in archival samples

DEFF Research Database (Denmark)

Svendsen, Claus Bo; Boye, Mette; Struve, Carsten

2009-01-01

A novel, sensitive and specific method for detecting Rickettsia spp. in archival samples is described. The method involves the use of fluorescently marked oligonucleotide probes for in situ hybridization. Specific hybridization of Ricekttsia was found without problems of cross-reactions with bact......A novel, sensitive and specific method for detecting Rickettsia spp. in archival samples is described. The method involves the use of fluorescently marked oligonucleotide probes for in situ hybridization. Specific hybridization of Ricekttsia was found without problems of cross...

In vivo tomographic imaging with fluorescence and MRI using tumor-targeted dual-labeled nanoparticles

Directory of Open Access Journals (Sweden)

Zhang Y

2013-12-01

Full Text Available Yue Zhang,1 Bin Zhang,1 Fei Liu,1,2 Jianwen Luo,1,3 Jing Bai1 1Department of Biomedical Engineering, School of Medicine, 2Tsinghua-Peking Center for Life Sciences, 3Center for Biomedical Imaging Research, Tsinghua University, Beijing, People's Republic of China Abstract: Dual-modality imaging combines the complementary advantages of different modalities, and offers the prospect of improved preclinical research. The combination of fluorescence imaging and magnetic resonance imaging (MRI provides cross-validated information and direct comparison between these modalities. Here, we report on the application of a novel tumor-targeted, dual-labeled nanoparticle (NP, utilizing iron oxide as the MRI contrast agent and near infrared (NIR dye Cy5.5 as the fluorescent agent. Results of in vitro experiments verified the specificity of the NP to tumor cells. In vivo tumor targeting and uptake of the NPs in a mouse model were visualized by fluorescence and MR imaging collected at different time points. Quantitative analysis was carried out to evaluate the efficacy of MRI contrast enhancement. Furthermore, tomographic images were also acquired using both imaging modalities and cross-validated information of tumor location and size between these two modalities was revealed. The results demonstrate that the use of dual-labeled NPs can facilitate the dual-modal detection of tumors, information cross-validation, and direct comparison by combing fluorescence molecular tomography (FMT and MRI. Keywords: dual-modality, fluorescence molecular tomography (FMT, magnetic resonance imaging (MRI, nanoparticle

Three-dimensional fluorescence analysis of chernozem humic acids and their electrophoretic fractions

Science.gov (United States)

Trubetskoi, O. A.; Trubetskaya, O. E.

2017-09-01

Polyacrylamide gel electrophoresis in combination with size-exclusion chromatography (SEC-PAGE) has been used to obtain stable electrophoretic fractions of different molecular size (MS) from chernozem humic acids (HAs). Three-dimensional fluorescence charts of chernozem HAs and their fractions have been obtained for the first time, and all fluorescence excitation-emission maxima have been identified in the excitation wavelength range of 250-500 nm. It has been found that fractionation by the SEC-PAGE method results in a nonuniform distribution of protein- and humin-like fluorescence of the original HA preparation among the electrophoretic fractions. The electrophoretic fractions of the highest and medium MSs have only the main protein-like fluorescence maximum and traces of humin-like fluorescence. In the electrophoretic fraction of the lowest MS, the intensity of protein-like fluorescence is low, but the major part of humin-like fluorescence is localized there. Relationships between the intensity of protein-like fluorescence and the weight distribution of amino acids have been revealed, as well as between the degree of aromaticity and the intensity of humin-like fluorescence in electrophoretic fractions of different MSs. The obtained relationships can be useful in the interpretation of the spatial structural organization and ecological functions of soil HAs.

Plasmonically amplified fluorescence bioassay with microarray format

Science.gov (United States)

Gogalic, S.; Hageneder, S.; Ctortecka, C.; Bauch, M.; Khan, I.; Preininger, Claudia; Sauer, U.; Dostalek, J.

2015-05-01

Plasmonic amplification of fluorescence signal in bioassays with microarray detection format is reported. A crossed relief diffraction grating was designed to couple an excitation laser beam to surface plasmons at the wavelength overlapping with the absorption and emission bands of fluorophore Dy647 that was used as a label. The surface of periodically corrugated sensor chip was coated with surface plasmon-supporting gold layer and a thin SU8 polymer film carrying epoxy groups. These groups were employed for the covalent immobilization of capture antibodies at arrays of spots. The plasmonic amplification of fluorescence signal on the developed microarray chip was tested by using interleukin 8 sandwich immunoassay. The readout was performed ex situ after drying the chip by using a commercial scanner with high numerical aperture collecting lens. Obtained results reveal the enhancement of fluorescence signal by a factor of 5 when compared to a regular glass chip.

Preparation of Rhodamine B Fluorescent Poly(methacrylic acid) Coated Gelatin Nanoparticles

OpenAIRE

Gan, Zhenhai; Ju, Jianhui; Zhang, Ting; Wu, Daocheng

2011-01-01

Poly(methacrylic acid) (PMAA)-coated gelatin nanoparticles encapsulated with fluorescent dye rhodamine B were prepared by the coacervation method with the aim to retard the release of rhodamine B from the gelatin matrix. With sodium sulfate as coacervation reagent for gelatin, a kind of biopolymer with excellent biocompatibility, the formed gelatin nanoparticles were cross-linked by formaldehyde followed by the polymerization of methacrylic acid coating. The fluorescent poly(methacrylic acid)...

Spatial correlation between weed species densities and soil properties

DEFF Research Database (Denmark)

Walter, Mette; Christensen, Svend; Simmelsgaard, Svend Erik

2002-01-01

The spatial cross-correlation between weed species densities and six soil properties within fields was analysed using cross-semivariograms. The survey was carried out in three successive years in two fields. The most consistent relationship between weed species density (numbers m−2) and soil...... properties was negative cross-correlation between the density of Viola arvensis Murray and clay content. This correlation was found in both fields; however, the range of spatial dependence varied between fields. In one of the fields, the density of Lamium purpureum L. was positively cross......-correlated with the phosphorus content in the soil in all years. The density of Veronica spp. and Poa annua L. was negatively cross-correlated with pH in all three years. Other spatial cross-correlations that were found in this study were inconsistent over time or field site. The densities of some of the weed species were...

A portable fluorescence microscopic imaging system for cholecystectomy

Science.gov (United States)

Ye, Jian; Yang, Chaoyu; Gan, Qi; Ma, Rong; Zhang, Zeshu; Chang, Shufang; Shao, Pengfei; Zhang, Shiwu; Liu, Chenhai; Xu, Ronald

2016-03-01

In this paper we proposed a portable fluorescence microscopic imaging system to prevent iatrogenic biliary injuries from occurring during cholecystectomy due to misidentification of the cystic structures. The system consisted of a light source module, a CMOS camera, a Raspberry Pi computer and a 5 inch HDMI LCD. Specifically, the light source module was composed of 690 nm and 850 nm LEDs, allowing the CMOS camera to simultaneously acquire both fluorescence and background images. The system was controlled by Raspberry Pi using Python programming with the OpenCV library under Linux. We chose Indocyanine green(ICG) as a fluorescent contrast agent and then tested fluorescence intensities of the ICG aqueous solution at different concentration levels by our fluorescence microscopic system compared with the commercial Xenogen IVIS system. The spatial resolution of the proposed fluorescence microscopic imaging system was measured by a 1951 USAF resolution target and the dynamic response was evaluated quantitatively with an automatic displacement platform. Finally, we verified the technical feasibility of the proposed system in mouse models of bile duct, performing both correct and incorrect gallbladder resection. Our experiments showed that the proposed system can provide clear visualization of the confluence between the cystic duct and common bile duct or common hepatic duct, suggesting that this is a potential method for guiding cholecystectomy. The proposed portable system only cost a total of $300, potentially promoting its use in resource-limited settings.

FLIMX: A Software Package to Determine and Analyze the Fluorescence Lifetime in Time-Resolved Fluorescence Data from the Human Eye.

Directory of Open Access Journals (Sweden)

Matthias Klemm

Full Text Available Fluorescence lifetime imaging ophthalmoscopy (FLIO is a new technique for measuring the in vivo autofluorescence intensity decays generated by endogenous fluorophores in the ocular fundus. Here, we present a software package called FLIM eXplorer (FLIMX for analyzing FLIO data. Specifically, we introduce a new adaptive binning approach as an optimal tradeoff between the spatial resolution and the number of photons required per pixel. We also expand existing decay models (multi-exponential, stretched exponential, spectral global analysis, incomplete decay to account for the layered structure of the eye and present a method to correct for the influence of the crystalline lens fluorescence on the retina fluorescence. Subsequently, the Holm-Bonferroni method is applied to FLIO measurements to allow for group comparisons between patients and controls on the basis of fluorescence lifetime parameters. The performance of the new approaches was evaluated in five experiments. Specifically, we evaluated static and adaptive binning in a diabetes mellitus patient, we compared the different decay models in a healthy volunteer and performed a group comparison between diabetes patients and controls. An overview of the visualization capabilities and a comparison of static and adaptive binning is shown for a patient with macular hole. FLIMX's applicability to fluorescence lifetime imaging microscopy is shown in the ganglion cell layer of a porcine retina sample, obtained by a laser scanning microscope using two-photon excitation.

FLIMX: A Software Package to Determine and Analyze the Fluorescence Lifetime in Time-Resolved Fluorescence Data from the Human Eye.

Science.gov (United States)

Klemm, Matthias; Schweitzer, Dietrich; Peters, Sven; Sauer, Lydia; Hammer, Martin; Haueisen, Jens

2015-01-01

Fluorescence lifetime imaging ophthalmoscopy (FLIO) is a new technique for measuring the in vivo autofluorescence intensity decays generated by endogenous fluorophores in the ocular fundus. Here, we present a software package called FLIM eXplorer (FLIMX) for analyzing FLIO data. Specifically, we introduce a new adaptive binning approach as an optimal tradeoff between the spatial resolution and the number of photons required per pixel. We also expand existing decay models (multi-exponential, stretched exponential, spectral global analysis, incomplete decay) to account for the layered structure of the eye and present a method to correct for the influence of the crystalline lens fluorescence on the retina fluorescence. Subsequently, the Holm-Bonferroni method is applied to FLIO measurements to allow for group comparisons between patients and controls on the basis of fluorescence lifetime parameters. The performance of the new approaches was evaluated in five experiments. Specifically, we evaluated static and adaptive binning in a diabetes mellitus patient, we compared the different decay models in a healthy volunteer and performed a group comparison between diabetes patients and controls. An overview of the visualization capabilities and a comparison of static and adaptive binning is shown for a patient with macular hole. FLIMX's applicability to fluorescence lifetime imaging microscopy is shown in the ganglion cell layer of a porcine retina sample, obtained by a laser scanning microscope using two-photon excitation.

In Vivo Imaging of Far-red Fluorescent Proteins after DNA Electrotransfer to Muscle Tissue

DEFF Research Database (Denmark)

Hojman, Pernille; Eriksen, Jens; Gehl, Julie

2009-01-01

DNA electrotransfer to muscle tissue yields long-term, high levels of gene expression; showing great promise for future gene therapy. We want to characterize the novel far-red fluorescent protein Katushka as a marker for gene expression using time domain fluorescence in vivo imaging. Highly...... weeks. Depth and 3D analysis proved that the expression was located in the target muscle. In vivo bio-imaging using the novel Katushka fluorescent protein enables excellent evaluation of the transfection efficacy, and spatial distribution, but lacks long-term stability....... efficient transgenic expression was observed after DNA electrotransfer with 100-fold increase in fluorescent intensity. The fluorescent signal peaked 1 week after transfection and returned to background level within 4 weeks. Katushka expression was not as stable as GFP expression, which was detectable for 8...

Nanostructural origin of blue fluorescence in the mineral karpatite.

Science.gov (United States)

Potticary, Jason; Jensen, Torsten T; Hall, Simon R

2017-08-29

The colour of crystals is a function of their atomic structure. In the case of organic crystals, it is the spatial relationships between molecules that determine the colour, so the same molecules in the same arrangement should produce crystals of the same colour, regardless of whether they arise geologically or synthetically. There is a naturally-occurring organic crystal known as karpatite which is prized for its beautiful blue fluorescence under ultra-violet illumination. When grown under laboratory conditions however, the crystals fluoresce with an intense green colour. For 20 years, this difference has been thought to be due to chemical impurities in the laboratory-grown material. Using electron microscopy coupled with fluorescence spectroscopy and X-Ray diffraction, we report here that this disparity is instead due to differences in the structure of the crystals at the nanoscale. The results show that in nature, karpatite has a nanotexture that is not present in the synthetic crystals, which enables different photonic pathways and therefore a blue, rather than green colour whilst undergoing fluorescence.

Neutron slowing down and transport in a medium of constant cross section. I. Spatial moments

International Nuclear Information System (INIS)

Cacuci, D.G.; Goldstein, H.

1977-01-01

Some aspects of the problem of neutron slowing down and transport have been investigated in an infinite medium consisting of a single nuclide scattering elastically and isotropically without absorption and with energy-independent cross sections. The method of singular eigenfunctions has been applied to the Boltzmann equation governing the Laplace transform (with respect to the lethargy variable) of the neutron flux. Formulas have been obtained for the lethargy dependent spatial moments of the scalar flux applicable in the limit of large lethargy. In deriving these formulas, use has been made of the well-known connection between the spatial moments of the Laplace-transformed scalar flux and the moments of the flux in the ''eigenvalue space.'' The calculations have been greatly aided by the construction of a closed general expression for these ''eigenvalue space'' moments. Extensive use has also been made of the methods of combinatorial analysis and of computer evaluation, via FORMAC, of complicated sequences of manipulations. It has been possible to obtain for materials of any atomic weight explicit corrections to the age theory formulas for the spatial moments M/sub 2n/(u), of the scalar flux, valid through terms of order of u -5 . Higher order correction terms could be obtained at the expense of additional computer time. The evaluation of the coefficients of the powers of n, as explicit functions of the nuclear mass, represent the end product of this investigation

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

Science.gov (United States)

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.

Calculation of the spatial resolution in two-photon absorption spectroscopy applied to plasma diagnosis

Energy Technology Data Exchange (ETDEWEB)

Garcia-Lechuga, M. [Departamento de Física Teórica, Atómica y Óptica, Universidad de Valladolid, 47011-Valladolid (Spain); Laser Processing Group, Instituto de Óptica “Daza de Valdés,” CSIC, 28006-Madrid (Spain); Fuentes, L. M. [Departamento de Física Aplicada, Universidad de Valladolid, 47011-Valladolid (Spain); Grützmacher, K.; Pérez, C., E-mail: concha@opt.uva.es; Rosa, M. I. de la [Departamento de Física Teórica, Atómica y Óptica, Universidad de Valladolid, 47011-Valladolid (Spain)

2014-10-07

We report a detailed characterization of the spatial resolution provided by two-photon absorption spectroscopy suited for plasma diagnosis via the 1S-2S transition of atomic hydrogen for optogalvanic detection and laser induced fluorescence (LIF). A precise knowledge of the spatial resolution is crucial for a correct interpretation of measurements, if the plasma parameters to be analysed undergo strong spatial variations. The present study is based on a novel approach which provides a reliable and realistic determination of the spatial resolution. Measured irradiance distribution of laser beam waists in the overlap volume, provided by a high resolution UV camera, are employed to resolve coupled rate equations accounting for two-photon excitation, fluorescence decay and ionization. The resulting three-dimensional yield distributions reveal in detail the spatial resolution for optogalvanic and LIF detection and related saturation due to depletion. Two-photon absorption profiles broader than the Fourier transform-limited laser bandwidth are also incorporated in the calculations. The approach allows an accurate analysis of the spatial resolution present in recent and future measurements.

Calculation of the spatial resolution in two-photon absorption spectroscopy applied to plasma diagnosis

International Nuclear Information System (INIS)

Garcia-Lechuga, M.; Fuentes, L. M.; Grützmacher, K.; Pérez, C.; Rosa, M. I. de la

2014-01-01

We report a detailed characterization of the spatial resolution provided by two-photon absorption spectroscopy suited for plasma diagnosis via the 1S-2S transition of atomic hydrogen for optogalvanic detection and laser induced fluorescence (LIF). A precise knowledge of the spatial resolution is crucial for a correct interpretation of measurements, if the plasma parameters to be analysed undergo strong spatial variations. The present study is based on a novel approach which provides a reliable and realistic determination of the spatial resolution. Measured irradiance distribution of laser beam waists in the overlap volume, provided by a high resolution UV camera, are employed to resolve coupled rate equations accounting for two-photon excitation, fluorescence decay and ionization. The resulting three-dimensional yield distributions reveal in detail the spatial resolution for optogalvanic and LIF detection and related saturation due to depletion. Two-photon absorption profiles broader than the Fourier transform-limited laser bandwidth are also incorporated in the calculations. The approach allows an accurate analysis of the spatial resolution present in recent and future measurements.

Laser induced fluorescence spectroscopy for FTU

International Nuclear Information System (INIS)

Hughes, T.P.

1995-07-01

Laser induced fluorescence spectroscopy (LIFS) is based on the absorption of a short pulse of tuned laser light by a group of atoms and the observation of the resulting fluorescence radiation from the excited state. Because the excitation is resonant it is very efficient, and the fluorescence can be many times brighter than the normal spontaneous emission, so low number densities of the selected atoms can be detected and measured. Good spatial resolution can be achieved by using a narrow laser beam. If the laser is sufficiently monochromatic, and it can be tuned over the absorption line profile of the selected atoms, information can also be obtained about the velocities of the atoms from the Doppler effect which can broaden and shift the line. In this report two topics are examined in detail. The first is the effect of high laser irradiance, which can cause 'power broadening' of the apparent absorption line profile. The second is the effect of the high magnetic field in FTU. Detailed calculations are given for LIFS of neutral iron and molybdenum atoms, including the Zeeman effect, and the implementation of LIFS for these atoms on FTU is discussed

PCR-free detection of genetically modified organisms using magnetic capture technology and fluorescence cross-correlation spectroscopy.

Directory of Open Access Journals (Sweden)

Xiaoming Zhou

2009-11-01

Full Text Available The safety of genetically modified organisms (GMOs has attracted much attention recently. Polymerase chain reaction (PCR amplification is a common method used in the identification of GMOs. However, a major disadvantage of PCR is the potential amplification of non-target DNA, causing false-positive identification. Thus, there remains a need for a simple, reliable and ultrasensitive method to identify and quantify GMO in crops. This report is to introduce a magnetic bead-based PCR-free method for rapid detection of GMOs using dual-color fluorescence cross-correlation spectroscopy (FCCS. The cauliflower mosaic virus 35S (CaMV35S promoter commonly used in transgenic products was targeted. CaMV35S target was captured by a biotin-labeled nucleic acid probe and then purified using streptavidin-coated magnetic beads through biotin-streptavidin linkage. The purified target DNA fragment was hybridized with two nucleic acid probes labeled respectively by Rhodamine Green and Cy5 dyes. Finally, FCCS was used to detect and quantify the target DNA fragment through simultaneously detecting the fluorescence emissions from the two dyes. In our study, GMOs in genetically engineered soybeans and tomatoes were detected, using the magnetic bead-based PCR-free FCCS method. A detection limit of 50 pM GMOs target was achieved and PCR-free detection of GMOs from 5 microg genomic DNA with magnetic capture technology was accomplished. Also, the accuracy of GMO determination by the FCCS method is verified by spectrophotometry at 260 nm using PCR amplified target DNA fragment from GM tomato. The new method is rapid and effective as demonstrated in our experiments and can be easily extended to high-throughput and automatic screening format. We believe that the new magnetic bead-assisted FCCS detection technique will be a useful tool for PCR-free GMOs identification and other specific nucleic acids.

Multi-color fluorescent DNA analysis in an integrated optofluidic lab on a chip

OpenAIRE

Dongre, C.

2010-01-01

Abstract: Sorting and sizing of DNA molecules within the human genome project has enabled the genetic mapping of various illnesses. Furthermore by employing tiny lab-on-a-chip device, integrated DNA sequencing and genetic diagnostics have become feasible. We present the combination of capillary electrophoresis with laser-induced fluorescence for optofluidic integration toward an on-chip bio-analysis tool. Integrated optical fluorescence excitation allows for a high spatial resolution (12 μm) ...

Two phase formation of massive elliptical galaxies: study through cross-correlation including spatial effect

Science.gov (United States)

Modak, Soumita; Chattopadhyay, Tanuka; Chattopadhyay, Asis Kumar

2017-11-01

Area of study is the formation mechanism of the present-day population of elliptical galaxies, in the context of hierarchical cosmological models accompanied by accretion and minor mergers. The present work investigates the formation and evolution of several components of the nearby massive early-type galaxies (ETGs) through cross-correlation function (CCF), using the spatial parameters right ascension (RA) and declination (DEC), and the intrinsic parameters mass (M_{*}) and size. According to the astrophysical terminology, here these variables, namely mass, size, RA and DEC are termed as parameters, whereas the unknown constants involved in the kernel function are called hyperparameters. Throughout this paper, the parameter size is used to represent the effective radius (Re). Following Huang et al. (2013a), each nearby ETG is divided into three parts on the basis of its Re value. We study the CCF between each of these three components of nearby massive ETGs and the ETGs in the high redshift range, 0.5conflict raised in a previous work (De et al. 2014) suggesting other possibilities for the formation of the outermost part. A probable cause of this improvement is the inclusion of the spatial effects in addition to the other parameters in the study.

A Fast Global Fitting Algorithm for Fluorescence Lifetime Imaging Microscopy Based on Image Segmentation

OpenAIRE

Pelet, S.; Previte, M.J.R.; Laiho, L.H.; So, P.T. C.

2004-01-01

Global fitting algorithms have been shown to improve effectively the accuracy and precision of the analysis of fluorescence lifetime imaging microscopy data. Global analysis performs better than unconstrained data fitting when prior information exists, such as the spatial invariance of the lifetimes of individual fluorescent species. The highly coupled nature of global analysis often results in a significantly slower convergence of the data fitting algorithm as compared with unconstrained ana...

Self-assembled fluorescent organic nanoparticles for live cell imaging

NARCIS (Netherlands)

Fischer, I.; Petkau, K.; Dorland, Y.L.; Schenning, A.P.H.J.; Brunsveld, L.

2013-01-01

Fluorescent, cell-permeable, organic nanoparticles based on self-assembled p-conjugated oligomers with high absorption cross-sections and high quantum yields have been developed. The nanoparticles are generated with a tuneable density of amino groups for charge-mediated cellular uptake by a

Fluorescence-enhanced gadolinium-doped zinc oxide quantum dots for magnetic resonance and fluorescence imaging.

Science.gov (United States)

Liu, Yanlan; Ai, Kelong; Yuan, Qinghai; Lu, Lehui

2011-02-01

We report here the development of Gd-doped ZnO quantum dots (QDs) as dual modal fluorescence and magnetic resonance imaging nanoprobes. They are fabricated in a simple, versatile and environmentally friendly method, not only decreasing the difficulty and complexity, but also avoiding the increase of particle's size brought about by silica coating procedure in the synthesis of nanoprobes reported previously. These nanoprobes, with exceptionally small size and enhanced fluorescence resulting from the Gd doping, can label successfully the HeLa cells in short time and present no evidence of toxicity or adverse affect on cell growth even at the concentration up to 1 mm. These results show that such nanoprobes have low toxicity, especially in comparison with the traditional PEGylated CdSe/ZnS or CdSe/CdS QDs. In MRI studies, they exert strong positive contrast effect with a large longitudinal relaxivity (r(1)) of water proton of 16 mm(-1) s(-1). Their capability of imaging HeLa cells with MRI implies that they have great potential as MRI contrast agents. Combining the high sensitivity of fluorescence imaging with high spatial resolution of MRI, We expect that the as-prepared Gd-doped Zno QDs can provide a better reliability of the collected data and find promising applications in biological, medical and other fields. Copyright © 2010 Elsevier Ltd. All rights reserved.

Self-Assembly of Fluorescent Hybrid Core-Shell Nanoparticles and Their Application.

Science.gov (United States)

Wang, Chun; Tang, Fu; Wang, Xiaoyu; Li, Lidong

2015-06-24

In this work, a fluorescent hybrid core-shell nanoparticle was prepared by coating a functional polymer shell onto silver nanoparticles via a facile one-pot method. The biomolecule poly-L-lysine (PLL) was chosen as the polymer shell and assembled onto the silver core via the amine-reactive cross-linker, 3,3'-dithiobis(sulfosuccinimidylpropionate). The fluorescent anticancer drug, doxorubicin, was incorporated into the PLL shell through the same linkage. As the cross-linker possesses a thiol-cleavable disulfide bond, disassembly of the PLL shell was observed in the presence of glutathione, leading to controllable doxorubicin release. The silver core there provided an easily modified surface to facilitate the shell coating and ensures the efficient separation of as-prepared nanoparticles from their reaction mixture through centrifugation. Cell assays show that the prepared hybrid fluorescent nanoparticles can internalize into cells possessing excellent biocompatibility prior to the release of doxorubicin, terminating cancer cells efficiently as the doxorubicin is released at the intracellular glutathione level. Such properties are important for designing smart containers for target drug delivery and cellular imaging.

Novel xenon calibration scheme for two-photon absorption laser induced fluorescence of hydrogen

Energy Technology Data Exchange (ETDEWEB)

Elliott, Drew; Scime, Earl; Short, Zachary, E-mail: zdshort@mix.wvu.edu [Department of Physics and Astronomy, West Virginia University, Morgantown, West Virginia 26056 (United States)

2016-11-15

Two photon absorption laser induced fluorescence (TALIF) measurements of neutral hydrogen and its isotopes are typically calibrated by performing TALIF measurements on krypton with the same diagnostic system and using the known ratio of the absorption cross sections [K. Niemi et al., J. Phys. D 34, 2330 (2001)]. Here we present the measurements of a new calibration method based on a ground state xenon scheme for which the fluorescent emission wavelength is nearly identical to that of hydrogen, thereby eliminating chromatic effects in the collection optics and simplifying detector calibration. We determine that the ratio of the TALIF cross sections of xenon and hydrogen is 0.024 ± 0.001.

Characterization of LiF-based soft X-ray imaging detectors by confocal fluorescence microscopy

International Nuclear Information System (INIS)

Bonfigli, F; Gaudio, P; Lupelli, I; Nichelatti, E; Richetta, M; Vincenti, M A; Montereali, R M

2010-01-01

X-ray microscopy represents a powerful tool to obtain images of samples with very high spatial resolution. The main limitation of this technique is represented by the poor spatial resolution of standard imaging detectors. We proposed an innovative high-performance X-ray imaging detector based on the visible photoluminescence of colour centres in lithium fluoride. In this work, a confocal microscope in fluorescence mode was used to characterize LiF-based imaging detectors measuring CC integrated visible fluorescence signals of LiF crystals and films (grown on several kinds of substrates) irradiated by soft X-rays produced by a laser plasma source in different exposure conditions. The results are compared with the CC photoluminescence spectra measured on the same samples and discussed.

Bulky Counterions: Enhancing the Two-Photon Excited Fluorescence of Gold Nanoclusters.

Science.gov (United States)

Bertorelle, Franck; Moulin, Christophe; Soleilhac, Antonin; Comby-Zerbino, Clothilde; Dugourd, Philippe; Russier-Antoine, Isabelle; Brevet, Pierre-François; Antoine, Rodolphe

2018-01-19

Increasing fluorescence quantum yields of ligand-protected gold nanoclusters has attracted wide research interest. The strategy consisting in using bulky counterions has been found to dramatically enhance the fluorescence. In this Communication, we push forward this concept to the nonlinear optical regime. We show that by an appropriate choice of bulky counterions and of solvent, a 30-fold increase in two-photon excited fluorescence (TPEF) signal at ≈600 nm for gold nanoclusters can be obtained. This would correspond to a TPEF cross-section in the range of 0.1 to 1 GM. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

APD detectors for biological fluorescence spectroscopy

International Nuclear Information System (INIS)

Mazeres, S.; Borrel, V.; Magenc, C.; Courrech, J.L.; Bazer-Bachi, R.

2006-01-01

Fluorescence spectroscopy is a very convenient and widely used method for studying the molecular background of biological processes [L. Salome, J.L. Cazeil, A. Lopez, J.F. Tocanne, Eur. Biophys. J. 27 (1998) 391-402]. Chromophores are included in the structure under study and a flash of laser light induces fluorescence (Fluorescence Recovery After Photo-bleaching), the decay of which yields information on the polarity, the speed of rotation, and the speed of diffusion as well as on the temporal and spatial evolution of interactions between molecular species. The method can even be used to study living cells [J.F. Tocanne, L. Cezanne, A. Lopez, Prog. Lipid Res. 33 (1994) 203-237, L. Cezanne, A. Lopez, F. Loste, G. Parnaud, O. Saurel, P. Demange, J.F. Tocanne, Biochemistry 38 (1999) 2779-2786]. This is classically performed with a PM-based system. For biological reasons a decrease of the excitation of the cells is highly desirable. Because the fluorescence response then becomes fainter a significant improvement in detector capability would be welcome. We present here results obtained with an Avalanche Photo Diode (APD)-based system. The small sensitive area of detection allows a very significant improvement in signal/noise ratio, improvement in gain, and the opening-up of a new parameter space. With these new detectors we can begin the study of information transmission between cells through morphine receptors. This work involves both electronics engineers and biophysicists, so results and techniques in both fields will be presented here

Measuring techniques for Compton, Rayleigh and fluorescence cross-sections excited by keV photons. Potential use for various applications

Energy Technology Data Exchange (ETDEWEB)

Rao, Donepudi V.; Takeda, Tohoru; Itai, Yuji [University of Tsukuba, Institute of Clinical Medicine, Tsukuba, Ibaraki (Japan); Akatsuka, Takao [Yamagata University, Faculty of Engineering, Yonezawa, Yamagata (Japan); Seltzer, Stephen M.; Hubbell, John H. [National Institute of Standards and Technology, Ionizing Radiation Division, Photon and Charged Particle Data Center, Gaithersburg, MD (United States); Cesareo, Roberto; Brunetti, Antonio [Universita di Sassari, Istituto di Matematica e Fisica, Sassari (Italy); Gigante, Giovanni E. [Universita di Rome, Dipartimento di Fisica, ' La Sapienza' , Roma (Italy)

2002-08-01

Compton, Rayleigh and fluorescence cross-sections for low, medium and high Z atoms are measured at low photon energies (<100 keV) using an X-ray tube with secondary target. A new approach is developed to estimate the solid angle approximation and geometrical efficiency for a system with experimental arrangement using X-ray tube and secondary target as excitation source. Using this arrangement basic interaction cross-section measurements are performed in vacuum and reasonable monochromacy is achieved. Our motivation is mainly based on fundamental radiation interactions and radiology in mind. The present X-ray system considering the geometrical effects of the measuring system is more advantageous as far as the reduction of incoherently scattered background and improvement of monochromacy is concerned. The variation of the solid angle is studied by changing the radius and length of the collimators towards and away from the source and sample. From these values the variation of the solid angle and geometrical efficiency is deduced and the optimum value is used for the experimental work. The experimental system is very simple in design, compact and economical. Experimental results based on these systems are compared with theoretical estimates and good agreement is observed in between them. (author)

Tomographic sensing and localization of fluorescently labeled circulating cells in mice in vivo

International Nuclear Information System (INIS)

Zettergren, Eric; Swamy, Tushar; Niedre, Mark; Runnels, Judith; Lin, Charles P

2012-01-01

Sensing and enumeration of specific types of circulating cells in small animals is an important problem in many areas of biomedical research. Microscopy-based fluorescence in vivo flow cytometry methods have been developed previously, but these are typically limited to sampling of very small blood volumes, so that very rare circulating cells may escape detection. Recently, we described the development of a ‘diffuse fluorescence flow cytometer’ (DFFC) that allows sampling of much larger blood vessels and therefore circulating blood volumes in the hindlimb, forelimb or tail of a mouse. In this work, we extend this concept by developing and validating a method to tomographically localize circulating fluorescently labeled cells in the cross section of a tissue simulating optical flow phantom and mouse limb. This was achieved using two modulated light sources and an array of six fiber-coupled detectors that allowed rapid, high-sensitivity acquisition of full tomographic data sets at 10 Hz. These were reconstructed into two-dimensional cross-sectional images using Monte Carlo models of light propagation and the randomized algebraic reconstruction technique. We were able to obtain continuous images of moving cells in the sample cross section with 0.5 mm accuracy or better. We first demonstrated this concept in limb-mimicking optical flow photons with up to four flow channels, and then in the tails of mice with fluorescently labeled multiple myeloma cells. This approach increases the overall diagnostic utility of our DFFC instrument. (paper)

Up-Conversion Intersystem Crossing Rates in Organic Emitters for Thermally Activated Delayed Fluorescence: Impact of the Nature of Singlet vs Triplet Excited States

KAUST Repository

Samanta, Pralok Kumar

2017-02-28

The rates for up-conversion intersystem crossing (UISC) from the T1 state to the S1 state are calculated for a series of organic emitters with an emphasis on thermally activated delayed fluorescence (TADF) materials. Both the spin-orbit coupling and the energy difference between the S1 and T1 states (ΔEST) are evaluated, at the density functional theory (DFT) and time-dependent DFT levels. The calculated UISC rates and ΔEST values are found to be in good agreement with available experimental data. Our results underline that small ΔEST values and sizable spin-orbit coupling matrix elements have to be simultaneously realized in order to facilitate UISC and ultimately TADF. Importantly, the spatial separation of the highest occupied and lowest unoccupied molecular orbitals of the emitter, a widely accepted strategy for the design of TADF molecules, does not necessarily lead to a sufficient reduction in ΔEST; in fact, either a significant charge-transfer (CT) contribution to the T1 state or a minimal energy difference between the local-excitation and charge-transfer triplet states is required to achieve a small ΔEST. Also, having S1 and T1 states of a different nature is found to strongly enhance spin-orbit coupling, which is consistent with the El-Sayed rule for ISC rates. Overall, our results indicate that having either similar energies for the local-excitation and charge-transfer triplet states or the right balance between a substantial CT contribution to T1 and somewhat different natures of the S1 and T1 states, paves the way toward UISC enhancement and thus TADF efficiency improvement.

Development of a new light collection and detection system optimized for ion beam induced fluorescence microscopy

International Nuclear Information System (INIS)

Vanga, Sudheer Kumar; Mi, Zhaohong; Koh, Long Cheng; Tao, Ye; Bettiol, Andrew A.; Watt, Frank

2015-01-01

Ion beam induced fluorescence microscopy is a new imaging technique which has the potential to achieve sub-50 nm spatial resolution fluorescence images. Currently the resolution of the technique has been limited to around 150 nm mainly because of inefficient collection and detection of emitted photons from the sample. To overcome this limitation, a new light collection system based on a custom made parabolic mirror is employed to enhance the fluorescence collection. The custom made mirror is designed so as to obtain both structural (scanning transmission ion microscopy) and ion beam induced fluorescence imaging simultaneously. The design and characterization of the parabolic mirror is discussed in detail

Laser-induced fluorescences due to quadrupole moment transition and Stark effect in a He glow discharge

International Nuclear Information System (INIS)

Sakai, Hisashi; Takiyama, Ken; Kimura, Masahiko; Yamasaki, Motokuni; Fujita, Toshiaki; Oda, Toshiatsu; Kawasaki, Ken.

1993-01-01

The electric quadrupole moment transition and the Stark effect are investigated in a He hollow cathode discharge with laser-induced fluorescence method. It is shown that the forbidden transition from 2 1 S to 3 1 D in the negative glow is dominantly due to the quadrupole moment transition. This absorption coefficient is obtained from the laser-induced fluorescence intensity measurement in which the collisional transfers are taken into account. The result agrees with the theoretical coefficient. In the cathode dark space the fluorescence due to the Stark effect is also observed. Spatial distribution of the fluorescence is discussed, compared with the electric field distribution in the dark space. (author)

Red fluorescence imaging for dental plaque detection and quantification: pilot study

Science.gov (United States)

Liu, Zhao; Gomez, Juliana; Khan, Soniya; Peru, Debbie; Ellwood, Roger

2017-09-01

The red fluorescence of dental plaque originating from porphyrins in oral bacteria may allow visualization, detection, and scoring of plaque without disclosing agents. Two studies were conducted. The first included 24 healthy participants who abstained from oral hygiene for 24 h. Dental plaque was collected from tooth surfaces, and a 10% solution was prepared. These were scanned by a molecular spectrometer to identify the optimum excitation and emission wavelengths of plaque for developing a red fluorescence imaging system. Fourteen healthy subjects completed the second study. After a washout period (1 week), participants had a prophylaxis at baseline and abstained from oral hygiene during the study. They were monitored using the fluorescence imaging system at baseline, 24 h, and 48 h. A dentist clinically assessed plaque after disclosing and on red fluorescence images. Three descriptors were extracted from images and a RUSBoost classifier derived computer fluorescence scores through cross-validation. Red fluorescence plaque levels increased during the 48-h accumulation. Plaque progression was identified by dentist assessment and computer analysis, presenting significant differences between visits at tooth and subject levels (phygiene assessment.

Singular value decomposition metrics show limitations of detector design in diffuse fluorescence tomography.

Science.gov (United States)

Leblond, Frederic; Tichauer, Kenneth M; Pogue, Brian W

2010-11-29

The spatial resolution and recovered contrast of images reconstructed from diffuse fluorescence tomography data are limited by the high scattering properties of light propagation in biological tissue. As a result, the image reconstruction process can be exceedingly vulnerable to inaccurate prior knowledge of tissue optical properties and stochastic noise. In light of these limitations, the optimal source-detector geometry for a fluorescence tomography system is non-trivial, requiring analytical methods to guide design. Analysis of the singular value decomposition of the matrix to be inverted for image reconstruction is one potential approach, providing key quantitative metrics, such as singular image mode spatial resolution and singular data mode frequency as a function of singular mode. In the present study, these metrics are used to analyze the effects of different sources of noise and model errors as related to image quality in the form of spatial resolution and contrast recovery. The image quality is demonstrated to be inherently noise-limited even when detection geometries were increased in complexity to allow maximal tissue sampling, suggesting that detection noise characteristics outweigh detection geometry for achieving optimal reconstructions.

Detecting thermal phase transitions in corneal stroma by fluorescence micro-imaging analysis

Science.gov (United States)

Matteini, P.; Rossi, F.; Ratto, F.; Bruno, I.; Nesi, P.; Pini, R.

2008-02-01

Thermal modifications induced in corneal stroma were investigated by the use of fluorescence microscopy. Freshly extracted porcine corneas were immersed for 5 minutes in a water bath at temperatures in the 35-90°C range and stored in formalin. The samples were then sliced in 200-μm-thick transversal sections and analyzed under a stereomicroscope to assess corneal shrinkage. Fluorescence images of the thermally treated corneal samples were acquired using a slow-scan cooled CCD camera, after staining the slices with Indocyanine Green (ICG) fluorescent dye which allowed to detect fluorescence signal from the whole tissue. All measurements were performed using an inverted epifluorescence microscope equipped with a mercury lamp. The thermally-induced modifications to the corneal specimens were evaluated by studying the grey level distribution in the fluorescence images. For each acquired image, Discrete Fourier Transform (DFT) and entropy analyses were performed. The spatial distribution of DFT absolute value indicated the spatial orientation of the lamellar planes, while entropy was used to study the image texture, correlated to the stromal structural transitions. As a result, it was possible to indicate a temperature threshold value (62°C) for high thermal damage, resulting in a disorganization of the lamellar planes and in full agreement with the measured temperature for corneal shrinkage onset. Analysis of the image entropy evidenced five strong modifications in stromal architecture at temperatures of ~45°C, 53°C, 57°C, 66°C, 75°C. The proposed procedure proved to be an effective micro-imaging method capable of detecting subtle changes in corneal tissue subjected to thermal treatment.

Li (i = 1-3) subshell X-ray production cross sections and fluorescence yields for some elements with 56 ≤ Z ≤ 68 at 22.6 keV

International Nuclear Information System (INIS)

Chauhan, Yogeshwar; Tiwari, M.K.; Puri, Sanjiv

2008-01-01

The L k (k = l, α, β 1,4 , β 3,6 , β 2,15,9,10,7 , γ 1,5 and γ 2,3,4 ) X-ray production (XRP) cross sections have been measured for six elements with 56 ≤ Z ≤ 68 at 22.6 keV incident photon energy using the EDXRF spectrometer. The incident photon intensity, detector efficiency and geometrical factors have been determined from the K X-ray yields emitted from elemental targets with 22 ≤ Z ≤ 42 in the same geometrical setup and from knowledge of the K XRP cross sections. The L 1 and L 2 subshell fluorescence yields have been deduced from the present measured L k XRP cross sections using the relativistic Hartree-Fock-Slater (HFS) model based photoionization cross sections. The present deduced ω 1 (exp) values have been found to be, on an average, higher by 15% and 20% than those based on the Dirac-Hartree-Slater (DHS) model and the semi-empirical values compiled by Krause, respectively, for elements with 60 ≤ Z ≤ 68

Estimation of presampling MTF in CR systems by using direct fluorescence and its problems

International Nuclear Information System (INIS)

Ono, Kitahei; Inatsu, Hiroshi; Harao, Mototsugu; Itonaga, Haruo; Miyamoto, Hideyuki

2001-01-01

We proposed a method for practical estimation of the presampling modulation transfer function (MTF) of a computed radiography (CR) system by using the MTFs of an imaging plate and the sampling aperture. The MTFs of three imaging plates (GP-25, ST-VN, and RP-1S) with different photostimulable phosphors were measured by using direct fluorescence (the light emitted instantaneously by x-ray exposure), and the presampling MTFs were estimated from these imaging plate MTFs and the sampling aperture MTF. Our results indicated that for imaging plate RP-1S the measured presampling MTF was significantly superior to the estimated presampling MTF at any spatial frequency. This was because the estimated presampling MTF was degraded by the diffusion of direct fluorescence in the protective layer of the imaging plate's surface. Therefore, when the presampling MTF of the imaging plate with a thick protective layer is estimated, correction for the thickness of the protective layer should be carried out. However, the estimated presampling MTF of imaging plates with a thin protective layer were almost the same as the measured presampling MTF, except in the high spatial frequency range. Therefore, we consider this estimation method to be useful and practical, because the spatial resolution property of a CR system can be obtained simply from the imaging plate MTF measured with direct fluorescence. (author)

Bessel beam fluorescence lifetime tomography of live embryos (Conference Presentation)

Science.gov (United States)

Xu, Dongli; Peng, Leilei

2016-03-01

Optical tomography allows isotropic 3D imaging of embryos. Scanning-laser optical tomography (SLOT) has superior light collecting efficiency than wide-field optical tomography, making it ideal for fluorescence imaging of live embryos. We previously reported an imaging system that combines SLOT with a novel Fourier-multiplexed fluorescence lifetime imaging (FmFLIM) technique named FmFLIM-SLOT. FmFLIM-SLOT performs multiplexed FLIM-FRET readout of multiple FRET sensors in live embryos. Here we report a recent effort on improving the spatial resolution of the FmFLIM-SLOT system in order to image complex biochemical processes in live embryos at the cellular level. Optical tomography has to compromise between resolution and the depth of view. In SLOT, the commonly-used focused Gaussian beam diverges quickly from the focal plane, making it impossible to achieve high resolution imaging in a large volume specimen. We thus introduce Bessel beam laser-scanning tomography, which illuminates the sample with a spatial-light-modulator-generated Bessel beam that has an extended focal depth. The Bessel beam is scanned across the whole specimen. Fluorescence projection images are acquired at equal angular intervals as the sample rotates. Reconstruction artifacts due to annular-rings of the Bessel beam are removed by a modified 3D filtered back projection algorithm. Furthermore, in combination of Fourier-multiplexing fluorescence lifetime imaging (FmFLIM) method, the Bessel FmFLIM-SLOT system is capable of perform 3D lifetime imaging of live embryos at cellular resolution. The system is applied to in-vivo imaging of transgenic Zebrafish embryos. Results prove that Bessel FmFLIM-SLOT is a promising imaging method in development biology research.

Excited-state absorption and fluorescence dynamics of Er3+:KY3F10

Science.gov (United States)

Labbé, C.; Doualan, J. L.; Moncorgé, R.; Braud, A.; Camy, P.

2018-05-01

We report here on a complete investigation of the excited-state absorption and fluorescence dynamics of Er3+ doped KY3F10 single crystals versus dopant concentrations and optical excitation conditions. Radiative and effective (including non-radiative relaxations) emission lifetimes and branching ratios are determined from a Judd-Ofelt analysis of the absorption spectra and via specific fluorescence experiments using wavelength selective laser excitations. Excited-state absorption and emission spectra are registered within seven spectral domains, i.e. 560 nm, 650 nm, 710 nm, 810 nm, 970 nm, 1550 nm and 2750 nm. A maximum gain cross-section of 0.93 × 10-21 cm2 is determined at the potential laser wavelength of 2.801 μm for a population ratio of 0.48. Saturation of fluorescence intensities and variations of population ratios versus pumping rates are registered and confronted with a rate equation model to derive the rates of the most important up-conversion and cross-relaxation energy transfers occurring at high dopant concentrations.

Cell-penetrating anti-GFAP VHH and corresponding fluorescent fusion protein VHH-GFP spontaneously cross the blood-brain barrier and specifically recognize astrocytes: application to brain imaging.

Science.gov (United States)

Li, Tengfei; Bourgeois, Jean-Pierre; Celli, Susanna; Glacial, Fabienne; Le Sourd, Anne-Marie; Mecheri, Salah; Weksler, Babette; Romero, Ignacio; Couraud, Pierre-Olivier; Rougeon, François; Lafaye, Pierre

2012-10-01

Antibodies normally do not cross the blood-brain barrier (BBB) and cannot bind an intracellular cerebral antigen. We demonstrate here for the first time that a new class of antibodies can cross the BBB without treatment. Camelids produce native homodimeric heavy-chain antibodies, the paratope being composed of a single-variable domain called VHH. Here, we used recombinant VHH directed against human glial fibrillary acidic protein (GFAP), a specific marker of astrocytes. Only basic VHHs (e.g., pI=9.4) were able to cross the BBB in vitro (7.8 vs. 0% for VHH with pI=7.7). By intracarotid and intravenous injections into live mice, we showed that these basic VHHs are able to cross the BBB in vivo, diffuse into the brain tissue, penetrate into astrocytes, and specifically label GFAP. To analyze their ability to be used as a specific transporter, we then expressed a recombinant fusion protein VHH-green fluorescent protein (GFP). These "fluobodies" specifically labeled GFAP on murine brain sections, and a basic variant (pI=9.3) of the fusion protein VHH-GFP was able to cross the BBB and to label astrocytes in vivo. The potential of VHHs as diagnostic or therapeutic agents in the central nervous system now deserves attention.

Hybrid phosphorescence and fluorescence native spectroscopy for breast cancer detection.

Science.gov (United States)

Alimova, Alexandra; Katz, A; Sriramoju, Vidyasagar; Budansky, Yuri; Bykov, Alexei A; Zeylikovich, Roman; Alfano, R R

2007-01-01

Fluorescence and phosphorescence measurements are performed on normal and malignant ex vivo human breast tissues using UV LED and xenon lamp excitation. Tryptophan (trp) phosphorescence intensity is higher in both normal glandular and adipose tissue when compared to malignant tissue. An algorithm based on the ratio of trp fluorescence intensity at 345 nm to phosphorescence intensity at 500 nm is successfully used to separate normal from malignant tissue types. Normal specimens consistently exhibited a low I(345)I(500) ratio (15). The ratio analysis correlates well with histopathology. Intensity ratio maps with a spatial resolution of 0.5 mm are generated in which local regions of malignancy could be identified.

Precision analysis for standard deviation measurements of immobile single fluorescent molecule images.

Science.gov (United States)

DeSantis, Michael C; DeCenzo, Shawn H; Li, Je-Luen; Wang, Y M

2010-03-29

Standard deviation measurements of intensity profiles of stationary single fluorescent molecules are useful for studying axial localization, molecular orientation, and a fluorescence imaging system's spatial resolution. Here we report on the analysis of the precision of standard deviation measurements of intensity profiles of single fluorescent molecules imaged using an EMCCD camera.We have developed an analytical expression for the standard deviation measurement error of a single image which is a function of the total number of detected photons, the background photon noise, and the camera pixel size. The theoretical results agree well with the experimental, simulation, and numerical integration results. Using this expression, we show that single-molecule standard deviation measurements offer nanometer precision for a large range of experimental parameters.

Laplacian manifold regularization method for fluorescence molecular tomography

Science.gov (United States)

He, Xuelei; Wang, Xiaodong; Yi, Huangjian; Chen, Yanrong; Zhang, Xu; Yu, Jingjing; He, Xiaowei

2017-04-01

Sparse regularization methods have been widely used in fluorescence molecular tomography (FMT) for stable three-dimensional reconstruction. Generally, ℓ1-regularization-based methods allow for utilizing the sparsity nature of the target distribution. However, in addition to sparsity, the spatial structure information should be exploited as well. A joint ℓ1 and Laplacian manifold regularization model is proposed to improve the reconstruction performance, and two algorithms (with and without Barzilai-Borwein strategy) are presented to solve the regularization model. Numerical studies and in vivo experiment demonstrate that the proposed Gradient projection-resolved Laplacian manifold regularization method for the joint model performed better than the comparative algorithm for ℓ1 minimization method in both spatial aggregation and location accuracy.

Fluorescent Pluronic nanodots for in vivo two-photon imaging

International Nuclear Information System (INIS)

Maurin, Mathieu; Vurth, Laeticia; Vial, Jean-Claude; Baldeck, Patrice; Stephan, Olivier; Marder, Seth R; Sanden, Boudewijn Van der

2009-01-01

We report the synthesis of new nanosized fluorescent probes based on bio-compatible polyethylene-polypropylene glycol (Pluronic) materials. In aqueous solution, mini-emulsification of Pluronic with a high fluorescent di-stryl benzene-modified derivative, exhibiting a two-photon absorption cross section as high as 2500 Goeppert-Mayer units at 800 nm, leads to nanoparticles exhibiting a hydrodynamic radius below 100 nm. We have demonstrated that these new probes with luminescence located in the spectral region of interest for bio-imaging (the yellow part of the visible spectrum) allow deep (500 μm) bio-imaging of the mice brain vasculature. The dose injected during our experiments is ten times lower when compared to the classical commercial rhodamine-B isothicyanate-Dextran system but gives similar results to homogeneous blood plasma staining. The mean fluorescent signal intensity stayed constant during more than 1 h.

An instrument for small-animal imaging using time-resolved diffuse and fluorescence optical methods

International Nuclear Information System (INIS)

Montcel, Bruno; Poulet, Patrick

2006-01-01

We describe time-resolved optical methods that use diffuse near-infrared photons to image the optical properties of tissues and their inner fluorescent probe distribution. The assembled scanner uses picosecond laser diodes at 4 wavelengths, an 8-anode photo-multiplier tube and time-correlated single photon counting. Optical absorption and reduced scattering images as well as fluorescence emission images are computed from temporal profiles of diffuse photons. This method should improve the spatial resolution and the quantification of fluorescence signals. We used the diffusion approximation of the radiation transport equation and the finite element method to solve the forward problem. The inverse problem is solved with an optimization algorithm such as ART or conjugate gradient. The scanner and its performances are presented, together with absorption, scattering and fluorescent images obtained with it

Parallel scan hyperspectral fluorescence imaging system and biomedical application for microarrays

International Nuclear Information System (INIS)

Liu Zhiyi; Ma Suihua; Liu Le; Guo Jihua; He Yonghong; Ji Yanhong

2011-01-01

Microarray research offers great potential for analysis of gene expression profile and leads to greatly improved experimental throughput. A number of instruments have been reported for microarray detection, such as chemiluminescence, surface plasmon resonance, and fluorescence markers. Fluorescence imaging is popular for the readout of microarrays. In this paper we develop a quasi-confocal, multichannel parallel scan hyperspectral fluorescence imaging system for microarray research. Hyperspectral imaging records the entire emission spectrum for every voxel within the imaged area in contrast to recording only fluorescence intensities of filter-based scanners. Coupled with data analysis, the recorded spectral information allows for quantitative identification of the contributions of multiple, spectrally overlapping fluorescent dyes and elimination of unwanted artifacts. The mechanism of quasi-confocal imaging provides a high signal-to-noise ratio, and parallel scan makes this approach a high throughput technique for microarray analysis. This system is improved with a specifically designed spectrometer which can offer a spectral resolution of 0.2 nm, and operates with spatial resolutions ranging from 2 to 30 μm . Finally, the application of the system is demonstrated by reading out microarrays for identification of bacteria.

Crossed molecular beams

International Nuclear Information System (INIS)

Lee, Y.T.

1976-01-01

Research activities with crossed molecular beams at Lawrence Berkeley Laboratory during 1976 are described. Topics covered include: scattering of Ar*, Kr*, with Xe; metastable rare gas interactions, He* + H 2 ; an atomic and molecular halogen beam source; a crossed molecular beam study of the Cl + Br 2 → BrCl + Br reaction; O( 3 P) reaction dynamics, development of the high pressure plasma beam source; energy randomization in the Cl + C 2 H 3 Br → Br + C 2 H 3 Cl reaction; high resolution photoionization studies of NO and ICl; photoionization of (H 2 O)/sub n/ and (NH 3 ) 2 ; photoionization mass spectroscopy of NH 3 + and O 3 + ; photo fragmentation of bromine; and construction of chemiluminescence-laser fluorescence crossed molecular beam machine

Near-IR Two-Photon Fluorescent Sensor for K(+) Imaging in Live Cells.

Science.gov (United States)

Sui, Binglin; Yue, Xiling; Kim, Bosung; Belfield, Kevin D

2015-08-19

A new two-photon excited fluorescent K(+) sensor is reported. The sensor comprises three moieties, a highly selective K(+) chelator as the K(+) recognition unit, a boron-dipyrromethene (BODIPY) derivative modified with phenylethynyl groups as the fluorophore, and two polyethylene glycol chains to afford water solubility. The sensor displays very high selectivity (>52-fold) in detecting K(+) over other physiological metal cations. Upon binding K(+), the sensor switches from nonfluorescent to highly fluorescent, emitting red to near-IR (NIR) fluorescence. The sensor exhibited a good two-photon absorption cross section, 500 GM at 940 nm. Moreover, it is not sensitive to pH in the physiological pH range. Time-dependent cell imaging studies via both one- and two-photon fluorescence microscopy demonstrate that the sensor is suitable for dynamic K(+) sensing in living cells.

Determination of experimental K-shell fluorescence yield for ...

Indian Academy of Sciences (India)

calcium compounds using a Si(Li) X-ray detector system (FWHM=5.96 keV at 160 eV). The samples were excited by 5.96 keV photons produced by a 55Fe radioisotope source. The experimental values are systematically lower than the theoretical values. Keywords. X-ray; fluorescence yield; cross-section and chemical ...

Mercury mass measurement in fluorescent lamps via neutron activation analysis

International Nuclear Information System (INIS)

Viererbl, L.; Vinš, M.; Lahodová, Z.; Fuksa, A.; Kučera, J.; Koleška, M.; Voljanskij, A.

2015-01-01

Mercury is an essential component of fluorescent lamps. Not all fluorescent lamps are recycled, resulting in contamination of the environment with toxic mercury, making measurement of the mercury mass used in fluorescent lamps important. Mercury mass measurement of lamps via instrumental neutron activation analysis (NAA) was tested under various conditions in the LVR-15 research reactor. Fluorescent lamps were irradiated in different positions in vertical irradiation channels and a horizontal channel in neutron fields with total fluence rates from 3×10 8 cm −2 s −1 to 10 14 cm −2 s −1 . The 202 Hg(n,γ) 203 Hg nuclear reaction was used for mercury mass evaluation. Activities of 203 Hg and others induced radionuclides were measured via gamma spectrometry with an HPGe detector at various times after irradiation. Standards containing an Hg 2 Cl 2 compound were used to determine mercury mass. Problems arise from the presence of elements with a large effective cross section in luminescent material (europium, antimony and gadolinium) and glass (boron). The paper describes optimization of the NAA procedure in the LVR-15 research reactor with particular attention to influence of neutron self-absorption in fluorescent lamps. - Highlights: • Mercury is an essential component of fluorescent lamps. • Fluorescent lamps were irradiated in neutron fields in research reactor. • 203 Hg induced radionuclide activity was measured using gamma spectrometry. • Mercury mass in fluorescent lamps can be measured by neutron activation analysis.

Spatially-resolved luminescence spectroscopy of CdSe quantum dots synthesized in ionic liquid crystal matrices

Energy Technology Data Exchange (ETDEWEB)

Magaryan, K.A., E-mail: xmagaros@gmail.com [Moscow State Pedagogical University, 29 Malaya Pirogovskaya Str., Moscow 119992 (Russian Federation); Mikhailov, M.A. [Moscow State Pedagogical University, 29 Malaya Pirogovskaya Str., Moscow 119992 (Russian Federation); Karimullin, K.R. [Moscow State Pedagogical University, 29 Malaya Pirogovskaya Str., Moscow 119992 (Russian Federation); Institute for Spectroscopy of RAS, 5 Fizicheskaya Str., Troitsk, Moscow 142190 (Russian Federation); E.K. Zavoyski Kazan Physical-Technical Institute of RAS, 10/7 Sibirski trakt Str., Kazan 420029 (Russian Federation); Knyazev, M.V.; Eremchev, I.Y. [Institute for Spectroscopy of RAS, 5 Fizicheskaya Str., Troitsk, Moscow 142190 (Russian Federation); Naumov, A.V. [Moscow State Pedagogical University, 29 Malaya Pirogovskaya Str., Moscow 119992 (Russian Federation); Institute for Spectroscopy of RAS, 5 Fizicheskaya Str., Troitsk, Moscow 142190 (Russian Federation); Vasilieva, I.A. [Moscow State Pedagogical University, 29 Malaya Pirogovskaya Str., Moscow 119992 (Russian Federation); Klimusheva, G.V. [Institute of Physics, NAS of Ukraine, 46 Prospect Nauki, Kiev 03028 (Ukraine)

2016-01-15

The paper is devoted to investigation of luminescence properties of new quantum dot (QD)-doped materials. We studied CdSe QDs (1.8 nm and 2.3 nm) grown inside of a liquid crystalline cadmium alcanoate matrix. Temperature dependence of parameters of fluorescence spectra obtained in a wide temperature range using epi-luminescence microscopy technique was analyzed. Spatially-resolved luminescence images were measured for the areas of the samples of 150×150 µm{sup 2}. Strong correlation between fluorescence spectra and sample structure was observed. - Highlights: • Glassy matrix with CdSe quantum dots inside fabricated in liquid crystalline mesophase. • Study of luminescence properties in a wide range of low temperatures. • Strong dependence of the luminescence spectra on spatial inhomogeneities. • Spatially-resolved luminescence imaging of quantum dots in liquid crystalline matrix.

Spatially-resolved luminescence spectroscopy of CdSe quantum dots synthesized in ionic liquid crystal matrices

International Nuclear Information System (INIS)

Magaryan, K.A.; Mikhailov, M.A.; Karimullin, K.R.; Knyazev, M.V.; Eremchev, I.Y.; Naumov, A.V.; Vasilieva, I.A.; Klimusheva, G.V.

2016-01-01

The paper is devoted to investigation of luminescence properties of new quantum dot (QD)-doped materials. We studied CdSe QDs (1.8 nm and 2.3 nm) grown inside of a liquid crystalline cadmium alcanoate matrix. Temperature dependence of parameters of fluorescence spectra obtained in a wide temperature range using epi-luminescence microscopy technique was analyzed. Spatially-resolved luminescence images were measured for the areas of the samples of 150×150 µm 2 . Strong correlation between fluorescence spectra and sample structure was observed. - Highlights: • Glassy matrix with CdSe quantum dots inside fabricated in liquid crystalline mesophase. • Study of luminescence properties in a wide range of low temperatures. • Strong dependence of the luminescence spectra on spatial inhomogeneities. • Spatially-resolved luminescence imaging of quantum dots in liquid crystalline matrix.

Challenges for Super-Resolution Localization Microscopy and Biomolecular Fluorescent Nano-Probing in Cancer Research

Science.gov (United States)

Ilić, Nataša; Pilarczyk, Götz; Lee, Jin-Ho; Logeswaran, Abiramy; Borroni, Aurora Paola; Krufczik, Matthias; Theda, Franziska; Waltrich, Nadine; Bestvater, Felix; Hildenbrand, Georg; Cremer, Christoph; Blank, Michael

2017-01-01

Understanding molecular interactions and regulatory mechanisms in tumor initiation, progression, and treatment response are key requirements towards advanced cancer diagnosis and novel treatment procedures in personalized medicine. Beyond decoding the gene expression, malfunctioning and cancer-related epigenetic pathways, investigations of the spatial receptor arrangements in membranes and genome organization in cell nuclei, on the nano-scale, contribute to elucidating complex molecular mechanisms in cells and tissues. By these means, the correlation between cell function and spatial organization of molecules or molecular complexes can be studied, with respect to carcinogenesis, tumor sensitivity or tumor resistance to anticancer therapies, like radiation or antibody treatment. Here, we present several new applications for bio-molecular nano-probes and super-resolution, laser fluorescence localization microscopy and their potential in life sciences, especially in biomedical and cancer research. By means of a tool-box of fluorescent antibodies, green fluorescent protein (GFP) tagging, or specific oligonucleotides, we present tumor relevant re-arrangements of Erb-receptors in membranes, spatial organization of Smad specific ubiquitin protein ligase 2 (Smurf2) in the cytosol, tumor cell characteristic heterochromatin organization, and molecular re-arrangements induced by radiation or antibody treatment. The main purpose of this article is to demonstrate how nano-scaled distance measurements between bio-molecules, tagged by appropriate nano-probes, can be applied to elucidate structures and conformations of molecular complexes which are characteristic of tumorigenesis and treatment responses. These applications open new avenues towards a better interpretation of the spatial organization and treatment responses of functionally relevant molecules, at the single cell level, in normal and cancer cells, offering new potentials for individualized medicine. PMID:28956810

Challenges for Super-Resolution Localization Microscopy and Biomolecular Fluorescent Nano-Probing in Cancer Research

Directory of Open Access Journals (Sweden)

Michael Hausmann

2017-09-01

Full Text Available Understanding molecular interactions and regulatory mechanisms in tumor initiation, progression, and treatment response are key requirements towards advanced cancer diagnosis and novel treatment procedures in personalized medicine. Beyond decoding the gene expression, malfunctioning and cancer-related epigenetic pathways, investigations of the spatial receptor arrangements in membranes and genome organization in cell nuclei, on the nano-scale, contribute to elucidating complex molecular mechanisms in cells and tissues. By these means, the correlation between cell function and spatial organization of molecules or molecular complexes can be studied, with respect to carcinogenesis, tumor sensitivity or tumor resistance to anticancer therapies, like radiation or antibody treatment. Here, we present several new applications for bio-molecular nano-probes and super-resolution, laser fluorescence localization microscopy and their potential in life sciences, especially in biomedical and cancer research. By means of a tool-box of fluorescent antibodies, green fluorescent protein (GFP tagging, or specific oligonucleotides, we present tumor relevant re-arrangements of Erb-receptors in membranes, spatial organization of Smad specific ubiquitin protein ligase 2 (Smurf2 in the cytosol, tumor cell characteristic heterochromatin organization, and molecular re-arrangements induced by radiation or antibody treatment. The main purpose of this article is to demonstrate how nano-scaled distance measurements between bio-molecules, tagged by appropriate nano-probes, can be applied to elucidate structures and conformations of molecular complexes which are characteristic of tumorigenesis and treatment responses. These applications open new avenues towards a better interpretation of the spatial organization and treatment responses of functionally relevant molecules, at the single cell level, in normal and cancer cells, offering new potentials for individualized medicine.

Laser-induced fluorescence imaging of acetone inside evaporating and burning fuel droplets

Science.gov (United States)

Shringi, D. S.; Shaw, B. D.; Dwyer, H. A.

2009-01-01

Laser-induced fluorescence was used to visualize acetone fields inside individual droplets of pure acetone as well as droplets composed of methanol or 1-propanol initially mixed with acetone. Droplets were supported on a horizontal wire and two vaporization conditions were investigated: (1) slow evaporation in room air and (2) droplet combustion, which leads to substantially faster droplet surface regression rates. Acetone was preferentially gasified, causing its concentration in droplets to drop in time with resultant decreases in acetone fluorescence intensities. Slowly vaporizing droplets did not exhibit large spatial variations of fluorescence within droplets, indicating that these droplets were relatively well mixed. Ignition of droplets led to significant variations in fluorescence intensities within droplets, indicating that these droplets were not well mixed. Ignited droplets composed of mixtures of 1-propanol and acetone showed large time-varying changes in shapes for higher acetone concentrations, suggesting that bubble formation was occurring in these droplets.

Quantitative analysis by X-ray fluorescence using first principles for matrix correction

International Nuclear Information System (INIS)

Hulett, L.D.; Dunn, H.W.; Tarter, J.G.

1978-01-01

The quantitative interpretation of X-ray fluorescence (XRF) data is often difficult because of matrix effects. The intensity of fluorescence measured for a given element is not only dependent on the element's concentration, but also on the mass absorption coefficients of the sample for the excitation and fluorescence radiation. Also, there are interelement effects in which high-energy fluorescence from heavier elements is absorbed by lighter elements with a resulting enhancement of their fluorescence. Recent theoretical treatments of this problem have shown that X-ray fluorescence data can be corrected for these matrix effects by calculations based on first principles. Fundamental constants, available in atomic physics data tables, are the only parameters needed. It is not necessary to make empirical calibrations. The application of this correctional procedure to alloys and alumina-supported catalysts is described. A description is given of a low-background spectrometer which uses monochromatic Ag Ksub(α) radiation for excitation. Matrix corrections by first principles can be easily applied to data from instruments of this type because fluorescence excitation cross-sections and mass absorption coefficients can be accurately defined for monochromatic radiation. (author)

Near-infrared II fluorescence for imaging hindlimb vessel regeneration with dynamic tissue perfusion measurement.

Science.gov (United States)

Hong, Guosong; Lee, Jerry C; Jha, Arshi; Diao, Shuo; Nakayama, Karina H; Hou, Luqia; Doyle, Timothy C; Robinson, Joshua T; Antaris, Alexander L; Dai, Hongjie; Cooke, John P; Huang, Ngan F

2014-05-01

Real-time vascular imaging that provides both anatomic and hemodynamic information could greatly facilitate the diagnosis of vascular diseases and provide accurate assessment of therapeutic effects. Here, we have developed a novel fluorescence-based all-optical method, named near-infrared II (NIR-II) fluorescence imaging, to image murine hindlimb vasculature and blood flow in an experimental model of peripheral arterial disease, by exploiting fluorescence in the NIR-II region (1000-1400 nm) of photon wavelengths. Because of the reduced photon scattering of NIR-II fluorescence compared with traditional NIR fluorescence imaging and thus much deeper penetration depth into the body, we demonstrated that the mouse hindlimb vasculature could be imaged with higher spatial resolution than in vivo microscopic computed tomography. Furthermore, imaging during 26 days revealed a significant increase in hindlimb microvascular density in response to experimentally induced ischemia within the first 8 days of the surgery (Pimaging make it a useful imaging tool for murine models of vascular disease. © 2014 American Heart Association, Inc.

MGlu5 antagonism impairs exploration and memory of spatial and non-spatial stimuli in rats

DEFF Research Database (Denmark)

Christoffersen, Gert Rene Juul; Simonyi, Agnes; Schachtman, Todd R.

2008-01-01

and it was found that: 1) Locomotion during exploration of spatial environments and exploration time at novel objects were reduced by i.p. but not by prelimbic administration of MPEP, 2) spatial short-term memory was impaired in cross-maze and object discrimination was reduced after both types of administration, 3......) long-term retention of spatial conditioning in the cross-maze was inhibited after i.p. applications which 4) also inhibited spontaneous alternation performance during maze-exploration. Reduced exploratory locomotion and exploration time after i.p. injections may have contributed to the observed......Metabotropic glutamate receptor subtype 5 (mGlu5) has been implicated in memory processing in some but not all learning tasks.  The reason why this receptor is involved in some tasks but not in others remains to be determined.  The present experiments using rats examined effects of the mGlu5...

Enhanced speed in fluorescence imaging using beat frequency multiplexing

Science.gov (United States)

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.

Synthesis and Fluorescence Spectra of Triazolylcoumarin Fluorescent Dyes

Institute of Scientific and Technical Information of China (English)

PENG Xian-fu; LI Hong-qi

2009-01-01

Much attention is devoted to fluorescent dyes especially those with potential in versatile applications. Reactions under "click" conditions between nonfluorescent 3 - azidocoumarins and terminal alkynes produced 3 -(1, 2, 3- triazol- 1 - yl)cournarins, a novel type of fluorescent dyes with intense fluorescence. The structures of the new coumarins were characterized by 1H NMR, MS, and IR spectra. Fluorescence spectra measurement demonstrated excellent fluorescence performance of the triazolylcoumarins and this click reaction is a promising candidate for bioconjugation and bioimaging applications since both azide and alkynes are quite inert to biological systems.

Broadband two-photon absorption cross sections of benzothiazole derivatives and benzobisthiazolium salts

Science.gov (United States)

Noskovičova, Eva; Lorenc, Dušan; Magdolen, Peter; Sigmundová, Ivica; Zahradník, Pavol; Velič, Dušan

2018-05-01

Two-photon absorption (TPA) cross sections of conjugated donor-π-acceptor dipolar structures containing benzothiazole or benzobisthiazolium moieties are determined in a broad spectral range from 700 nm to 1000 nm using two-photon induced fluorescence technique. The TPA cross section values range from 150 GM to 4600 GM. The largest values are observed in near-infrared region. The dipolar derivative of benzothiazole has the largest TPA cross section of 4600 GM at wavelength of 890 nm. A combination of the large TPA in the near-infrared region and the high emission quantum yield makes these compounds excellent candidates for two-photon fluorescence microscopy.

Computer Reconstruction of Plant Growth and Chlorophyll Fluorescence Emission in Three Spatial Dimensions

Czech Academy of Sciences Publication Activity Database

Bellasio, Ch.; Olejníčková, Julie; Tesař, R.; Šebela, David; Nedbal, Ladislav

2012-01-01

Roč. 12, č. 1 (2012), s. 1052-1071 ISSN 1424-8220 R&D Projects: GA ČR GA522/09/1565; GA MŠk(CZ) ED1.1.00/02.0073 Institutional support: RVO:67179843 Keywords : 3D reconstruction * chlorophyll fluorescence imaging * leaf area * leaf angle * plant growth * coded light Subject RIV: EH - Ecology, Behaviour Impact factor: 1.953, year: 2012

Development of laser-induced fluorescence for precombustion diagnostics in spark-ignition engines

Energy Technology Data Exchange (ETDEWEB)

Neij, H.

1998-11-01

Motivated by a desire to understand and optimize combustion in spark-ignition (SI) engines, laser techniques have been developed for measurement of fuel and residual gas, respectively, in the precombustion mixture of an operating SI engine. The primary objective was to obtain two-dimensional, quantitative data in the vicinity of the spark gap at the time of ignition. A laser-induced fluorescence (LIF) technique was developed for fuel visualization in engine environments. Since the fluorescence signal from any commercial gasoline fuel would be unknown to its origin, with an unpredictable dependence on collisional partners, pressure and temperature, a non-fluorescent base fuel - isooctane - was used. For LIF detection, a fluorescent species was added to the fuel. An additive not commonly used in this context - 3-pentanone - was chosen based on its suitable vaporization characteristics and fluorescent properties. The LIF technique was applied to an optically accessible research engine. By calibration, the fluorescence signal from the additive was converted to fuel-to-air equivalence ratio ({phi}). The accuracy and precision of the acquired data were assessed. A statistical evaluation revealed that the spatially averaged equivalence ratio around the spark plug had a significant impact on the combustion event. The strong correlation between these two quantities suggested that the early combustion was sensitive to large-scale inhomogeneities in the precombustion mixture. A similar LIF technique, using acetone as a fluorescent additive in methane, was applied to a combustion cell for ion current evaluation. The local equivalence ratio around the spark gap at the time of ignition was extracted from LIF data. Useful relations were identified between different ion current parameters and the local equivalence ratio, although the impact of the flow field, the fuel type, and the electrode geometry were identified as areas for future research. A novel fuel - dimethyl ether (DME

X-ray microtome by fluorescence tomography

CERN Document Server

Simionovici, A S; Guenzler, F; Schrör, C; Snigirev, A; Snigireva, I; Tümmler, J; Weitkamp, T

2001-01-01

The X-ray fluorescence microtomography method is presented, which is capable of virtually slicing samples to obtain cross-sections of their inner structure. High precision experimental results of fluo-tomography in 'pencil-beam' geometry with up to 1.2 mu m resolution are described. Image reconstructions are based on either a simplified algebraic reconstruction method (ART) or the filtered back-projection method (FBP). Phantoms of inhomogeneous test objects as well as biological samples are successfully analyzed.

Mercury mass measurement in fluorescent lamps via neutron activation analysis

Science.gov (United States)

Viererbl, L.; Vinš, M.; Lahodová, Z.; Fuksa, A.; Kučera, J.; Koleška, M.; Voljanskij, A.

2015-11-01

Mercury is an essential component of fluorescent lamps. Not all fluorescent lamps are recycled, resulting in contamination of the environment with toxic mercury, making measurement of the mercury mass used in fluorescent lamps important. Mercury mass measurement of lamps via instrumental neutron activation analysis (NAA) was tested under various conditions in the LVR-15 research reactor. Fluorescent lamps were irradiated in different positions in vertical irradiation channels and a horizontal channel in neutron fields with total fluence rates from 3×108 cm-2 s-1 to 1014 cm-2 s-1. The 202Hg(n,γ)203Hg nuclear reaction was used for mercury mass evaluation. Activities of 203Hg and others induced radionuclides were measured via gamma spectrometry with an HPGe detector at various times after irradiation. Standards containing an Hg2Cl2 compound were used to determine mercury mass. Problems arise from the presence of elements with a large effective cross section in luminescent material (europium, antimony and gadolinium) and glass (boron). The paper describes optimization of the NAA procedure in the LVR-15 research reactor with particular attention to influence of neutron self-absorption in fluorescent lamps.

Considerations in the use of fluorescence in situ hybridization (FISH) and confocal laser scanning microscopy to characterize rumen methanogens and define their spatial distributions.

Science.gov (United States)

Valle, Edith R; Henderson, Gemma; Janssen, Peter H; Cox, Faith; Alexander, Trevor W; McAllister, Tim A

2015-06-01

In this study, methanogen-specific coenzyme F420 autofluorescence and confocal laser scanning microscopy were used to identify rumen methanogens and define their spatial distribution in free-living, biofilm-, and protozoa-associated microenvironments. Fluorescence in situ hybridization (FISH) with temperature-controlled hybridization was used in an attempt to describe methanogen diversity. A heat pretreatment (65 °C, 1 h) was found to be a noninvasive method to increase probe access to methanogen RNA targets. Despite efforts to optimize FISH, 16S rRNA methanogen-specific probes, including Arch915, bound to some cells that lacked F420, possibly identifying uncharacterized Methanomassiliicoccales or reflecting nonspecific binding to other members of the rumen bacterial community. A probe targeting RNA from the methanogenesis-specific methyl coenzyme M reductase (mcr) gene was shown to detect cultured Methanosarcina cells with signal intensities comparable to those of 16S rRNA probes. However, the probe failed to hybridize with the majority of F420-emitting rumen methanogens, possibly because of differences in cell wall permeability among methanogen species. Methanogens were shown to integrate into microbial biofilms and to exist as ecto- and endosymbionts with rumen protozoa. Characterizing rumen methanogens and defining their spatial distribution may provide insight into mitigation strategies for ruminal methanogenesis.

Quantification and localization of trace metals in natural plankton using a synchrotron x-ray fluorescence microprobe

International Nuclear Information System (INIS)

Twining, B. S.; Baines, S. B.; Fisher, N. S.; Jacobsen, C.; Maser, J.; State Univ. of New York at Stony Brook

2003-01-01

The accumulation of trace metals by planktonic protists influences the growth of primary producers, metal biogeochemical cycling, and metal bioaccumulation in aquatic food chains. Despite their importance, unequivocal measurements of trace element concentrations in individual plankton cells have not been possible to date. We have used the 2-ID-E side-branch hard x-ray microprobe at the Advanced Photon Source to measure trace elements in individual marine plankton cells. This microprobe employs zoneplate optics to produce the sub-micron spatial resolution and low background fluorescence required to produce trace element maps of planktonic protist cells ranging in size from 3 to >50 (micro)m. We have developed preservation, rinsing, and mounting protocols that remove most of the salt from our marine samples, thus simplifying the identification of unknown cells and reducing high Cl-related background fluorescence. We have also developed spectral modeling techniques that account for the frequent overlap of adjacent fluorescence peaks and non-uniform detector response. Finally, we have used parallel soft x-ray transmission and epifluorescence microscopy images to estimate C normalized trace element concentrations, identify functional cell types (e.g., photosynthetic vs. non-photosynthetic), and correlate cell structures with spatial patterns in trace element fluorescence

Quantification and localization of trace metals in natural plancton using a synchrotron x-ray fluorescence microprobe.

Energy Technology Data Exchange (ETDEWEB)

Twining, B. S.; Baines, S. B.; Fisher, N. S.; Jacobsen, C.; Maser, J.; State Univ. of New York at Stony Brook

2003-03-01

The accumulation of trace metals by planktonic protists influences the growth of primary producers, metal biogeochemical cycling, and metal bioaccumulation in aquatic food chains. Despite their importance, unequivocal measurements of trace element concentrations in individual plankton cells have not been possible to date. We have used the 2-ID-E side-branch hard x-ray microprobe at the Advanced Photon Source to measure trace elements in individual marine plankton cells. This microprobe employs zoneplate optics to produce the sub-micron spatial resolution and low background fluorescence required to produce trace element maps of planktonic protist cells ranging in size from 3 to >50 {micro}m. We have developed preservation, rinsing, and mounting protocols that remove most of the salt from our marine samples, thus simplifying the identification of unknown cells and reducing high Cl-related background fluorescence. We have also developed spectral modeling techniques that account for the frequent overlap of adjacent fluorescence peaks and non-uniform detector response. Finally, we have used parallel soft x-ray transmission and epifluorescence microscopy images to estimate C normalized trace element concentrations, identify functional cell types (e.g., photosynthetic vs. non-photosynthetic), and correlate cell structures with spatial patterns in trace element fluorescence.

Preservation of three-dimensional spatial structure in the gut microbiome.

Directory of Open Access Journals (Sweden)

Yuko Hasegawa

Full Text Available Preservation of three-dimensional structure in the gut is necessary in order to analyze the spatial organization of the gut microbiota and gut luminal contents. In this study, we evaluated preparation methods for mouse gut with the goal of preserving micron-scale spatial structure while performing fluorescence imaging assays. Our evaluation of embedding methods showed that commonly used media such as Tissue-Tek Optimal Cutting Temperature (OCT compound, paraffin, and polyester waxes resulted in redistribution of luminal contents. By contrast, a hydrophilic methacrylate resin, Technovit H8100, preserved three-dimensional organization. Our mouse intestinal preparation protocol optimized using the Technovit H8100 embedding method was compatible with microbial fluorescence in situ hybridization (FISH and other labeling techniques, including immunostaining and staining with both wheat germ agglutinin (WGA and 4', 6-diamidino-2-phenylindole (DAPI. Mucus could be visualized whether the sample was fixed with paraformaldehyde (PFA or with Carnoy's fixative. The protocol optimized in this study enabled simultaneous visualization of micron-scale spatial patterns formed by microbial cells in the mouse intestines along with biogeographical landmarks such as host-derived mucus and food particles.

X-ray fluorescence analysis of thin films at glancing-incident and -takeoff angles

International Nuclear Information System (INIS)

Tsuji, K.; Sato, S.; Hirokawa, K.

1995-01-01

We have developed a new analytical method, Glancing-Incidence and -Takeoff X-Ray Fluorescence (GIT-XRF) method for the first time. Here, we present an idea for a thin-film analysis and a surface analysis by the GIT-XRF method. In this method, the dependence of the fluorescent x-ray intensity on takeoff angle is measured at various incident angles of the primary x-ray. Compared with a total reflection x-ray fluorescence method, the GIT-XRF method allows a detailed thin-film analysis, because the thin film is cross-checked by many experimental curves. Moreover, a surface-sensitive analysis is also possible by the GIT-XRF method. (author)

Proton and electron impact on molecular and atomic oxygen: I. High resolution fluorescence spectra in the visible and VUV spectral range and emission cross-sections for dissociative ionisation and excitation of O2

International Nuclear Information System (INIS)

Wilhelmi, O.; Schartner, K.H.

2000-01-01

For pt.II see ibid., vol.11, p.45-58, 2000. Molecular oxygen O 2 was dissociated in collisions with protons and electrons in the intermediate velocity range (p + -energies: 17-800 keV, e - -energies: 0.2-2 keV). Fluorescence from excited atomic and singly ionised fragments and from singly ionised molecules was detected in the VUV and in the visible and near UV spectral range. Highly resolved spectra are presented for the VUV (46-131 nm) and the near UV/visible (340-605 nm) spectral range. Absolute emission cross-sections have been determined for dissociative ionisation and excitation leading to fluorescence in the VUV. Results are compared with published data. (orig.)

Preparation of Rhodamine B Fluorescent Poly(methacrylic acid Coated Gelatin Nanoparticles

Directory of Open Access Journals (Sweden)

Zhenhai Gan

2011-01-01

Full Text Available Poly(methacrylic acid (PMAA-coated gelatin nanoparticles encapsulated with fluorescent dye rhodamine B were prepared by the coacervation method with the aim to retard the release of rhodamine B from the gelatin matrix. With sodium sulfate as coacervation reagent for gelatin, a kind of biopolymer with excellent biocompatibility, the formed gelatin nanoparticles were cross-linked by formaldehyde followed by the polymerization of methacrylic acid coating. The fluorescent poly(methacrylic acid coated gelatin (FPMAAG nanoparticles had a uniform spherical shape and a size distribution of 60±5 nm. Infrared spectral analysis confirmed the formation of PMAA coating on the gelatin nanoparticles. Based on UV-Vis spectra, the loading efficiency of rhodamine B for the FPMAAG nanoparticles was 0.26 μg per mg nanoparticles. The encapsulated rhodamine B could sustain for two weeks. Favorable fluorescence property and fluorescence imaging of cells confirmed that the FPMAAG nanoparticles have promising biochemical, bioanalytical, and biomedical applications.

L{sub {iota}}, L{sub {alpha}}, L{sub {beta}} and L{sub {gamma}} X-ray fluorescence cross-sections of heavy elements for the exciting photons energy 38.18, 43.95, 50.21 and 59.5 keV

Energy Technology Data Exchange (ETDEWEB)

Bastug, Arif [Department of Physics Education, College of Education, Erzincan University, 24030 Erzincan (Turkey)], E-mail: abastug40@hotmail.com

2008-03-15

The cross-sections for the production of L{sub {iota}}, L{sub {alpha}}, L{sub {beta}} and L{sub {gamma}} X-ray fluorescence (XRF) in Er, Ta, W, Au, Hg, Tl, Pb and Bi by photons with energies in the range 38-59.5 keV have been measured, using a standard doublereflection experimental set-up. Measurements have been performed using an annular 241 Am primary source and X-ray emitting secondary-exciter system. Experimental cross-sections have been compared with the theoretically calculated values of L X-ray cross-sections and fairly good agreement is observed between the experimental and theoretical values.

Fluorescence Image Segmentation by using Digitally Reconstructed Fluorescence Images

OpenAIRE

Blumer, Clemens; Vivien, Cyprien; Oertner, Thomas G; Vetter, Thomas

2011-01-01

In biological experiments fluorescence imaging is used to image living and stimulated neurons. But the analysis of fluorescence images is a difficult task. It is not possible to conclude the shape of an object from fluorescence images alone. Therefore, it is not feasible to get good manual segmented nor ground truth data from fluorescence images. Supervised learning approaches are not possible without training data. To overcome this issues we propose to synthesize fluorescence images and call...

Site-specific multipoint fluorescence measurement system with end-capped optical fibers.

Science.gov (United States)

Song, Woosub; Moon, Sucbei; Lee, Byoung-Cheol; Park, Chul-Seung; Kim, Dug Young; Kwon, Hyuk Sang

2011-07-10

We present the development and implementation of a spatially and spectrally resolved multipoint fluorescence correlation spectroscopy (FCS) system utilizing multiple end-capped optical fibers and an inexpensive laser source. Specially prepared end-capped optical fibers placed in an image plane were used to both collect fluorescence signals from the sample and to deliver signals to the detectors. The placement of independently selected optical fibers on the image plane was done by monitoring the end-capped fiber tips at the focus using a CCD, and fluorescence from specific positions of a sample were collected by an end-capped fiber, which could accurately represent light intensities or spectral data without incurring any disturbance. A fast multipoint spectroscopy system with a time resolution of ∼1.5 ms was then implemented using a prism and an electron multiplying charge coupled device with a pixel binning for the region of interest. The accuracy of our proposed system was subsequently confirmed by experimental results, based on an FCS analysis of microspheres in distilled water. We expect that the proposed multipoint site-specific fluorescence measurement system can be used as an inexpensive fluorescence measurement tool to study many intracellular and molecular dynamics in cell biology. © 2011 Optical Society of America

Infrared images of reflection nebulae and Orion's bar: Fluorescent molecular hydrogen and the 3.3 micron feature

International Nuclear Information System (INIS)

Burton, M.G.; Moorhouse, A.; Brand, P.W.J.L.; Roche, P.F.; Geballe, T.R.

1989-01-01

Images were obtained of the (fluorescent) molecular hydrogen 1-0 S(1) line, and of the 3.3 micron emission feature, in Orion's Bar and three reflection nebulae. The emission from these species appears to come from the same spatial locations in all sources observed. This suggests that the 3.3 micron feature is excited by the same energetic UV-photons which cause the molecular hydrogen to fluoresce

Synchrotron X-ray fluorescence analysis in environmental and earth sciences

Directory of Open Access Journals (Sweden)

Adams F.

2010-12-01

Full Text Available Compared to other microscopic analytical tools X-ray microscopy techniques have the advantage that the large penetration depth of X-rays in matter allows one to investigate the interior of an object without destructive sample preparation. In combination with X-ray fluorescence tomography, analytical information from inside of a specimen can be obtained. Different X-ray analytical techniques can be used to produce contrast, X-ray absorption, fluorescence, and diffraction, to yield chemical, elemental, and structural information about the sample. Scanning microscopy on the basis of various lens systems in synchrotron radiation sources provides a routine spatial resolution of now about 100 nanometer but in the foreseeable future a 10–20 nanometer spatial resolution can be expected. X-ray absorption spectrometry can also provide chemical (speciation information on the sample. All this makes X-ray microscopy attractive to many fields of science. In this paper the techniques are briefly reviewed and a number of applications in the earth, planetary and cosmos sciences are illustrated with state-of-the art examples, while applications in the environmental sciences and biology are also briefly discussed.

Direct Evidence of Intrinsic Blue Fluorescence from Oligomeric Interfaces of Human Serum Albumin.

Science.gov (United States)

Bhattacharya, Arpan; Bhowmik, Soumitra; Singh, Amit K; Kodgire, Prashant; Das, Apurba K; Mukherjee, Tushar Kanti

2017-10-10

The molecular origin behind the concentration-dependent intrinsic blue fluorescence of human serum albumin (HSA) is not known yet. This unusual blue fluorescence is believed to be a characteristic feature of amyloid-like fibrils of protein/peptide and originates due to the delocalization of peptide bond electrons through the extended hydrogen bond networks of cross-β-sheet structure. Herein, by combining the results of spectroscopy, size exclusion chromatography, native gel electrophoresis, and confocal microscopy, we have shown that the intrinsic blue fluorescence of HSA exclusively originates from oligomeric interfaces devoid of any amyloid-like fibrillar structure. Our study suggests that this low energy fluorescence band is not due to any particular residue/sequence, but rather it is a common feature of self-assembled peptide bonds. The present findings of intrinsic blue fluorescence from oligomeric interfaces pave the way for future applications of this unique visual phenomenon for early stage detection of various protein aggregation related human diseases.

Bioconjugated fluorescent silica nanoparticles for the rapid detection of Entamoeba histolytica.

Science.gov (United States)

Hemadi, Ahmad; Ekrami, Alireza; Oormazdi, Hormozd; Meamar, Ahmad Reza; Akhlaghi, Lame; Samarbaf-Zadeh, Ali Reza; Razmjou, Elham

2015-05-01

Rapid detection of Entamoeba histolytica based on fluorescent silica nanoparticle (FSNP) indirect immunofluorescence microscopy was evaluated. Silica nanoparticles were synthesized using Stöber's method, with their surface activated to covalently bind to, and immobilize, protein A. For biolabeling, FSNP was added to conjugated E. histolytica trophozoites with monoclonal anti-E. histolytica IgG1 for microscopic observation of fluorescence. Fluorescent silica nanoparticle sensitivity was determined with axenically cultured E. histolytica serially diluted to seven concentrations. Specificity was evaluated using other intestinal protozoa. Fluorescent silica nanoparticles detected E. histolytica at the lowest tested concentration with no cross-reaction with Entamoeba dispar, Entamoeba moshkovskii, Blastocystis sp., or Giardia lamblia. Visualization of E. histolytica trophozoites with anti-E. histolytica antibody labeled with fluorescein isothiocyanate (FITC) was compared with that using anti-E. histolytica antibody bioconjugated FSNP. Although FITC and FSNP produced similar results, the amount of specific antibody required for FITC to induce fluorescence of similar intensity was fivefold that for FSNP. Fluorescent silica nanoparticles delivered a rapid, simple, cost-effective, and highly sensitive and specific method of detecting E. histolytica. Further study is needed before introducing FSNP for laboratory diagnosis of amoebiasis. Copyright © 2015 Elsevier B.V. All rights reserved.

Circular dichroism probed by two-photon fluorescence microscopy in enantiopure chiral polyfluorene thin films

NARCIS (Netherlands)

Savoini, M.; Wu, Xiaofei; Celebrano, M.; Ziegler, J.; Biagioni, P.; Meskers, S.C.J.; Duo, L.; Hecht, B.; Finazzi, M.

2012-01-01

Two-photon fluorescence scanning confocal microscopy sensitive to circular dichroism with a diffraction-limited resolution well below 500 nm is demonstrated. With this method, the spatial variation of the circular dichroism of thermally annealed chiral polyfluorene thin films has been imaged. We

Two-Photon Absorption Properties of Gold Fluorescent Protein: A Combined Molecular Dynamics and Quantum Chemistry Study.

Science.gov (United States)

Simsek, Yusuf; Brown, Alex

2018-05-09

Molecular dynamic (MD) simulations were carried out to obtain the conformational changes of the chromophore in the gold fluorescent protein (PDB ID: 1OXF). To obtain two-photon absorption (TPA) cross-sections, time dependent density functional theory (TD-DFT) computations were performed for chromophore geometries sampled along the trajectory. The TD-DFT computations used the CAM-B3LYP functional and 6-31+G(d) basis set with the conductor-like polarizable continuum model (PCM) with parameters for water. Results showed that two dihedral angles change remarkably over the simulation time. TPA cross-sections were found to average 20 GM for the excitation to S1 between 430 and 460 nm; however, the maximal and minimal values were 35GM and 5GM, respectively. Besides the effects of the dihedrals on the spectroscopic properties, some bond lengths affected the excitation energies and the TPA cross-sections significantly (up to ±25-30%) while the effects of bond angles were smaller (±5%). Overall the present results provide insight in the effects of conformational exibility on TPA (with gold fluorescent protein as a specific example) and suggest that further experimental measurements of TPA for gold fluorescent protein should be undertaken.

Stabilization of structure in near-infrared fluorescent proteins by binding of biliverdin chromophore

Science.gov (United States)

Stepanenko, Olesya V.; Stepanenko, Olga V.; Bublikov, G. S.; Kuznetsova, I. M.; Verkhusha, V. V.; Turoverov, K. K.

2017-07-01

Near-infrared fluorescent proteins (NIR FPs) engineered from bacterial phytochromes and their mutants with different location of Cys residues, which able to bind a biliverdin chromophore, or without these Cys residues were studied using intrinsic tryptophan fluorescence, NIR fluorescence and circular dichroism. It was shown that a covalent binding of the biliverdin chromophore to a Cys residue via thioether group substantially stabilizes the spatial structure of NIR FPs. The stability of the protein structure and the chromophore association strength strongly depends on the location of Cys residues and decreases in the following order: a protein with Cys residues in both domains, a protein with Cys in PAS domains, and a protein with Cys in GAF domains. NIR FPs without Cys residues capable to covalently attach biliverdin have the lowest stability, comparable to NIR FP apoforms.

Use of multiphoton tomography and fluorescence lifetime imaging to investigate skin pigmentation in vivo

Science.gov (United States)

Dancik, Yuri; Favre, Amandine; Loy, Chong Jin; Zvyagin, Andrei V.; Roberts, Michael S.

2013-02-01

There is a growing body of literature showing the usefulness of multiphoton tomography (MPT) and fluorescence lifetime imaging for in situ characterization of skin constituents and the ensuing development of noninvasive diagnostic tools against skin diseases. Melanin and pigmentation-associated skin cancers constitute some of the major applications. We show that MPT and fluorescence lifetime imaging can be used to measure changes in cutaneous melanin concentration and that these can be related to the visible skin color. Melanin in the skin of African, Indian, Caucasian, and Asian volunteers is detected on the basis of its emission wavelength and fluorescence lifetimes in solution and in a melanocyte-keratinocyte cell culture. Fluorescence intensity is used to characterize the melanin content and distribution as a function of skin type and depth into the skin (stratum granulosum and stratum basale). The measured fluorescence intensities in given skin types agree with melanin amounts reported by others using biopsies. Our results suggest that spatial distribution of melanin in skin can be studied using MPT and fluorescence lifetime imaging, but further studies are needed to ascertain that the method can resolve melanin amount in smaller depth intervals.

Multiphoton fluorescence lifetime imaging of chemotherapy distribution in solid tumors

Science.gov (United States)

Carlson, Marjorie; Watson, Adrienne L.; Anderson, Leah; Largaespada, David A.; Provenzano, Paolo P.

2017-11-01

Doxorubicin is a commonly used chemotherapeutic employed to treat multiple human cancers, including numerous sarcomas and carcinomas. Furthermore, doxorubicin possesses strong fluorescent properties that make it an ideal reagent for modeling drug delivery by examining its distribution in cells and tissues. However, while doxorubicin fluorescence and lifetime have been imaged in live tissue, its behavior in archival samples that frequently result from drug and treatment studies in human and animal patients, and murine models of human cancer, has to date been largely unexplored. Here, we demonstrate imaging of doxorubicin intensity and lifetimes in archival formalin-fixed paraffin-embedded sections from mouse models of human cancer with multiphoton excitation and multiphoton fluorescence lifetime imaging microscopy (FLIM). Multiphoton excitation imaging reveals robust doxorubicin emission in tissue sections and captures spatial heterogeneity in cells and tissues. However, quantifying the amount of doxorubicin signal in distinct cell compartments, particularly the nucleus, often remains challenging due to strong signals in multiple compartments. The addition of FLIM analysis to display the spatial distribution of excited state lifetimes clearly distinguishes between signals in distinct compartments such as the cell nuclei versus cytoplasm and allows for quantification of doxorubicin signal in each compartment. Furthermore, we observed a shift in lifetime values in the nuclei of transformed cells versus nontransformed cells, suggesting a possible diagnostic role for doxorubicin lifetime imaging to distinguish normal versus transformed cells. Thus, data here demonstrate that multiphoton FLIM is a highly sensitive platform for imaging doxorubicin distribution in normal and diseased archival tissues.

Imaging performance of a hybrid x-ray computed tomography-fluorescence molecular tomography system using priors.

Science.gov (United States)

Ale, Angelique; Schulz, Ralf B; Sarantopoulos, Athanasios; Ntziachristos, Vasilis

2010-05-01

The performance is studied of two newly introduced and previously suggested methods that incorporate priors into inversion schemes associated with data from a recently developed hybrid x-ray computed tomography and fluorescence molecular tomography system, the latter based on CCD camera photon detection. The unique data set studied attains accurately registered data of high spatially sampled photon fields propagating through tissue along 360 degrees projections. Approaches that incorporate structural prior information were included in the inverse problem by adding a penalty term to the minimization function utilized for image reconstructions. Results were compared as to their performance with simulated and experimental data from a lung inflammation animal model and against the inversions achieved when not using priors. The importance of using priors over stand-alone inversions is also showcased with high spatial sampling simulated and experimental data. The approach of optimal performance in resolving fluorescent biodistribution in small animals is also discussed. Inclusion of prior information from x-ray CT data in the reconstruction of the fluorescence biodistribution leads to improved agreement between the reconstruction and validation images for both simulated and experimental data.

Photoacoustic tomography of human hepatic malignancies using intraoperative indocyanine green fluorescence imaging.

Directory of Open Access Journals (Sweden)

Akinori Miyata

Full Text Available Recently, fluorescence imaging following the preoperative intravenous injection of indocyanine green has been used in clinical settings to identify hepatic malignancies during surgery. The aim of this study was to evaluate the ability of photoacoustic tomography using indocyanine green as a contrast agent to produce representative fluorescence images of hepatic tumors by visualizing the spatial distribution of indocyanine green on ultrasonographic images. Indocyanine green (0.5 mg/kg, intravenous was preoperatively administered to 9 patients undergoing hepatectomy. Intraoperatively, photoacoustic tomography was performed on the surface of the resected hepatic specimens (n = 10 under excitation with an 800 nm pulse laser. In 4 hepatocellular carcinoma nodules, photoacoustic imaging identified indocyanine green accumulation in the cancerous tissue. In contrast, in one hepatocellular carcinoma nodule and five adenocarcinoma foci (one intrahepatic cholangiocarcinoma and 4 colorectal liver metastases, photoacoustic imaging delineated indocyanine green accumulation not in the cancerous tissue but rather in the peri-cancerous hepatic parenchyma. Although photoacoustic tomography enabled to visualize spatial distribution of ICG on ultrasonographic images, which was consistent with fluorescence images on cut surfaces of the resected specimens, photoacoustic signals of ICG-containing tissues decreased approximately by 40% even at 4 mm depth from liver surfaces. Photoacoustic tomography using indocyanine green also failed to identify any hepatocellular carcinoma nodules from the body surface of model mice with non-alcoholic steatohepatitis. In conclusion, photoacoustic tomography has a potential to enhance cancer detectability and differential diagnosis by ultrasonographic examinations and intraoperative fluorescence imaging through visualization of stasis of bile-excreting imaging agents in and/or around hepatic tumors. However, further technical

Photoacoustic tomography of human hepatic malignancies using intraoperative indocyanine green fluorescence imaging.

Science.gov (United States)

Miyata, Akinori; Ishizawa, Takeaki; Kamiya, Mako; Shimizu, Atsushi; Kaneko, Junichi; Ijichi, Hideaki; Shibahara, Junji; Fukayama, Masashi; Midorikawa, Yutaka; Urano, Yasuteru; Kokudo, Norihiro

2014-01-01

Recently, fluorescence imaging following the preoperative intravenous injection of indocyanine green has been used in clinical settings to identify hepatic malignancies during surgery. The aim of this study was to evaluate the ability of photoacoustic tomography using indocyanine green as a contrast agent to produce representative fluorescence images of hepatic tumors by visualizing the spatial distribution of indocyanine green on ultrasonographic images. Indocyanine green (0.5 mg/kg, intravenous) was preoperatively administered to 9 patients undergoing hepatectomy. Intraoperatively, photoacoustic tomography was performed on the surface of the resected hepatic specimens (n = 10) under excitation with an 800 nm pulse laser. In 4 hepatocellular carcinoma nodules, photoacoustic imaging identified indocyanine green accumulation in the cancerous tissue. In contrast, in one hepatocellular carcinoma nodule and five adenocarcinoma foci (one intrahepatic cholangiocarcinoma and 4 colorectal liver metastases), photoacoustic imaging delineated indocyanine green accumulation not in the cancerous tissue but rather in the peri-cancerous hepatic parenchyma. Although photoacoustic tomography enabled to visualize spatial distribution of ICG on ultrasonographic images, which was consistent with fluorescence images on cut surfaces of the resected specimens, photoacoustic signals of ICG-containing tissues decreased approximately by 40% even at 4 mm depth from liver surfaces. Photoacoustic tomography using indocyanine green also failed to identify any hepatocellular carcinoma nodules from the body surface of model mice with non-alcoholic steatohepatitis. In conclusion, photoacoustic tomography has a potential to enhance cancer detectability and differential diagnosis by ultrasonographic examinations and intraoperative fluorescence imaging through visualization of stasis of bile-excreting imaging agents in and/or around hepatic tumors. However, further technical advances are needed

Three-dimensional super-resolution imaging for fluorescence emission difference microscopy

Energy Technology Data Exchange (ETDEWEB)

You, Shangting; Kuang, Cuifang, E-mail: cfkuang@zju.edu.cn; Li, Shuai; Liu, Xu; Ding, Zhihua [State key laboratory of modern optical instrumentations, Zhejiang University, Hangzhou 310027 (China)

2015-08-15

We propose a method theoretically to break the diffraction limit and to improve the resolution in all three dimensions for fluorescence emission difference microscopy. We produce two kinds of hollow focal spot by phase modulation. By incoherent superposition, these two kinds of focal spot yield a 3D hollow focal spot. The optimal proportion of these two kinds of spot is given in the paper. By employing 3D hollow focal spot, super-resolution image can be yielded by means of fluorescence emission difference microscopy, with resolution enhanced both laterally and axially. According to computation result, size of point spread function of three-dimensional super-resolution imaging is reduced by about 40% in all three spatial directions with respect to confocal imaging.

Trade-off between angular and spatial resolutions in in vivo fiber tractography

OpenAIRE

Vos, Sjoerd B.; Aksoy, Murat; Han, Zhaoying; Holdsworth, Samantha J.; Maclaren, Julian; Viergever, Max A.; Leemans, Alexander; Bammer, Roland

2016-01-01

Tractography is becoming an increasingly popular method to reconstruct white matter connections in vivo. The diffusion MRI data that tractography is based on requires a high angular resolution to resolve crossing fibers whereas high spatial resolution is required to distinguish kissing from crossing fibers. However, scan time increases with increasing spatial and angular resolutions, which can become infeasible in clinical settings. Here we investigated the trade-off between spatial and angul...

The Threshold Hypothesis Applied to Spatial Skill and Mathematics

Science.gov (United States)

Freer, Daniel

2017-01-01

This cross-sectional study assessed the relation between spatial skills and mathematics in 854 participants across kindergarten, third grade, and sixth grade. Specifically, the study probed for a threshold for spatial skills when performing mathematics, above which spatial scores and mathematics scores would be significantly less related. This…

Mesoporous silica for drug delivery: Interactions with model fluorescent lipid vesicles and live cells.

Science.gov (United States)

Bardhan, Munmun; Majumdar, Anupa; Jana, Sayantan; Ghosh, Tapas; Pal, Uttam; Swarnakar, Snehasikta; Senapati, Dulal

2018-01-01

Formulated mesoporous silica nanoparticle (MSN) systems offer the best possible drug delivery system through the release of drug molecules from the accessible pores. In the present investigation, steady state and time resolved fluorescence techniques along with the fluorescence imaging were applied to investigate the interactions of dye loaded MSN with fluorescent unilamellar vesicles and live cells. Here 1,2-dimyristoyl-sn-glycero-3-phospocholine (DMPC) was used to prepare Small Unilamellar Vesicles (SUVs) as the model membrane with fluorescent 1,6-diphenyl-1,3,5-hexatriene (DPH) molecule incorporated inside the lipid bilayer. The interaction of DPH incorporated DMPC membrane with Fluorescein loaded MSN lead to the release of Fluorescein (Fl) dye from the interior pores of MSN systems. The extent of release of Fl and spatial distribution of the DPH molecule has been explored by monitoring steady-state fluorescence intensity and fluorescence lifetime at physiological condition. To investigate the fate of drug molecule released from MSN, fluorescence anisotropy has been used. The drug delivery efficiency of the MSN as a carrier for doxorubicin (DOX), a fluorescent chemotherapeutic drug, has also been investigated at physiological conditions. The study gives a definite confirmation for high uptake and steady release of DOX in primary oral mucosal non-keratinized squamous cells in comparison to naked DOX treatment. Copyright © 2017 Elsevier B.V. All rights reserved.

Two-Photon Fluorescence Microscope for Microgravity Research

Science.gov (United States)

Fischer, David G.; Zimmerli, Gregory A.; Asipauskas, Marius

2005-01-01

A two-photon fluorescence microscope has been developed for the study of biophysical phenomena. Two-photon microscopy is a novel form of laser-based scanning microscopy that enables three-dimensional imaging without many of the problems inherent in confocal microscopy. Unlike one-photon optical microscopy, two-photon microscopy utilizes the simultaneous nonlinear absorption of two near-infrared photons. However, the efficiency of two-photon absorption is much lower than that of one-photon absorption, so an ultra-fast pulsed laser source is typically employed. On the other hand, the critical energy threshold for two-photon absorption leads to fluorophore excitation that is intrinsically localized to the focal volume. Consequently, two-photon microscopy enables optical sectioning and confocal performance without the need for a signal-limiting pinhole. In addition, there is a reduction (relative to one-photon optical microscopy) in photon-induced damage because of the longer excitation wavelength. This reduction is especially advantageous for in vivo studies. Relative to confocal microscopy, there is also a reduction in background fluorescence, and, because of a reduction in Rayleigh scattering, there is a 4 increase of penetration depth. The prohibitive cost of a commercial two-photon fluorescence-microscope system, as well as a need for modularity, has led to the construction of a custom-built system (see Figure 1). This system includes a coherent mode-locked titanium: sapphire laser emitting 120-fs-duration pulses at a repetition rate of 80 MHz. The pulsed laser has an average output power of 800 mW and a wavelength tuning range of 700 to 980 nm, enabling the excitation of a variety of targeted fluorophores. The output from the laser is attenuated, spatially filtered, and then directed into a confocal scanning head that has been modified to provide for side entry of the laser beam. The laser output coupler has been replaced with a dichroic filter that reflects the

Rotational multispectral fluorescence lifetime imaging and intravascular ultrasound: bimodal system for intravascular applications

Science.gov (United States)

Ma, Dinglong; Bec, Julien; Yankelevich, Diego R.; Gorpas, Dimitris; Fatakdawala, Hussain; Marcu, Laura

2014-01-01

Abstract. We report the development and validation of a hybrid intravascular diagnostic system combining multispectral fluorescence lifetime imaging (FLIm) and intravascular ultrasound (IVUS) for cardiovascular imaging applications. A prototype FLIm system based on fluorescence pulse sampling technique providing information on artery biochemical composition was integrated with a commercial IVUS system providing information on artery morphology. A customized 3-Fr bimodal catheter combining a rotational side-view fiberoptic and a 40-MHz IVUS transducer was constructed for sequential helical scanning (rotation and pullback) of tubular structures. Validation of this bimodal approach was conducted in pig heart coronary arteries. Spatial resolution, fluorescence detection efficiency, pulse broadening effect, and lifetime measurement variability of the FLIm system were systematically evaluated. Current results show that this system is capable of temporarily resolving the fluorescence emission simultaneously in multiple spectral channels in a single pullback sequence. Accurate measurements of fluorescence decay characteristics from arterial segments can be obtained rapidly (e.g., 20 mm in 5 s), and accurate co-registration of fluorescence and ultrasound features can be achieved. The current finding demonstrates the compatibility of FLIm instrumentation with in vivo clinical investigations and its potential to complement conventional IVUS during catheterization procedures. PMID:24898604

The one-sample PARAFAC approach reveals molecular size distributions of fluorescent components in dissolved organic matter

DEFF Research Database (Denmark)

Wünsch, Urban; Murphy, Kathleen R.; Stedmon, Colin

2017-01-01

Molecular size plays an important role in dissolved organic matter (DOM) biogeochemistry, but its relationship with the fluorescent fraction of DOM (FDOM) remains poorly resolved. Here high-performance size exclusion chromatography (HPSEC) was coupled to fluorescence emission-excitation (EEM...... but not their spectral properties. Thus, in contrast to absorption measurements, bulk fluorescence is unlikely to reliably indicate the average molecular size of DOM. The one-sample approach enables robust and independent cross-site comparisons without large-scale sampling efforts and introduces new analytical...... opportunities for elucidating the origins and biogeochemical properties of FDOM...

Application of synchrotron radiation to x-ray fluorescence analysis of trace elements

International Nuclear Information System (INIS)

Gordon, B.M.; Jones, K.W.; Hanson, A.L.

1986-08-01

The development of synchrotron radiation x-ray sources has provided the means to greatly extend the capabilities of x-ray fluorescence analysis for determinations of trace element concentrations. A brief description of synchrotron radiation properties provides a background for a discussion of the improved detection limits compared to existing x-ray fluorescence techniques. Calculated detection limits for x-ray microprobes with micrometer spatial resolutions are described and compared with experimental results beginning to appear from a number of laboratories. The current activities and future plans for a dedicated x-ray microprobe beam line at the National Synchrotron Light Source (NSLS) of Brookhaven National Laboratory are presented

The role of secondary electrons in some experiments determining fluorescence emission from nitrogen C3Πu levels

International Nuclear Information System (INIS)

Blanco, F.; Arqueros, F.

2005-01-01

The processes involved in fluorescence emission from C 3 Π u levels of N 2 molecule by electron impact are studied. Secondary electrons are shown to play an important role in typical experiments for the measurement of emission cross sections and fluorescence yields, dominating at high impact energies. A simple model is proposed which accounts for fluorescence measurements in a wide range of experimental conditions, and in particular for some recent results up to 1 GeV energies

Fluorescence spectroscopy as a tool for quality assessment of humic substances

Science.gov (United States)

Boguta, Patrycja

2016-04-01

*The studies were partly carried out within the framework of a research project. The project was financed from funds of National Science Center on the base of decision number DEC-2013/11/D/NZ9/02545. Fluorescence spectroscopy belongs to modern, non-destructive, rapid and relatively cheap methods, as well as for many years it was successfully used in studies of organic compounds in the fields of medicine, biology and chemistry. On the other hand, soil organic matter is a group of compounds with a complex spatial structure showing a large number of groups with different kinds of fluorophores. This could suggest the possibility of application of fluorescence spectroscopy in assessing the quality of humic substances as well as in monitoring of their chemical transformations. The aim of study was chemical description of humic and fulvic acids based on fluorescence spectra, as well as an attempt of evaluation of changes occurring under the influence of different pH and during interactions with various concentrations of metal. The humic and fulvic acids were isolated from chemically different soils. The measurements were carried out on Hitachi fluorescence spectrometer in solutions with a concentration of humic acids 40mg dm-3, at pH from 3 to 7, and for the evaluation of the metal impact: with increasing Zn concentrations (0-50mg dm-3). The fluorescence spectra were recorded in the form of synchronous and emission-excitation matrices (EEM). Studies have shown the presence of different groups of fluorophores. Synchronous spectra were characterized by a well-separated bands showing fluorescence in the area of low, medium and high wavelengths, suggesting the presence of structures, both weakly and strongly humified. EEM spectra revealed map of fluorophores within wide ranges of emission and excitation. Fluorophores differed in both position and intensity. The highest intensity was observed for compounds with the lowest humification degree which might be due to high amount

Two-photon-excited fluorescence spectroscopy of atomic fluorine at 170 nm

Science.gov (United States)

Herring, G. C.; Dyer, Mark J.; Jusinski, Leonard E.; Bischel, William K.

1988-01-01

Two-photon-excited fluorescence spectroscopy of atomic fluorine is reported. A doubled dye laser at 286-nm is Raman shifted in H2 to 170 nm (sixth anti-Stokes order) to excite ground-state 2P(0)J fluorine atoms to the 2D(0)J level. The fluorine atoms are detected by one of two methods: observing the fluorescence decay to the 2PJ level or observing F(+) production through the absorption of an additional photon by the excited atoms. Relative two-photon absorption cross sections to and the radiative lifetimes of the 2D(0)J states are measured.

X-ray fluorescence in some medium-Z elements excited by 59.5 keV photons

International Nuclear Information System (INIS)

Han, I.; Shahin, M.; Demir, L.; Narmanli, E.

2010-01-01

K X-ray fluorescence parameters cross sections and average shell fluorescence yields) for selected ten elements in the atomic range 42 ≤ Z ≤ 66 have been experimentally determined at photon excitation energy of 59.5 keV. K X-rays emitted from the samples have been counted by a Si (Li) detector. The K spectra for investigated elements have been derived from the measured K shell X-ray spectra by peak fitting process. Experimental results of K X-ray fluorescence parameters have been compared with theory. In general there is an agreement within the standard uncertainties of the experimental and theoretical values

[Rapid identification of hogwash oil by using synchronous fluorescence spectroscopy].

Science.gov (United States)

Sun, Yan-Hui; An, Hai-Yang; Jia, Xiao-Li; Wang, Juan

2012-10-01

To identify hogwash oil quickly, the characteristic delta lambda of hogwash oil was analyzed by three dimensional fluorescence spectroscopy with parallel factor analysis, and the model was built up by using synchronous fluorescence spectroscopy with support vector machines (SVM). The results showed that the characteristic delta lambda of hogwash oil was 60 nm. Collecting original spectrum of different samples under the condition of characteristic delta lambda 60 nm, the best model was established while 5 principal components were selected from original spectrum and the radial basis function (RBF) was used as the kernel function, and the optimal penalty factor C and kernel function g were 512 and 0.5 respectively obtained by the grid searching and 6-fold cross validation. The discrimination rate of the model was 100% for both training sets and prediction sets. Thus, it is quick and accurate to apply synchronous fluorescence spectroscopy to identification of hogwash oil.

Assessing implicit odor localization in humans using a cross-modal spatial cueing paradigm.

Science.gov (United States)

Moessnang, Carolin; Finkelmeyer, Andreas; Vossen, Alexandra; Schneider, Frank; Habel, Ute

2011-01-01

Navigation based on chemosensory information is one of the most important skills in the animal kingdom. Studies on odor localization suggest that humans have lost this ability. However, the experimental approaches used so far were limited to explicit judgements, which might ignore a residual ability for directional smelling on an implicit level without conscious appraisal. A novel cueing paradigm was developed in order to determine whether an implicit ability for directional smelling exists. Participants performed a visual two-alternative forced choice task in which the target was preceded either by a side-congruent or a side-incongruent olfactory spatial cue. An explicit odor localization task was implemented in a second experiment. No effect of cue congruency on mean reaction times could be found. However, a time by condition interaction emerged, with significantly slower responses to congruently compared to incongruently cued targets at the beginning of the experiment. This cueing effect gradually disappeared throughout the course of the experiment. In addition, participants performed at chance level in the explicit odor localization task, thus confirming the results of previous research. The implicit cueing task suggests the existence of spatial information processing in the olfactory system. Response slowing after a side-congruent olfactory cue is interpreted as a cross-modal attentional interference effect. In addition, habituation might have led to a gradual disappearance of the cueing effect. It is concluded that under immobile conditions with passive monorhinal stimulation, humans are unable to explicitly determine the location of a pure odorant. Implicitly, however, odor localization seems to exert an influence on human behaviour. To our knowledge, these data are the first to show implicit effects of odor localization on overt human behaviour and thus support the hypothesis of residual directional smelling in humans. © 2011 Moessnang et al.

Assessing implicit odor localization in humans using a cross-modal spatial cueing paradigm.

Directory of Open Access Journals (Sweden)

Carolin Moessnang

Full Text Available Navigation based on chemosensory information is one of the most important skills in the animal kingdom. Studies on odor localization suggest that humans have lost this ability. However, the experimental approaches used so far were limited to explicit judgements, which might ignore a residual ability for directional smelling on an implicit level without conscious appraisal.A novel cueing paradigm was developed in order to determine whether an implicit ability for directional smelling exists. Participants performed a visual two-alternative forced choice task in which the target was preceded either by a side-congruent or a side-incongruent olfactory spatial cue. An explicit odor localization task was implemented in a second experiment.No effect of cue congruency on mean reaction times could be found. However, a time by condition interaction emerged, with significantly slower responses to congruently compared to incongruently cued targets at the beginning of the experiment. This cueing effect gradually disappeared throughout the course of the experiment. In addition, participants performed at chance level in the explicit odor localization task, thus confirming the results of previous research.The implicit cueing task suggests the existence of spatial information processing in the olfactory system. Response slowing after a side-congruent olfactory cue is interpreted as a cross-modal attentional interference effect. In addition, habituation might have led to a gradual disappearance of the cueing effect. It is concluded that under immobile conditions with passive monorhinal stimulation, humans are unable to explicitly determine the location of a pure odorant. Implicitly, however, odor localization seems to exert an influence on human behaviour. To our knowledge, these data are the first to show implicit effects of odor localization on overt human behaviour and thus support the hypothesis of residual directional smelling in humans.

Exciton-controlled fluorescence: application to hybridization-sensitive fluorescent DNA probe.

Science.gov (United States)

Okamoto, Akimitsu; Ikeda, Shuji; Kubota, Takeshi; Yuki, Mizue; Yanagisawa, Hiroyuki

2009-01-01

A hybridization-sensitive fluorescent probe has been designed for nucleic acid detection, using the concept of fluorescence quenching caused by the intramolecular excitonic interaction of fluorescence dyes. We synthesized a doubly thiazole orange-labeled nucleotide showing high fluorescence intensity for a hybrid with the target nucleic acid and effective quenching for the single-stranded state. This exciton-controlled fluorescent probe was applied to living HeLa cells using microinjection to visualize intracellular mRNA localization. Immediately after injection of the probe into the cell, fluorescence was observed from the probe hybridizing with the target RNA. This fluorescence rapidly decreased upon addition of a competitor DNA. Multicoloring of this probe resulted in the simple simultaneous detection of plural target nucleic acid sequences. This probe realized a large, rapid, reversible change in fluorescence intensity in sensitive response to the amount of target nucleic acid, and facilitated spatiotemporal monitoring of the behavior of intracellular RNA.

Dual-channel (green and red) fluorescence microendoscope with subcellular resolution

Science.gov (United States)

de Paula D'Almeida, Camila; Fortunato, Thereza Cury; Teixeira Rosa, Ramon Gabriel; Romano, Renan Arnon; Moriyama, Lilian Tan; Pratavieira, Sebastião.

2018-02-01

Usually, tissue images at cellular level need biopsies to be done. Considering this, diagnostic devices, such as microendoscopes, have been developed with the purpose of do not be invasive. This study goal is the development of a dual-channel microendoscope, using two fluorescent labels: proflavine and protoporphyrin IX (PpIX), both approved by Food and Drug Administration. This system, with the potential to perform a microscopic diagnosis and to monitor a photodynamic therapy (PDT) session, uses a halogen lamp and an image fiber bundle to perform subcellular image. Proflavine fluorescence indicates the nuclei of the cell, which is the reference for PpIX localization on image tissue. Preliminary results indicate the efficacy of this optical technique to detect abnormal tissues and to improve the PDT dosimetry. This was the first time, up to our knowledge, that PpIX fluorescence was microscopically observed in vivo, in real time, combined to other fluorescent marker (Proflavine), which allowed to simultaneously observe the spatial localization of the PpIX in the mucosal tissue. We believe this system is very promising tool to monitor PDT in mucosa as it happens. Further experiments have to be performed in order to validate the system for PDT monitoring.

Picosecond wide-field time-correlated single photon counting fluorescence microscopy with a delay line anode detector

Energy Technology Data Exchange (ETDEWEB)

Hirvonen, Liisa M.; Le Marois, Alix; Suhling, Klaus, E-mail: klaus.suhling@kcl.ac.uk [Department of Physics, King' s College London, Strand, London WC2R 2LS (United Kingdom); Becker, Wolfgang; Smietana, Stefan [Becker & Hickl GmbH, Nahmitzer Damm 30, 12277 Berlin (Germany); Milnes, James; Conneely, Thomas [Photek Ltd., 26 Castleham Rd, Saint Leonards-on-Sea TN38 9NS (United Kingdom); Jagutzki, Ottmar [Institut für Kernphysik, Max-von-Laue-Str. 1, 60438 Frankfurt (Germany)

2016-08-15

We perform wide-field time-correlated single photon counting-based fluorescence lifetime imaging (FLIM) with a crossed delay line anode image intensifier, where the pulse propagation time yields the photon position. This microchannel plate-based detector was read out with conventional fast timing electronics and mounted on a fluorescence microscope with total internal reflection (TIR) illumination. The picosecond time resolution of this detection system combines low illumination intensity of microwatts with wide-field data collection. This is ideal for fluorescence lifetime imaging of cell membranes using TIR. We show that fluorescence lifetime images of living HeLa cells stained with membrane dye di-4-ANEPPDHQ exhibit a reduced lifetime near the coverslip in TIR compared to epifluorescence FLIM.

On the influence of spatial discretization on cross section preparation with HELIOS 1.9

International Nuclear Information System (INIS)

Merk, B.; Koch, R.

2008-01-01

function'' [ 2]. This approximation requires a careful spatial discretization to produce reliable results. The problem of discretization for the solution of the diffusion equation by finite difference methods has been studied in the past [ 3]. The influence of the spatial discretization strategy on the infinite multiplication facto k inf , the neutron flux distribution and the prepared two group cross sections in a single cell will be investigated here for the collision probability method of the code system HELIOS 1.9. (orig.)

Fluorescence spectroscopy

DEFF Research Database (Denmark)

Bagatolli, Luis

2016-01-01

Fluorescence spectroscopy is a powerful experimental tool used by scientists from many disciplines. During the last decades there have been important developments on distinct fluorescence methods, particularly those related to the study of biological phenomena. This chapter discusses the foundati......Fluorescence spectroscopy is a powerful experimental tool used by scientists from many disciplines. During the last decades there have been important developments on distinct fluorescence methods, particularly those related to the study of biological phenomena. This chapter discusses...

Ultrasensitive near-infrared fluorescence-enhanced probe for in vivo nitroreductase imaging.

Science.gov (United States)

Li, Yuhao; Sun, Yun; Li, Jiachang; Su, Qianqian; Yuan, Wei; Dai, Yu; Han, Chunmiao; Wang, Qiuhong; Feng, Wei; Li, Fuyou

2015-05-20

Nitroreductase (NTR) can be overexpressed in hypoxic tumors, thus the selective and efficient detection of NTR is of great importance. To date, although a few optical methods have been reported for the detection of NTR in solution, an effective optical probe for NTR monitoring in vivo is still lacking. Therefore, it is necessary to develop a near-infrared (NIR) fluorescent detection probe for NTR. In this study, five NIR cyanine dyes with fluorescence reporting structure decorated with different nitro aromatic groups, Cy7-1-5, have been designed and explored for possible rapid detection of NTR. Our experimental results presented that only a para-nitro benzoate group modified cyanine probe (Cy7-1) could serve as a rapid NIR fluorescence-enhanced probe for monitoring and bioimaging of NTR. The structure-function relationship has been revealed by theoretical study. The linker connecting the detecting and fluorescence reporting groups and the nitro group position is a key factor for the formation of hydrogen bonds and spatial structure match, inducing the NTR catalytic ability enhancement. The in vitro response and mechanism of the enzyme-catalyzed reduction of Cy7-1 have been investigated through kinetic optical studies and other methods. The results have indicated that an electro-withdrawing group induced electron-transfer process becomes blocked when Cy7-1 is catalytically reduced to Cy7-NH2 by NTR, which is manifested in enhanced fluorescence intensity during the detection process. Confocal fluorescence imaging of hypoxic A549 cells has confirmed the NTR detection ability of Cy7-1 at the cellular level. Importantly, Cy7-1 can detect tumor hypoxia in a murine hypoxic tumor model, showing a rapid and significant enhancement of its NIR fluorescence characteristics suitable for fluorescence bioimaging. This method may potentially be used for tumor hypoxia diagnosis.

On-chip multiplexed solid-phase nucleic acid hybridization assay using spatial profiles of immobilized quantum dots and fluorescence resonance energy transfer.

Science.gov (United States)

Noor, M Omair; Tavares, Anthony J; Krull, Ulrich J

2013-07-25

A microfluidic based solid-phase assay for the multiplexed detection of nucleic acid hybridization using quantum dot (QD) mediated fluorescence resonance energy transfer (FRET) is described herein. The glass surface of hybrid glass-polydimethylsiloxane (PDMS) microfluidic channels was chemically modified to assemble the biorecognition interface. Multiplexing was demonstrated using a detection system that was comprised of two colors of immobilized semi-conductor QDs and two different oligonucleotide probe sequences. Green-emitting and red-emitting QDs were paired with Cy3 and Alexa Fluor 647 (A647) labeled oligonucleotides, respectively. The QDs served as energy donors for the transduction of dye labeled oligonucleotide targets. The in-channel assembly of the biorecognition interface and the subsequent introduction of oligonucleotide targets was accomplished within minutes using a combination of electroosmotic flow and electrophoretic force. The concurrent quantification of femtomole quantities of two target sequences was possible by measuring the spatial coverage of FRET sensitized emission along the length of the channel. In previous reports, multiplexed QD-FRET hybridization assays that employed a ratiometric method for quantification had challenges associated with lower analytical sensitivity arising from both donor and acceptor dilution that resulted in reduced energy transfer pathways as compared to single-color hybridization assays. Herein, a spatial method for quantification that is based on in-channel QD-FRET profiles provided higher analytical sensitivity in the multiplexed assay format as compared to single-color hybridization assays. The selectivity of the multiplexed hybridization assays was demonstrated by discrimination between a fully-complementary sequence and a 3 base pair sequence at a contrast ratio of 8 to 1. Copyright © 2013 Elsevier B.V. All rights reserved.

The Mapping X-ray Fluorescence Spectrometer (MapX)

Science.gov (United States)

Sarrazin, P.; Blake, D. F.; Marchis, F.; Bristow, T.; Thompson, K.

2017-12-01

Many planetary surface processes leave traces of their actions as features in the size range 10s to 100s of microns. The Mapping X-ray Fluorescence Spectrometer (MapX) will provide elemental imaging at 100 micron spatial resolution, yielding elemental chemistry at a scale where many relict physical, chemical, or biological features can be imaged and interpreted in ancient rocks on planetary bodies and planetesimals. MapX is an arm-based instrument positioned on a rock or regolith with touch sensors. During an analysis, an X-ray source (tube or radioisotope) bombards the sample with X-rays or alpha-particles / gamma-rays, resulting in sample X-ray Fluorescence (XRF). X-rays emitted in the direction of an X-ray sensitive CCD imager pass through a 1:1 focusing lens (X-ray micro-pore Optic (MPO)) that projects a spatially resolved image of the X-rays onto the CCD. The CCD is operated in single photon counting mode so that the energies and positions of individual X-ray photons are recorded. In a single analysis, several thousand frames are both stored and processed in real-time. Higher level data products include single-element maps with a lateral spatial resolution of 100 microns and quantitative XRF spectra from ground- or instrument- selected Regions of Interest (ROI). XRF spectra from ROI are compared with known rock and mineral compositions to extrapolate the data to rock types and putative mineralogies. When applied to airless bodies and implemented with an appropriate radioisotope source for alpha-particle excitation, MapX will be able to analyze biogenic elements C, N, O, P, S, in addition to the cations of the rock-forming elements >Na, accessible with either X-ray or gamma-ray excitation. The MapX concept has been demonstrated with a series of lab-based prototypes and is currently under refinement and TRL maturation.

Fluorescence enhancement in large-scale self-assembled gold nanoparticle double arrays

International Nuclear Information System (INIS)

Chekini, M.; Bierwagen, J.; Cunningham, A.; Bürgi, T.; Filter, R.; Rockstuhl, C.

2015-01-01

Localized surface plasmon resonances excited in metallic nanoparticles confine and enhance electromagnetic fields at the nanoscale. This is particularly pronounced in dimers made from two closely spaced nanoparticles. When quantum emitters, such as dyes, are placed in the gap of those dimers, their absorption and emission characteristics can be modified. Both processes have to be considered when aiming to enhance the fluorescence from the quantum emitters. This is particularly challenging for dimers, since the electromagnetic properties and the enhanced fluorescence sensitively depend on the distance between the nanoparticles. Here, we use a layer-by-layer method to precisely control the distances in such systems. We consider a dye layer deposited on top of an array of gold nanoparticles or integrated into a central position of a double array of gold nanoparticles. We study the effect of the spatial arrangement and the average distance on the plasmon-enhanced fluorescence. We found a maximum of a 99-fold increase in the fluorescence intensity of the dye layer sandwiched between two gold nanoparticle arrays. The interaction of the dye layer with the plasmonic system also causes a spectral shift in the emission wavelengths and a shortening of the fluorescence life times. Our work paves the way for large-scale, high throughput, and low-cost self-assembled functionalized plasmonic systems that can be used as efficient light sources

Deep-tissue reporter-gene imaging with fluorescence and optoacoustic tomography: a performance overview.

Science.gov (United States)

Deliolanis, Nikolaos C; Ale, Angelique; Morscher, Stefan; Burton, Neal C; Schaefer, Karin; Radrich, Karin; Razansky, Daniel; Ntziachristos, Vasilis

2014-10-01

A primary enabling feature of near-infrared fluorescent proteins (FPs) and fluorescent probes is the ability to visualize deeper in tissues than in the visible. The purpose of this work is to find which is the optimal visualization method that can exploit the advantages of this novel class of FPs in full-scale pre-clinical molecular imaging studies. Nude mice were stereotactically implanted with near-infrared FP expressing glioma cells to from brain tumors. The feasibility and performance metrics of FPs were compared between planar epi-illumination and trans-illumination fluorescence imaging, as well as to hybrid Fluorescence Molecular Tomography (FMT) system combined with X-ray CT and Multispectral Optoacoustic (or Photoacoustic) Tomography (MSOT). It is shown that deep-seated glioma brain tumors are possible to visualize both with fluorescence and optoacoustic imaging. Fluorescence imaging is straightforward and has good sensitivity; however, it lacks resolution. FMT-XCT can provide an improved rough resolution of ∼1 mm in deep tissue, while MSOT achieves 0.1 mm resolution in deep tissue and has comparable sensitivity. We show imaging capacity that can shift the visualization paradigm in biological discovery. The results are relevant not only to reporter gene imaging, but stand as cross-platform comparison for all methods imaging near infrared fluorescent contrast agents.

A fluorescent screen + CCD system for quality assurance of therapeutic scanned ion beams

Energy Technology Data Exchange (ETDEWEB)

Takeshita, E., E-mail: eriuli@nirs.go.jp [National Institute of Radiological Sciences, Chiba (Japan); Furukawa, T., E-mail: t_furu@nirs.go.jp [National Institute of Radiological Sciences, Chiba (Japan); Inaniwa, T., E-mail: taku@nirs.go.jp [National Institute of Radiological Sciences, Chiba (Japan); Sato, S., E-mail: shin_s@nirs.go.jp [National Institute of Radiological Sciences, Chiba (Japan); Himukai, T., E-mail: himukai@nirs.go.jp [National Institute of Radiological Sciences, Chiba (Japan); Shirai, T., E-mail: t_shirai@nirs.go.jp [National Institute of Radiological Sciences, Chiba (Japan); Noda, K., E-mail: noda_k@nirs.go.jp [National Institute of Radiological Sciences, Chiba (Japan)

2011-12-15

A fluorescent screen + a charge coupled device (CCD) system were developed to verify the performance of scanned ion beams at the HIMAC. The fluorescent light from the screen is observed by the CCD camera. Two-dimensional fields, produced by the scanning process, i.e., the position and the size of the beam for each scan, represent of the important issues in scanning irradiation. In the developed system, the two-dimensional relative fluence and the flatness of the irradiation field were measured in a straightforward technique from the luminance distribution on the screen. The position and the size of the beams were obtained from centroid computation results of the brightness. By the good sensitivity and spatial resolution of the fluorescent screen + CCD system, the scanned ion beams were verified as the measurements at the HIMAC prototype scanning system.

A fluorescent screen + CCD system for quality assurance of therapeutic scanned ion beams

Science.gov (United States)

Takeshita, E.; Furukawa, T.; Inaniwa, T.; Sato, S.; Himukai, T.; Shirai, T.; Noda, K.

2011-12-01

A fluorescent screen + a charge coupled device (CCD) system were developed to verify the performance of scanned ion beams at the HIMAC. The fluorescent light from the screen is observed by the CCD camera. Two-dimensional fields, produced by the scanning process, i.e., the position and the size of the beam for each scan, represent of the important issues in scanning irradiation. In the developed system, the two-dimensional relative fluence and the flatness of the irradiation field were measured in a straightforward technique from the luminance distribution on the screen. The position and the size of the beams were obtained from centroid computation results of the brightness. By the good sensitivity and spatial resolution of the fluorescent screen + CCD system, the scanned ion beams were verified as the measurements at the HIMAC prototype scanning system.

Fluorescent modification and orientation of myosin sulfhydryl 2 in skeletal muscle fibers

International Nuclear Information System (INIS)

Ajtai, K.; Burghardt, T.P.

1989-01-01

The authors describe a protocol for the selective covalent labeling of the sulfhydryl 2 (SH2) on the myosin cross-bridge in glycerinated muscle fibers using the sulfhydryl-selective label 4-[N-[(iodoacetoxy)ethyl]-N-methylamino]-7-nitrobenz-2-oxa-1,3-diazole (IANBD). The protocol promotes the specificity of IANBD by using the ability to protect sulfhydryl 1 (SH1) from modification by binding the cross-bridge to the actin filament and using cross-bridge-bound MgADP to promote the accessibility of SH2. They determined the specificity of the probe using fluorescence gel scanning of fiber-extracted proteins to isolate the probe on myosin subfragment 1 (S1), limited proteolysis of the purified S1 to isolate the probe on the 20-kilodalton fragment of S1, and titration of the free SH1's on purified S1 using the radiolabeled SH1-specific reagent [ 14 C]iodoacetamide or enzymatic activity measurements. They characterized the angular distribution of the IANBD on cross-bridges in fibers when the fibers are in rigor, in relaxation, in the presence of MgADP, and in isometric contraction using wavelength-dependent fluorescence polarization. They find that the SH2 probe distinguishes the different states of the fiber such that rigor and MgADP are ordered and maintain a similar orientation throughout the excitation wavelength domain. The relaxed cross-bridge is ordered and has an orientation that is distinct from the orientation of the cross-bridge in rigor and MgADP over the entire wavelength domain. The active isometric cross-bridge is also oriented differently from the other states, suggesting the presence of a predominant actin-bound cross-bridge state that precedes the power stroke during muscle contraction

Isotopic imaging via nuclear resonance fluorescence with laser-based Thomson radiation

Science.gov (United States)

Barty, Christopher P. J. [Hayward, CA; Hartemann, Frederic V [San Ramon, CA; McNabb, Dennis P [Alameda, CA; Pruet, Jason A [Brentwood, CA

2009-07-21

The present invention utilizes novel laser-based, high-brightness, high-spatial-resolution, pencil-beam sources of spectrally pure hard x-ray and gamma-ray radiation to induce resonant scattering in specific nuclei, i.e., nuclear resonance fluorescence. By monitoring such fluorescence as a function of beam position, it is possible to image in either two dimensions or three dimensions, the position and concentration of individual isotopes in a specific material configuration. Such methods of the present invention material identification, spatial resolution of material location and ability to locate and identify materials shielded by other materials, such as, for example, behind a lead wall. The foundation of the present invention is the generation of quasimonochromatic high-energy x-ray (100's of keV) and gamma-ray (greater than about 1 MeV) radiation via the collision of intense laser pulses from relativistic electrons. Such a process as utilized herein, i.e., Thomson scattering or inverse-Compton scattering, produces beams having diameters from about 1 micron to about 100 microns of high-energy photons with a bandwidth of .DELTA.E/E of approximately 10E.sup.-3.

Efficient triplet application in exciplex delayed-fluorescence OLEDs using a reverse intersystem crossing mechanism based on a ΔES-T of around zero.

Science.gov (United States)

Zhang, Tianyou; Chu, Bei; Li, Wenlian; Su, Zisheng; Peng, Qi Ming; Zhao, Bo; Luo, Yongshi; Jin, Fangming; Yan, Xingwu; Gao, Yuan; Wu, Hairuo; Zhang, Feng; Fan, Di; Wang, Junbo

2014-08-13

We demonstrate highly efficient exciplex delayed-fluorescence organic light-emitting diodes (OLEDs) in which 4,4',4″-tris[3-methylphenyl(phenyl)aminotriphenylamine (m-MTDATA) and 4,7-diphenyl-1,10-phenanthroline (Bphen) were selected as donor and acceptor components, respectively. Our m-MTDATA:Bphen exciplex electroluminescence (EL) mechanism is based on reverse intersystem crossing (RISC) from the triplet to singlet excited states. As a result, an external quantum efficiency (EQE) of 7.79% at 10 mA/cm(2) was observed, which increases by 3.2 and 1.5 times over that reported in Nat. Photonics 2012, 6, 253 and Appl. Phys. Lett. 2012, 101, 023306, respectively. The high EQE would be attributed to a very easy RISC process because the energy difference between the singlet and triplet excited states is almost around zero. The verdict was proven by photoluminescence (PL) rate analysis at different temperatures and time-resolved spectral analysis. Besides, the study of the transient PL process indicates that the presence of an unbalanced charge in exciplex EL devices is responsible for the low EQE and high-efficiency roll-off. When the exciplex devices were placed in a 100 mT magnetic field, the permanently positive magnetoelectroluminescence and magnetoconductivity were observed. The magnetic properties confirm that the efficient exciplex EL only originates from delayed fluorescence via RISC processes but is not related to the triplet-triplet annihilation process.

In vivo multiphoton tomography and fluorescence lifetime imaging of human brain tumor tissue.

Science.gov (United States)

Kantelhardt, Sven R; Kalasauskas, Darius; König, Karsten; Kim, Ella; Weinigel, Martin; Uchugonova, Aisada; Giese, Alf

2016-05-01

High resolution multiphoton tomography and fluorescence lifetime imaging differentiates glioma from adjacent brain in native tissue samples ex vivo. Presently, multiphoton tomography is applied in clinical dermatology and experimentally. We here present the first application of multiphoton and fluorescence lifetime imaging for in vivo imaging on humans during a neurosurgical procedure. We used a MPTflex™ Multiphoton Laser Tomograph (JenLab, Germany). We examined cultured glioma cells in an orthotopic mouse tumor model and native human tissue samples. Finally the multiphoton tomograph was applied to provide optical biopsies during resection of a clinical case of glioblastoma. All tissues imaged by multiphoton tomography were sampled and processed for conventional histopathology. The multiphoton tomograph allowed fluorescence intensity- and fluorescence lifetime imaging with submicron spatial resolution and 200 picosecond temporal resolution. Morphological fluorescence intensity imaging and fluorescence lifetime imaging of tumor-bearing mouse brains and native human tissue samples clearly differentiated tumor and adjacent brain tissue. Intraoperative imaging was found to be technically feasible. Intraoperative image quality was comparable to ex vivo examinations. To our knowledge we here present the first intraoperative application of high resolution multiphoton tomography and fluorescence lifetime imaging of human brain tumors in situ. It allowed in vivo identification and determination of cell density of tumor tissue on a cellular and subcellular level within seconds. The technology shows the potential of rapid intraoperative identification of native glioma tissue without need for tissue processing or staining.

Optofluidic fluorescent imaging cytometry on a cell phone.

Science.gov (United States)

Zhu, Hongying; Mavandadi, Sam; Coskun, Ahmet F; Yaglidere, Oguzhan; Ozcan, Aydogan

2011-09-01

Fluorescent microscopy and flow cytometry are widely used tools in biomedical sciences. Cost-effective translation of these technologies to remote and resource-limited environments could create new opportunities especially for telemedicine applications. Toward this direction, here we demonstrate the integration of imaging cytometry and fluorescent microscopy on a cell phone using a compact, lightweight, and cost-effective optofluidic attachment. In this cell-phone-based optofluidic imaging cytometry platform, fluorescently labeled particles or cells of interest are continuously delivered to our imaging volume through a disposable microfluidic channel that is positioned above the existing camera unit of the cell phone. The same microfluidic device also acts as a multilayered optofluidic waveguide and efficiently guides our excitation light, which is butt-coupled from the side facets of our microfluidic channel using inexpensive light-emitting diodes. Since the excitation of the sample volume occurs through guided waves that propagate perpendicular to the detection path, our cell-phone camera can record fluorescent movies of the specimens as they are flowing through the microchannel. The digital frames of these fluorescent movies are then rapidly processed to quantify the count and the density of the labeled particles/cells within the target solution of interest. We tested the performance of our cell-phone-based imaging cytometer by measuring the density of white blood cells in human blood samples, which provided a decent match to a commercially available hematology analyzer. We further characterized the imaging quality of the same platform to demonstrate a spatial resolution of ~2 μm. This cell-phone-enabled optofluidic imaging flow cytometer could especially be useful for rapid and sensitive imaging of bodily fluids for conducting various cell counts (e.g., toward monitoring of HIV+ patients) or rare cell analysis as well as for screening of water quality in

Determination of K shell absorption jump factors and jump ratios of 3d transition metals by measuring K shell fluorescence parameters.

Science.gov (United States)

Kaçal, Mustafa Recep; Han, İbrahim; Akman, Ferdi

2015-01-01

Energy dispersive X-ray fluorescence technique (EDXRF) has been employed for measuring K-shell absorption jump factors and jump ratios for Ti, Cr, Fe, Co, Ni and Cu elements. The jump factors and jump ratios for these elements were determined by measuring K shell fluorescence parameters such as the Kα X-ray production cross-sections, K shell fluorescence yields, Kβ-to-Kα X-rays intensity ratios, total atomic absorption cross sections and mass attenuation coefficients. The measurements were performed using a Cd-109 radioactive point source and an Si(Li) detector in direct excitation and transmission experimental geometry. The measured values for jump factors and jump ratios were compared with theoretically calculated and the ones available in the literature. Copyright © 2014 Elsevier Ltd. All rights reserved.

Effect of cocoon fluorescence, silkworm hybrid and gender on sericin content of Bombyx mori L. silk thread

Directory of Open Access Journals (Sweden)

M. Panayotov

2016-06-01

Full Text Available Abstract. The goal of the present study was to determine the influence of the ultraviolet fluorescence of cocoons, the hybrid, the sex and the interaction among them on the sericin content in silk threads. The study was performed with 3 di- and 2 tetra-cross silkmoth (Bombyx mori L. hybrids, differentiated in three groups – with violet, intermediate and yellow fluorescence of the cocoons. The examined factors had a significant effect (p≤0.001 on the sericin content. The highest sericin content was detected in the silk threads of the violet-fluorescent and the lowest – in the yellow-fluorescent group. The analysed di-hybrids were distinguished by better characteristics in terms of sericin content, compared to the tetra-hybrids, most obvious for the yellow-fluorescent fraction.

Cross sections for double K-shell ionization in slow heavy ion-atom collisions, ch. 4

International Nuclear Information System (INIS)

Hoogkamer, T.P.; Woerlee, P.H.; Saris, F.W.

1977-01-01

Cross sections for Ksub(αα) X-ray production have been measured in collisions of 200 keV N + → NH 3 , N 2 , H 2 O; O + → NH 3 , H 2 O and 200, 400, 600 keV Ne 2+ → Ne. Fluorescence yields for Ksub(αα) two-electron one-photon transitions have been estimated for N and O, whereas for Ne, theoretical values for the fluorescence yield are available. We have used these to obtain double K-shell ionizzation cross sections which are compared to calculated cross section based on the theory for rotational coupling in the transient quasimolecular state

Fluorescence Intrinsic Characterization of Excitation-Emission Matrix Using Multi-Dimensional Ensemble Empirical Mode Decomposition

Directory of Open Access Journals (Sweden)

Tzu-Chien Hsiao

2013-11-01

Full Text Available Excitation-emission matrix (EEM fluorescence spectroscopy is a noninvasive method for tissue diagnosis and has become important in clinical use. However, the intrinsic characterization of EEM fluorescence remains unclear. Photobleaching and the complexity of the chemical compounds make it difficult to distinguish individual compounds due to overlapping features. Conventional studies use principal component analysis (PCA for EEM fluorescence analysis, and the relationship between the EEM features extracted by PCA and diseases has been examined. The spectral features of different tissue constituents are not fully separable or clearly defined. Recently, a non-stationary method called multi-dimensional ensemble empirical mode decomposition (MEEMD was introduced; this method can extract the intrinsic oscillations on multiple spatial scales without loss of information. The aim of this study was to propose a fluorescence spectroscopy system for EEM measurements and to describe a method for extracting the intrinsic characteristics of EEM by MEEMD. The results indicate that, although PCA provides the principal factor for the spectral features associated with chemical compounds, MEEMD can provide additional intrinsic features with more reliable mapping of the chemical compounds. MEEMD has the potential to extract intrinsic fluorescence features and improve the detection of biochemical changes.

Crash rates analysis in China using a spatial panel model

Directory of Open Access Journals (Sweden)

Wonmongo Lacina Soro

2017-10-01

Full Text Available The consideration of spatial externalities in traffic safety analysis is of paramount importance for the success of road safety policies. Yet, the quasi-totality of spatial dependence studies on crash rates is performed within the framework of single-equation spatial cross-sectional studies. The present study extends the spatial cross-sectional scheme to a spatial fixed-effects panel model estimated using the maximum likelihood method. The spatial units are the 31 administrative regions of mainland China over the period 2004–2013. The presence of neighborhood effects is evidenced through the Moran's I statistic. Consistent with previous studies, the analysis reveals that omitting the spatial effects in traffic safety analysis is likely to bias the estimation results. The spatial and error lags are all positive and statistically significant suggesting similarities of crash rates pattern in neighboring regions. Some other explanatory variables, such as freight traffic, the length of paved roads and the populations of age 65 and above are related to higher rates while the opposite trend is observed for the Gross Regional Product, the urban unemployment rate and passenger traffic.

Spatial distribution of arsenic and heavy metals in willow roots from a contaminated floodplain soil measured by X-ray fluorescence spectroscopy

International Nuclear Information System (INIS)

Zimmer, Dana; Kruse, Jens; Baum, Christel; Borca, Camelia; Laue, Michael; Hause, Gerd; Meissner, Ralph; Leinweber, Peter

2011-01-01

Under changing redox conditions some plants create plaques at their root surface, which may affect the mobility and uptake of As and heavy metals but it is unknown to what extent this also holds true for willows in contaminated floodplain soils. Therefore, willow roots were sampled from a phytoremediation trial in the contaminated floodplain of the river Elbe (Germany), cryofixed, freeze-dried, and cross sections were mapped for the distribution of As, Ca, Cu, Fe, K, Mn, Ni, S and Zn by synchrotron based X-ray fluorescence spectroscopy. The elements Ca, Cu, Ni, S and Zn were concentrated in the aerenchymatic tissue, and not associated with Fe and Mn. Mixed Fe-Mn plaques covered the surface of the willow roots and As was accumulated in these plaques. The observed association pattern between As and Fe was explained by the different sorption/desorption properties of As(III) and As(V). The Cu and Zn intensities were not associated with the intensity of Fe in the plaque, which seems to be a willow-specific difference compared to other wetland plants. These results suggested that willows are especially suited to stabilize low-phytoextractable elements like Cu and As in their roots and rhizosphere. Thus, short rotation coppicing of willows may be a practical approach to mitigate the adverse effects of floodplain soil contamination. - Research highlights: → Elemental distributions were mapped on willow roots for the first time by synchrotron-based X-ray fluorescence. → Ca, Cu, Ni, S and Zn were enriched in the aerenchyma but As, Fe and Mn formed root plaques. → The Cu and Zn enrichments in aerenchyma but absence in plaques appeared to be willow-specific. → In the plaques were three groups of pixels which strongly differed in the As to Fe and As to Mn ratios. → This indicated different species of these redox-sensitive elements.

Spatial distribution of arsenic and heavy metals in willow roots from a contaminated floodplain soil measured by X-ray fluorescence spectroscopy

Energy Technology Data Exchange (ETDEWEB)

Zimmer, Dana, E-mail: dana.zimmer@uni-rostock.de [Soil Science, University of Rostock, Justus-von-Liebig-Weg 6, D-18051 Rostock (Germany); Kruse, Jens; Baum, Christel [Soil Science, University of Rostock, Justus-von-Liebig-Weg 6, D-18051 Rostock (Germany); Borca, Camelia [Paul Scherrer Institute, Swiss Light Source, CH-5232 Villigen PSI (Switzerland); Laue, Michael [Electron Microscopy Centre, University of Rostock, Medical Faculty, Strempelstr. 14, D-18057 Rostock (Germany); Hause, Gerd [Microscopy Unit, Biocenter of the University of Halle, Weinbergweg 22, D-06120 Halle/Saale (Germany); Meissner, Ralph [UFZ-Helmholtz Centre for Environmental Research, Department of Soil Physics, Lysimeter Station, Dorfstrasse 55, D-39615 Falkenberg (Germany); Leinweber, Peter [Soil Science, University of Rostock, Justus-von-Liebig-Weg 6, D-18051 Rostock (Germany)

2011-09-01

Under changing redox conditions some plants create plaques at their root surface, which may affect the mobility and uptake of As and heavy metals but it is unknown to what extent this also holds true for willows in contaminated floodplain soils. Therefore, willow roots were sampled from a phytoremediation trial in the contaminated floodplain of the river Elbe (Germany), cryofixed, freeze-dried, and cross sections were mapped for the distribution of As, Ca, Cu, Fe, K, Mn, Ni, S and Zn by synchrotron based X-ray fluorescence spectroscopy. The elements Ca, Cu, Ni, S and Zn were concentrated in the aerenchymatic tissue, and not associated with Fe and Mn. Mixed Fe-Mn plaques covered the surface of the willow roots and As was accumulated in these plaques. The observed association pattern between As and Fe was explained by the different sorption/desorption properties of As(III) and As(V). The Cu and Zn intensities were not associated with the intensity of Fe in the plaque, which seems to be a willow-specific difference compared to other wetland plants. These results suggested that willows are especially suited to stabilize low-phytoextractable elements like Cu and As in their roots and rhizosphere. Thus, short rotation coppicing of willows may be a practical approach to mitigate the adverse effects of floodplain soil contamination. - Research highlights: {yields} Elemental distributions were mapped on willow roots for the first time by synchrotron-based X-ray fluorescence. {yields} Ca, Cu, Ni, S and Zn were enriched in the aerenchyma but As, Fe and Mn formed root plaques. {yields} The Cu and Zn enrichments in aerenchyma but absence in plaques appeared to be willow-specific. {yields} In the plaques were three groups of pixels which strongly differed in the As to Fe and As to Mn ratios. {yields} This indicated different species of these redox-sensitive elements.

Laser induced fluorescence in atmospheric pressure discharges

International Nuclear Information System (INIS)

Dilecce, G; De Benedictis, S; Martini, L M; Tosi, P; Scotoni, M

2015-01-01

This paper offers an outline of laser induced fluorescence (LIF) diagnostics and practical recommendations for its use in atmospheric pressure discharges. LIF principles, technical requirements and rationalization of experimental outcomes by modelling are addressed. Important issues that are particularly relevant to small scale, spatially inhomogeneous discharges, like plasma-jets, are emphasized. For the first time, all collision processes and the spatial non-homogeneity of the laser beam are together accounted for in the LIF model. Saturation characteristics are discussed and used for the assessment of model parameters. A calibration procedure is discussed and implemented. Gas temperature measurements by LIF are also addressed. The whole description of the technique is given, without loss of generality, through the example of its application to the OH radical. Notes on other diatomic radicals, CH, NO and CN, are given along the paper. Some results in a RF plasma-jet are presented as an example of application in a discharge system where all the concepts developed in the paper are applied. (paper)

Cross-Sensory Transfer of Reference Frames in Spatial Memory

Science.gov (United States)

Kelly, Jonathan W.; Avraamides, Marios N.

2011-01-01

Two experiments investigated whether visual cues influence spatial reference frame selection for locations learned through touch. Participants experienced visual cues emphasizing specific environmental axes and later learned objects through touch. Visual cues were manipulated and haptic learning conditions were held constant. Imagined perspective…

Statistical filtering in fluorescence microscopy and fluorescence correlation spectroscopy

Czech Academy of Sciences Publication Activity Database

Macháň, Radek; Kapusta, Peter; Hof, Martin

Roč. 406 , č. 20 (2014), s. 4797-4813 ISSN 1618-2642 R&D Projects: GA ČR GBP208/12/G016 Institutional support: RVO:61388955 Keywords : Filtered fluorescence correlation spectroscopy * Fluorescence lifetime correlation spectroscopy * Fluorescence spectral correlation spectroscopy Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 3.436, year: 2014

Use of a portable fluorescence imaging device to facilitate cleaning of deli slicers

Science.gov (United States)

Cross-contamination is a recognized food safety concern for deli departments, and deli slicers remain a particular concern. Our laboratory previously demonstrated that deli commodity residues can be detected using fluorescence imaging, and that the efficacy of cleaning and sanitation in produce proc...

Sensitized fluorescence in thallium induced in collisions with Hg(6/sup 3/P/sub 1/) atoms

Energy Technology Data Exchange (ETDEWEB)

Wade, M K; Czajkowski, M; Krause, L [Windsor Univ., Ontario (Canada). Dept. of Physics

1978-07-01

The transfer of excitation from excited mercury atoms to ground-state thallium atoms was investigated using techniques of sensitized fluorescence. A Hg-Tl vapor mixture contained in a quartz cell was irradiated with Hg 2537 A resonance radiation which caused the mercury atoms to become excited to the 6/sup 3/P/sub 1/ state. Subsequent collisions between the Hg(6/sup 3/P/sub 1/) and Tl(6/sup 2/Psub(1/2)) atoms resulted in the population of the 8/sup 2/Ssub(1/2), 6/sup 2/D, and 7/sup 2/Ssub(1/2) thallium states, whose decay gave rise to sensitized fluorescence of wavelengths 3231, 3520, 3776, and 5352 A. Intensity measurements on the sensitized fluorescence and on the Hg 2537 A resonance fluorescence, observed at right angles to the direction of excitation, yielded cross sections of 3.0, 0.3, and 0.05 A/sup 2/ for collisional excitation transfer from Hg(6/sup 3/P/sub 1/) to the 8/sup 2/Ssub(1/2), 6/sup 2/D, and 7/sup 2/Ssub(1/2) states in thallium, respectively. The results are fully consistent with previously determined cross sections for excitation transfer in other binary metallic vapor systems.

A spirobifluorene-based two-photon fluorescence probe for mercury ions and its applications in living cells

Energy Technology Data Exchange (ETDEWEB)

Xiao, Haibo, E-mail: xiaohb@shnu.edu.cn; Zhang, Yanzhen; Zhang, Wu; Li, Shaozhi; Tan, Jingjing; Han, Zhongying

2017-05-01

A novel spirobifluorene derivative SPF-TMS, which containing dithioacetal groups and triphenylamine units, was synthesized. The probing behaviors toward various metal ions were investigated via UV/Vis absorption spectra as well as one-photon fluorescence changes. The results indicated that SPF-TMS exhibits high sensitivity and selectivity for mercury ions. The detection limit was at least 8.6 × 10{sup −8}M, which is excellent comparing with other optical sensors for Hg{sup 2+}. When measured by two-photon excited fluorescence technique in THF at 800 nm, the two-photon cross-section of SPF-TMS is 272 GM. Especially, upon reaction with mercury species, SPF-TMS yielded another two-photon dye SPF-DA. Both SPF-TMS and SPF-DA emit strong two-photon induced fluorescence and can be applied in cell imaging by two-photon microscopy. - Highlights: • We report a spirobifluorene-based molecule as two-photon fluorescent probe with large two-photon cross-section. • The molecule has exclusive selectivity and sensitivity for mercury species. • The molecule has large two-photon emission changes before and after addition of Hg{sup 2+}. • Both the probe and the mercury ion-promoted reaction product can be applied in cell imaging by two-photon microscopy.

Enhancement of uranyl fluorescence using trimesic acid: Ligand sensitization and co-fluorescence

Energy Technology Data Exchange (ETDEWEB)

Maji, S. [Chemistry Group, Materials Chemistry Division, Indira Gandhi Centre for Atomic Research, Kalpakkam 603102 (India); Viswanathan, K.S., E-mail: vish@igcar.gov.in [Chemistry Group, Materials Chemistry Division, Indira Gandhi Centre for Atomic Research, Kalpakkam 603102 (India)

2011-09-15

Trimesic acid (TMA) was shown to sensitize and enhance uranyl fluorescence in aqueous medium, with the enhancement being a maximum at pH 5.0. Fluorescence spectra and lifetime data together suggest that TMA complexes with uranyl (UO{sub 2}{sup 2+}). The fluorescence of UO{sub 2}{sup 2+} in its acid complex is further enhanced by more than two orders of magnitude following the addition of Y{sup 3+}; a process referred to as co-fluorescence, leading to the possibility of detecting uranium at sub ng/mL level. The present study demonstrates, for the first time, fluorescence enhancement of the uranyl species due to co-fluorescence. - Highlights: > Trimesic acid was shown to sensitize and enhance the fluorescence of uranium in aqueous medium. > This ligand also exhibited co-fluorescence of uranium with Y{sup 3+}. > To the best of our knowledge this is the first report of co-fluorescence in uranium. > The enhancement of uranium fluorescence, resulted in detection limits in the ng/mL regime.

Let's Exploit Available Knowledge on Vegetation Fluorescence

Science.gov (United States)

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

2014-01-01

. 2).We know, however, that the ratio is not constant, but changes widely in response to light, CO2, stomatal limitations, and extreme stress (4, 5). What’s more, we can make sense of such changes, thus extracting valuable information from the very scatter that is apparent in their data. However, this process will require the availability of more tailored instruments, such as the one planned for the FLEX mission. As already stressed by Guanter et al. (2), the spatial resolution of the Global Ozone Monitoring Experiment-2 sensor (40 × 80 km) makes it difficult to compare meaningfully the fluorescence signal with ground measurements, when only 60–70% of the footprint consists of the desired land-cover type (table S1 in ref. 2), suggesting that this could be largely responsible for the low signals observed in European grasslands. Moreover, the overpass time of the MetOp-A satellite (9:30 AM) implies that fluorescence is generally measured under light-limiting conditions, when fluorescence is only marginally affected by stomatal closure even under stress conditions. This result could explain the seasonal mismatch with daily GPP observed in natural ecosystems in the absence of irrigation (figure 4 in ref. 2). We hope, therefore, that this welcome contribution to this fast-advancing field will help demonstrate the potential of the new technique, and pave the way for more refined studies under both a technological and scientific point of view.

Anomalous scattering, transport, and spatial distribution of X-ray fluorescence at the exit of polycapillary structures

Energy Technology Data Exchange (ETDEWEB)

Mazuritskiy, M. I., E-mail: mazurmik@gmail.com; Lerer, A. M.; Makhno, P. V. [Southern Federal University (Russian Federation)

2016-12-15

The angular distribution of the X-ray intensity at the exit of microchannel plates at grazing incidence of monochromatic radiation on the walls of microcapillaries has been investigated. The angles and energies of the primary radiation quanta at which the synchrotron beam excites X-ray fluorescence propagating inside polycapillary structures have been determined. The angular dependences of the intensity distribution of X-rays transmitted through the microcapillaries have been studied theoretically and experimentally for energies corresponding to the region of anomalous dispersion near the L{sub 2,3} absorption edges of silicon. The propagation of waves in hollow polycapillary waveguides, the excitation of X-ray fluorescence, and the X-ray diffraction at the exit of microchannel plates have been modeled mathematically. The mathematical model takes into account the presence of a transition layer on the microchannel surface.

Novel silicone compatible cross-linkers for controlled functionalization of PDMS networks

DEFF Research Database (Denmark)

Madsen, Frederikke Bahrt; Daugaard, Anders Egede; Hvilsted, Søren

2013-01-01

. In order to improve the dielectric properties of PDMS a novel system is developed where push-pull dipoles are grafted to a new silicone compatible cross-linker. The grafted cross-linkers are prepared by reaction of two different push-pull dipole alkynes as well as a fluorescent alkyne with the new azide...

High-resolution imaging of redox signaling in live cells through an oxidation-sensitive yellow fluorescent protein

DEFF Research Database (Denmark)

Maulucci, Giuseppe; Labate, Valentina; Mele, Marina

2008-01-01

We present the application of a redox-sensitive mutant of the yellow fluorescent protein (rxYFP) to image, with elevated sensitivity and high temporal and spatial resolution, oxidative responses of eukaryotic cells to pathophysiological stimuli. The method presented, based on the ratiometric...... quantitation of the distribution of fluorescence by confocal microscopy, allows us to draw real-time "redox maps" of adherent cells and to score subtle changes in the intracellular redox state, such as those induced by overexpression of redox-active proteins. This strategy for in vivo imaging of redox...

Photophysical characterization of pyrromethene 597 laser dye in cross-linked silicon-containing organic copolymers

Energy Technology Data Exchange (ETDEWEB)

Tyagi, A.; Agua, D. del [Institut II - Experimentelle und Angewandte Physik, Universitaet Regensburg, Universitaetsstrasse 31, 93053 Regensburg (Germany); Penzkofer, A. [Institut II - Experimentelle und Angewandte Physik, Universitaet Regensburg, Universitaetsstrasse 31, 93053 Regensburg (Germany)], E-mail: alfons.penzkofer@physik.uni-regensburg.de; Garcia, O.; Sastre, R. [Instituto de Ciencia y Tecnologia de Polimeros, CSIC, Juan de la Cierva 3, 28006 Madrid (Spain); Costela, A.; Garcia-Moreno, I. [Instituto de Quimica Fisica ' Rocasolano' , CSIC, Serrano 119, 28006 Madrid (Spain)

2007-12-06

Samples of the dipyrromethene-BF{sub 2} dye PM597 incorporated in copolymers of 3-trimethoxysilylpropyl 2-methylprop-2-enoate (TMSPMA, number of polymerizable CC double bonds: {kappa} = 1) with 2-(2-methylprop-2-enoyloxy)ethyl 2-methylprop-2-enoate (EGDMA, {kappa} = 2), [2-(hydroxymethyl)-3-prop-2-(prop-2-enoyloxymethyl)propyl] prop-2-enoate (PETA, {kappa} = 3), and [3-prop-2-enoyloxy-2,2-bis(prop-2-2-enoyloxymethyl)propyl]prop-2-enoate (PETRA, {kappa} = 4) are characterized. The fluorescence quantum distributions, fluorescence quantum yields, degrees of fluorescence polarization, and fluorescence lifetimes are measured. The radiative lifetimes are calculated from fluorescence lifetime and quantum yield. Absorption coefficient spectra are determined from transmission measurements. Absolute absorption cross-section spectra and dye concentrations are obtained by calibration to the radiative lifetimes and to saturable absorptions. Excited-state absorption cross-sections at 527 nm are determined by saturable absorption measurements. The photo-degradation is studied under cw laser excitation conditions and quantum yields of photo-degradation are extracted. The excited-state absorption cross-sections were found to be rather small, and the photo-stability turned out to be high (up to 3 million excitation cycles before degradation) making this class of dipyrromethene dye-doped polymers attractive active laser media. Structural and thermo-mechanical properties of the materials have been determined by Fourier transform infrared spectroscopy, differential scanning calorimetry, thermogravimetry, densitometry, and refractometry. They improve with increasing inter-crossing (copolymerization of TMSPMA with PETA and PETRA). The laser properties of the PM597 doped copolymers were evaluated by transverse pumping with 6 ns laser pulses at 532 nm. The best laser materials resulted to be the 7:3 and 9:1 TMSPMA-monomer copolymers.

Dual-detection confocal fluorescence microscopy: fluorescence axial imaging without axial scanning.

Science.gov (United States)

Lee, Dong-Ryoung; Kim, Young-Duk; Gweon, Dae-Gab; Yoo, Hongki

2013-07-29

We propose a new method for high-speed, three-dimensional (3-D) fluorescence imaging, which we refer to as dual-detection confocal fluorescence microscopy (DDCFM). In contrast to conventional beam-scanning confocal fluorescence microscopy, where the focal spot must be scanned either optically or mechanically over a sample volume to reconstruct a 3-D image, DDCFM can obtain the depth of a fluorescent emitter without depth scanning. DDCFM comprises two photodetectors, each with a pinhole of different size, in the confocal detection system. Axial information on fluorescent emitters can be measured by the axial response curve through the ratio of intensity signals. DDCFM can rapidly acquire a 3-D fluorescent image from a single two-dimensional scan with less phototoxicity and photobleaching than confocal fluorescence microscopy because no mechanical depth scans are needed. We demonstrated the feasibility of the proposed method by phantom studies.

Simulation study of two-energy X-ray fluorescence holograms reconstruction algorithm to remove twin images

International Nuclear Information System (INIS)

Xie Honglan; Hu Wen; Luo Hongxin; Deng Biao; Du Guohao; Xue Yanling; Chen Rongchang; Shi Shaomeng; Xiao Tiqiao

2008-01-01

Unlike traditional outside-source holography, X-ray fluorescence holography is carded out with fluorescent atoms in a sample as source light for holographic imaging. With the method, three-dimensional arrangement of atoms into crystals can be observed obviously. However, just like traditional outside-source holography, X-ray fluorescence holography suffers from the inherent twin-image problem, too. With a 27-Fe-atoms cubic lattice as model, we discuss in this paper influence of the photon energy of incident source in removing twin images in reconstructed atomic images by numerical simulation and reconstruction with two-energy X-ray fluorescence holography. The results indicate that incident X-rays of nearer energies have better effect of removing twin images. In the detector of X-ray holography, minimum difference of the two incident energies depends on energy resolution of the monochromator and detector, and for inside source X-ray holography, minimum difference of the two incident energies depends on difference of two neighboring fluorescent energies emitting from the element and energy resolution of detector. The spatial resolution of atomic images increases with the incident energies. This is important for experiments of X-ray fluorescence holography, which is being developed on Shanghai Synchrotron Radiation Facility. (authors)

Estimating phytoplankton photosynthesis by active fluorescence

Energy Technology Data Exchange (ETDEWEB)

Falkowski, P.G.; Kolber, Z.

1992-01-01

Photosynthesis can be described by target theory, At low photon flux densities, photosynthesis is a linear function of irradiance (I), The number of reaction centers (n), their effective absorption capture cross section {sigma}, and a quantum yield {phi}. As photosynthesis becomes increasingly light saturated, an increased fraction of reaction centers close. At light saturation the maximum photosynthetic rate is given as the product of the number of reaction centers (n) and their maximum electron transport rate (I/{tau}). Using active fluorometry it is possible to measure non-destructively and in real time the fraction of open or closed reaction centers under ambient irradiance conditions in situ, as well as {sigma} and {phi} {tau} can be readily, calculated from knowledge of the light saturation parameter, I{sub k} (which can be deduced by in situ by active fluorescence measurements) and {sigma}. We built a pump and probe fluorometer, which is interfaced with a CTD. The instrument measures the fluorescence yield of a weak probe flash preceding (f{sub 0}) and succeeding (f{sub 0}) a saturating pump flash. Profiles of the these fluorescence yields are used to derive the instantaneous rate of gross photosynthesis in natural phytoplankton communities without any incubation. Correlations with short-term simulated in situ radiocarbon measurements are extremely high. The average slope between photosynthesis derived from fluorescence and that measured by radiocarbon is 1.15 and corresponds to the average photosynthetic quotient. The intercept is about 15% of the maximum radiocarbon uptake and corresponds to the average net community respiration. Profiles of photosynthesis and sections showing the variability in its composite parameters reveal a significant effect of nutrient availability on biomass specific rates of photosynthesis in the ocean.

Estimating phytoplankton photosynthesis by active fluorescence

Energy Technology Data Exchange (ETDEWEB)

Falkowski, P.G.; Kolber, Z.

1992-10-01

Photosynthesis can be described by target theory, At low photon flux densities, photosynthesis is a linear function of irradiance (I), The number of reaction centers (n), their effective absorption capture cross section {sigma}, and a quantum yield {phi}. As photosynthesis becomes increasingly light saturated, an increased fraction of reaction centers close. At light saturation the maximum photosynthetic rate is given as the product of the number of reaction centers (n) and their maximum electron transport rate (I/{tau}). Using active fluorometry it is possible to measure non-destructively and in real time the fraction of open or closed reaction centers under ambient irradiance conditions in situ, as well as {sigma} and {phi} {tau} can be readily, calculated from knowledge of the light saturation parameter, I{sub k} (which can be deduced by in situ by active fluorescence measurements) and {sigma}. We built a pump and probe fluorometer, which is interfaced with a CTD. The instrument measures the fluorescence yield of a weak probe flash preceding (f{sub 0}) and succeeding (f{sub 0}) a saturating pump flash. Profiles of the these fluorescence yields are used to derive the instantaneous rate of gross photosynthesis in natural phytoplankton communities without any incubation. Correlations with short-term simulated in situ radiocarbon measurements are extremely high. The average slope between photosynthesis derived from fluorescence and that measured by radiocarbon is 1.15 and corresponds to the average photosynthetic quotient. The intercept is about 15% of the maximum radiocarbon uptake and corresponds to the average net community respiration. Profiles of photosynthesis and sections showing the variability in its composite parameters reveal a significant effect of nutrient availability on biomass specific rates of photosynthesis in the ocean.

Biomolecule-to-fluorescent-color encoder: modulation of fluorescence emission via DNA structural changes

Science.gov (United States)

Nishimura, Takahiro; Ogura, Yusuke; Yamada, Kenji; Ohno, Yuko; Tanida, Jun

2014-01-01

A biomolecule-to-fluorescent-color (B/F) encoder for optical readout of biomolecular information is proposed. In the B/F encoder, a set of fluorescence wavelengths and their intensity levels are used for coding of a biomolecular signal. A hybridization chain reaction of hairpin DNAs labeled with fluorescent reporters was performed to generate the fluorescence color codes. The fluorescence is modulated via fluorescence resonance energy transfer, which is controlled by DNA structural changes. The results demonstrate that fluorescent color codes can be configured based on two wavelengths and five intensities using the B/F encoder, and the assigned codes can be retrieved via fluorescence measurements. PMID:25071950

Evaluating the peak-to-valley dose ratio of synchrotron microbeams using PRESAGE fluorescence

International Nuclear Information System (INIS)

Annabell, N.; Yagi, N.; Umetani, K.; Wong, C.; Geso, M.

2012-01-01

The peak-to-valley dose ratio of a microbeam array can be measured by fluorescence of PRESAGE dosimeters. Peak-to-valley dose ratios are calculated using this new technique and also by EBT2 film. Synchrotron-generated microbeam radiotherapy holds great promise for future treatment, but the high dose gradients present conventional dosimetry with a challenge. Measuring the important peak-to-valley dose ratio (PVDR) of a microbeam-collimated synchrotron source requires both a dosimeter and an analysis method capable of exceptional spatial resolution. The PVDR is of great interest since it is the limiting factor for potential application of the microbeam radiation therapy technique clinically for its tissue-sparing properties (i.e. the valley dose should be below the tolerance of normal tissue). In this work a new method of measuring the dose response of PRESAGE dosimeters is introduced using the fluorescence from a 638 nm laser on a confocal laser-scanning microscope. This fluorescent microscopy method produces dosimetry data at a pixel size as low as 78 nm, giving a much better spatial resolution than optical computed tomography, which is normally used for scanning PRESAGE dosimeters. Using this technique the PVDR of the BL28B2 microbeam at the SPring-8 synchrotron in Japan is estimated to be approximately 52:1 at a depth of 2.5 mm. The PVDR was also estimated with EBT2 GAFchromic films as 30.5:1 at the surface in order to compare the PRESAGE fluorescent results with a more established dosimetry system. This estimation is in good agreement with previously measured ratios using other dosimeters and Monte Carlo simulations. This means that it is possible to use PRESAGE dosimeters with confocal microscopy for the determination of PVDR

Determination of fission products in irradiated fuel by X-ray fluorescence

International Nuclear Information System (INIS)

Mogensen, M.; Als-Nielsen, J.; Hessel Andersen, N.

1986-08-01

X-ray fluorescence is a well established analytical tool for measuring elemental composition of fairly large (approximately 5 cm 2 ) ''cold'' samples. A version of this technique has been developed for analysis of radial distribution of fission products Xe, Cs and Ba in irradiated UO 2 fuel samples. About 0.1 mm thin slices of fuel pellets (full cross sections) are irradiated by 50 keV X-rays. The intensity of the Xe (Cs, Ba) K α fluorescence radiation generated is measured by means of a Ge detector fitted with a collimator. The slit is 0.5 mm wide in the scanning direction and 2 mm long. The measured Xe K α X-ray intensities are converted to absolute concentrations by comparing to the intensity from a Xe gas standard. In the case of Cs and Ba solid standards may be used. The X-ray fluorescence analysis is compared to other techniques used to obtain radial fission product profiles. It is shown how a combination of X-ray fluorescence and electron probe micro analysis is able to reveal the amount of Xe in the grain boundary porosities. (author)

Light-sheet fluorescence imaging to localize cardiac lineage and protein distribution

Science.gov (United States)

Ding, Yichen; Lee, Juhyun; Ma, Jianguo; Sung, Kevin; Yokota, Tomohiro; Singh, Neha; Dooraghi, Mojdeh; Abiri, Parinaz; Wang, Yibin; Kulkarni, Rajan P.; Nakano, Atsushi; Nguyen, Thao P.; Fei, Peng; Hsiai, Tzung K.

2017-02-01

Light-sheet fluorescence microscopy (LSFM) serves to advance developmental research and regenerative medicine. Coupled with the paralleled advances in fluorescence-friendly tissue clearing technique, our cardiac LSFM enables dual-sided illumination to rapidly uncover the architecture of murine hearts over 10 by 10 by 10 mm3 in volume; thereby allowing for localizing progenitor differentiation to the cardiomyocyte lineage and AAV9-mediated expression of exogenous transmembrane potassium channels with high contrast and resolution. Without the steps of stitching image columns, pivoting the light-sheet and sectioning the heart mechanically, we establish a holistic strategy for 3-dimentional reconstruction of the “digital murine heart” to assess aberrant cardiac structures as well as the spatial distribution of the cardiac lineages in neonates and ion-channels in adults.

The ethics of Google Earth: crossing thresholds from spatial data to landscape visualisation.

Science.gov (United States)

Sheppard, Stephen R J; Cizek, Petr

2009-05-01

'Virtual globe' software systems such as Google Earth are growing rapidly in popularity as a way to visualise and share 3D environmental data. Scientists and environmental professionals, many of whom are new to 3D modeling and visual communications, are beginning routinely to use such techniques in their work. While the appeal of these techniques is evident, with unprecedented opportunities for public access to data and collaborative engagement over the web, are there nonetheless risks in their widespread usage when applied in areas of the public interest such as planning and policy-making? This paper argues that the Google Earth phenomenon, which features realistic imagery of places, cannot be dealt with only as a question of spatial data and geographic information science. The virtual globe type of visualisation crosses several key thresholds in communicating scientific and environmental information, taking it well beyond the realm of conventional spatial data and geographic information science, and engaging more complex dimensions of human perception and aesthetic preference. The realism, perspective views, and social meanings of the landscape visualisations embedded in virtual globes invoke not only cognition but also emotional and intuitive responses, with associated issues of uncertainty, credibility, and bias in interpreting the imagery. This paper considers the types of risks as well as benefits that may exist with participatory uses of virtual globes by experts and lay-people. It is illustrated with early examples from practice and relevant themes from the literature in landscape visualisation and related disciplines such as environmental psychology and landscape planning. Existing frameworks and principles for the appropriate use of environmental visualisation methods are applied to the special case of widely accessible, realistic 3D and 4D visualisation systems such as Google Earth, in the context of public awareness-building and agency decision

The application of a microstrip gas counter to energy-dispersive x-ray fluorescence analysis

International Nuclear Information System (INIS)

Veloso, J.F.C.A.; Santos, J.M.F. dos; Conde, C.A.N.

1996-01-01

Performance characteristics of a microstrip gas counter operated as a x-ray fluorescence spectrometer are reported. Gas amplification as a function of microstrip anode-cathode voltage was measured, and the breakdown threshold voltage was determined in pure xenon. The detector temporal stability and the effect of gas purity were assessed. Energy resolution and linearity, detection efficiency, and uniformity of spatial response in the 2- to 60-keV x-ray energy range were determined from the pulse-height distributions of the fluorescence x-ray spectra induced in a variety of single- and multi-element sample materials. Energy resolution similar to conventional proportional counters was achieved at 6 keV

Microbialite Biosignature Analysis by Mesoscale X-ray Fluorescence (μXRF) Mapping.

Science.gov (United States)

Tice, Michael M; Quezergue, Kimbra; Pope, Michael C

2017-11-01

As part of its biosignature detection package, the Mars 2020 rover will carry PIXL, the Planetary Instrument for X-ray Lithochemistry, a spatially resolved X-ray fluorescence (μXRF) spectrometer. Understanding the types of biosignatures detectable by μXRF and the rock types μXRF is most effective at analyzing is therefore an important goal in preparation for in situ Mars 2020 science and sample selection. We tested mesoscale chemical mapping for biosignature interpretation in microbialites. In particular, we used μXRF to identify spatial distributions and associations between various elements ("fluorescence microfacies") to infer the physical, biological, and chemical processes that produced the observed compositional distributions. As a test case, elemental distributions from μXRF scans of stromatolites from the Mesoarchean Nsuze Group (2.98 Ga) were analyzed. We included five fluorescence microfacies: laminated dolostone, laminated chert, clotted dolostone and chert, stromatolite clast breccia, and cavity fill. Laminated dolostone was formed primarily by microbial mats that trapped and bound loose sediment and likely precipitated carbonate mud at a shallow depth below the mat surface. Laminated chert was produced by the secondary silicification of microbial mats. Clotted dolostone and chert grew as cauliform, cryptically laminated mounds similar to younger thrombolites and was likely formed by a combination of mat growth and patchy precipitation of early-formed carbonate. Stromatolite clast breccias formed as lag deposits filling erosional scours and interstromatolite spaces. Cavities were filled by microquartz, Mn-rich dolomite, and partially dolomitized calcite. Overall, we concluded that μXRF is effective for inferring genetic processes and identifying biosignatures in compositionally heterogeneous rocks. Key Words: Stromatolites-Biosignatures-Spectroscopy-Archean. Astrobiology 17, 1161-1172.

Fluorescent Reporters and Biosensors for Probing the Dynamic Behavior of Protein Kinases

Directory of Open Access Journals (Sweden)

Juan A. González-Vera

2015-11-01

Full Text Available Probing the dynamic activities of protein kinases in real-time in living cells constitutes a major challenge that requires specific and sensitive tools tailored to meet the particular demands associated with cellular imaging. The development of genetically-encoded and synthetic fluorescent biosensors has provided means of monitoring protein kinase activities in a non-invasive fashion in their native cellular environment with high spatial and temporal resolution. Here, we review existing technologies to probe different dynamic features of protein kinases and discuss limitations where new developments are required to implement more performant tools, in particular with respect to infrared and near-infrared fluorescent probes and strategies which enable improved signal-to-noise ratio and controlled activation of probes.

Laser-Induced Photofragmentation Fluorescence Imaging of Alkali Compounds in Flames.

Science.gov (United States)

Leffler, Tomas; Brackmann, Christian; Aldén, Marcus; Li, Zhongshan

2017-06-01

Laser-induced photofragmentation fluorescence has been investigated for the imaging of alkali compounds in premixed laminar methane-air flames. An ArF excimer laser, providing pulses of wavelength 193 nm, was used to photodissociate KCl, KOH, and NaCl molecules in the post-flame region and fluorescence from the excited atomic alkali fragment was detected. Fluorescence emission spectra showed distinct lines of the alkali atoms allowing for efficient background filtering. Temperature data from Rayleigh scattering measurements together with simulations of potassium chemistry presented in literature allowed for conclusions on the relative contributions of potassium species KOH and KCl to the detected signal. Experimental approaches for separate measurements of these components are discussed. Signal power dependence and calculated fractions of dissociated molecules indicate the saturation of the photolysis process, independent on absorption cross-section, under the experimental conditions. Quantitative KCl concentrations up to 30 parts per million (ppm) were evaluated from the fluorescence data and showed good agreement with results from ultraviolet absorption measurements. Detection limits for KCl photofragmentation fluorescence imaging of 0.5 and 1.0 ppm were determined for averaged and single-shot data, respectively. Moreover, simultaneous imaging of KCl and NaCl was demonstrated using a stereoscope with filters. The results indicate that the photofragmentation method can be employed for detailed studies of alkali chemistry in laboratory flames for validation of chemical kinetic mechanisms crucial for efficient biomass fuel utilization.

A vacuum-UV laser-induced fluorescence experiment for measurement of rotationally and vibrationally excited H2

NARCIS (Netherlands)

Vankan, P.J.W.; Heil, S.B.S.; Mazouffre, S.; Engeln, R.A.H.; Schram, D.C.; Döbele, H.F.

2004-01-01

An experimental setup is built to detect spatially resolved rovibrationally excited hydrogen molecules via laser-induced fluorescence. To excite the hydrogen molecules, laser radiation is produced in the vacuum UV part of the spectrum. The laser radiation is tunable between 120 nm and 230 nm and has

A rapid-screening approach to detect and quantify microplastics based on fluorescent tagging with Nile Red

Science.gov (United States)

Maes, Thomas; Jessop, Rebecca; Wellner, Nikolaus; Haupt, Karsten; Mayes, Andrew G.

2017-03-01

A new approach is presented for analysis of microplastics in environmental samples, based on selective fluorescent staining using Nile Red (NR), followed by density-based extraction and filtration. The dye adsorbs onto plastic surfaces and renders them fluorescent when irradiated with blue light. Fluorescence emission is detected using simple photography through an orange filter. Image-analysis allows fluorescent particles to be identified and counted. Magnified images can be recorded and tiled to cover the whole filter area, allowing particles down to a few micrometres to be detected. The solvatochromic nature of Nile Red also offers the possibility of plastic categorisation based on surface polarity characteristics of identified particles. This article details the development of this staining method and its initial cross-validation by comparison with infrared (IR) microscopy. Microplastics of different sizes could be detected and counted in marine sediment samples. The fluorescence staining identified the same particles as those found by scanning a filter area with IR-microscopy.

Detection of organic residues on poultry processing equipment surfaces by LED-induced fluorescence imaging

Science.gov (United States)

Organic residues on equipment surfaces in poultry processing plants can generate cross- contamination and increase the risk of unsafe food for consumers. This research was aimed to investigate the potential of LED-induced fluorescence imaging technique for rapid inspection of stainless steel proces...

Hybrid diffusion–transport spatial homogenization method

International Nuclear Information System (INIS)

Kooreman, Gabriel; Rahnema, Farzad

2014-01-01

Highlights: • A new hybrid diffusion–transport homogenization method. • An extension of the consistent spatial homogenization (CSH) transport method. • Auxiliary cross section makes homogenized diffusion consistent with heterogeneous diffusion. • An on-the-fly re-homogenization in transport. • The method is faster than fine-mesh transport by 6–8 times. - Abstract: A new hybrid diffusion–transport homogenization method has been developed by extending the consistent spatial homogenization (CSH) transport method to include diffusion theory. As in the CSH method, an “auxiliary cross section” term is introduced into the source term, making the resulting homogenized diffusion equation consistent with its heterogeneous counterpart. The method then utilizes an on-the-fly re-homogenization in transport theory at the assembly level in order to correct for core environment effects on the homogenized cross sections and the auxiliary cross section. The method has been derived in general geometry and tested in a 1-D boiling water reactor (BWR) core benchmark problem for both controlled and uncontrolled configurations. The method has been shown to converge to the reference solution with less than 1.7% average flux error in less than one third the computational time as the CSH method – 6 to 8 times faster than fine-mesh transport

Co-localized or randomly distributed? Pair cross correlation of in vivo grown subgingival biofilm bacteria quantified by digital image analysis.

Directory of Open Access Journals (Sweden)

Claudia Schillinger

Full Text Available The polymicrobial nature of periodontal diseases is reflected by the diversity of phylotypes detected in subgingival plaque and the finding that consortia of suspected pathogens rather than single species are associated with disease development. A number of these microorganisms have been demonstrated in vitro to interact and enhance biofilm integration, survival or even pathogenic features. To examine the in vivo relevance of these proposed interactions, we extended the spatial arrangement analysis tool of the software daime (digital image analysis in microbial ecology. This modification enabled the quantitative analysis of microbial co-localization in images of subgingival biofilm species, where the biomass was confined to fractions of the whole-image area, a situation common for medical samples. Selected representatives of the disease-associated red and orange complexes that were previously suggested to interact with each other in vitro (Tannerella forsythia with Fusobacterium nucleatum and Porphyromonas gingivalis with Prevotella intermedia were chosen for analysis and labeled with specific fluorescent probes via fluorescence in situ hybridization. Pair cross-correlation analysis of in vivo grown biofilms revealed tight clustering of F. nucleatum/periodonticum and T. forsythia at short distances (up to 6 µm with a pronounced peak at 1.5 µm. While these results confirmed previous in vitro observations for F. nucleatum and T. forsythia, random spatial distribution was detected between P. gingivalis and P. intermedia in the in vivo samples. In conclusion, we successfully employed spatial arrangement analysis on the single cell level in clinically relevant medical samples and demonstrated the utility of this approach for the in vivo validation of in vitro observations by analyzing statistically relevant numbers of different patients. More importantly, the culture-independent nature of this approach enables similar quantitative analyses for "as

The feasibility study on 3-dimensional fluorescent x-ray computed tomography using the pinhole effect for biomedical applications.

Science.gov (United States)

Sunaguchi, Naoki; Yuasa, Tetsuya; Hyodo, Kazuyuki; Zeniya, Tsutomu

2013-01-01

We propose a 3-dimensional fluorescent x-ray computed tomography (CT) pinhole collimator, aimed at providing molecular imaging with quantifiable measures and sub-millimeter spatial resolution. In this study, we demonstrate the feasibility of this concept and investigate imaging properties such as spatial resolution, contrast resolution and quantifiable measures, by imaging physical phantoms using a preliminary imaging system developed with monochromatic synchrotron x rays constructed at the BLNE-7A experimental line at KEK, Japan.

Non-invasive imaging of skin cancer with fluorescence lifetime imaging using two photon tomography

Science.gov (United States)

Patalay, Rakesh; Talbot, Clifford; Alexandrov, Yuriy; Munro, Ian; Breunig, Hans Georg; König, Karsten; Warren, Sean; Neil, Mark A. A.; French, Paul M. W.; Chu, Anthony; Stamp, Gordon W.; Dunsby, Christopher

2011-07-01

Multispectral fluorescence lifetime imaging (FLIM) using two photon microscopy as a non-invasive technique for the diagnosis of skin lesions is described. Skin contains fluorophores including elastin, keratin, collagen, FAD and NADH. This endogenous contrast allows tissue to be imaged without the addition of exogenous agents and allows the in vivo state of cells and tissues to be studied. A modified DermaInspect® multiphoton tomography system was used to excite autofluorescence at 760 nm in vivo and on freshly excised ex vivo tissue. This instrument simultaneously acquires fluorescence lifetime images in four spectral channels between 360-655 nm using time-correlated single photon counting and can also provide hyperspectral images. The multispectral fluorescence lifetime images were spatially segmented and binned to determine lifetimes for each cell by fitting to a double exponential lifetime model. A comparative analysis between the cellular lifetimes from different diagnoses demonstrates significant diagnostic potential.

2D/3D cryo x-ray fluorescence imaging at the bionanoprobe at the advanced photon source

International Nuclear Information System (INIS)

Chen, S.; Vine, D. J.; Lai, B.; Paunesku, T.; Yuan, Y.; Woloschak, G. E.; Deng, J.; Jin, Q.; Hong, Y. P.; Flachenecker, C.; Hornberger, B.; Brister, K.; Jacobsen, C.; Vogt, S.

2016-01-01

Trace elements, particularly metals, play very important roles in biological systems. Synchrotron-based hard X-ray fluorescence microscopy offers the most suitable capabilities to quantitatively study trace metals in thick biological samples, such as whole cells and tissues. In this manuscript, we have demonstrated X-ray fluorescence imaging of frozen-hydrated whole cells using the recent developed Bionanoprobe (BNP). The BNP provides spatial resolution down to 30 nm and cryogenic capabilities. Frozen-hydrated biological cells have been directly examined on a sub-cellular level at liquid nitrogen temperatures with minimal sample preparation

Reviews in fluorescence 2010

CERN Document Server

Geddes, Chris D

2011-01-01

""Reviews in Fluorescence 2010"", the seventh volume of the book serial from Springer, serves as a comprehensive collection of current trends and emerging hot topics in the field of fluorescence and closely related disciplines. It summarizes the year's progress in fluorescence and its applications, with authoritative analytical reviews specialized enough to be attractive to professional researchers, yet also appealing to the wider audience of scientists in related disciplines of fluorescence. ""Reviews in Fluorescence"" offers an essential reference material for any lab working in the fluoresc

Preparation of dual-responsive hybrid fluorescent nano probe based on graphene oxide and boronic acid/BODIPY-conjugated polymer for cell imaging

Energy Technology Data Exchange (ETDEWEB)

Khoerunnisa [Department of IT Convergence, Korea National University of Transportation, Chungju 380–702 (Korea, Republic of); Kang, Eun Bi [Department of Chemical and Biological Engineering, Korea National University of Transportation, Chungju 380–702 (Korea, Republic of); Mazrad, Zihnil Adha Islamy [Department of IT Convergence, Korea National University of Transportation, Chungju 380–702 (Korea, Republic of); Lee, Gibaek [Department of Chemical and Biological Engineering, Korea National University of Transportation, Chungju 380–702 (Korea, Republic of); In, Insik [Department of IT Convergence, Korea National University of Transportation, Chungju 380–702 (Korea, Republic of); Department of Polymer Science and Engineering, Korea National University of Transportation, Chungju 380–702 (Korea, Republic of); Park, Sung Young, E-mail: parkchem@ut.ac.kr [Department of IT Convergence, Korea National University of Transportation, Chungju 380–702 (Korea, Republic of); Department of Chemical and Biological Engineering, Korea National University of Transportation, Chungju 380–702 (Korea, Republic of)

2017-02-01

Here, we report a pH- and thermo-responsive fluorescent nanomaterial of functionalized reduced graphene oxide (rGO) with cross-linked polymer produced via catechol-boronate diol binding mechanism. When conjugated with the hydrophobic dye boron dipyrromethane (BODIPY), this material can act as a dual-responsive nanoplatform for cells imaging. 2-Chloro-3′,4′-dihydroxyacetophenone (CCDP)-quaternized-poly(dimethylaminoethyl methacrylate-co-N-isopropylacrylamide) [C-PDN] was cross-linked with BODIPY and 4-chlorophenyl boronic acid (BA)-quaternized-poly(ethylene glycol)-g-poly(dimethylaminoethyl methacrylate-co-N-isopropylacrylamide) [BB-PPDN]. The GO was then reduced by the catechol group in the cross-linked polymer to synthesize rGO nanoparticles, which able to stabilize the quenching mechanism. This nanoplatform exhibits intense fluorescence at acidic pH and low fluorescence at physiological pH. Confocal laser scanning microscopy (CLSM) images shows bright fluorescence at lysosomal pH and total quench at physiological pH. Therefore, we have successfully developed a promising sensitive bio-imaging probe for identifying cancer cells. - Graphical abstract: [BB-PPDN]-[C-PDN]/rGO nanoparticles with boronic acid-catechol cis-diol binding mechanism toward change in pH demonstrated good biocompatibility and effective quenching for cancer cell detection. - Highlights: • Dual responsive (pH- and thermo) fluorescent nano probe was proposed for cells imaging. • The mechanism was based on cis-diol binding mechanism of boronic acid and catechol. • Reduced graphene oxide was used as quencher on nano-platform. • Detection was controlled dependent on pH based on diol compound of boron chemistry.

Tools for the quantitative analysis of sedimentation boundaries detected by fluorescence optical analytical ultracentrifugation.

Directory of Open Access Journals (Sweden)

Huaying Zhao

Full Text Available Fluorescence optical detection in sedimentation velocity analytical ultracentrifugation allows the study of macromolecules at nanomolar concentrations and below. This has significant promise, for example, for the study of systems of high-affinity protein interactions. Here we describe adaptations of the direct boundary modeling analysis approach implemented in the software SEDFIT that were developed to accommodate unique characteristics of the confocal fluorescence detection system. These include spatial gradients of signal intensity due to scanner movements out of the plane of rotation, temporal intensity drifts due to instability of the laser and fluorophores, and masking of the finite excitation and detection cone by the sample holder. In an extensive series of experiments with enhanced green fluorescent protein ranging from low nanomolar to low micromolar concentrations, we show that the experimental data provide sufficient information to determine the parameters required for first-order approximation of the impact of these effects on the recorded data. Systematic deviations of fluorescence optical sedimentation velocity data analyzed using conventional sedimentation models developed for absorbance and interference optics are largely removed after these adaptations, resulting in excellent fits that highlight the high precision of fluorescence sedimentation velocity data, thus allowing a more detailed quantitative interpretation of the signal boundaries that is otherwise not possible for this system.

Spatial ecology across scales.

Science.gov (United States)

Hastings, Alan; Petrovskii, Sergei; Morozov, Andrew

2011-04-23

The international conference 'Models in population dynamics and ecology 2010: animal movement, dispersal and spatial ecology' took place at the University of Leicester, UK, on 1-3 September 2010, focusing on mathematical approaches to spatial population dynamics and emphasizing cross-scale issues. Exciting new developments in scaling up from individual level movement to descriptions of this movement at the macroscopic level highlighted the importance of mechanistic approaches, with different descriptions at the microscopic level leading to different ecological outcomes. At higher levels of organization, different macroscopic descriptions of movement also led to different properties at the ecosystem and larger scales. New developments from Levy flight descriptions to the incorporation of new methods from physics and elsewhere are revitalizing research in spatial ecology, which will both increase understanding of fundamental ecological processes and lead to tools for better management.

Fluorescence studies on the aggregation behaviors of collagen modified with NHS-activated poly(γ-glutamic acid).

Science.gov (United States)

Zhang, Min; Yang, Junhui; Yang, Qili; Huang, Liulian; Wu, Hui; Chen, Lihui; Ding, Cuicui

2018-06-01

The poly(γ-glutamic acid)-NHS (γ-PGA-NHS) esters were used to endow collagen with both of excellent water-solubility and thermal stability via cross-linking reaction between γ-PGA-NHS and collagen. In the present work, the effect of γ-PGA-NHS on the aggregation of collagen molecules was studied by fluorescence techniques. The fluorescence emission spectra of pyrene in collagen solutions and the intrinsic fluorescence emission spectra of collagen suggested different effects of γ-PGA-NHS on collagen molecules: inhibiting aggregation below critical aggregation concentration (CAC) and promoting aggregation above CAC. The two-dimensional (2D) fluorescence correlation spectra indicated that the intermolecular hydrogen bonding and cross-linking between γ-PGA-NHS and collagen would influence the aggregation of collagen molecules. By the ultra-sensitive differential scanning calorimeter (VP-DSC), it was found that the main denaturational transition temperature (T m2 ) of modified collagen increased, while its calorimetric enthalpy changes (ΔH 2 ) decreased compared to those of native collagen, further indicating that the modification of γ-PGA-NHS influenced the aggregation of collagen molecules. The study provide useful information for the utilizing and or the processing of water-soluble collagen in aqueous solution in the fields such as cosmetics, health care products, tissue engineering and biomedical materials, etc. Copyright © 2018 Elsevier B.V. All rights reserved.

Integrating spatial and biomass planning for the United States

International Nuclear Information System (INIS)

Wang, Sicong; Wang, Shifeng

2016-01-01

Biomass is low-carbon energy and has tremendous potential as an alternative to fossil fuels. However, the significant role of biomass in future low-carbon energy portfolio depends heavily on its consumption. The paper presents a first attempt to examine the spatial-temporal patterns of biomass consumption in the United States (US), using a novel method-spatial Seemingly Unrelated Regression (SUR) model, in order to strengthen the link between energy planning and spatial planning. In order to obtain the robust parameters of spatial SUR models and estimate the parameters efficiently, an iterative maximum likelihood method, which takes full advantage of the stationary characteristic of maximum likelihood estimation, has been developed. The robust parameters of models can help draw a proper inference for biomass consumption. Then the spatial-temporal patterns of biomass consumption in the US at the state level are investigated using the spatial SUR models with the estimation method developed and data covering the period of 2000–2012. Results show that there are spatial dependences among biomass consumption. The presence of spatial dependence in biomass consumption has informative implications for making sustainable biomass polices. It suggests new efforts to adding a cross-state dimension to state-level energy policy and coordinating some elements of energy policy across states are still needed. In addition, results consistent with classic economic theory further proves the correctness of applying the spatial SUR models to investigate the spatial-temporal patterns of biomass consumption. - Highlights: • A spatial model is suggested as framework to investigate biomass consumption. • A new estimation method is developed to obtain the robust parameters of model. • There are spatial dependences among biomass consumption. • The spatial dependence can contribute to making sustainable biomass policies. • Efforts to adding cross-state dimension to state

Drought is Coming: Monitoring Vegetation Response to Water Scarcity through Variable Chlorophyll a Fluorescence

Science.gov (United States)

Guadagno, C. R.; Beverly, D.; Pleban, J. R.; Speckman, H. N.; Ewers, B. E.; Weinig, C.

2017-12-01

are limited in scale, chlorophyll fluorescence comprises an indicator of drought stress across multiple spatial scales, from leaf to ecosystem level.

Fluorescence excitation-emission matrix spectroscopy of vitiligo skin in vivo (Conference Presentation)

Science.gov (United States)

Zhao, Jianhua; Richer, Vincent; Al Jasser, Mohammed; Zandi, Soodabeh; Kollias, Nikiforos; Kalia, Sunil; Zeng, Haishan; Lui, Harvey

2016-02-01

Fluorescence signals depend on the intensity of the exciting light, the absorption properties of the constituent molecules, and the efficiency with which the absorbed photons are converted to fluorescence emission. The optical features and appearance of vitiligo have been explained primarily on the basis of reduced epidermal pigmentation, which results in abnormal white patches on the skin. The objective of this study is to explore the fluorescence properties of vitiligo and its adjacent normal skin using fluorescence excitation-emission matrix (EEM) spectroscopy. Thirty five (35) volunteers with vitiligo were acquired using a double-grating spectrofluorometer with excitation and emission wavelengths of 260-450 nm and 300-700 nm respectively. As expected, the most pronounced difference between the spectra obtained from vitiligo lesions compared to normally pigmented skin was that the overall fluorescence was much higher in vitiligo; these differences increased at shorter wavelengths, thus matching the characteristic spectral absorption of epidermal melanin. When comparing the fluorescence spectra from vitiligo to normal skin we detected three distinct spectral bands centered at 280nm, 310nm, and 335nm. The 280nm band may possibly be related to inflammation, whereas the 335 nm band may arise from collagen or keratin cross links. The source of the 310 nm band is uncertain; it is interesting to note its proximity to the 311 nm UV lamps used for vitiligo phototherapy. These differences are accounted for not only by changes in epidermal pigment content, but also by other optically active cutaneous biomolecules.

Cross-cultural variation of speech-accompanying gesture : a review

OpenAIRE

Kita, Sotaro

2009-01-01

This article reviews the literature on cross-cultural variation of gestures. Four factors governing the variation were identified. The first factor is the culture-specific convention for form-meaning associations. This factor is involved in well-known cross-cultural differences in emblem gestures (e.g., the OK-sign), as well as pointing gestures. The second factor is culture-specific spatial cognition. Representational gestures (i.e., iconic and deictic gestures) that express spatial contents...

Open source tools for fluorescent imaging.

Science.gov (United States)

Hamilton, Nicholas A

2012-01-01

As microscopy becomes increasingly automated and imaging expands in the spatial and time dimensions, quantitative analysis tools for fluorescent imaging are becoming critical to remove both bottlenecks in throughput as well as fully extract and exploit the information contained in the imaging. In recent years there has been a flurry of activity in the development of bio-image analysis tools and methods with the result that there are now many high-quality, well-documented, and well-supported open source bio-image analysis projects with large user bases that cover essentially every aspect from image capture to publication. These open source solutions are now providing a viable alternative to commercial solutions. More importantly, they are forming an interoperable and interconnected network of tools that allow data and analysis methods to be shared between many of the major projects. Just as researchers build on, transmit, and verify knowledge through publication, open source analysis methods and software are creating a foundation that can be built upon, transmitted, and verified. Here we describe many of the major projects, their capabilities, and features. We also give an overview of the current state of open source software for fluorescent microscopy analysis and the many reasons to use and develop open source methods. Copyright © 2012 Elsevier Inc. All rights reserved.

Quantitative structural modeling on the wavelength interval (Δλ) in synchronous fluorescence spectroscopy

Science.gov (United States)

Samari, Fayezeh; Yousefinejad, Saeed

2017-11-01

Emission fluorescence spectroscopy has an extremely restricted scope of application to analyze of complex mixtures since its selectivity is reduced by the extensive spectral overlap. Synchronous fluorescence spectroscopy (SFS) is a technique enables us to analyze complex mixtures with overlapped emission and/or excitation spectra. The difference of excitation and emission wavelength of compounds (interval wavelength or Δλ) is an important characteristic in SFS. Thus a multi-parameter model was constructed to predict Δλ in 63 fluorescent compounds and the regression coefficient in training set, cross validation and test set were 0.88, 0.85 and 0.91 respectively. Furthermore, the applicability and validity of model were evaluated using different statistical methods such as y-scrambling and applicability domain. It was concluded that increasing average valence connectivity, number of Al2-NH functional group and Geary autocorrelation (lag 4) with electronegative weights can lead to increasing Δλ in the fluorescent compounds. The current study obtained an insight into the structural properties of compounds effective on their Δλ as an important parameter in SFS.

The E. coli immunosorbent as used in serodiagnosis of Legionella infections studied by crossed immunoelectrophoresis

DEFF Research Database (Denmark)

Bangsborg, Jette Marie; Friis-Møller, A; Rechnitzer, C

1988-01-01

In this study we investigated an immunosorbent, E. coli blocking fluid (BF), proposed for use in the Legionella Indirect Fluorescent Antibody Test (IFA). With crossed immunoelectrophoresis (CIE) of clinically relevant Legionella species, only one heat-stable antigen (no. 1) cross...

Principles of fluorescence techniques

CERN Document Server

2016-01-01

Fluorescence techniques are being used and applied increasingly in academics and industry. The Principles of Fluorescence Techniques course will outline the basic concepts of fluorescence techniques and the successful utilization of the currently available commercial instrumentation. The course is designed for students who utilize fluorescence techniques and instrumentation and for researchers and industrial scientists who wish to deepen their knowledge of fluorescence applications. Key scientists in the field will deliver theoretical lectures. The lectures will be complemented by the direct utilization of steady-state and lifetime fluorescence instrumentation and confocal microscopy for FLIM and FRET applications provided by leading companies.

Visualizing Escherichia coli sub-cellular structure using sparse deconvolution Spatial Light Interference Tomography.

Directory of Open Access Journals (Sweden)

Mustafa Mir

Full Text Available Studying the 3D sub-cellular structure of living cells is essential to our understanding of biological function. However, tomographic imaging of live cells is challenging mainly because they are transparent, i.e., weakly scattering structures. Therefore, this type of imaging has been implemented largely using fluorescence techniques. While confocal fluorescence imaging is a common approach to achieve sectioning, it requires fluorescence probes that are often harmful to the living specimen. On the other hand, by using the intrinsic contrast of the structures it is possible to study living cells in a non-invasive manner. One method that provides high-resolution quantitative information about nanoscale structures is a broadband interferometric technique known as Spatial Light Interference Microscopy (SLIM. In addition to rendering quantitative phase information, when combined with a high numerical aperture objective, SLIM also provides excellent depth sectioning capabilities. However, like in all linear optical systems, SLIM's resolution is limited by diffraction. Here we present a novel 3D field deconvolution algorithm that exploits the sparsity of phase images and renders images with resolution beyond the diffraction limit. We employ this label-free method, called deconvolution Spatial Light Interference Tomography (dSLIT, to visualize coiled sub-cellular structures in E. coli cells which are most likely the cytoskeletal MreB protein and the division site regulating MinCDE proteins. Previously these structures have only been observed using specialized strains and plasmids and fluorescence techniques. Our results indicate that dSLIT can be employed to study such structures in a practical and non-invasive manner.

In-situ optical and acoustical measurements of the buoyant cyanobacterium p. Rubescens: spatial and temporal distribution patterns.

Directory of Open Access Journals (Sweden)

Hilmar Hofmann

Full Text Available Optical (fluorescence and acoustic in-situ techniques were tested in their ability to measure the spatial and temporal distribution of plankton in freshwater ecosystems with special emphasis on the harmful and buoyant cyanobacterium P. rubescens. Fluorescence was measured with the multi-spectral FluoroProbe (Moldaenke FluoroProbe, MFP and a Seapoint Chlorophyll Fluorometer (SCF. In-situ measurements of the acoustic backscatter strength (ABS were conducted with three different acoustic devices covering multiple acoustic frequencies (614 kHz ADCP, 2 MHz ADP, and 6 MHz ADV. The MFP provides a fast and reliable technique to measure fluorescence at different wavelengths in situ, which allows discriminating between P. rubescens and other phytoplankton species. All three acoustic devices are sensitive to P. rubescens even if other scatterers, e.g., zooplankton or suspended sediment, are present in the water column, because P. rubescens containing gas vesicles has a strong density difference and hence acoustic contrast to the ambient water and other scatterers. After calibration, the combination of optical and acoustical measurements not only allows qualitative and quantitative observation of P. rubescens, but also distinction between P. rubescens, other phytoplankton, and zooplankton. As the measuring devices can sample in situ at high rates they enable assessment of plankton distributions at high temporal (minutes and spatial (decimeters resolution or covering large temporal (seasonal and spatial (basin scale scales.

Efficient L1 regularization-based reconstruction for fluorescent molecular tomography using restarted nonlinear conjugate gradient.

Science.gov (United States)

Shi, Junwei; Zhang, Bin; Liu, Fei; Luo, Jianwen; Bai, Jing

2013-09-15

For the ill-posed fluorescent molecular tomography (FMT) inverse problem, the L1 regularization can protect the high-frequency information like edges while effectively reduce the image noise. However, the state-of-the-art L1 regularization-based algorithms for FMT reconstruction are expensive in memory, especially for large-scale problems. An efficient L1 regularization-based reconstruction algorithm based on nonlinear conjugate gradient with restarted strategy is proposed to increase the computational speed with low memory consumption. The reconstruction results from phantom experiments demonstrate that the proposed algorithm can obtain high spatial resolution and high signal-to-noise ratio, as well as high localization accuracy for fluorescence targets.

Fluorescent optical position sensor

Science.gov (United States)

Weiss, Jonathan D.

2005-11-15

A fluorescent optical position sensor and method of operation. A small excitation source side-pumps a localized region of fluorescence at an unknown position along a fluorescent waveguide. As the fluorescent light travels down the waveguide, the intensity of fluorescent light decreases due to absorption. By measuring with one (or two) photodetectors the attenuated intensity of fluorescent light emitted from one (or both) ends of the waveguide, the position of the excitation source relative to the waveguide can be determined by comparing the measured light intensity to a calibrated response curve or mathematical model. Alternatively, excitation light can be pumped into an end of the waveguide, which generates an exponentially-decaying continuous source of fluorescent light along the length of the waveguide. The position of a photodetector oriented to view the side of the waveguide can be uniquely determined by measuring the intensity of the fluorescent light emitted radially at that location.

Total Internal Reflection Fluorescence Microscopy Imaging-Guided Confocal Single-Molecule Fluorescence Spectroscopy

OpenAIRE

Zheng, Desheng; Kaldaras, Leonora; Lu, H. Peter

2013-01-01

We have developed an integrated spectroscopy system combining total internal reflection fluorescence microscopy imaging with confocal single-molecule fluorescence spectroscopy for two-dimensional interfaces. This spectroscopy approach is capable of both multiple molecules simultaneously sampling and in situ confocal fluorescence dynamics analyses of individual molecules of interest. We have demonstrated the calibration with fluorescent microspheres, and carried out single-molecule spectroscop...

Scanning fluorescent microscopy is an alternative for quantitative fluorescent cell analysis.

Science.gov (United States)

Varga, Viktor Sebestyén; Bocsi, József; Sipos, Ferenc; Csendes, Gábor; Tulassay, Zsolt; Molnár, Béla

2004-07-01

Fluorescent measurements on cells are performed today with FCM and laser scanning cytometry. The scientific community dealing with quantitative cell analysis would benefit from the development of a new digital multichannel and virtual microscopy based scanning fluorescent microscopy technology and from its evaluation on routine standardized fluorescent beads and clinical specimens. We applied a commercial motorized fluorescent microscope system. The scanning was done at 20 x (0.5 NA) magnification, on three channels (Rhodamine, FITC, Hoechst). The SFM (scanning fluorescent microscopy) software included the following features: scanning area, exposure time, and channel definition, autofocused scanning, densitometric and morphometric cellular feature determination, gating on scatterplots and frequency histograms, and preparation of galleries of the gated cells. For the calibration and standardization Immuno-Brite beads were used. With application of shading compensation, the CV of fluorescence of the beads decreased from 24.3% to 3.9%. Standard JPEG image compression until 1:150 resulted in no significant change. The change of focus influenced the CV significantly only after +/-5 microm error. SFM is a valuable method for the evaluation of fluorescently labeled cells. Copyright 2004 Wiley-Liss, Inc.

Simulating fluorescence light-canopy interaction in support of laser-induced fluorescence measurements

International Nuclear Information System (INIS)

Rosema, A.; Verhoef, W.; Schroote, J.; Snel, J.F.H.

1991-01-01

In the Netherlands an operational field instrument for the measurement of laser induced fluorescence of vegetation (LEAF) is developed. In addition, plant physiological and remote sensing research is done to support this new remote sensing instrument. This paper presents a general introduction on the subject of laser-induced fluorescence, including the relation between chlorophyll fluorescence and photosynthesis, spectral characteristics, and previous research. Also the LEAF system is briefly described. Subsequently, the development of a leaf fluorescence model (KMF) and a canopy fluorescence model (FLSAIL) are reported. With these simulation models a sensitivity study is carried out. Fluorescence of 685 nm appears to be most suitable to obtain information on photosynthesis and stress, but is also influenced by canopy structure. Separation of these two effects is studied

UV-Vis Ratiometric Resonance Synchronous Spectroscopy for Determination of Nanoparticle and Molecular Optical Cross Sections.

Science.gov (United States)

Nettles, Charles B; Zhou, Yadong; Zou, Shengli; Zhang, Dongmao

2016-03-01

Demonstrated herein is a UV-vis Ratiometric Resonance Synchronous Spectroscopic (R2S2, pronounced as "R-two-S-two" for simplicity) technique where the R2S2 spectrum is obtained by dividing the resonance synchronous spectrum of a NP-containing solution by the solvent resonance synchronous spectrum. Combined with conventional UV-vis measurements, this R2S2 method enables experimental quantification of the absolute optical cross sections for a wide range of molecular and nanoparticle (NP) materials that range optically from pure photon absorbers or scatterers to simultaneous photon absorbers and scatterers, simultaneous photon absorbers and emitters, and all the way to simultaneous photon absorbers, scatterers, and emitters in the UV-vis wavelength region. Example applications of this R2S2 method were demonstrated for quantifying the Rayleigh scattering cross sections of solvents including water and toluene, absorption and resonance light scattering cross sections for plasmonic gold nanoparticles, and absorption, scattering, and on-resonance fluorescence cross sections for semiconductor quantum dots (Qdots). On-resonance fluorescence quantum yields were quantified for the model molecular fluorophore Eosin Y and fluorescent Qdots CdSe and CdSe/ZnS. The insights and methodology presented in this work should be of broad significance in physical and biological science research that involves photon/matter interactions.

Functional inverted Wishart for Bayesian multivariate spatial modeling with application to regional climatology model data.

Science.gov (United States)

Duan, L L; Szczesniak, R D; Wang, X

2017-11-01

Modern environmental and climatological studies produce multiple outcomes at high spatial resolutions. Multivariate spatial modeling is an established means to quantify cross-correlation among outcomes. However, existing models typically suffer from poor computational efficiency and lack the flexibility to simultaneously estimate auto- and cross-covariance structures. In this article, we undertake a novel construction of covariance by utilizing spectral convolution and by imposing an inverted Wishart prior on the cross-correlation structure. The cross-correlation structure with this functional inverted Wishart prior flexibly accommodates not only positive but also weak or negative associations among outcomes while preserving spatial resolution. Furthermore, the proposed model is computationally efficient and produces easily interpretable results, including the individual autocovariances and full cross-correlation matrices, as well as a partial cross-correlation matrix reflecting the outcome correlation after excluding the effects caused by spatial convolution. The model is examined using simulated data sets under different scenarios. It is also applied to the data from the North American Regional Climate Change Assessment Program, examining long-term associations between surface outcomes for air temperature, pressure, humidity, and radiation, on the land area of the North American West Coast. Results and predictive performance are compared with findings from approaches using convolution only or coregionalization.

Functional inverted Wishart for Bayesian multivariate spatial modeling with application to regional climatology model data

Science.gov (United States)

Duan, L. L.; Szczesniak, R. D.; Wang, X.

2018-01-01

Modern environmental and climatological studies produce multiple outcomes at high spatial resolutions. Multivariate spatial modeling is an established means to quantify cross-correlation among outcomes. However, existing models typically suffer from poor computational efficiency and lack the flexibility to simultaneously estimate auto- and cross-covariance structures. In this article, we undertake a novel construction of covariance by utilizing spectral convolution and by imposing an inverted Wishart prior on the cross-correlation structure. The cross-correlation structure with this functional inverted Wishart prior flexibly accommodates not only positive but also weak or negative associations among outcomes while preserving spatial resolution. Furthermore, the proposed model is computationally efficient and produces easily interpretable results, including the individual autocovariances and full cross-correlation matrices, as well as a partial cross-correlation matrix reflecting the outcome correlation after excluding the effects caused by spatial convolution. The model is examined using simulated data sets under different scenarios. It is also applied to the data from the North American Regional Climate Change Assessment Program, examining long-term associations between surface outcomes for air temperature, pressure, humidity, and radiation, on the land area of the North American West Coast. Results and predictive performance are compared with findings from approaches using convolution only or coregionalization. PMID:29576735

Research progress and hotspot analysis of spatial interpolation

Science.gov (United States)

Jia, Li-juan; Zheng, Xin-qi; Miao, Jin-li

2018-02-01

In this paper, the literatures related to spatial interpolation between 1982 and 2017, which are included in the Web of Science core database, are used as data sources, and the visualization analysis is carried out according to the co-country network, co-category network, co-citation network, keywords co-occurrence network. It is found that spatial interpolation has experienced three stages: slow development, steady development and rapid development; The cross effect between 11 clustering groups, the main convergence of spatial interpolation theory research, the practical application and case study of spatial interpolation and research on the accuracy and efficiency of spatial interpolation. Finding the optimal spatial interpolation is the frontier and hot spot of the research. Spatial interpolation research has formed a theoretical basis and research system framework, interdisciplinary strong, is widely used in various fields.

Silica encapsulation of fluorescent nanodiamonds for colloidal stability and facile surface functionalization.

Science.gov (United States)

Bumb, Ambika; Sarkar, Susanta K; Billington, Neil; Brechbiel, Martin W; Neuman, Keir C

2013-05-29

Fluorescent nanodiamonds (FNDs) emit in the near-IR and do not photobleach or photoblink. These properties make FNDs better suited for numerous imaging applications compared with commonly used fluorescence agents such as organic dyes and quantum dots. However, nanodiamonds do not form stable suspensions in aqueous buffer, are prone to aggregation, and are difficult to functionalize. Here we present a method for encapsulating nanodiamonds with silica using an innovative liposome-based encapsulation process that renders the particle surface biocompatible, stable, and readily functionalized through routine linking chemistries. Furthermore, the method selects for a desired particle size and produces a monodisperse agent. We attached biotin to the silica-coated FNDs and tracked the three-dimensional motion of a biotinylated FND tethered by a single DNA molecule with high spatial and temporal resolution.

Kr II laser-induced fluorescence for measuring plasma acceleration.

Science.gov (United States)

Hargus, W A; Azarnia, G M; Nakles, M R

2012-10-01

We present the application of laser-induced fluorescence of singly ionized krypton as a diagnostic technique for quantifying the electrostatic acceleration within the discharge of a laboratory cross-field plasma accelerator also known as a Hall effect thruster, which has heritage as spacecraft propulsion. The 728.98 nm Kr II transition from the metastable 5d(4)D(7/2) to the 5p(4)P(5/2)(∘) state was used for the measurement of laser-induced fluorescence within the plasma discharge. From these measurements, it is possible to measure velocity as krypton ions are accelerated from near rest to approximately 21 km/s (190 eV). Ion temperature and the ion velocity distributions may also be extracted from the fluorescence data since available hyperfine splitting data allow for the Kr II 5d(4)D(7/2)-5p(4)P(5/2)(∘) transition lineshape to be modeled. From the analysis, the fluorescence lineshape appears to be a reasonable estimate for the relatively broad ion velocity distributions. However, due to an apparent overlap of the ion creation and acceleration regions within the discharge, the distributed velocity distributions increase ion temperature determination uncertainty significantly. Using the most probable ion velocity as a representative, or characteristic, measure of the ion acceleration, overall propellant energy deposition, and effective electric fields may be calculated. With this diagnostic technique, it is possible to nonintrusively characterize the ion acceleration both within the discharge and in the plume.

Fluorescence molecular tomography in the presence of background fluorescence

International Nuclear Information System (INIS)

Soubret, Antoine; Ntziachristos, Vasilis

2006-01-01

Fluorescence molecular tomography is an emerging imaging technique that resolves the bio-distribution of engineered fluorescent probes developed for in vivo reporting of specific cellular and sub-cellular targets. The method can detect fluorochromes in picomole amounts or less, imaged through entire animals, but the detection sensitivity and imaging performance drop in the presence of background, non-specific fluorescence. In this study, we carried out a theoretical and an experimental investigation on the effect of background fluorescence on the measured signal and on the tomographic reconstruction. We further examined the performance of three subtraction methods based on physical models of photon propagation, using experimental data on phantoms and small animals. We show that the data pre-processing with subtraction schemes can improve image quality and quantification when non-specific background florescence is present

Detection of rhodopsin dimerization in situ by PIE-FCCS, a time-resolved fluorescence spectroscopy.

Science.gov (United States)

Smith, Adam W

2015-01-01

Rhodopsin self-associates in the plasma membrane. At low concentrations, the interactions are consistent with a monomer-dimer equilibrium (Comar et al., J Am Chem Soc 136(23):8342-8349, 2014). At high concentrations in native tissue, higher-order clusters have been observed (Fotiadis et al., Nature 421:127-128, 2003). The physiological role of rhodopsin dimerization is still being investigated, but it is clear that a quantitative assessment is essential to determining the function of rhodopsin clusters in vision. To quantify rhodopsin interactions, I will outline the theory and methodology of a specialized time-resolved fluorescence spectroscopy for measuring membrane protein-protein interactions called pulsed-interleaved excitation fluorescence cross-correlation spectroscopy (PIE-FCCS). The strength of this technique is its ability to quantify rhodopsin interactions in situ (i.e., a live cell plasma membrane). There are two reasons for restricting the scope to live cell membranes. First, the compositional heterogeneity of the plasma membrane creates a complex milieu with thousands of lipid, protein, and carbohydrate species. This makes it difficult to infer quaternary interactions from detergent solubilized samples or construct a model phospholipid bilayer that recapitulates all of the interactions present in native membranes. Second, organizational structure and dynamics is a key feature of the plasma membrane, and fixation techniques like formaldehyde cross-linking and vitrification will modulate the interactions. PIE-FCCS is based on two-color fluorescence imaging with time-correlated single-photon counting (TCSPC) (Becker et al., Rev Sci Instrum 70:1835-1841, 1999). By time-tagging every detected photon, the data can be analyzed as a fluorescence intensity distribution, fluorescence lifetime histogram, or fluorescence (cross-)correlation spectra (FCS/FCCS) (Becker, Advanced time-correlated single-photon counting techniques, Springer, Berlin, 2005). These

Measuring Photosynthetic Response to Drought Stress using Active and Passive Fluorescence

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Helm, L.; Lerdau, M.; Wang, W.; Yang, X.

2017-12-01

Photosynthesis, the endothermic reactions involving the absorption of light and fixation and reduction of carbon dioxide by plants, plays important roles in carbon and water cycles, food security, and even weather and climate patterns. Solar radiation provides the energy for photosynthesis, but often plants absorb more solar energy than they can use to reduce carbon dioxide. This excess energy, which is briefly stored as high-energy electrons in the chloroplast, must be removed or damage to the leaf's photosynthetic machinery will occur. One important energy dissipation pathway is for the high energy electrons to return to their lower valance state and, in doing so, release radiation (fluorescence). This fluorescence (known as solar induced fluorescence (SIF) has been found to strongly correlate with gross photosynthesis. Recent advances in the remote sensing of SIF allow for large-scale real-time estimation of photosynthesis. In a warming climate with more frequent stress, remote sensing is necessary for measuring the spatial and temporal variability of photosynthesis. However, the mechanisms that link SIF and photosynthesis are unclear, particularly how the relationship may or may not change under stress. We present data from leaf-level measurements of gas exchange, pulse amplitude modulation (PAM) fluorescence, and SIF in two major tree species in North America. Water-stressed and well-watered plants were compared to determine how SIF and carbon dioxide exchange are modulated by drought diurnally and seasonally. Secondly, photosynthesis and fluorescence under high and low oxygen concentrations were compared to determine how photorespiration alters the relationship between SIF and gross photosynthesis. We find a strong correlation between SIF and steady-state fluorescence measured with conventional PAM fluorometry. Our results also indicate that drought-stress modulates the SIF-photosynthesis relationship, and this may be driven by drought-induced changes in

Background fluorescence estimation and vesicle segmentation in live cell imaging with conditional random fields.

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Pécot, Thierry; Bouthemy, Patrick; Boulanger, Jérôme; Chessel, Anatole; Bardin, Sabine; Salamero, Jean; Kervrann, Charles

2015-02-01

Image analysis applied to fluorescence live cell microscopy has become a key tool in molecular biology since it enables to characterize biological processes in space and time at the subcellular level. In fluorescence microscopy imaging, the moving tagged structures of interest, such as vesicles, appear as bright spots over a static or nonstatic background. In this paper, we consider the problem of vesicle segmentation and time-varying background estimation at the cellular scale. The main idea is to formulate the joint segmentation-estimation problem in the general conditional random field framework. Furthermore, segmentation of vesicles and background estimation are alternatively performed by energy minimization using a min cut-max flow algorithm. The proposed approach relies on a detection measure computed from intensity contrasts between neighboring blocks in fluorescence microscopy images. This approach permits analysis of either 2D + time or 3D + time data. We demonstrate the performance of the so-called C-CRAFT through an experimental comparison with the state-of-the-art methods in fluorescence video-microscopy. We also use this method to characterize the spatial and temporal distribution of Rab6 transport carriers at the cell periphery for two different specific adhesion geometries.

A Naturally Fluorescent Mgp Transgenic Mouse for Angiogenesis and Glaucoma Longitudinal Studies.

Science.gov (United States)

Asokan, Priyadarsini; Mitra, Rajendra N; Periasamy, Ramesh; Han, Zongchao; Borrás, Teresa

2018-02-01

Our goal was to generate and characterize a new mouse model in which only angiogenesis- and glaucoma-relevant tissues would be naturally fluorescent. The Matrix Gla (MGP) gene is highly expressed in vascular smooth muscle cells (VSMC) and trabecular meshwork (TM). We sought to direct our Mgp-Cre.KI mouse recombinase to VSMC/TM cells to produce their longitudinal fluorescent profiles. Homozygous Mgp-Cre.KI mice were crossed with Ai9 homozygous reporter mice harboring a loxP-flanked STOP cassette preventing transcription of a DsRed fluorescent protein (tdTomato). The F1 double-heterozygous (Mgp-tdTomato) was examined by direct fluorescence, whole mount, histology, and fundus photography. Custom-made filters had 554/23 emission and 609/54 exciter nanometer wavelengths. Proof of concept of the model's usefulness was conducted by inducing guided imaging laser burns. Evaluation of a vessel's leakage and proliferation was followed by noninvasive angiography. The Mgp-tdTomato mouse was viable, fertile, with normal IOP and ERG. Its phenotype exhibited red paws and snout (cartilage expression), which precluded genotyping. A fluorescent red ring was seen at the limbus and confirmed to be TM expression by histology. The entire retinal vasculature was red fluorescent (VSMC) and directly visualized by fundus photography. Laser burns on the Mgp-tdTomato allowed separation of leakiness and neovascularization evaluation parameters. The availability of a transgenic mouse naturally fluorescent in glaucoma-relevant tissues and retinal vasculature brings the unique opportunity to study a wide spectrum of single and combined glaucomatous conditions in vivo. Moreover, the Mgp-tdTomato mouse provides a new tool to study mechanisms and therapeutics of retinal angiogenesis longitudinally.

A Naturally Fluorescent Mgp Transgenic Mouse for Angiogenesis and Glaucoma Longitudinal Studies

Science.gov (United States)

Asokan, Priyadarsini; Mitra, Rajendra N.; Periasamy, Ramesh; Han, Zongchao

2018-01-01

Purpose Our goal was to generate and characterize a new mouse model in which only angiogenesis- and glaucoma-relevant tissues would be naturally fluorescent. The Matrix Gla (MGP) gene is highly expressed in vascular smooth muscle cells (VSMC) and trabecular meshwork (TM). We sought to direct our Mgp-Cre.KI mouse recombinase to VSMC/TM cells to produce their longitudinal fluorescent profiles. Methods Homozygous Mgp-Cre.KI mice were crossed with Ai9 homozygous reporter mice harboring a loxP-flanked STOP cassette preventing transcription of a DsRed fluorescent protein (tdTomato). The F1 double-heterozygous (Mgp-tdTomato) was examined by direct fluorescence, whole mount, histology, and fundus photography. Custom-made filters had 554/23 emission and 609/54 exciter nanometer wavelengths. Proof of concept of the model's usefulness was conducted by inducing guided imaging laser burns. Evaluation of a vessel's leakage and proliferation was followed by noninvasive angiography. Results The Mgp-tdTomato mouse was viable, fertile, with normal IOP and ERG. Its phenotype exhibited red paws and snout (cartilage expression), which precluded genotyping. A fluorescent red ring was seen at the limbus and confirmed to be TM expression by histology. The entire retinal vasculature was red fluorescent (VSMC) and directly visualized by fundus photography. Laser burns on the Mgp-tdTomato allowed separation of leakiness and neovascularization evaluation parameters. Conclusions The availability of a transgenic mouse naturally fluorescent in glaucoma-relevant tissues and retinal vasculature brings the unique opportunity to study a wide spectrum of single and combined glaucomatous conditions in vivo. Moreover, the Mgp-tdTomato mouse provides a new tool to study mechanisms and therapeutics of retinal angiogenesis longitudinally. PMID:29392320

Cross-Sectional Transport Imaging in a Multijunction Solar Cell

Energy Technology Data Exchange (ETDEWEB)

Haegel, Nancy M.; Ke, Chi-Wen; Taha, Hesham; Guthrey, Harvey; Fetzer, C. M.; King, Richard

2015-06-14

Combining highly localized electron-beam excitation at a point with the spatial resolution capability of optical near-field imaging, we have imaged carrier transport in a cross-sectioned multijunction (GaInP/GaInAs/Ge) solar cell. We image energy transport associated with carrier diffusion throughout the full width of the middle (GaInAs) cell and luminescent coupling from point excitation in the top cell GaInP to the middle cell. Supporting cathodoluminescence and near-field photoluminescence measurements demonstrate excitation-dependent Fermi level splitting effects that influence cross-sectioned spectroscopy results as well as transport limitations on the spatial resolution of cross-sectional measurements.

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

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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.

Precision automation of cell type classification and sub-cellular fluorescence quantification from laser scanning confocal images

Directory of Open Access Journals (Sweden)

Hardy Craig Hall

2016-02-01

Full Text Available While novel whole-plant phenotyping technologies have been successfully implemented into functional genomics and breeding programs, the potential of automated phenotyping with cellular resolution is largely unexploited. Laser scanning confocal microscopy has the potential to close this gap by providing spatially highly resolved images containing anatomic as well as chemical information on a subcellular basis. However, in the absence of automated methods, the assessment of the spatial patterns and abundance of fluorescent markers with subcellular resolution is still largely qualitative and time-consuming. Recent advances in image acquisition and analysis, coupled with improvements in microprocessor performance, have brought such automated methods within reach, so that information from thousands of cells per image for hundreds of images may be derived in an experimentally convenient time-frame. Here, we present a MATLAB-based analytical pipeline to 1 segment radial plant organs into individual cells, 2 classify cells into cell type categories based upon random forest classification, 3 divide each cell into sub-regions, and 4 quantify fluorescence intensity to a subcellular degree of precision for a separate fluorescence channel. In this research advance, we demonstrate the precision of this analytical process for the relatively complex tissues of Arabidopsis hypocotyls at various stages of development. High speed and robustness make our approach suitable for phenotyping of large collections of stem-like material and other tissue types.

Melanin-originated carbonaceous dots for triple negative breast cancer diagnosis by fluorescence and photoacoustic dual-mode imaging.

Science.gov (United States)

Xiao, Wei; Li, Yuan; Hu, Chuan; Huang, Yuan; He, Qin; Gao, Huile

2017-07-01

Carbonaceous dots exhibit increasing applications in diagnosis and drug delivery due to excellent photostability and biocompatibility properties. However, relative short excitation and emission of melanin carbonaceous dots (MCDs) limit the applicability in fluorescence bioimaging. Furthermore, the generally poor spatial resolution of fluorescence imaging limits potential in vivo applications. Due to a variety of beneficial properties, in this study, MCDs were prepared exhibiting great potential in fluorescence and photoacoustic dual-mode bioimaging. The MCDs exhibited a long excitation peak at 615nm and emission peak at 650nm, further highlighting the applicability in fluorescence imaging, while the absorbance peak at 633nm renders MCDs suitable for photoacoustic imaging. In vivo, the photoacoustic signal of MCDs was linearly correlated with the concentration of MCDs. Moreover, the MCDs were shown to be taken up into triple negative breast cancer cell line 4T1 in both a time- and concentration-dependent manner. In vivo fluorescence and photoacoustic imaging of subcutaneous 4T1 tumor demonstrated that MCDs could passively target triple negative breast cancer tissue by enhanced permeability and retention effects and may therefore be used for tumor dual-mode imaging. Furthermore, fluorescence distribution in tissue slices suggested that MCDs may distribute in 4T1 tumor with high efficacy. In conclusion, the MCDs studied offer potential application in fluorescence and photoacoustic dual-mode imaging. Copyright © 2017 Elsevier Inc. All rights reserved.

Investigation of spatial resolution characteristics of an in vivo microcomputed tomography system

Energy Technology Data Exchange (ETDEWEB)

Ghani, Muhammad U. [Center for Biomedical engineering and School of Electrical and Computer Engineering, University of Oklahoma, Norman, OK 73019 (United States); Zhou, Zhongxing [Center for Biomedical engineering and School of Electrical and Computer Engineering, University of Oklahoma, Norman, OK 73019 (United States); School of Precision and Optoelectronics Engineering, Tianjin University, Tianjin 300072 (China); Ren, Liqiang; Wong, Molly; Li, Yuhua; Zheng, Bin [Center for Biomedical engineering and School of Electrical and Computer Engineering, University of Oklahoma, Norman, OK 73019 (United States); Yang, Kai [Department of Radiology, Massachusetts General Hospital, 55 Fruit Street, Boston, MA 02114 (United States); Liu, Hong, E-mail: liu@ou.edu [Center for Biomedical engineering and School of Electrical and Computer Engineering, University of Oklahoma, Norman, OK 73019 (United States)

2016-01-21

The spatial resolution characteristics of an in vivo microcomputed tomography (CT) system was investigated in the in-plane (x–y), cross plane (z) and projection imaging modes. The microCT system utilized in this study employs a flat panel detector with a 127 µm pixel pitch, a microfocus x-ray tube with a focal spot size ranging from 5–30 µm, and accommodates three geometric magnifications (M) of 1.72, 2.54 and 5.10. The in-plane modulation transfer function (MTF) curves were measured as a function of the number of projections, geometric magnification (M), detector binning and reconstruction magnification (M{sub Recon}). The in plane cutoff frequency (10% MTF) ranged from 2.31 lp/mm (M=1.72, 2×2 binning) to 12.56 lp/mm (M=5.10, 1×1 binning) and a bar pattern phantom validated those measurements. A slight degradation in the spatial resolution was observed when comparing the image reconstruction with 511 and 918 projections, whose effect was visible at the lower frequencies. Small value of M{sub Recon} has little or no impact on the in-plane spatial resolution owning to a stable system. Large value of M{sub Recon} has implications on the spatial resolution and it was evident when comparing the bar pattern images reconstructed with M{sub Recon}=1.25 and 2.5. The cross plane MTF curves showed that the spatial resolution increased as the slice thickness decreased. The cutoff frequencies in the projection imaging mode yielded slightly higher values as compared to the in-plane and cross plane modes at all the geometric magnifications (M). At M=5.10, the cutoff resolution of the projection and cross plane on an ultra-high contrast resolution bar chip phantom were 14.9 lp/mm and 13–13.5 lp/mm. Due to the finite focal spot size of the x-ray tube, the detector blur and the reconstruction kernel functions, the system's spatial resolution does not reach the limiting spatial resolution as defined by the Nyquist's detector criteria with an ideal point source

Spatial cognition in apes and humans.

Science.gov (United States)

Gentner, Dedre

2007-05-01

The debate on whether language influences cognition is sometimes seen as a simple dichotomy: cognitive development is governed either by innate predispositions or by influences of language and culture. In two recent papers on spatial cognition, Haun and colleagues break new ground in bringing together a comparative cognition approach with a cross-linguistic framework to arrive at a third position: that humans begin with the same spatial reference frames as our near relatives, the great apes, and diverge later owing to the influence of language and culture.

Atomic-fluorescence spectrophotometry

International Nuclear Information System (INIS)

Bakhturova, N.F.; Yudelevich, I.G.

1975-01-01

Atomic-fluorescence spectrophotometry, a comparatively new method for the analysis of trace quantities, has developed rapidly in the past ten years. Theoretical and experimental studies by many workers have shown that atomic-fluorescence spectrophotometry (AFS) is capable of achieving a better limit than atomic absorption for a large number of elements. The present review examines briefly the principles of atomic-fluorescence spectrophotometry and the types of fluorescent transition. The excitation sources, flame and nonflame atomizers, used in AFS are described. The limits of detection achieved up to the present, using flame and nonflame methods of atomization are given

A cross-sectional ecological study of spatial scale and geographic inequality in access to drinking-water and sanitation.

Science.gov (United States)

Yu, Weiyu; Bain, Robert E S; Mansour, Shawky; Wright, Jim A

2014-11-26

Measuring inequality in access to safe drinking-water and sanitation is proposed as a component of international monitoring following the expiry of the Millennium Development Goals. This study aims to evaluate the utility of census data in measuring geographic inequality in access to drinking-water and sanitation. Spatially referenced census data were acquired for Colombia, South Africa, Egypt, and Uganda, whilst non-spatially referenced census data were acquired for Kenya. Four variants of the dissimilarity index were used to estimate geographic inequality in access to both services using large and small area units in each country through a cross-sectional, ecological study. Inequality was greatest for piped water in South Africa in 2001 (based on 53 areas (N) with a median population (MP) of 657,015; D = 0.5599) and lowest for access to an improved water source in Uganda in 2008 (N = 56; MP = 419,399; D = 0.2801). For sanitation, inequality was greatest for those lacking any facility in Kenya in 2009 (N = 158; MP = 216,992; D = 0.6981), and lowest for access to an improved facility in Uganda in 2002 (N = 56; MP = 341,954; D = 0.3403). Although dissimilarity index values were greater for smaller areal units, when study countries were ranked in terms of inequality, these ranks remained unaffected by the choice of large or small areal units. International comparability was limited due to definitional and temporal differences between censuses. This five-country study suggests that patterns of inequality for broad regional units do often reflect inequality in service access at a more local scale. This implies household surveys designed to estimate province-level service coverage can provide valuable insights into geographic inequality at lower levels. In comparison with household surveys, censuses facilitate inequality assessment at different spatial scales, but pose challenges in harmonising water and sanitation typologies across countries.

Detection of SiO2 nanoparticles in lung tissue by ToF-SIMS imaging and fluorescence microscopy.

Science.gov (United States)

Veith, Lothar; Vennemann, Antje; Breitenstein, Daniel; Engelhard, Carsten; Wiemann, Martin; Hagenhoff, Birgit

2017-07-10

The direct detection of nanoparticles in tissues at high spatial resolution is a current goal in nanotoxicology. Time-of-Flight Secondary Ion Mass Spectrometry (ToF-SIMS) is widely used for the direct detection of inorganic and organic substances with high spatial resolution but its capability to detect nanoparticles in tissue sections is still insufficiently explored. To estimate the applicability of this technique for nanotoxicological questions, comparative studies with established techniques on the detection of nanoparticles can offer additional insights. Here, we compare ToF-SIMS imaging data with sub-micrometer spatial resolution to fluorescence microscopy imaging data to explore the usefulness of ToF-SIMS for the detection of nanoparticles in tissues. SiO 2 nanoparticles with a mean diameter of 25 nm, core-labelled with fluorescein isothiocyanate, were intratracheally instilled into rat lungs. Subsequently, imaging of lung cryosections was performed with ToF-SIMS and fluorescence microscopy. Nanoparticles were successfully detected with ToF-SIMS in 3D microanalysis mode based on the lateral distribution of SiO 3 - (m/z 75.96), which was co-localized with the distribution pattern that was obtained from nanoparticle fluorescence. In addition, the lateral distribution of protein (CN - , m/z 26.00) and phosphate based signals (PO 3 - , m/z 78.96) originating from the tissue material could be related to the SiO 3 - lateral distribution. In conclusion, ToF-SIMS is suitable to directly detect and laterally resolve SiO 2 nanomaterials in biological tissue at sufficient intensity levels. At the same time, information about the chemical environment of the nanoparticles in the lung tissue sections is obtained.

Development of a wide-field fluorescence imaging system for evaluation of wound re-epithelialization

Science.gov (United States)

Franco, Walfre; Gutierrez-Herrera, Enoch; Purschke, Martin; Wang, Ying; Tam, Josh; Anderson, R. Rox; Doukas, Apostolos

2013-03-01

Normal skin barrier function depends on having a viable epidermis, an epithelial layer formed by keratinocytes. The transparent epidermis, which is less than a 100 mum thick, is nearly impossible to see. Thus, the clinical evaluation of re-epithelialization is difficult, which hinders selecting appropriate therapy for promoting wound healing. An imaging system was developed to evaluate epithelialization by detecting endogenous fluorescence emissions of cellular proliferation over a wide field of view. A custom-made 295 nm ultraviolet (UV) light source was used for excitation. Detection was done by integrating a near-UV camera with sensitivity down to 300 nm, a 12 mm quartz lens with iris and focus lock for the UV regime, and a fluorescence bandpass filter with 340 nm center wavelength. To demonstrate that changes in fluorescence are related to cellular processes, the epithelialization of a skin substitute was monitored in vitro. The skin substitute or construct was made by embedding microscopic live human skin tissue columns, 1 mm in diameter and spaced 1 mm apart, in acellular porcine dermis. Fluorescence emissions clearly delineate the extent of lateral surface migration of keratinocytes and the total surface covered by the new epithelium. The fluorescence image of new epidermis spatially correlates with the corresponding color image. A simple, user-friendly way of imaging the presence of skin epithelium would improve wound care in civilian burns, ulcers and surgeries.

Chromenoquinoline-based thiol probes: a study on the quencher position for controlling fluorescent Off-On characteristics.

Science.gov (United States)

Kand, Dnyaneshwar; Kalle, Arunasree Marasanapalli; Talukdar, Pinaki

2013-02-13

The design, synthesis and thiol sensing ability of chromenoquinoline-based fluorescent probes 4, 5 and 6 and are reported here. The relative position of the maleimide moiety was varied along the chromenoquinoline fluorophore to decrease the background fluorescence. Lower background fluorescence in probes 4 and 6 was rationalized by the smaller k(r)/k(nr) values compared to that of probe 5. An intramolecular charge transfer (ICT) mechanism was proposed for quenching and the extent was dependent on the position of the maleimide quencher. Fluorescent Off-On characteristics were evaluated by theoretical calculations. All probes were selective only towards thiol containing amino acids. Thiol sensing by probes 4 and 6 were much better compared to 5. Probe 4 displayed a better fluorescence response for less hindered thiol (185-, 223- and 156-fold for Hcy, Cys and GSH, respectively), while for probe 6, a higher enhancement in fluorescence was observed with more hindered thiols (180-, 205- and 245-fold for Hcy, Cys and GSH, respectively). The better response to bulkier thiol, GSH by probe 6 was attributed to the steric crowding at the C-4 position and bulkiness of the GSH group which force the succinimide unit to be in a nearly orthogonal conformation. This spatial arrangement was important in reducing the fluorescence quenching ability of the succinimide moiety. The application of probes 4, 5 and 6 was demonstrated by naked eye detection thiols using a 96-well plate system as well as by live-cell imaging.

Fluorescence excitation studies of molecular photoionization in external electric fields

International Nuclear Information System (INIS)

Poliakoff, E.D.; Dehmer, J.L.; Parr, A.C.; Leroi, G.E.

1985-01-01

Using molecular nitrogen as an example, we show that fluorescence excitation spectroscopy can be used to measure partial photoionization cross sections of free molecules in external electric fields. The production of the N 2 + (B 2 Σ/sub u/ + ) state was studied and the threshold for this process was found to shift linearly with the square root of the applied field. This behavior is compared with the hydrogenic case and with previously studied systems

Activated barrier crossing dynamics in the non-radiative decay of NADH and NADPH

Energy Technology Data Exchange (ETDEWEB)

Blacker, Thomas S., E-mail: t.blacker@ucl.ac.uk [Centre for Mathematics and Physics in the Life Sciences and Experimental Biology (CoMPLEX), University College London, London WC1E 6BT (United Kingdom); Department of Physics and Astronomy, University College London, London WC1E 6BT (United Kingdom); Research Department of Cell and Developmental Biology, University College London, London WC1E 6BT (United Kingdom); Marsh, Richard J., E-mail: richard.marsh@ucl.ac.uk [Department of Physics and Astronomy, University College London, London WC1E 6BT (United Kingdom); Duchen, Michael R., E-mail: m.duchen@ucl.ac.uk [Research Department of Cell and Developmental Biology, University College London, London WC1E 6BT (United Kingdom); Bain, Angus J., E-mail: a.bain@ucl.ac.uk [Centre for Mathematics and Physics in the Life Sciences and Experimental Biology (CoMPLEX), University College London, London WC1E 6BT (United Kingdom); Department of Physics and Astronomy, University College London, London WC1E 6BT (United Kingdom)

2013-08-30

Highlights: ► NADH and NADPH have a high rate of non-radiative excited state decay. ► Conformational relaxation is shown to be a significant non-radiative pathway. ► The Kramers equation describes the barrier crossing dynamics of the relaxation. ► Conformational restriction upon enzyme binding will alter NAD(P)H lifetimes. - Abstract: In live tissue, alterations in metabolism induce changes in the fluorescence decay of the biological coenzyme NAD(P)H, the mechanism of which is not well understood. In this work, the fluorescence and anisotropy decay dynamics of NADH and NADPH were investigated as a function of viscosity in a range of water–glycerol solutions. The viscosity dependence of the non-radiative decay is well described by Kramers and Kramers–Hubbard models of activated barrier crossing over a wide viscosity range. Our combined lifetime and anisotropy analysis indicates common mechanisms of non-radiative relaxation in the two emitting states (conformations) of both molecules. The low frequencies associated with barrier crossing suggest that non-radiative decay is mediated by small scale motion (e.g. puckering) of the nicotinamide ring. Variations in the fluorescence lifetimes of NADH and NADPH when bound to different enzymes may therefore be attributed to differing levels of conformational restriction upon binding.