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Sample records for determine fluorescence lifetimes

  1. Three-dimensional fluorescence lifetime tomography

    International Nuclear Information System (INIS)

    Godavarty, Anuradha; Sevick-Muraca, Eva M.; Eppstein, Margaret J.

    2005-01-01

    Near-infrared fluorescence tomography using molecularly targeted lifetime-sensitive, fluorescent contrast agents have applications for early-stage cancer diagnostics. Yet, although the measurement of fluorescent lifetime imaging microscopy (FLIM) is extensively used in microscopy and spectroscopy applications, demonstration of fluorescence lifetime tomography for medical imaging is limited to two-dimensional studies. Herein, the feasibility of three-dimensional fluorescence-lifetime tomography on clinically relevant phantom volumes is established, using (i) a gain-modulated intensified charge coupled device (CCD) and modulated laser diode imaging system, (ii) two fluorescent contrast agents, e.g., Indocyanine green and 3-3'-Diethylthiatricarbocyanine iodide differing in their fluorescence lifetime by 0.62 ns, and (iii) a two stage approximate extended Kalman filter reconstruction algorithm. Fluorescence measurements of phase and amplitude were acquired on the phantom surface under different target to background fluorescence absorption (70:1, 100:1) and fluorescence lifetime (1:1, 2.1:1) contrasts at target depths of 1.4-2 cm. The Bayesian tomography algorithm was employed to obtain three-dimensional images of lifetime and absorption owing to the fluorophores

  2. Refractive index sensing using Fluorescence Lifetime Imaging (FLIM)

    International Nuclear Information System (INIS)

    Jones, Carolyn; Suhling, Klaus

    2006-01-01

    The fluorescence lifetime is a function of the refractive index of the fluorophore's environment, for example in the case of the biologically important green fluorescent protein (GFP). In order to address the question whether this effect can be exploited to image the local environment of specific proteins in cell biology, we need to determine the distance over which the fluorophore's lifetime is sensitive to the refractive index. To this end, we employ Fluorescence Lifetime Imaging (FLIM) of fluorescein in NaOH buffer at an interface. This approach allows us to map the fluorescence lifetime as a function of distance from a buffer/air and buffer/oil interface. Preliminary data show that the fluorescence lifetime of fluorescein increases near a buffer/air interface and decreases near a buffer/oil interface. The range over which this fluorescence lifetime change occurs is found to be of the order several μm which is consistent with a theoretical model based on the full width at half maximum of the emission spectrum proposed by Toptygin

  3. Fluorescence lifetime imaging of skin cancer

    Science.gov (United States)

    Patalay, Rakesh; Talbot, Clifford; Munro, Ian; Breunig, Hans Georg; König, Karsten; Alexandrov, Yuri; Warren, Sean; Neil, Mark A. A.; French, Paul M. W.; Chu, Anthony; Stamp, Gordon W.; Dunsby, Chris

    2011-03-01

    Fluorescence intensity imaging and fluorescence lifetime imaging microscopy (FLIM) using two photon microscopy (TPM) have been used to study tissue autofluorescence in ex vivo skin cancer samples. A commercially available system (DermaInspect®) was modified to collect fluorescence intensity and lifetimes in two spectral channels using time correlated single photon counting and depth-resolved steady state measurements of the fluorescence emission spectrum. Uniquely, image segmentation has been used to allow fluorescence lifetimes to be calculated for each cell. An analysis of lifetime values obtained from a range of pigmented and non-pigmented lesions will be presented.

  4. Quantitative analysis of fluorescence lifetime measurements of the macula using the fluorescence lifetime imaging ophthalmoscope in healthy subjects.

    Science.gov (United States)

    Dysli, Chantal; Quellec, Gwénolé; Abegg, Mathias; Menke, Marcel N; Wolf-Schnurrbusch, Ute; Kowal, Jens; Blatz, Johannes; La Schiazza, Olivier; Leichtle, Alexander B; Wolf, Sebastian; Zinkernagel, Martin S

    2014-04-03

    Fundus autofluorescence (FAF) cannot only be characterized by the intensity or the emission spectrum, but also by its lifetime. As the lifetime of a fluorescent molecule is sensitive to its local microenvironment, this technique may provide more information than fundus autofluorescence imaging. We report here the characteristics and repeatability of FAF lifetime measurements of the human macula using a new fluorescence lifetime imaging ophthalmoscope (FLIO). A total of 31 healthy phakic subjects were included in this study with an age range from 22 to 61 years. For image acquisition, a fluorescence lifetime ophthalmoscope based on a Heidelberg Engineering Spectralis system was used. Fluorescence lifetime maps of the retina were recorded in a short- (498-560 nm) and a long- (560-720 nm) spectral channel. For quantification of fluorescence lifetimes a standard ETDRS grid was used. Mean fluorescence lifetimes were shortest in the fovea, with 208 picoseconds for the short-spectral channel and 239 picoseconds for the long-spectral channel, respectively. Fluorescence lifetimes increased from the central area to the outer ring of the ETDRS grid. The test-retest reliability of FLIO was very high for all ETDRS areas (Spearman's ρ = 0.80 for the short- and 0.97 for the long-spectral channel, P macula in healthy subjects. By using a custom-built software, we were able to quantify fluorescence lifetimes within the ETDRS grid. Establishing a clinically accessible standard against which to measure FAF lifetimes within the retina is a prerequisite for future studies in retinal disease.

  5. Azadioxatriangulenium: exploring the effect of a 20 ns fluorescence lifetime in fluorescence anisotropy measurements

    Science.gov (United States)

    Bogh, Sidsel A.; Bora, Ilkay; Rosenberg, Martin; Thyrhaug, Erling; Laursen, Bo W.; Just Sørensen, Thomas

    2015-12-01

    Azaoxatriangulenium (ADOTA) has been shown to be highly emissive despite a moderate molar absorption coefficient of the primary electronic transition. As a result, the fluorescence lifetime is ~20 ns, longer than all commonly used red fluorescent organic probes. The electronic transitions in ADOTA are highly polarised (r 0  =  0.38), which in combination with the long fluorescence lifetime extents the size-range of biomolecular weights that can be detected in fluorescence polarisation-based experiments. Here, the rotational dynamics of bovine serum albumin (BSA) are monitored with three different ADOTA derivatives, differing only in constitution of the reactive linker. A detailed study of the degree of labelling, the steady-state anisotropy, and the time-resolved anisotropy of the three different ADOTA-BSA conjugates are reported. The fluorescence quantum yields (ϕ fl) of the free dyes in PBS solution are determined to be ~55%, which is reduced to ~20% in the ADOTA-BSA conjugates. Despite the reduction in ϕ fl, a ~20 ns intensity averaged lifetime is maintained, allowing for the rotational dynamics of BSA to be monitored for up to 100 ns. Thus, ADOTA can be used in fluorescence polarisation assays to fill the gap between commonly used organic dyes and the long luminescence lifetime transition metal complexes. This allows for efficient steady-state fluorescence polarisation assays for detecting binding of analytes with molecular weights of up to 100 kDa.

  6. Measuring and sorting cell populations expressing isospectral fluorescent proteins with different fluorescence lifetimes.

    Directory of Open Access Journals (Sweden)

    Bryan Sands

    Full Text Available Study of signal transduction in live cells benefits from the ability to visualize and quantify light emitted by fluorescent proteins (XFPs fused to different signaling proteins. However, because cell signaling proteins are often present in small numbers, and because the XFPs themselves are poor fluorophores, the amount of emitted light, and the observable signal in these studies, is often small. An XFP's fluorescence lifetime contains additional information about the immediate environment of the fluorophore that can augment the information from its weak light signal. Here, we constructed and expressed in Saccharomyces cerevisiae variants of Teal Fluorescent Protein (TFP and Citrine that were isospectral but had shorter fluorescence lifetimes, ∼ 1.5 ns vs ∼ 3 ns. We modified microscopic and flow cytometric instruments to measure fluorescence lifetimes in live cells. We developed digital hardware and a measure of lifetime called a "pseudophasor" that we could compute quickly enough to permit sorting by lifetime in flow. We used these abilities to sort mixtures of cells expressing TFP and the short-lifetime TFP variant into subpopulations that were respectively 97% and 94% pure. This work demonstrates the feasibility of using information about fluorescence lifetime to help quantify cell signaling in living cells at the high throughput provided by flow cytometry. Moreover, it demonstrates the feasibility of isolating and recovering subpopulations of cells with different XFP lifetimes for subsequent experimentation.

  7. Fluorescence lifetime imaging of oxygen in dental biofilm

    Science.gov (United States)

    Gerritsen, Hans C.; de Grauw, Cees J.

    2000-12-01

    Dental biofilm consists of micro-colonies of bacteria embedded in a matrix of polysaccharides and salivary proteins. pH and oxygen concentration are of great importance in dental biofilm. Both can be measured using fluorescence techniques. The imaging of dental biofilm is complicated by the thickness of the biofilms that can be up to several hundred micrometers thick. Here, we employed a combination of two-photon excitation microscopy with fluorescence lifetime imaging to quantify the oxygen concentration in dental biofilm. Collisional quenching of fluorescent probes by molecular oxygen leads to a reduction of the fluorescence lifetime of the probe. We employed this mechanism to measure the oxygen concentration distribution in dental biofilm by means of fluorescence lifetime imaging. Here, TRIS Ruthenium chloride hydrate was used as an oxygen probe. A calibration procedure on buffers was use to measure the lifetime response of this Ruthenium probe. The results are in agreement with the Stern-Volmer equation. A linear relation was found between the ratio of the unquenched and the quenched lifetime and the oxygen concentration. The biofilm fluorescence lifetime imaging results show a strong oxygen gradient at the buffer - biofilm interface and the average oxygen concentration in the biofilm amounted to 50 μM.

  8. 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.

  9. Modulated electron-multiplied fluorescence lifetime imaging microscope: all-solid-state camera for fluorescence lifetime imaging.

    Science.gov (United States)

    Zhao, Qiaole; Schelen, Ben; Schouten, Raymond; van den Oever, Rein; Leenen, René; van Kuijk, Harry; Peters, Inge; Polderdijk, Frank; Bosiers, Jan; Raspe, Marcel; Jalink, Kees; Geert Sander de Jong, Jan; van Geest, Bert; Stoop, Karel; Young, Ian Ted

    2012-12-01

    We have built an all-solid-state camera that is directly modulated at the pixel level for frequency-domain fluorescence lifetime imaging microscopy (FLIM) measurements. This novel camera eliminates the need for an image intensifier through the use of an application-specific charge coupled device design in a frequency-domain FLIM system. The first stage of evaluation for the camera has been carried out. Camera characteristics such as noise distribution, dark current influence, camera gain, sampling density, sensitivity, linearity of photometric response, and optical transfer function have been studied through experiments. We are able to do lifetime measurement using our modulated, electron-multiplied fluorescence lifetime imaging microscope (MEM-FLIM) camera for various objects, e.g., fluorescein solution, fixed green fluorescent protein (GFP) cells, and GFP-actin stained live cells. A detailed comparison of a conventional microchannel plate (MCP)-based FLIM system and the MEM-FLIM system is presented. The MEM-FLIM camera shows higher resolution and a better image quality. The MEM-FLIM camera provides a new opportunity for performing frequency-domain FLIM.

  10. Fluorescence lifetime imaging using light emitting diodes

    Energy Technology Data Exchange (ETDEWEB)

    Kennedy, Gordon T; Munro, Ian; Poher, Vincent; French, Paul M W; Neil, Mark A A [Blackett Laboratory, Imperial College London, South Kensington Campus, London SW7 2AZ (United Kingdom); Elson, Daniel S [Institute of Biomedical Engineering, Imperial College London, South Kensington Campus, London SW7 2AZ (United Kingdom); Hares, Jonathan D [Kentech Instruments Ltd, Unit 9, Hall Farm Workshops, South Moreton, Didcot, Oxfordshire, OX11 9AG (United Kingdom)], E-mail: gordon.kennedy@imperial.ac.uk

    2008-05-07

    We demonstrate flexible use of low cost, high-power light emitting diodes as illumination sources for fluorescence lifetime imaging (FLIM). Both time-domain and frequency-domain techniques have been implemented at wavelengths spanning the range 450-640 nm. Additionally, we demonstrate optically sectioned fluorescence lifetime imaging by combining structured illumination with frequency-domain FLIM.

  11. Azadioxatriangulenium: a long fluorescence lifetime fluorophore for large biomolecule binding assay

    International Nuclear Information System (INIS)

    Sørensen, Thomas Just; Thyrhaug, Erling; Szabelski, Mariusz; Gryczynski, Ignacy; Gryczynski, Zygmunt; Luchowski, Rafal; Laursen, Bo W

    2013-01-01

    Of the many optical bioassays available, sensing by fluorescence anisotropy has great advantages as it provides a sensitive, instrumentally simple, ratiometric method of detection. However, it is hampered by a severe limitation, as the emission lifetime of the label needs to be comparable to the correlation lifetime (tumbling time) of the biomolecule which is labelled. For proteins of moderate size this is on the order of 20–200 ns, which due to practical issues currently limits the choice of labels to the dansyl-type dyes and certain aromatic dyes. These have the significant drawback of UV/blue absorption and emission as well as an often significant solvent sensitivity. Here, we report the synthesis and characterization of a new fluorescent label for high molecular weight biomolecule assay based on the azadioxatriangulenium motif. The NHS ester of the long fluorescence lifetime, red-emitting fluorophore: azadioxatriangulenium (ADOTA-NHS) was conjugated to anti-rabbit Immunoglobulin G (antiIgG). The long fluorescence lifetime was exploited to determine the correlation time of the high molecular weight antibody and its complex with rabbit Immunoglobulin G (IgG) with steady-state fluorescence anisotropy and time-resolved methods: solution phase immuno-assay was performed following either steady-state or time-resolved fluorescence anisotropy. By performing a variable temperature experiment it was determined that the binding of the ligand resulted in an increase in correlation time of more than 75%, and an increase in the steady-state anisotropy of 18%. The results show that the triangulenium class of dyes can be used in anisotropy assay to detect binding events involving biomolecules of far larger size than what is possible with most other red-emitting organic dyes. (paper)

  12. Azadioxatriangulenium: a long fluorescence lifetime fluorophore for large biomolecule binding assay

    Science.gov (United States)

    Just Sørensen, Thomas; Thyrhaug, Erling; Szabelski, Mariusz; Luchowski, Rafal; Gryczynski, Ignacy; Gryczynski, Zygmunt; Laursen, Bo W.

    2013-06-01

    Of the many optical bioassays available, sensing by fluorescence anisotropy has great advantages as it provides a sensitive, instrumentally simple, ratiometric method of detection. However, it is hampered by a severe limitation, as the emission lifetime of the label needs to be comparable to the correlation lifetime (tumbling time) of the biomolecule which is labelled. For proteins of moderate size this is on the order of 20-200 ns, which due to practical issues currently limits the choice of labels to the dansyl-type dyes and certain aromatic dyes. These have the significant drawback of UV/blue absorption and emission as well as an often significant solvent sensitivity. Here, we report the synthesis and characterization of a new fluorescent label for high molecular weight biomolecule assay based on the azadioxatriangulenium motif. The NHS ester of the long fluorescence lifetime, red-emitting fluorophore: azadioxatriangulenium (ADOTA-NHS) was conjugated to anti-rabbit Immunoglobulin G (antiIgG). The long fluorescence lifetime was exploited to determine the correlation time of the high molecular weight antibody and its complex with rabbit Immunoglobulin G (IgG) with steady-state fluorescence anisotropy and time-resolved methods: solution phase immuno-assay was performed following either steady-state or time-resolved fluorescence anisotropy. By performing a variable temperature experiment it was determined that the binding of the ligand resulted in an increase in correlation time of more than 75%, and an increase in the steady-state anisotropy of 18%. The results show that the triangulenium class of dyes can be used in anisotropy assay to detect binding events involving biomolecules of far larger size than what is possible with most other red-emitting organic dyes.

  13. 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.

  14. 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.

  15. 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.

  16. Clinical results of fluorescence lifetime imaging in ophthalmology

    Science.gov (United States)

    Schweitzer, D.; Quick, S.; Klemm, M.; Hammer, M.; Jentsch, S.; Dawczynski, J.; Becker, W.

    2009-07-01

    A laser scanner ophthalmoscope was developed for in vivo fluorescence lifetime measurements at the human retina. Measurements were performed in 30 degree fundus images. The fundus was excited by pulses of 75 ps (FWHM). The dynamic fluorescence was detected in two spectral channels K1(490-560nm), K2(560-700 nm) by time-correlated single photon counting. The decay of fluorescence was three-exponentially. Local and global alterations in lifetimes were found between healthy subjects and patients suffering from age-related macular degeneration, diabetic retinopathy, and vessel occlusion. The lifetimes T1, T2, and T3 in both channels are changed to longer values in AMD and diabetic retinopathy in comparison with healthy subjects. The lifetime T2 in K1 is most sensitive to metabolic alterations in branch arterial vessel occlusion.

  17. Generation of a new spectral format, the lifetime synchronous spectrum (LiSS), using phase-resolved fluorescence spectroscopy

    International Nuclear Information System (INIS)

    Shaver, J.M.; McGown, L.B.

    1994-01-01

    A new fluorescence spectral format is introduced in which fluorescence lifetime is shown as a function of synchronously scanned wavelength to generate a Lifetime Synchronous Spectrum (LiSS). Lifetimes are determined in the frequency domain with the use of Phase-Resolved Fluorescence Spectroscopy (PRFS) to obtain the phase of the fluorescence signal. Theory and construction of the LiSS are presented and experimental results are shown for solutions of single components and simple binary and ternary mixtures. These results show how the lifetime information in the LiSS augments the steady-state intensity information of a standard synchronous spectrum, providing unique information for identification of components and resolution of overlapping spectral peaks. The LiSS technique takes advantage of noise reduction inherent in the extraction of lifetime from PRFS in addition to standard spectral smoothing techniques. The precision of phase determination through PRFS is found to be comparable to that of direct phase measurements at normal fluorescence intensities and superior for low-intensity signals

  18. Fluorescence lifetime measurements to determine the core-shell nanostructure of FITC-doped silica nanoparticles: An optical approach to evaluate nanoparticle photostability

    International Nuclear Information System (INIS)

    Santra, Swadeshmukul; Liesenfeld, Bernd; Bertolino, Chiara; Dutta, Debamitra; Cao Zehui; Tan Weihong; Moudgil, Brij M.; Mericle, Robert A.

    2006-01-01

    In this paper, we described a novel fluorescence lifetime-based approach to determine the core-shell nanostructure of FITC-(fluorescein isothiocyanate, isomer I) doped fluorescent silica nanoparticles (FSNPs). Because of phase homogeneity between the core and the shell, electron microscopic technique could not be used to characterize such core-shell nanostructure. Our optical approach not only revealed the core-shell nanostructure of FSNPs but also evaluated photobleaching of FSNPs both in the solvated and non-solvated (dry) states. The FSNPs were produced via Stoeber's method by hydrolysis and co-condensation reaction of tetraethylorthosilicate (TEOS) and fluorescein linked (3-aminopropyl)triethoxysilane (FITC-APTS conjugate) in the presence of ammonium hydroxide catalyst. To obtain a pure silica surface coating, FSNPs were then post-coated with TEOS. The average particle size was 135 nm as determined by TEM (transmission electron microscope) measurements. Fluorescence excitation and emission spectral data demonstrated successful doping of FITC dye molecules in FSNPs. Fluorescence lifetime data revealed that approximately 62% of dye molecules remained in the solvated silica shell, while 38% of dye molecules remained in the non-solvated (dry) silica core. Photobleaching experiments of FSNPs were conducted both in DI water (solution state) and in air (dry state). Severe photobleaching of FSNPs was observed in air. However, FSNPs were moderately photostable in the solution state. Photostability of FSNPs in both solution and dry states was explained on the basis of fluorescence lifetime data

  19. 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.

  20. Fluorescence lifetime imaging microscopy using near-infrared contrast agents.

    Science.gov (United States)

    Nothdurft, R; Sarder, P; Bloch, S; Culver, J; Achilefu, S

    2012-08-01

    Although single-photon fluorescence lifetime imaging microscopy (FLIM) is widely used to image molecular processes using a wide range of excitation wavelengths, the captured emission of this technique is confined to the visible spectrum. Here, we explore the feasibility of utilizing near-infrared (NIR) fluorescent molecular probes with emission >700 nm for FLIM of live cells. The confocal microscope is equipped with a 785 nm laser diode, a red-enhanced photomultiplier tube, and a time-correlated single photon counting card. We demonstrate that our system reports the lifetime distributions of NIR fluorescent dyes, cypate and DTTCI, in cells. In cells labelled separately or jointly with these dyes, NIR FLIM successfully distinguishes their lifetimes, providing a method to sort different cell populations. In addition, lifetime distributions of cells co-incubated with these dyes allow estimate of the dyes' relative concentrations in complex cellular microenvironments. With the heightened interest in fluorescence lifetime-based small animal imaging using NIR fluorophores, this technique further serves as a bridge between in vitro spectroscopic characterization of new fluorophore lifetimes and in vivo tissue imaging. © 2012 The Author Journal of Microscopy © 2012 Royal Microscopical Society.

  1. 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.

  2. Fluorescence Lifetime Imaging in Stargardt Disease: Potential Marker for Disease Progression

    OpenAIRE

    Dysli Chantal; Wolf Sebastian; Hatz Katja; Zinkernagel Martin

    2016-01-01

    PURPOSE The purpose of this study was to describe autofluorescence lifetime characteristics in Stargardt disease (STGD) using fluorescence lifetime imaging ophthalmoscopy (FLIO) and to investigate potential prognostic markers for disease activity and progression. METHODS Fluorescence lifetime data of 16 patients with STGD (mean age, 40 years; range, 22-56 years) and 15 age-matched controls were acquired using a fluorescence lifetime imaging ophthalmoscope based on a Heidelberg Eng...

  3. Accurate Rapid Lifetime Determination on Time-Gated FLIM Microscopy with Optical Sectioning.

    Science.gov (United States)

    Silva, Susana F; Domingues, José Paulo; Morgado, António Miguel

    2018-01-01

    Time-gated fluorescence lifetime imaging microscopy (FLIM) is a powerful technique to assess the biochemistry of cells and tissues. When applied to living thick samples, it is hampered by the lack of optical sectioning and the need of acquiring many images for an accurate measurement of fluorescence lifetimes. Here, we report on the use of processing techniques to overcome these limitations, minimizing the acquisition time, while providing optical sectioning. We evaluated the application of the HiLo and the rapid lifetime determination (RLD) techniques for accurate measurement of fluorescence lifetimes with optical sectioning. HiLo provides optical sectioning by combining the high-frequency content from a standard image, obtained with uniform illumination, with the low-frequency content of a second image, acquired using structured illumination. Our results show that HiLo produces optical sectioning on thick samples without degrading the accuracy of the measured lifetimes. We also show that instrument response function (IRF) deconvolution can be applied with the RLD technique on HiLo images, improving greatly the accuracy of the measured lifetimes. These results open the possibility of using the RLD technique with pulsed diode laser sources to determine accurately fluorescence lifetimes in the subnanosecond range on thick multilayer samples, providing that offline processing is allowed.

  4. Theoretical lifetimes and fluorescence yields for multiply-ionized fluorine

    International Nuclear Information System (INIS)

    Tunnell, T.W.; Can, C.; Bhalla, C.P.

    1978-01-01

    Theoretical lifetimes and multiplet partial fluorescence yields for various fluorine ions with a single K-shell vacancy were calculated. For few-electron systems, the lifetimes and line fluorescence yields were computed in the intermediate coupling scheme with the inclusion of the effects arising from configuration interactions. 6 references

  5. Azadioxatriangulenium (ADOTA+): A long fluorescence lifetime fluorophore for large biomolecule binding assay

    Science.gov (United States)

    Sørensen, Thomas Just; Thyrhaug, Erling; Szabelski, Mariusz; Luchowski, Rafal; Gryczynski, Ignacy; Gryczynski, Zygmunt; Laursen, Bo W.

    2013-01-01

    Of the many optical bioassays available, sensing by fluorescence anisotropy have great advantages as it provides a sensitive, instrumentally simple, ratiometric method of detection. However, it is hampered by a severe limitation as the emission lifetime of the label needs to be comparable to the correlation lifetime (tumbling time) of the biomolecule which is labelled. For proteins of moderate size this is in the order of 20–200 ns, which due to practical issues currently limits the choice of labels to the dansyl-type dyes and certain aromatics dyes. These have the significant drawback of UV/blue absorption and emission as well as an often significant solvent sensitivity. Here, we report the synthesis and characterization of a new fluorescent label for high molecular weight biomolecules assay based on the azadioxatriangulenium motif. The NHS ester of the long fluorescence lifetime, red emitting fluorophore: azadioxatriangulenium (ADOTA-NHS) was conjugated to anti-rabbit Immunoglobulin G (antiIgG). The long fluorescence lifetime was exploited to determine the correlation time of the high molecular weight antibody and its complex with rabbit Immuniglobulin G (IgG) with steady-state fluorescence anisotropy and time-resolved methods: solution phase immuno-assay was performed following either steady-state or time-resolved fluorescence anisotropy. By performing a variable temperature experiment it was determined that the binding of the ligand resulted in an increase in correlation time by more than 75 %, and a change in the steady-state anisotropy increase of 18%. The results show that the triangulenium class of dyes can be used in anisotropy assay for detecting binding events involving biomolecules of far larger size than what is possible with the other red emitting organic dyes. PMID:24058730

  6. Fluorescence lifetime evaluation of whole soils from the Amazon rainforest.

    Science.gov (United States)

    Nicolodelli, Gustavo; Tadini, Amanda Maria; Nogueira, Marcelo Saito; Pratavieira, Sebastião; Mounier, Stephane; Huaman, Jose Luis Clabel; Dos Santos, Cléber Hilário; Montes, Célia Regina; Milori, Débora Marcondes Bastos Pereira

    2017-08-20

    Time-resolved fluorescence spectroscopy (TRFS) is a new tool that can be used to investigate processes of interaction between metal ions and organic matter (OM) in soils, providing a specific analysis of the structure and dynamics of macromolecules. To the best of our knowledge, there are no studies in the literature reporting the use of this technique applied to whole/non-fractionated soil samples, making it a potential method for use in future studies. This work describes the use of TRFS to evaluate the fluorescence lifetimes of OM of whole soils from the Amazon region. Analysis was made of pellets of soils from an oxisol-spodosol system, collected in São Gabriel da Cachoeira (Amazonas, Brazil). The fluorescence lifetimes in the oxisol-spodosol system were attributed to two different fluorophores. One was related to complexation of an OM fraction with metals, resulting in a shorter fluorophore lifetime. A short fluorescence lifetime (2-12 ns) could be associated with simpler structures of the OM, while a long lifetime (19-66 ns) was associated with more complex OM structures. This new TRFS technique for analysis of the fluorescence lifetime in whole soil samples complies with the principles of green chemistry.

  7. Time variation of fluorescence lifetime in enhanced cyan fluorescence protein

    International Nuclear Information System (INIS)

    Lee, Soonhyouk; Kim, Soo Yong; Park, Kyoungsook; Jeong, Jinyoung; Chung, Bong Hyun; Kim, Sok Won

    2010-01-01

    The lifetime variations of enhanced cyan fluorescence protein (ECFP) in relatively short integration time bins were studied via time-correlated single photon counting (TCSPC) measurement. We observed that minimum photon counts are necessary for the lifetime estimation to achieve a certain range of variance. The conditions to decrease the variance of lifetime were investigated and the channel width of the measurement of TCSPC data was found to be another important factor for the variance of lifetime. Though the lifetime of ECFP is best fit by a double exponential, a mono exponential fit for the same integration time is more stable. The results may be useful in the analysis of photophysical dynamics for ensemble molecules in short measurement time windows.

  8. Measurements of fluorescence lifetime of group III metalo-8-quinolinolates and their use in analytical chemistry

    Energy Technology Data Exchange (ETDEWEB)

    Nishikawa, Y; Hiraki, K; Morishige, K; Takahashi, K [Kinki Univ., Higashi-Osaka, Osaka (Japan). Faculty of Science and Technology; Shigematsu, T

    1976-07-01

    8-Quinolinolates of aluminum, gallium, and indium in chloroform exhibit strong yellowish green fluorescence with an emission maximum at 510, 526, and 528 nm, respectively. The time resolved fluorescence spectra and the fluorescence lifetime properties of these chelates were measured with a time-resolved spectrofluorometer. The fluorescence intensity of these chelates decays exponentially with time t, and obeys the following equation: F=F/sub 0/e-t/tau=F/sub 0/e-k sub(f).t where F/sub 0/ and F are the fluorescence intensity when the exciting light is irradiating and shut off, respectively; tau and k sub(f) being the lifetime and the rate constant for the process of fluorescence emission. The lifetimes of these chelates in chloroform solution at the ordinary temperature were 17.8, 10.1, and 8.4 ns for Al(C/sub 9/H/sub 6/ON)/sub 3/, Ga(C/sub 9/H/sub 6/ON)/sub 3/, and In(C/sub 9/H/sub 6/ON)/sub 3/, respectively. Thus, 8-quinolinolates of group III metals emit the same type radiation with different lifetimes. Between Al-chelate and In-chelate, there were significant difference in the lifetime by 9.4 ns. Then, the logarithmic plot of the composite fluorescence intensity against time is the overlap of some straight lines with different slopes which indicate k sub(f) of various decay processes. The linear portion of the logarithmic plot of the composite fluorescence intensity corresponded to the longer lifetime component (Al-chelate), and by substracting this component from the whole one, the straight line due to the shorter lifetime component (In-chelate) is obtained. Aluminum and indium contents were then determined by comparing the fluorescence intensity of the sample with that of the standard at a definite time (extrapolated to t=0). By using this composite decay curve, the composition of mixtures of nx10/sup -4/ mol/l of Al and In-chelates in chloroform could be determined.

  9. Ns-scaled time-gated fluorescence lifetime imaging for forensic document examination

    Science.gov (United States)

    Zhong, Xin; Wang, Xinwei; Zhou, Yan

    2018-01-01

    A method of ns-scaled time-gated fluorescence lifetime imaging (TFLI) is proposed to distinguish different fluorescent substances in forensic document examination. Compared with Video Spectral Comparator (VSC) which can examine fluorescence intensity images only, TFLI can detect questioned documents like falsification or alteration. TFLI system can enhance weak signal by accumulation method. The two fluorescence intensity images of the interval delay time tg are acquired by ICCD and fitted into fluorescence lifetime image. The lifetimes of fluorescence substances are represented by different colors, which make it easy to detect the fluorescent substances and the sequence of handwritings. It proves that TFLI is a powerful tool for forensic document examination. Furthermore, the advantages of TFLI system are ns-scaled precision preservation and powerful capture capability.

  10. Fluorophore:dendrimer ratio impacts cellular uptake and intracellular fluorescence lifetime.

    Science.gov (United States)

    Dougherty, Casey A; Vaidyanathan, Sriram; Orr, Bradford G; Banaszak Holl, Mark M

    2015-02-18

    G5-NH2-TAMRAn (n = 1-4, 5+, and 1.5(avg)) were prepared with n = 1-4 as a precise dye:dendrimer ratio, 5+ as a mixture of dendrimers with 5 or more dye per dendrimer, and 1.5(avg) as a Poisson distribution of dye:dendrimer ratios with a mean of 1.5 dye per dendrimer. The absorption intensity increased sublinearly with n whereas the fluorescence emission and lifetime decreased with an increasing number of dyes per dendrimer. Flow cytometry was employed to quantify uptake into HEK293A cells. Dendrimers with 2-4 dyes were found to have greater uptake than dendrimer with a single dye. Fluorescence lifetime imaging microscopy (FLIM) showed that the different dye:dendrimer ratio alone was sufficient to change the fluorescence lifetime of the material observed inside cells. We also observed that the lifetime of G5-NH2-TAMRA5+ increased when present in the cell as compared to solution. However, cells treated with G5-NH2-TAMRA1.5(avg) did not exhibit the high lifetime components present in G5-NH2-TAMRA1 and G5-NH2-TAMRA5+. In general, the effects of the dye:dendrimer ratio on fluorescence lifetime were of similar magnitude to environmentally induced lifetime shifts.

  11. Two-dimensional fluorescence lifetime correlation spectroscopy. 2. Application.

    Science.gov (United States)

    Ishii, Kunihiko; Tahara, Tahei

    2013-10-03

    In the preceding article, we introduced the theoretical framework of two-dimensional fluorescence lifetime correlation spectroscopy (2D FLCS). In this article, we report the experimental implementation of 2D FLCS. In this method, two-dimensional emission-delay correlation maps are constructed from the photon data obtained with the time-correlated single photon counting (TCSPC), and then they are converted to 2D lifetime correlation maps by the inverse Laplace transform. We develop a numerical method to realize reliable transformation, employing the maximum entropy method (MEM). We apply the developed actual 2D FLCS to two real systems, a dye mixture and a DNA hairpin. For the dye mixture, we show that 2D FLCS is experimentally feasible and that it can identify different species in an inhomogeneous sample without any prior knowledge. The application to the DNA hairpin demonstrates that 2D FLCS can disclose microsecond spontaneous dynamics of biological molecules in a visually comprehensible manner, through identifying species as unique lifetime distributions. A FRET pair is attached to the both ends of the DNA hairpin, and the different structures of the DNA hairpin are distinguished as different fluorescence lifetimes in 2D FLCS. By constructing the 2D correlation maps of the fluorescence lifetime of the FRET donor, the equilibrium dynamics between the open and the closed forms of the DNA hairpin is clearly observed as the appearance of the cross peaks between the corresponding fluorescence lifetimes. This equilibrium dynamics of the DNA hairpin is clearly separated from the acceptor-missing DNA that appears as an isolated diagonal peak in the 2D maps. The present study clearly shows that newly developed 2D FLCS can disclose spontaneous structural dynamics of biological molecules with microsecond time resolution.

  12. Fluorescence lifetime assays: current advances and applications in drug discovery.

    Science.gov (United States)

    Pritz, Stephan; Doering, Klaus; Woelcke, Julian; Hassiepen, Ulrich

    2011-06-01

    Fluorescence lifetime assays complement the portfolio of established assay formats available in drug discovery, particularly with the recent advances in microplate readers and the commercial availability of novel fluorescent labels. Fluorescence lifetime assists in lowering complexity of compound screening assays, affording a modular, toolbox-like approach to assay development and yielding robust homogeneous assays. To date, materials and procedures have been reported for biochemical assays on proteases, as well as on protein kinases and phosphatases. This article gives an overview of two assay families, distinguished by the origin of the fluorescence signal modulation. The pharmaceutical industry demands techniques with a robust, integrated compound profiling process and short turnaround times. Fluorescence lifetime assays have already helped the drug discovery field, in this sense, by enhancing productivity during the hit-to-lead and lead optimization phases. Future work will focus on covering other biochemical molecular modifications by investigating the detailed photo-physical mechanisms underlying the fluorescence signal.

  13. Fluorescence intensity and lifetime-based cyanide sensitive probes for physiological safeguard

    International Nuclear Information System (INIS)

    Badugu, Ramachandram; Lakowicz, Joseph R.; Geddes, Chris D.

    2004-01-01

    We characterize six new fluorescent probes that show both intensity and lifetime changes in the presence of free uncomplexed aqueous cyanide, allowing for fluorescence based cyanide sensing up to physiological safeguard levels, i.e. 2 to the anionic R-B - (CN) 3 form, a new cyanide binding mechanism which we have recently reported. The presence of an electron deficient quaternary heterocyclic nitrogen nucleus, and the electron rich cyanide bound form, provides for the intensity changes observed. We have determined the disassociation constants of the probes to be in the range ∼15-84 μM 3 . In addition we have synthesized control compounds which do not contain the boronic acid moiety, allowing for a rationale of the cyanide responses between the probe isomers to be made. The lifetime of the cyanide bound probes are significantly shorter than the free R-B(OH) 2 probe forms, providing for the opportunity of lifetime based cyanide sensing up to physiologically lethal levels. Finally, while fluorescent probes containing the boronic acid moiety have earned a well-deserved reputation for monosaccharide sensing, we show that strong bases such as CN - and OH - preferentially bind as compared to glucose, enabling the potential use of these probes for cyanide safeguard and determination in physiological fluids, especially given that physiologies do not experience any notable changes in pH

  14. Fluorescence lifetime measurement of radical ions

    International Nuclear Information System (INIS)

    Ichinose, Nobuyuki; Kinugasa, Jun-ichiro; Hagiri, Masahide; Nakayama, Toshihiro; Murakami, Hiroshi; Kishimoto, Maki; Daido, Hiroyuki

    2004-01-01

    One-photonic excitation of a charge transfer complex of hexamethoxybenzene (HMB) and nitrosonium tetrafluoroborate (NO + BF 4 - ) in acetonitrile afforded fluorescences emission from excited radical cation of HMB (HMB + *). Lifetime of the excited radical ion species was measured to be 7 ps by the pump-probe transient absorption technique. The lifetime was much shorter than that of free radical ion (63 ps), indicating the presence of an interaction between HMB + * and NO in the excited complex. (author)

  15. The enhanced cyan fluorescent protein: a sensitive pH sensor for fluorescence lifetime imaging.

    Science.gov (United States)

    Poëa-Guyon, Sandrine; Pasquier, Hélène; Mérola, Fabienne; Morel, Nicolas; Erard, Marie

    2013-05-01

    pH is an important parameter that affects many functions of live cells, from protein structure or function to several crucial steps of their metabolism. Genetically encoded pH sensors based on pH-sensitive fluorescent proteins have been developed and used to monitor the pH of intracellular compartments. The quantitative analysis of pH variations can be performed either by ratiometric or fluorescence lifetime detection. However, most available genetically encoded pH sensors are based on green and yellow fluorescent proteins and are not compatible with multicolor approaches. Taking advantage of the strong pH sensitivity of enhanced cyan fluorescent protein (ECFP), we demonstrate here its suitability as a sensitive pH sensor using fluorescence lifetime imaging. The intracellular ECFP lifetime undergoes large changes (32 %) in the pH 5 to pH 7 range, which allows accurate pH measurements to better than 0.2 pH units. By fusion of ECFP with the granular chromogranin A, we successfully measured the pH in secretory granules of PC12 cells, and we performed a kinetic analysis of intragranular pH variations in living cells exposed to ammonium chloride.

  16. Portable fluorescence lifetime spectroscopy system for in-situ interrogation of biological tissues

    Science.gov (United States)

    Saito Nogueira, Marcelo; Cosci, Alessandro; Teixeira Rosa, Ramon Gabriel; Salvio, Ana Gabriela; Pratavieira, Sebastião; Kurachi, Cristina

    2017-12-01

    Fluorescence spectroscopy and lifetime techniques are potential methods for optical diagnosis and characterization of biological tissues with an in-situ, fast, and noninvasive interrogation. Several diseases may be diagnosed due to differences in the fluorescence spectra of targeted fluorophores, when, these spectra are similar, considering steady-state fluorescence, others may be detected by monitoring their fluorescence lifetime. Despite this complementarity, most of the current fluorescence lifetime systems are not robust and portable, and not being feasible for clinical applications. We describe the assembly of a fluorescence lifetime spectroscopy system in a suitcase, its characterization, and validation with clinical measurements of skin lesions. The assembled system is all encased and robust, maintaining its mechanical, electrical, and optical stability during transportation, and is feasible for clinical measurements. The instrument response function measured was about 300 ps, and the system is properly calibrated. At the clinical study, the system showed to be reliable, and the achieved spectroscopy results support its potential use as an auxiliary tool for skin diagnostics.

  17. Remote UV Fluorescence Lifetime Spectrometer, Phase II

    Data.gov (United States)

    National Aeronautics and Space Administration — The goal of this project is to develop, demonstrate, and deliver to NASA an innovative, portable, and power efficient Remote UV Fluorescence Lifetime Spectrometer...

  18. Quantitative fluorescence lifetime spectroscopy in turbid media: comparison of theoretical, experimental and computational methods

    International Nuclear Information System (INIS)

    Vishwanath, Karthik; Mycek, Mary-Ann; Pogue, Brian

    2002-01-01

    A Monte Carlo model developed to simulate time-resolved fluorescence propagation in a semi-infinite turbid medium was validated against previously reported theoretical and computational results. Model simulations were compared to experimental measurements of fluorescence spectra and lifetimes on tissue-simulating phantoms for single and dual fibre-optic probe geometries. Experiments and simulations using a single probe revealed that scattering-induced artefacts appeared in fluorescence emission spectra, while fluorescence lifetimes were unchanged. Although fluorescence lifetime measurements are generally more robust to scattering artefacts than are measurements of fluorescence spectra, in the dual-probe geometry scattering-induced changes in apparent lifetime were predicted both from diffusion theory and via Monte Carlo simulation, as well as measured experimentally. In all cases, the recovered apparent lifetime increased with increasing scattering and increasing source-detector separation. Diffusion theory consistently underestimated the magnitude of these increases in apparent lifetime (predicting a maximum increase of ∼15%), while Monte Carlo simulations and experiment were closely matched (showing increases as large as 30%). These results indicate that quantitative simulations of time-resolved fluorescence propagation in turbid media will be important for accurate recovery of fluorophore lifetimes in biological spectroscopy and imaging applications. (author)

  19. Portable fluorescence lifetime spectroscopy system for in-situ interrogation of biological tissues.

    Science.gov (United States)

    Saito Nogueira, Marcelo; Cosci, Alessandro; Teixeira Rosa, Ramon Gabriel; Salvio, Ana Gabriela; Pratavieira, Sebastião; Kurachi, Cristina

    2017-10-01

    Fluorescence spectroscopy and lifetime techniques are potential methods for optical diagnosis and characterization of biological tissues with an in-situ, fast, and noninvasive interrogation. Several diseases may be diagnosed due to differences in the fluorescence spectra of targeted fluorophores, when, these spectra are similar, considering steady-state fluorescence, others may be detected by monitoring their fluorescence lifetime. Despite this complementarity, most of the current fluorescence lifetime systems are not robust and portable, and not being feasible for clinical applications. We describe the assembly of a fluorescence lifetime spectroscopy system in a suitcase, its characterization, and validation with clinical measurements of skin lesions. The assembled system is all encased and robust, maintaining its mechanical, electrical, and optical stability during transportation, and is feasible for clinical measurements. The instrument response function measured was about 300 ps, and the system is properly calibrated. At the clinical study, the system showed to be reliable, and the achieved spectroscopy results support its potential use as an auxiliary tool for skin diagnostics. (2017) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE).

  20. Xanthophyll cycle-dependent quenching of photosystem II chlorophyll a fluorescence: Formation of a quenching complex with a short fluorescence lifetime

    Energy Technology Data Exchange (ETDEWEB)

    Gilmore, A.M.; Hazlett, T.L.; Govindjee [Univ. of Illinois, Urbana, IL (United States)

    1995-03-14

    Excess light triggers protective nonradiative dissipation of excitation energy in photosystem II through the formation of a trans-thylakoid pH gradient that in turn stimulates formation of zeaxanthin and antheraxanthin. These xanthophylls when combined with protonation of antenna pigment-protein complexes may increase nonradiative dissipation and, thus, quench chlorophyll a fluorescence. Here we measured, in parallel, the chlorophyll a fluorescence lifetime and intensity to understand the mechanism of this process. Increasing the xanthophyll concentration in the presence of a pH gradient (quenched conditions) decreases the fractional intensity of a fluorescence lifetime component centered at {approx}2 ns and increases a component at {approx}0.4 ns. Uncoupling the pH gradient (unquenched conditions) eliminates the 0.4-ns component. Changes in the xanthophyll concentration do not significantly affect the fluorescence lifetimes in either the quenched or unquenched sample conditions. However, there are differences in fluorescence lifetimes between the quenched and unquenched states that are due to pH-related, but nonxanthophyll-related, processes. Quenching of the maximal fluorescence intensity correlates with both the xanthophyll concentration and the fractional intensity of the 0.4-ns component. The unchanged fluorescence lifetimes and the proportional quenching of the maximal and dark-level fluorescence intensities indicate that the xanthophyllact on antenna, not reaction center processes. Further, the fluorescence quenching is interpreted as the combined effect of the pH gradient and xanthophyll concentration, resulting in the formation of a quenching complex with a short ({approx}0.4 ns) fluorescence lifetime. 33 refs., 6 figs., 2 tabs.

  1. Mesh adaptation technique for Fourier-domain fluorescence lifetime imaging

    International Nuclear Information System (INIS)

    Soloviev, Vadim Y.

    2006-01-01

    A novel adaptive mesh technique in the Fourier domain is introduced for problems in fluorescence lifetime imaging. A dynamical adaptation of the three-dimensional scheme based on the finite volume formulation reduces computational time and balances the ill-posed nature of the inverse problem. Light propagation in the medium is modeled by the telegraph equation, while the lifetime reconstruction algorithm is derived from the Fredholm integral equation of the first kind. Stability and computational efficiency of the method are demonstrated by image reconstruction of two spherical fluorescent objects embedded in a tissue phantom

  2. The fluorescence lifetime of BRI1-GFP as probe for the noninvasive determination of the membrane potential in living cells

    Science.gov (United States)

    Elgass, K.; Caesar, K.; Schleifenbaum, F.; Meixner, A. J.; Harter, K.

    2010-02-01

    As the excited state lifetime of a fluorescent molecule depends on its environment, it is possible to use it as a probe for physico-chemical parameters of the surrounding medium. Whereas this is well known for many solid guest/host systems, only few reports of quantitative, temporal resolved in vivo studies to monitor the nano-environment for a protein-coupled chromophore such as GFP are known from literature. Here we present a novel approach to determine the membrane potential of living (plant) cells based on the fluorescence lifetime (FLT) analysis of membrane-located GFP. By using confocal sample scanning microscopy (CSSM) combined with fluorescence lifetime imaging microscopy, we recently showed that the phytohormone brassinolide (BL) induces cell wall expansion and a decrease in the FLT of the BRI1-GFP in living cells of Arabidopsis thaliana seedlings. BRI1 is the dominant functional receptor for BL in Arabidopsis and locates to the plasma membrane. Although the dependence of the FLT of GFP on its physico-chemical environment such as pH-value, refractive index and pressure has been reported, the observed FLT decrease of BRI1-GFP in response to BL application could not be explained by these parameters. However, our in vivo FLT and CSSM analyses indicate that the BLinduced change in the FLT of BRI1-GFP is caused by hyperpolarisation of the plasma membrane (Em). Thus, our results indicate that BRI1-GFP serves as sensitive and non-invasive probe for recording the Em of the plasma membrane in living plant cells with high spatio-temporal resolution.

  3. 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

  4. 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.

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

    OpenAIRE

    Elson, DS; Jo, JA; Marcu, L

    2007-01-01

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

  6. In vivo fluorescence lifetime imaging for monitoring the efficacy of the cancer treatment.

    Science.gov (United States)

    Ardeshirpour, Yasaman; Chernomordik, Victor; Hassan, Moinuddin; Zielinski, Rafal; Capala, Jacek; Gandjbakhche, Amir

    2014-07-01

    Advances in tumor biology created a foundation for targeted therapy aimed at inactivation of specific molecular mechanisms responsible for cell malignancy. In this paper, we used in vivo fluorescence lifetime imaging with HER2-targeted fluorescent probes as an alternative imaging method to investigate the efficacy of targeted therapy with 17-DMAG (an HSP90 inhibitor) on tumors with high expression of HER2 receptors. HER2-specific Affibody, conjugated to Alexafluor 750, was injected into nude mice bearing HER2-positive tumor xenograft. The fluorescence lifetime was measured before treatment and monitored after the probe injections at 12 hours after the last treatment dose, when the response to the 17-DMAG therapy was the most pronounced as well as a week after the last treatment when the tumors grew back almost to their pretreatment size. Imaging results showed significant difference between the fluorescence lifetimes at the tumor and the contralateral site (∼0.13 ns) in the control group (before treatment) and 7 days after the last treatment when the tumors grew back to their pretreatment dimensions. However, at the time frame that the treatment had its maximum effect (12 hours after the last treatment), the difference between the fluorescence lifetime at the tumor and contralateral site decreased to 0.03 ns. The results showed a good correlation between fluorescence lifetime and the efficacy of the treatment. These findings show that in vivo fluorescence lifetime imaging can be used as a promising molecular imaging tool for monitoring the treatment outcome in preclinical models and potentially in patients. ©2014 American Association for Cancer Research.

  7. In-vivo fluorescence lifetime imaging for monitoring the efficacy of the cancer treatment

    Science.gov (United States)

    Ardeshirpour, Yasaman; Chernomordik, Victor; Hassan, Moinuddin; Zielinski, Rafal; Capala, Jacek; Gandjbakhche, Amir

    2015-01-01

    Purpose Advances in tumor biology created a foundation for targeted therapy aimed at inactivation of specific molecular mechanisms responsible for cell malignancy. In this paper, we used in-vivo fluorescence lifetime imaging with HER2 targeted fluorescent probes as an alternative imaging method to investigate the efficacy of targeted therapy with 17-DMAG (an HSP90 inhibitor) on tumors with high expression of HER2 receptors. Experimental Design HER2-specific Affibody, conjugated to Alexafluor 750, was injected into nude mice, bearing HER2-positive tumor xenograft. The fluorescence lifetime was measured before treatment and monitored after the probe injections at 12 hours after the last treatment dose, when the response to the 17-DMAG therapy was the most pronounced as well as a week after the last treatment when the tumors grew back almost to their pre-treatment size. Results Imaging results showed significant difference between the fluorescence lifetimes at the tumor and the contralateral site (~0.13ns) in the control group (before treatment) and 7 days after the last treatment when the tumors grew back to their pretreatment dimensions. However, at the time frame that the treatment had its maximum effect (12 hours after the last treatment) the difference between the fluorescence lifetime at the tumor and contralateral site decreased to 0.03ns. Conclusions The results showed a good correlation between fluorescence lifetime and the efficacy of the treatment. These findings show that in-vivo fluorescence lifetime imaging can be used as a promising molecular imaging tool for monitoring the treatment outcome in preclinical models and potentially in patients. PMID:24671949

  8. 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.

  9. Fluorescence lifetime imaging of endogenous molecules in live mouse cancer models (Conference Presentation)

    Science.gov (United States)

    Svindrych, Zdenek; Wang, Tianxiong; Hu, Song; Periasamy, Ammasi

    2017-02-01

    NADH and FAD are important endogenous fluorescent coenzymes participating in key enzymatic reactions of cellular metabolism. While fluorescence intensities of NADH and FAD have been used to determine the redox state of cells and tissues, this simple approach breaks down in the case of deep-tissue intravital imaging due to depth- and wavelength-dependent light absorption and scattering. To circumvent this limitation, our research focuses on fluorescence lifetimes of two-photon excited NADH and FAD emission to study the metabolic state of live tissues. In our custom-built scanning microscope we combine tunable femtosecond Ti:sapphire laser (operating at 740 nm for NADH excitation and 890 nm for FAD excitation), two GaAsP hybrid detectors for registering individual fluorescence photons and two Becker and Hickl time correlator boards for high precision lifetime measurements. Together with our rigorous FLIM analysis approach (including image segmentation, multi-exponential decay fitting and detailed statistical analysis) we are able to detect metabolic changes in cancer xenografts (human pancreatic cancer MPanc96 cells injected subcutaneously into the ear of an immunodeficient nude mouse), relative to surrounding healthy tissue. Advantageously, with the same instrumentation we can also take high-resolution and high-contrast images of second harmonic signal (SHG) originating from collagen fibers of both the healthy skin and the growing tumor. The combination of metabolic measurements (NADH and FAD lifetime) and morphological information (collagen SHG) allows us to follow the tumor growth in live mouse model and the changes in tumor microenvironment.

  10. Plasmonic-based instrument response function for time-resolved fluorescence: toward proper lifetime analysis

    Energy Technology Data Exchange (ETDEWEB)

    Szlazak, Radoslaw; Tutaj, Krzysztof; Grudzinski, Wojciech; Gruszecki, Wieslaw I.; Luchowski, Rafal, E-mail: rafal.luchowski@umcs.pl [Maria Curie-Sklodowska University, Department of Biophysics, Institute of Physics (Poland)

    2013-06-15

    In this report, we investigated the so-called plasmonic platforms prepared to target ultra-short fluorescence and accurate instrumental response function in a time-domain spectroscopy and microscopy. The interaction of metallic nanoparticles with nearby fluorophores results in the increase of the dye fluorescence quantum yield, photostability and decrease of the lifetime parameter. The mentioned properties of platforms were applied to achieve a picosecond fluorescence lifetime (21 ps) of erythrosin B, used later as a better choice for deconvolution of fluorescence decays measured with 'color' sensitive photo-detectors. The ultra-short fluorescence standard based on combination of thin layers of silver film, silver colloidal nanoparticles (about 60 nm in diameter), and top layer of erythrosin B embedded in 0.2 % poly(vinyl) alcohol. The response functions were monitored on two photo-detectors; microchannel plate photomultiplier and single photon avalanche photodiode as a Rayleigh scattering and ultra-short fluorescence. We demonstrated that use of the plasmonic base fluorescence standard as an instrumental response function results in the absence of systematic error in lifetime measurements and analysis.

  11. Chlorophyll fluorescence lifetime imaging provides new insight into the chlorosis induced by plant virus infection.

    Science.gov (United States)

    Lei, Rong; Jiang, Hongshan; Hu, Fan; Yan, Jin; Zhu, Shuifang

    2017-02-01

    Leaf chlorosis induced by plant virus infection has a short fluorescence lifetime, which reflects damaged photosynthetic complexes and degraded chloroplasts. Plant viruses often induce chlorosis and necrosis, which are intimately related to photosynthetic functions. Chlorophyll fluorescence lifetime measurement is a valuable noninvasive tool for analyzing photosynthetic processes and is a sensitive indicator of the environment surrounding the fluorescent molecules. In this study, our central goal was to explore the effect of viral infection on photosynthesis by employing chlorophyll fluorescence lifetime imaging (FLIM), steady-state fluorescence, non-photochemical quenching (NPQ), transmission electron microscopy (TEM), and pigment analysis. The data indicated that the chlorophyll fluorescence lifetime of chlorotic leaves was significantly shorter than that of healthy control leaves, and the fitted short lifetime component of chlorophyll fluorescence of chlorotic leaves was dominant. This dominant short lifetime component may result from damage to the structure of thylakoid, which was confirmed by TEM. The NPQ value of chlorotic leaves was slightly higher than that of healthy green leaves, which can be explained by increased neoxanthin, lutein and violaxanthin content relative to chlorophyll a. The difference in NPQ is slight, but FLIM can provide simple and direct characterization of PSII structure and photosynthetic function. Therefore, this technique shows great potential as a simple and rapid method for studying mechanisms of plant virus infection.

  12. Refractive index sensing of green fluorescent proteins in living cells using fluorescence lifetime imaging microscopy

    NARCIS (Netherlands)

    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 gp91(phox), which are both subunits of the phagocyte NADPH

  13. Nanoantenna array-induced fluorescence enhancement and reduced lifetimes

    DEFF Research Database (Denmark)

    Bakker, R. M.; Drachev, V. P.; Liu, Z.

    2008-01-01

    Enhanced fluorescence is observed from dye molecules interacting with optical nanoantenna arrays. Elliptical gold dimers form individual nanoantennae with tunable plasmon resonances depending upon the geometry of the two particles and the size of the gap between them. A fluorescent dye, Rhodamine...... 800, is uniformly embedded in a dielectric host that coats the nanoantennae. The nanoantennae act to enhance the dye absorption. In turn, emission from the dye drives the plasmon resonance of the antennae; the nanoantennae act to enhance the fluorescence signal and change the angular distribution...... of emission. These effects depend upon the overlap of the plasmon resonance with the excitation wavelength and the fluorescence emission band. A decreased fluorescence lifetime is observed along with highly polarized emission that displays the characteristics of the nanoantenna's dipole mode. Being able...

  14. Fluorescence Lifetime Readouts of Troponin-C-Based Calcium FRET Sensors: A Quantitative Comparison of CFP and mTFP1 as Donor Fluorophores

    Science.gov (United States)

    Laine, Romain; Stuckey, Daniel W.; Manning, Hugh; Warren, Sean C.; Kennedy, Gordon; Carling, David

    2012-01-01

    We have compared the performance of two Troponin-C-based calcium FRET sensors using fluorescence lifetime read-outs. The first sensor, TN-L15, consists of a Troponin-C fragment inserted between CFP and Citrine while the second sensor, called mTFP-TnC-Cit, was realized by replacing CFP in TN-L15 with monomeric Teal Fluorescent Protein (mTFP1). Using cytosol preparations of transiently transfected mammalian cells, we have measured the fluorescence decay profiles of these sensors at controlled concentrations of calcium using time-correlated single photon counting. These data were fitted to discrete exponential decay models using global analysis to determine the FRET efficiency, fraction of donor molecules undergoing FRET and calcium affinity of these sensors. We have also studied the decay profiles of the donor fluorescent proteins alone and determined the sensitivity of the donor lifetime to temperature and emission wavelength. Live-cell fluorescence lifetime imaging (FLIM) of HEK293T cells expressing each of these sensors was also undertaken. We confirmed that donor fluorescence of mTFP-TnC-Cit fits well to a two-component decay model, while the TN-L15 lifetime data was best fitted to a constrained four-component model, which was supported by phasor analysis of the measured lifetime data. If the constrained global fitting is employed, the TN-L15 sensor can provide a larger dynamic range of lifetime readout than the mTFP-TnC-Cit sensor but the CFP donor is significantly more sensitive to changes in temperature and emission wavelength compared to mTFP and, while the mTFP-TnC-Cit solution phase data broadly agreed with measurements in live cells, this was not the case for the TN-L15 sensor. Our titration experiment also indicates that a similar precision in determination of calcium concentration can be achieved with both FRET biosensors when fitting a single exponential donor fluorescence decay model to the fluorescence decay profiles. We therefore suggest that m

  15. 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.

  16. Standard reference for instrument response function in fluorescence lifetime measurements in visible and near infrared

    International Nuclear Information System (INIS)

    Chib, Rahul; Shah, Sunil; Gryczynski, Zygmunt; Fudala, Rafal; Borejdo, Julian; Gryczynski, Ignacy; Zelent, Bogumil; Corradini, Maria G; Ludescher, Richard D

    2016-01-01

    Allura red (AR) fluorophore, a common dye in the food industry, displays a broad emission spectrum in water (visible-to-near infrared region of the electromagnetic spectrum) and has a remarkably short fluorescence lifetime of about 10 ps. This short lifetime does not depend on the emission (observation) wavelength. We examined time responses of AR fluorescence across emission wavelengths from 550 nm to 750 nm and found that it is an ideal candidate for impulse response functions in fluorescence lifetime measurements. (technical note)

  17. Fluorescence lifetime spectroscopy in multiple-scattering environments: an application to biotechnology

    Science.gov (United States)

    Cerussi, Albert E.; Gratton, Enrico; Fantini, Sergio

    1999-07-01

    Over the past few years, there has been significant research activity devoted to the application of fluorescence spectroscopy to strongly scattering media, where photons propagate diffusely. Much of this activity focused on fluorescence as a source of contrast enhancement in optical tomography. Our efforts have emphasized the quantitative recovery of fluorescence parameters for spectroscopy. Using a frequency-domain diffusion-based model, we have successfully recovered the lifetime, the absolute quantum yield, the fluorophore concentration, and the emission spectrum of the fluorophore, as well as the absorption and the reduced scattering coefficients at the emission wavelength of the medium in different measurements. In this contribution, we present a sensitive monitor of the binding between ethidium bromide and bovine cells in fresh milk. The spectroscopic contrast was the approximately tenfold increase in the ethidium bromide lifetime upon binding to DNA. The measurement clearly demonstrated that we could quantitatively measure the density of cells in the milk, which is an application vital to the tremendous economic burden of bovine subclinical mastitis detection. Furthermore, we may in principle use the spirit of this technique as a quantitative monitor of the binding of fluorescent drugs inside tissues. This is a first step towards lifetime spectroscopy in tissues.

  18. CVD grown 2D MoS{sub 2} layers: A photoluminescence and fluorescence lifetime imaging study

    Energy Technology Data Exchange (ETDEWEB)

    Oezden, Ayberk; Madenoglu, Buesra [Department of Materials Science and Engineering, Faculty of Engineering, Anadolu University, Eskisehir (Turkey); Sar, Hueseyin; Ay, Feridun; Perkgoez, Nihan Kosku [Department of Electrical and Electronics Engineering, Faculty of Engineering, Anadolu University, Eskisehir (Turkey); Yeltik, Aydan [Department of Physics, UNAM Institute of Materials Science and Nanotechnology, Bilkent University, Ankara (Turkey); Sevik, Cem [Department of Mechanical Engineering, Faculty of Engineering, Anadolu University, Eskisehir (Turkey)

    2016-11-15

    In this letter, we report on the fluorescence lifetime imaging and accompanying photoluminescence properties of a chemical vapour deposition (CVD) grown atomically thin material, MoS{sub 2}. μ-Raman, μ-photoluminescence (PL) and fluorescence lifetime imaging microscopy (FLIM) are utilized to probe the fluorescence lifetime and photoluminescence properties of individual flakes of MoS{sub 2} films. Usage of these three techniques allows identification of the grown layers, grain boundaries, structural defects and their relative effects on the PL and fluorescence lifetime spectra. Our investigation on individual monolayer flakes reveals a clear increase of the fluorescence lifetime from 0.3 ns to 0.45 ns at the edges with respect to interior region. On the other hand, investigation of the film layer reveals quenching of PL intensity and lifetime at the grain boundaries. These results could be important for applications where the activity of edges is important such as in photocatalytic water splitting. Finally, it has been demonstrated that PL mapping and FLIM are viable techniques for the investigation of the grain-boundaries. (copyright 2016 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

  19. Parallel excitation-emission multiplexed fluorescence lifetime confocal microscopy for live cell imaging.

    Science.gov (United States)

    Zhao, Ming; Li, Yu; Peng, Leilei

    2014-05-05

    We present a novel excitation-emission multiplexed fluorescence lifetime microscopy (FLIM) method that surpasses current FLIM techniques in multiplexing capability. The method employs Fourier multiplexing to simultaneously acquire confocal fluorescence lifetime images of multiple excitation wavelength and emission color combinations at 44,000 pixels/sec. The system is built with low-cost CW laser sources and standard PMTs with versatile spectral configuration, which can be implemented as an add-on to commercial confocal microscopes. The Fourier lifetime confocal method allows fast multiplexed FLIM imaging, which makes it possible to monitor multiple biological processes in live cells. The low cost and compatibility with commercial systems could also make multiplexed FLIM more accessible to biological research community.

  20. Two-photon excitation with pico-second fluorescence lifetime imaging to detect nuclear association of flavanols

    Energy Technology Data Exchange (ETDEWEB)

    Mueller-Harvey, Irene, E-mail: i.mueller-harvey@reading.ac.uk [Chemistry and Biochemistry Laboratory, Food Production and Quality Research Division, School of Agriculture, Policy and Development, University of Reading, P O Box 236, Reading RG6 6AT (United Kingdom); Feucht, Walter, E-mail: walter.feucht@gmail.com [Department of Plant Sciences, Technical University of Munich (TUM), Wissenschaftszentrum Weihenstephan (WZW), D-85354 Freising (Germany); Polster, Juergen, E-mail: j.polster@wzw.tum.de [Department of Physical Biochemistry, Technical University of Munich (TUM), Wissenschaftszentrum Weihenstephan (WZW), D-85354 Freising (Germany); Trnkova, Lucie, E-mail: lucie.trnkova@uhk.cz [University of Hradec Kralove, Faculty of Science, Department of Chemistry, Rokitanskeho 62, 50003 Hradec Kralove (Czech Republic); Burgos, Pierre, E-mail: pierre.burgos@stfc.ac.uk [Central Laser Facility, Research Complex at Harwell, Science and Technology Facilities Council, Rutherford Appleton Laboratory, Harwell-Oxford, Didcot, Oxfordshire, OX11 0QX (United Kingdom); Parker, Anthony W., E-mail: tony.parker@stfc.ac.uk [Central Laser Facility, Research Complex at Harwell, Science and Technology Facilities Council, Rutherford Appleton Laboratory, Harwell-Oxford, Didcot, Oxfordshire, OX11 0QX (United Kingdom); Botchway, Stanley W., E-mail: stan.botchway@stfc.ac.uk [Central Laser Facility, Research Complex at Harwell, Science and Technology Facilities Council, Rutherford Appleton Laboratory, Harwell-Oxford, Didcot, Oxfordshire, OX11 0QX (United Kingdom)

    2012-03-16

    Highlights: Black-Right-Pointing-Pointer This fluorescence lifetime imaging microscopy (FLIM) technique for flavanols overcomes autofluorescence interference in cells. Black-Right-Pointing-Pointer Plant flavanols differed in their lifetimes. Black-Right-Pointing-Pointer Dissolved and bound flavanols revealed contrasting lifetime changes. Black-Right-Pointing-Pointer This technique will allow studying of flavanol trafficking in live cells. - Abstract: Two-photon excitation enabled for the first time the observation and measurement of excited state fluorescence lifetimes from three flavanols in solution, which were {approx}1.0 ns for catechin and epicatechin, but <45 ps for epigallocatechin gallate (EGCG). The shorter lifetime for EGCG is in line with a lower fluorescence quantum yield of 0.003 compared to catechin (0.015) and epicatechin (0.018). In vivo experiments with onion cells demonstrated that tryptophan and quercetin, which tend to be major contributors of background fluorescence in plant cells, have sufficiently low cross sections for two-photon excitation at 630 nm and therefore do not interfere with detection of externally added or endogenous flavanols in Allium cepa or Taxus baccata cells. Applying two-photon excitation to flavanols enabled 3-D fluorescence lifetime imaging microscopy and showed that added EGCG penetrated the whole nucleus of onion cells. Interestingly, EGCG and catechin showed different lifetime behaviour when bound to the nucleus: EGCG lifetime increased from <45 to 200 ps, whilst catechin lifetime decreased from 1.0 ns to 500 ps. Semi-quantitative measurements revealed that the relative ratios of EGCG concentrations in nucleoli associated vesicles: nucleus: cytoplasm were ca. 100:10:1. Solution experiments with catechin, epicatechin and histone proteins provided preliminary evidence, via the appearance of a second lifetime ({tau}{sub 2} = 1.9-3.1 ns), that both flavanols may be interacting with histone proteins. We conclude that there

  1. The LB Films of Dansyl Chloride Labeled Octadecylamine and Its Fluorescence Lifetime

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Octadecylamine was derivatized with dansyl chloride (5-dimethylaminonaphthalene-1-sulfonyl chloride) in order to simplify and understand the LB films of fluorescent probe labeling proteins.Its monolayer and multilayers in the absence and presence of stearic acid were deposited by LB technique.Fluorescence spectra and lifetimes of the fluorescent products were studied to elucidate the microenvironment of molecules in the LB films.

  2. Fluorescence lifetime imaging of lipids during 3T3-L1 cell differentiation

    Science.gov (United States)

    Song, Young Sik; Won, Young Jae; Lee, Sang-Hak; Kim, Dug Young

    2014-03-01

    Obesity is becoming a big health problem in these days. Since increased body weight is due to increased number and size of the triglyceride-storing adipocytes, many researchers are working on differentiation conditions and processes of adipocytes. Adipocytes also work as regulators of whole-body energy homeostasis by secreting several proteins that regulate processes as diverse as haemostasis, blood pressure, immune function, angiogenesis and energy balance. 3T3-L1 cells are widely used cell line for studying adipogenesis because it can differentiate into an adipocyte-like phenotype under appropriate conditions. In this paper, we propose an effective fluorescence lifetime imaging technique which can easily distinguish lipids in membrane and those in lipid droplets. Nile red dyes are attached to lipids in 3T3-L1 cells. Fluorescence lifetime images were taken for 2 week during differentiation procedure of 3T3-L1 cells into adipocytes. We used 488 nm pulsed laser with 5MHz repetition rate and emission wavelength is 520 nm of Nile Red fluorescent dye. Results clearly show that the lifetime of Nile red in lipid droplets are smaller than those in cell membrane. Our results suggest that fluorescence lifetime imaging can be a very powerful tool to monitor lipid droplet formation in adipocytes from 3T3-L1 cells.

  3. 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.

  4. In Vivo Fluorescence Lifetime Imaging Monitors Binding of Specific Probes to Cancer Biomarkers

    Science.gov (United States)

    Ardeshirpour, Yasaman; Chernomordik, Victor; Zielinski, Rafal; Capala, Jacek; Griffiths, Gary; Vasalatiy, Olga; Smirnov, Aleksandr V.; Knutson, Jay R.; Lyakhov, Ilya; Achilefu, Samuel; Gandjbakhche, Amir; Hassan, Moinuddin

    2012-01-01

    One of the most important factors in choosing a treatment strategy for cancer is characterization of biomarkers in cancer cells. Particularly, recent advances in Monoclonal Antibodies (MAB) as primary-specific drugs targeting tumor receptors show that their efficacy depends strongly on characterization of tumor biomarkers. Assessment of their status in individual patients would facilitate selection of an optimal treatment strategy, and the continuous monitoring of those biomarkers and their binding process to the therapy would provide a means for early evaluation of the efficacy of therapeutic intervention. In this study we have demonstrated for the first time in live animals that the fluorescence lifetime can be used to detect the binding of targeted optical probes to the extracellular receptors on tumor cells in vivo. The rationale was that fluorescence lifetime of a specific probe is sensitive to local environment and/or affinity to other molecules. We attached Near-InfraRed (NIR) fluorescent probes to Human Epidermal Growth Factor 2 (HER2/neu)-specific Affibody molecules and used our time-resolved optical system to compare the fluorescence lifetime of the optical probes that were bound and unbound to tumor cells in live mice. Our results show that the fluorescence lifetime changes in our model system delineate HER2 receptor bound from the unbound probe in vivo. Thus, this method is useful as a specific marker of the receptor binding process, which can open a new paradigm in the “image and treat” concept, especially for early evaluation of the efficacy of the therapy. PMID:22384092

  5. In vivo fluorescence lifetime imaging monitors binding of specific probes to cancer biomarkers.

    Directory of Open Access Journals (Sweden)

    Yasaman Ardeshirpour

    Full Text Available One of the most important factors in choosing a treatment strategy for cancer is characterization of biomarkers in cancer cells. Particularly, recent advances in Monoclonal Antibodies (MAB as primary-specific drugs targeting tumor receptors show that their efficacy depends strongly on characterization of tumor biomarkers. Assessment of their status in individual patients would facilitate selection of an optimal treatment strategy, and the continuous monitoring of those biomarkers and their binding process to the therapy would provide a means for early evaluation of the efficacy of therapeutic intervention. In this study we have demonstrated for the first time in live animals that the fluorescence lifetime can be used to detect the binding of targeted optical probes to the extracellular receptors on tumor cells in vivo. The rationale was that fluorescence lifetime of a specific probe is sensitive to local environment and/or affinity to other molecules. We attached Near-InfraRed (NIR fluorescent probes to Human Epidermal Growth Factor 2 (HER2/neu-specific Affibody molecules and used our time-resolved optical system to compare the fluorescence lifetime of the optical probes that were bound and unbound to tumor cells in live mice. Our results show that the fluorescence lifetime changes in our model system delineate HER2 receptor bound from the unbound probe in vivo. Thus, this method is useful as a specific marker of the receptor binding process, which can open a new paradigm in the "image and treat" concept, especially for early evaluation of the efficacy of the therapy.

  6. Label-free identification of macrophage phenotype by fluorescence lifetime imaging microscopy

    Science.gov (United States)

    Alfonso-García, Alba; Smith, Tim D.; Datta, Rupsa; Luu, Thuy U.; Gratton, Enrico; Potma, Eric O.; Liu, Wendy F.

    2016-04-01

    Macrophages adopt a variety of phenotypes that are a reflection of the many functions they perform as part of the immune system. In particular, metabolism is a phenotypic trait that differs between classically activated, proinflammatory macrophages, and alternatively activated, prohealing macrophages. Inflammatory macrophages have a metabolism based on glycolysis while alternatively activated macrophages generally rely on oxidative phosphorylation to generate chemical energy. We employ this shift in metabolism as an endogenous marker to identify the phenotype of individual macrophages via live-cell fluorescence lifetime imaging microscopy (FLIM). We demonstrate that polarized macrophages can be readily discriminated with the aid of a phasor approach to FLIM, which provides a fast and model-free method for analyzing fluorescence lifetime images.

  7. 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...

  8. Fluorescence lifetime, dipole orientation and bilayer polymer films

    Science.gov (United States)

    Ho, Xuan Long; Chen, Po-Jui; Woon, Wei-Yen; White, Jonathon David

    2017-10-01

    Bilayer films consisting of the optically transparent polymers, polystyrene (PS) and poly(methyl methacrylate) (PMMA) were spin-cast on glass substrates. The upper 13.5 nm layer (PS) was lightly doped with Rhodamine-6 G (RH6G) or MEH-PPV. While the fluorescence of MEH-PPV was independent of PMMA thickness, the lifetime of RH6G increased 3-fold as the underlying PMMA thickness increased from 0 to 500 nm while the collected flux decreased suggesting a reorientation of the smaller molecule's dipole with respect to the air-polymer interface with PMMA thickness. This suggests lifetime may find application for nondestructive thickness measurements of transparent films with sub-micron lateral resolution and large range.

  9. Towards sensitive, high-throughput, biomolecular assays based on fluorescence lifetime

    Science.gov (United States)

    Ioanna Skilitsi, Anastasia; Turko, Timothé; Cianfarani, Damien; Barre, Sophie; Uhring, Wilfried; Hassiepen, Ulrich; Léonard, Jérémie

    2017-09-01

    Time-resolved fluorescence detection for robust sensing of biomolecular interactions is developed by implementing time-correlated single photon counting in high-throughput conditions. Droplet microfluidics is used as a promising platform for the very fast handling of low-volume samples. We illustrate the potential of this very sensitive and cost-effective technology in the context of an enzymatic activity assay based on fluorescently-labeled biomolecules. Fluorescence lifetime detection by time-correlated single photon counting is shown to enable reliable discrimination between positive and negative control samples at a throughput as high as several hundred samples per second.

  10. Actin cytoskeleton-dependent Rab GTPase-regulated angiotensin type I receptor lysosomal degradation studied by fluorescence lifetime imaging microscopy

    Science.gov (United States)

    Li, Hewang; Yu, Peiying; Sun, Yuansheng; Felder, Robin A.; Periasamy, Ammasi; Jose, Pedro A.

    2010-09-01

    The dynamic regulation of the cellular trafficking of human angiotensin (Ang) type 1 receptor (AT1R) is not well understood. Therefore, we investigated the cellular trafficking of AT1R-enhanced green fluorescent protein (EGFP) (AT1R-EGFP) heterologously expressed in HEK293 cells by determining the change in donor lifetime (AT1R-EGFP) in the presence or absence of acceptor(s) using fluorescence lifetime imaging-fluorescence resonance energy transfer (FRET) microscopy. The average lifetime of AT1R-EGFP in our donor-alone samples was ~2.33 ns. The basal state lifetime was shortened slightly in the presence of Rab5 (2.01+/-0.10 ns) or Rab7 (2.11+/-0.11 ns) labeled with Alexa 555, as the acceptor fluorophore. A 5-min Ang II treatment markedly shortened the lifetime of AT1R-EGFP in the presence of Rab5-Alexa 555 (1.78+/-0.31 ns) but was affected minimally in the presence of Rab7-Alexa 555 (2.09+/-0.37 ns). A 30-min Ang II treatment further decreased the AT1R-EGFP lifetime in the presence of both Rab5- and Rab7-Alexa 555. Latrunculin A but not nocodazole pretreatment blocked the ability of Ang II to shorten the AT1R-EGFP lifetime. The occurrence of FRET between AT1R-EGFP (donor) and LAMP1-Alexa 555 (acceptor) with Ang II stimulation was impaired by photobleaching the acceptor. These studies demonstrate that Ang II-induced AT1R lysosomal degradation through its association with LAMP1 is regulated by Rab5/7 via mechanisms that are dependent on intact actin cytoskeletons.

  11. Assessment of post-implantation integration of engineered tissues using fluorescence lifetime spectroscopy

    Science.gov (United States)

    Elahi, Sakib F.; Lee, Seung Y.; Lloyd, William R.; Chen, Leng-Chun; Kuo, Shiuhyang; Zhou, Ying; Kim, Hyungjin M.; Kennedy, Robert; Marcelo, Cynthia; Feinberg, Stephen E.; Mycek, Mary-Ann

    2018-02-01

    Clinical translation of engineered tissue constructs requires noninvasive methods to assess construct health and viability after implantation in patients. However, current practices to monitor post-implantation construct integration are either qualitative (visual assessment) or destructive (tissue histology). As label-free fluorescence lifetime sensing can noninvasively characterize pre-implantation construct viability, we employed a handheld fluorescence lifetime spectroscopy probe to quantitatively and noninvasively assess tissue constructs that were implanted in a murine model. We designed the system to be suitable for intravital measurements: portability, localization with precise maneuverability, and rapid data acquisition. Our model tissue constructs were manufactured from primary human cells to simulate patient variability and were stressed to create a range of health states. Secreted amounts of three cytokines that relate to cellular viability were measured in vitro to assess pre-implantation construct health. In vivo optical sensing assessed tissue integration of constructs at one-week and three-weeks post-implantation. At one-week post-implantation, optical parameters correlated with in vitro pre-implantation secretion levels of all three cytokines (p clinical optical diagnostic tools based on label-free fluorescence lifetime sensing of endogenous tissue fluorophores could noninvasively monitor post-implantation integration of engineered tissues.

  12. Time gated fluorescence lifetime imaging and micro-volume spectroscopy using two-photon excitation

    NARCIS (Netherlands)

    Sytsma, J.; Vroom, J.M.; de Grauw, C.J.; Gerritsen, H.C.

    A scanning microscope utilizing two-photon excitation in combination with fluorescence lifetime contrast is presented. The microscope makes use of a tunable femtosecond titanium:sapphire laser enabling the two-photon excitation of a broad range of fluorescent molecules, including UV probes.

  13. Multiphoton Laser Microscopy and Fluorescence Lifetime Imaging for the Evaluation of the Skin

    Directory of Open Access Journals (Sweden)

    Stefania Seidenari

    2012-01-01

    Full Text Available Multiphoton laser microscopy is a new, non-invasive technique providing access to the skin at a cellular and subcellular level, which is based both on autofluorescence and fluorescence lifetime imaging. Whereas the former considers fluorescence intensity emitted by epidermal and dermal fluorophores and by the extra-cellular matrix, fluorescence lifetime imaging (FLIM, is generated by the fluorescence decay rate. This innovative technique can be applied to the study of living skin, cell cultures and ex vivo samples. Although still limited to the clinical research field, the development of multiphoton laser microscopy is thought to become suitable for a practical application in the next few years: in this paper, we performed an accurate review of the studies published so far, considering the possible fields of application of this imaging method and providing high quality images acquired in the Department of Dermatology of the University of Modena.

  14. Enhancement of early cervical cancer diagnosis with epithelial layer analysis of fluorescence lifetime images.

    Directory of Open Access Journals (Sweden)

    Jun Gu

    Full Text Available This work reports the use of layer analysis to aid the fluorescence lifetime diagnosis of cervical intraepithelial neoplasia (CIN from H&E stained cervical tissue sections. The mean and standard deviation of lifetimes in single region of interest (ROI of cervical epithelium were previously shown to correlate to the gold standard histopathological classification of early cervical cancer. These previously defined single ROIs were evenly divided into layers for analysis. A 10-layer model revealed a steady increase in fluorescence lifetime from the inner to the outer epithelial layers of healthy tissue sections, suggesting a close association with cellular maturity. The shorter lifetime and minimal lifetime increase towards the epithelial surface of CIN-affected regions are in good agreement with the absence of cellular maturation in CIN. Mean layer lifetimes in the top-half cervical epithelium were used as feature vectors for extreme learning machine (ELM classifier discriminations. It was found that the proposed layer analysis technique greatly improves the sensitivity and specificity to 94.6% and 84.3%, respectively, which can better supplement the traditional gold standard cervical histopathological examinations.

  15. A fast global fitting algorithm for fluorescence lifetime imaging microscopy based on image segmentation.

    Science.gov (United States)

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

    2004-10-01

    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 analysis. Convergence speed can be greatly accelerated by providing appropriate initial guesses. Realizing that the image morphology often correlates with fluorophore distribution, a global fitting algorithm has been developed to assign initial guesses throughout an image based on a segmentation analysis. This algorithm was tested on both simulated data sets and time-domain lifetime measurements. We have successfully measured fluorophore distribution in fibroblasts stained with Hoechst and calcein. This method further allows second harmonic generation from collagen and elastin autofluorescence to be differentiated in fluorescence lifetime imaging microscopy images of ex vivo human skin. On our experimental measurement, this algorithm increased convergence speed by over two orders of magnitude and achieved significantly better fits. Copyright 2004 Biophysical Society

  16. Diagnosis of basal cell carcinoma by two photon excited fluorescence combined with lifetime imaging

    Science.gov (United States)

    Fan, Shunping; Peng, Xiao; Liu, Lixin; Liu, Shaoxiong; Lu, Yuan; Qu, Junle

    2014-02-01

    Basal cell carcinoma (BCC) is the most common type of human skin cancer. The traditional diagnostic procedure of BCC is histological examination with haematoxylin and eosin staining of the tissue biopsy. In order to reduce complexity of the diagnosis procedure, a number of noninvasive optical methods have been applied in skin examination, for example, multiphoton tomography (MPT) and fluorescence lifetime imaging microscopy (FLIM). In this study, we explored two-photon optical tomography of human skin specimens using two-photon excited autofluorescence imaging and FLIM. There are a number of naturally endogenous fluorophores in skin sample, such as keratin, melanin, collagen, elastin, flavin and porphyrin. Confocal microscopy was used to obtain structures of the sample. Properties of epidermic and cancer cells were characterized by fluorescence emission spectra, as well as fluorescence lifetime imaging. Our results show that two-photon autofluorescence lifetime imaging can provide accurate optical biopsies with subcellular resolution and is potentially a quantitative optical diagnostic method in skin cancer diagnosis.

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

    Directory of Open Access Journals (Sweden)

    Robert K. Henderson

    2012-05-01

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

  18. 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

  19. Sensitive determination of nucleic acids using organic nanoparticle fluorescence probes

    Science.gov (United States)

    Zhou, Yunyou; Bian, Guirong; Wang, Leyu; Dong, Ling; Wang, Lun; Kan, Jian

    2005-06-01

    This paper describes the preparation of organic nanoparticles by reprecipitation method under sonication and vigorous stirring. Transmission electron microscopy (TEM) was used to characterize the size and size distribution of the luminescent nanoparticles. Their average diameter was about 25 nm with a size variation of ±18%. The fluorescence decay lifetime of the nanoparticles also was determined on a self-equipped fluorospectrometer with laser light source. The lifetime (˜0.09 μs) of nanoparticles is about three times long as that of the monomer. The nanoparticles were in abundant of hydrophilic groups, which increased their miscibility in aqueous solution. These organic nanoparticles have high photochemical stability, excellent resistance to chemical degradation and photodegradation, and a good fluorescence quantum yield (25%). The fluorescence can be efficiently quenched by nucleic acids. Based on the fluorescence quenching of nanoparticles, a fluorescence quenching method was developed for determination of microamounts of nucleic acids by using the nanoparticles as a new fluorescent probe. Under optimal conditions, maximum fluorescence quenching is produced, with maximum excitation and emission wavelengths of 345 and 402 nm, respectively. Under optimal conditions, the calibration graphs are linear over the range 0.4-19.0 μg ml -1 for calf thymus DNA (ct-DNA) and 0.3-19.0 μg ml -1 for fish sperm DNA (fs-DNA). The corresponding detection limits are 0.25 μg ml -1 for ct-DNA and 0.17 μg ml -1 for fs-DNA. The relative standard deviation of six replicate measurements is 1.3-2.1%. The method is simple, rapid and sensitive with wide linear range. The recovery and relative standard deviation are very satisfactory.

  20. Study on the effect of deposition rate and concentration of Eu on the fluorescent lifetime of CsI: Tl thin film

    Energy Technology Data Exchange (ETDEWEB)

    Xie, Yijun; Guo, Lina [University of Electronic Science and Technology of China, State Key Laboratory of Electronic Thin Films and Integrated Devices, School of Optoelectronic Information, Chengdu 610054 (China); Liu, Shuang, E-mail: shuangliu@uestc.edu.cn [University of Electronic Science and Technology of China, State Key Laboratory of Electronic Thin Films and Integrated Devices, School of Optoelectronic Information, Chengdu 610054 (China); Wang, Qianfeng; Zhang, Shangjian; Liu, Yong [University of Electronic Science and Technology of China, State Key Laboratory of Electronic Thin Films and Integrated Devices, School of Optoelectronic Information, Chengdu 610054 (China); Zhong, Zhiyong [University of Electronic Science and Technology of China, State Key Laboratory of Electronic Thin Films and Integrated Devices, Chengdu 610054 (China)

    2017-06-21

    Although there are many new scintillators being developed recently, CsI: Tl is still very efficient among them. The fluorescent lifetime is a very important parameter of CsI: Tl thin film and two series of experiments have been conducted to learn about it. Our experiments, however, have demonstrated that the deposition rate and the codoping of Eu{sup 2+} will significantly influence its fluorescent lifetime. In order to increase the efficiency of the imaging system, we intend to obtain a higher fluorescent lifetime for CsI: Tl thin film by controlling these two conditions. - Highlights: • We used vacuum vapor deposition method to grow the high-quality thin films. • The relationship between the deposition rate and the fluorescent lifetime of CsI: Tl thin film was tested. • Concentration of Eu on fluorescent lifetime of the CsI: Tl thin film was studied.

  1. Community detection for fluorescent lifetime microscopy image segmentation

    Science.gov (United States)

    Hu, Dandan; Sarder, Pinaki; Ronhovde, Peter; Achilefu, Samuel; Nussinov, Zohar

    2014-03-01

    Multiresolution community detection (CD) method has been suggested in a recent work as an efficient method for performing unsupervised segmentation of fluorescence lifetime (FLT) images of live cell images containing fluorescent molecular probes.1 In the current paper, we further explore this method in FLT images of ex vivo tissue slices. The image processing problem is framed as identifying clusters with respective average FLTs against a background or "solvent" in FLT imaging microscopy (FLIM) images derived using NIR fluorescent dyes. We have identified significant multiresolution structures using replica correlations in these images, where such correlations are manifested by information theoretic overlaps of the independent solutions ("replicas") attained using the multiresolution CD method from different starting points. In this paper, our method is found to be more efficient than a current state-of-the-art image segmentation method based on mixture of Gaussian distributions. It offers more than 1:25 times diversity based on Shannon index than the latter method, in selecting clusters with distinct average FLTs in NIR FLIM images.

  2. Fluorescence lifetime microscopy for monitoring cell adhesion using metal induced energy transfer

    Science.gov (United States)

    Hwang, Wonsang; Seo, JinWon; Song, Jun ho; Kim, DongEun; Won, YoungJae; Choi, In-Hong; Yoo, Kyung-Hwa; Kim, Dug Young

    2018-02-01

    A precise control and a reliable monitoring tool for the adhesion properties of a cell are very important in atherosclerosis studies. If endothelial cells in contact with the intracellular membrane are not attached securely, low-density lipoprotein (LDL) particles can enter into the inner membrane. It is therefore necessary to measure conditions under which endothelial cell detachment occurs. When a cell is attached to a metal thin film, the lifetime of a fluorescence probe attached to the membrane of the cell is reduced by the metal-induced energy transfer (MIET). Fluorescence lifetime imaging microscopy (FLIM) is used to monitor the attachment condition of a cell to a metal surface using FRET. However, this requires high numerical aperture (NA) objective lens because axial confocal resolution must be smaller than the cell thickness. This requirement limits the field of view of the measurement specimen. In this study we provides a new method which can measure adhesion properties of endothelial cells even with a low NA objective lens by resolving two lifetime components in FLIM.

  3. Monitoring macular pigment changes in macular holes using fluorescence lifetime imaging ophthalmoscopy.

    Science.gov (United States)

    Sauer, Lydia; Peters, Sven; Schmidt, Johanna; Schweitzer, Dietrich; Klemm, Matthias; Ramm, Lisa; Augsten, Regine; Hammer, Martin

    2017-08-01

    To investigate the impact of macular pigment (MP) on fundus autofluorescence (FAF) lifetimes in vivo by characterizing full-thickness idiopathic macular holes (MH) and macular pseudo-holes (MPH). A total of 37 patients with MH and 52 with MPH were included. Using the fluorescence lifetime imaging ophthalmoscope (FLIO), based on a Heidelberg Engineering Spectralis system, a 30° retinal field was investigated. FAF decays were detected in a short (498-560 nm; ch1) and long (560-720 nm; ch2) wavelength channel. τ m , the mean fluorescence lifetime, was calculated from a three-exponential approximation of the FAF decays. Macular coherence tomography scans were recorded, and macular pigment's optical density (MPOD) was measured (one-wavelength reflectometry). Two MH subgroups were analysed according to the presence or absence of an operculum above the MH. A total of 17 healthy fellow eyes were included. A longitudinal FAF decay examination was conducted in nine patients, which were followed up after surgery and showed a closed MH. In MH without opercula, significant τ m differences (p hole area (MHa) and surrounding areas (MHb) (ch1: MHa 238 ± 64 ps, MHb 181 ± 78 ps; ch2: MHa 275 ± 49 ps, MHb 223 ± 48 ps), as well as between MHa and healthy eyes or closed MH. Shorter τ m , adjacent to the hole, can be assigned to areas with equivalently higher MPOD. Opercula containing MP also show short τ m . In MPH, the intactness of the Hele fibre layer is associated with shortest τ m . Shortest τ m originates from MP-containing retinal layers, especially from the Henle fibre layer. Fluorescence lifetime imaging ophthalmoscope (FLIO) provides information on the MP distribution, the pathogenesis and topology of MH. Macular pigment (MP) fluorescence may provide a biomarker for monitoring pathological changes in retinal diseases. © 2016 Acta Ophthalmologica Scandinavica Foundation. Published by John Wiley & Sons Ltd.

  4. Time-Resolved Fluorescence Spectroscopy and Fluorescence Lifetime Imaging Microscopy for Characterization of Dendritic Polymer Nanoparticles and Applications in Nanomedicine

    Directory of Open Access Journals (Sweden)

    Alexander Boreham

    2016-12-01

    Full Text Available The emerging field of nanomedicine provides new approaches for the diagnosis and treatment of diseases, for symptom relief and for monitoring of disease progression. One route of realizing this approach is through carefully constructed nanoparticles. Due to the small size inherent to the nanoparticles a proper characterization is not trivial. This review highlights the application of time-resolved fluorescence spectroscopy and fluorescence lifetime imaging microscopy (FLIM for the analysis of nanoparticles, covering aspects ranging from molecular properties to particle detection in tissue samples. The latter technique is particularly important as FLIM allows for distinguishing of target molecules from the autofluorescent background and, due to the environmental sensitivity of the fluorescence lifetime, also offers insights into the local environment of the nanoparticle or its interactions with other biomolecules. Thus, these techniques offer highly suitable tools in the fields of particle development, such as organic chemistry, and in the fields of particle application, such as in experimental dermatology or pharmaceutical research.

  5. Time-Resolved Fluorescence Spectroscopy and Fluorescence Lifetime Imaging Microscopy for Characterization of Dendritic Polymer Nanoparticles and Applications in Nanomedicine.

    Science.gov (United States)

    Boreham, Alexander; Brodwolf, Robert; Walker, Karolina; Haag, Rainer; Alexiev, Ulrike

    2016-12-24

    The emerging field of nanomedicine provides new approaches for the diagnosis and treatment of diseases, for symptom relief and for monitoring of disease progression. One route of realizing this approach is through carefully constructed nanoparticles. Due to the small size inherent to the nanoparticles a proper characterization is not trivial. This review highlights the application of time-resolved fluorescence spectroscopy and fluorescence lifetime imaging microscopy (FLIM) for the analysis of nanoparticles, covering aspects ranging from molecular properties to particle detection in tissue samples. The latter technique is particularly important as FLIM allows for distinguishing of target molecules from the autofluorescent background and, due to the environmental sensitivity of the fluorescence lifetime, also offers insights into the local environment of the nanoparticle or its interactions with other biomolecules. Thus, these techniques offer highly suitable tools in the fields of particle development, such as organic chemistry, and in the fields of particle application, such as in experimental dermatology or pharmaceutical research.

  6. Fluorescence lifetime measurements of native and glycated human serum albumin and bovine serum albumin

    Science.gov (United States)

    Joshi, Narahari V.; Joshi, Virgina O. d.; Contreras, Silvia; Gil, Herminia; Medina, Honorio; Siemiarczuk, Aleksander

    1999-05-01

    Nonenzymatic glycation, also known as Maillard reaction, plays an important role in the secondary complications of the diabetic pathology and aging, therefore, human serum albumin (HSA) and bovine serum albumin (BSA) were glycated by a conventional method in our laboratory using glucose as the glycating agent. Fluorescence lifetime measurements were carried out with a laser strobe fluorometer equipped with a nitrogen/dye laser and a frequency doubler as a pulsed excitation source. The samples were excited at 295 nm and the emission spectra were recorded at 345 nm. The obtained decay curves were tried for double and triple exponential functions. It has been found that the shorter lifetime increases for glycated proteins as compared with that of the native ones. For example, in the case of glycated BSA the lifetime increased from 1.36 ns to 2.30 ns. Similarly, for HSA, the lifetime increases from 1.58 ns to 2.26 ns. Meanwhile, the longer lifetime changed very slightly for both proteins (from 6.52 ns to 6.72 ns). The increase in the lifetime can be associated with the environmental effect; originated from the attachment of glucose to some lysine residues. A good example is Trp 214 which is in the cage of Lys 225, Lys 212, Lys 233, Lys 205, Lys 500, Lys 199 and Lys 195. If fluorescence lifetime technique is calibrated and properly used it could be employed for assessing glycation of proteins.

  7. 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

  8. Fluorescence lifetime microscopy of NADH distinguishes alterations in cerebral metabolism in vivo.

    Science.gov (United States)

    Yaseen, Mohammad A; Sutin, Jason; Wu, Weicheng; Fu, Buyin; Uhlirova, Hana; Devor, Anna; Boas, David A; Sakadžić, Sava

    2017-05-01

    Evaluating cerebral energy metabolism at microscopic resolution is important for comprehensively understanding healthy brain function and its pathological alterations. Here, we resolve specific alterations in cerebral metabolism in vivo in Sprague Dawley rats utilizing minimally-invasive 2-photon fluorescence lifetime imaging (2P-FLIM) measurements of reduced nicotinamide adenine dinucleotide (NADH) fluorescence. Time-resolved fluorescence lifetime measurements enable distinction of different components contributing to NADH autofluorescence. Ostensibly, these components indicate different enzyme-bound formulations of NADH. We observed distinct variations in the relative proportions of these components before and after pharmacological-induced impairments to several reactions involved in glycolytic and oxidative metabolism. Classification models were developed with the experimental data and used to predict the metabolic impairments induced during separate experiments involving bicuculline-induced seizures. The models consistently predicted that prolonged focal seizure activity results in impaired activity in the electron transport chain, likely the consequence of inadequate oxygen supply. 2P-FLIM observations of cerebral NADH will help advance our understanding of cerebral energetics at a microscopic scale. Such knowledge will aid in our evaluation of healthy and diseased cerebral physiology and guide diagnostic and therapeutic strategies that target cerebral energetics.

  9. A Compact Fluorescence Lifetime Excitation-Emission Spectrometer (FLEXEMS) for Detecting Trace Organics, Phase II

    Data.gov (United States)

    National Aeronautics and Space Administration — In this Small Business Innovative Research (SBIR) effort, Leiden Measurement Technology (LMT) proposes to design and build the Fluorescence Lifetime Excitation...

  10. A Compact Fluorescence Lifetime Excitation-Emission Spectrometer (FLEXEMS) for Detecting Trace Organics, Phase I

    Data.gov (United States)

    National Aeronautics and Space Administration — In this Small Business Innovative Research (SBIR) effort, Leiden Measurement Technology (LMT) proposes to design and build the Fluorescence Lifetime Excitation...

  11. Using non-empirically tuned range-separated functionals with simulated emission bands to model fluorescence lifetimes.

    Science.gov (United States)

    Wong, Z C; Fan, W Y; Chwee, T S; Sullivan, Michael B

    2017-08-09

    Fluorescence lifetimes were evaluated using TD-DFT under different approximations for the emitting molecule and various exchange-correlation functionals, such as B3LYP, BMK, CAM-B3LYP, LC-BLYP, M06, M06-2X, M11, PBE0, ωB97, ωB97X, LC-BLYP*, and ωB97X* where the range-separation parameters in the last two functionals were tuned in a non-empirical fashion. Changes in the optimised molecular geometries between the ground and electronically excited states were found to affect the quality of the calculated lifetimes significantly, while the inclusion of vibronic features led to further improvements over the assumption of a vertical electronic transition. The LC-BLYP* functional was found to return the most accurate fluorescence lifetimes with unsigned errors that are mostly within 1.5 ns of experimental values.

  12. Alterations in cerebral metabolism observed in living rodents using fluorescence lifetime microscopy of intrinsic NADH (Conference Presentation)

    Science.gov (United States)

    Yaseen, Mohammad A.; Sakadžić, Sava; Sutin, Jason; Wu, Weicheng; Fu, Buyin; Boas, David A.

    2017-02-01

    Monitoring cerebral energy metabolism at a cellular level is essential to improve our understanding of healthy brain function and its pathological alterations. In this study, we resolve specific alterations in cerebral metabolism utilizing minimally-invasive 2-Photon fluorescence lifetime imaging (2P-FLIM) measurements of reduced nicotinamide adenine dinucleotide (NADH) fluorescence, collected in vivo from anesthetized rats and mice. Time-resolved lifetime measurements enables distinction of different components contributing to NADH autofluorescence. These components reportedly represent different enzyme-bound formulations of NADH. Our observations from this study confirm the hypothesis that NADH FLIM can identify specific alterations in cerebral metabolism. Using time-correlated single photon counting (TCSPC) equipment and a custom-built multimodal imaging system, 2-photon fluorescence lifetime imaging (FLIM) was performed in cerebral tissue with high spatial and temporal resolution. Multi-exponential fits for NADH fluorescence lifetimes indicate 4 distinct components, or 'species.' We observed distinct variations in the relative proportions of these components before and after pharmacological-induced impairments to several reactions involved in anaerobic glycolysis and aerobic oxidative metabolism. Classification models developed with experimental data correctly predict the metabolic impairments associated with bicuculline-induced focal seizures in separate experiments. Compared to traditional intensity-based NADH measurements, lifetime imaging of NADH is less susceptible to the adverse effects of overlying blood vessels. Evaluating NADH measurements will ultimately lead to a deeper understanding of cerebral energetics and its pathology-related alterations. Such knowledge will likely aid development of therapeutic strategies for neurodegenerative diseases such as Alzheimer's Disease, Parkinson's disease, and stroke.

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

    Science.gov (United States)

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

    2018-03-01

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

  14. A high-throughput direct fluorescence resonance energy transfer-based assay for analyzing apoptotic proteases using flow cytometry and fluorescence lifetime measurements.

    Science.gov (United States)

    Suzuki, Miho; Sakata, Ichiro; Sakai, Takafumi; Tomioka, Hiroaki; Nishigaki, Koichi; Tramier, Marc; Coppey-Moisan, Maïté

    2015-12-15

    Cytometry is a versatile and powerful method applicable to different fields, particularly pharmacology and biomedical studies. Based on the data obtained, cytometric studies are classified into high-throughput (HTP) or high-content screening (HCS) groups. However, assays combining the advantages of both are required to facilitate research. In this study, we developed a high-throughput system to profile cellular populations in terms of time- or dose-dependent responses to apoptotic stimulations because apoptotic inducers are potent anticancer drugs. We previously established assay systems involving protease to monitor live cells for apoptosis using tunable fluorescence resonance energy transfer (FRET)-based bioprobes. These assays can be used for microscopic analyses or fluorescence-activated cell sorting. In this study, we developed FRET-based bioprobes to detect the activity of the apoptotic markers caspase-3 and caspase-9 via changes in bioprobe fluorescence lifetimes using a flow cytometer for direct estimation of FRET efficiencies. Different patterns of changes in the fluorescence lifetimes of these markers during apoptosis were observed, indicating a relationship between discrete steps in the apoptosis process. The findings demonstrate the feasibility of evaluating collective cellular dynamics during apoptosis. Copyright © 2015 Elsevier Inc. All rights reserved.

  15. Fluorescence life-time imaging and steady state polarization for examining binding of fluorophores to gold nanoparticles.

    Science.gov (United States)

    Schwartz, Shmulik; Fixler, Dror; Popovtzer, Rachela; Shefi, Orit

    2015-11-01

    Nanocomposites as multifunctional agents are capable of combing imaging and cell biology technologies. The conventional methods used for validation of the conjugation process of nanoparticles (NPs) to fluorescent molecules such as spectroscopy analysis and surface potential measurements, are not sufficient. In this paper we present a new and highly sensitive procedure that uses the combination of (1) fluorescence spectrum, (2) fluorescence lifetime, and (3) steady state fluorescence polarization measurements. We characterize and analyze gold NPs with Lucifer yellow (LY) surface coating as a model. We demonstrate the ability to differentiate between LY-GNP (the conjugated complex) and a mixture of coated NP and free dyes. We suggest the approach for neuroscience applications where LY is used for detecting and labeling cells, studying morphology and intracellular communications. Histograms of Fluorescence lifetime imaging (FLIM) of free LY dye (Left) in comparison to the conjugated dye to gold nanoparticles, LY-GNP (Middle) enable the differentiation between LY-GNP (the conjugated complex) and a mixture of coated NP and free dyes (Right). © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. Online multispectral fluorescence lifetime values estimation and overlay onto tissue white-light video frames

    Science.gov (United States)

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

    2016-03-01

    Fluorescence lifetime imaging has been shown to be a robust technique for biochemical and functional characterization of tissues and to present great potential for intraoperative tissue diagnosis and guidance of surgical procedures. We report a technique for real-time mapping of fluorescence parameters (i.e. lifetime values) onto the location from where the fluorescence measurements were taken. This is achieved by merging a 450 nm aiming beam generated by a diode laser with the excitation light in a single delivery/collection fiber and by continuously imaging the region of interest with a color CMOS camera. The interrogated locations are then extracted from the acquired frames via color-based segmentation of the aiming beam. Assuming a Gaussian profile of the imaged aiming beam, the segmentation results are fitted to ellipses that are dynamically scaled at the full width of three automatically estimated thresholds (50%, 75%, 90%) of the Gaussian distribution's maximum value. This enables the dynamic augmentation of the white-light video frames with the corresponding fluorescence decay parameters. A fluorescence phantom and fresh tissue samples were used to evaluate this method with motorized and hand-held scanning measurements. At 640x512 pixels resolution the area of interest augmented with fluorescence decay parameters can be imaged at an average 34 frames per second. The developed method has the potential to become a valuable tool for real-time display of optical spectroscopy data during continuous scanning applications that subsequently can be used for tissue characterization and diagnosis.

  17. Single pulse two photon fluorescence lifetime imaging (SP-FLIM) with MHz pixel rate.

    Science.gov (United States)

    Eibl, Matthias; Karpf, Sebastian; Weng, Daniel; Hakert, Hubertus; Pfeiffer, Tom; Kolb, Jan Philip; Huber, Robert

    2017-07-01

    Two-photon-excited fluorescence lifetime imaging microscopy (FLIM) is a chemically specific 3-D sensing modality providing valuable information about the microstructure, composition and function of a sample. However, a more widespread application of this technique is hindered by the need for a sophisticated ultra-short pulse laser source and by speed limitations of current FLIM detection systems. To overcome these limitations, we combined a robust sub-nanosecond fiber laser as the excitation source with high analog bandwidth detection. Due to the long pulse length in our configuration, more fluorescence photons are generated per pulse, which allows us to derive the lifetime with a single excitation pulse only. In this paper, we show high quality FLIM images acquired at a pixel rate of 1 MHz. This approach is a promising candidate for an easy-to-use and benchtop FLIM system to make this technique available to a wider research community.

  18. Long-term fluorescence lifetime imaging of a genetically encoded sensor for caspase-3 activity in mouse tumor xenografts

    Science.gov (United States)

    Zherdeva, Victoria; Kazachkina, Natalia I.; Shcheslavskiy, Vladislav; Savitsky, Alexander P.

    2018-03-01

    Caspase-3 is known for its role in apoptosis and programmed cell death regulation. We detected caspase-3 activation in vivo in tumor xenografts via shift of mean fluorescence lifetimes of a caspase-3 sensor. We used the genetically encoded sensor TR23K based on the red fluorescent protein TagRFP and chromoprotein KFP linked by 23 amino acid residues (TagRFP-23-KFP) containing a specific caspase cleavage DEVD motif to monitor the activity of caspase-3 in tumor xenografts by means of fluorescence lifetime imaging-Forster resonance energy transfer. Apoptosis was induced by injection of paclitaxel for A549 lung adenocarcinoma and etoposide and cisplatin for HEp-2 pharynx adenocarcinoma. We observed a shift in lifetime distribution from 1.6 to 1.9 ns to 2.1 to 2.4 ns, which indicated the activation of caspase-3. Even within the same tumor, the lifetime varied presumably due to the tumor heterogeneity and the different depth of tumor invasion. Thus, processing time-resolved fluorescence images allows detection of both the cleaved and noncleaved states of the TR23K sensor in real-time mode during the course of several weeks noninvasively. This approach can be used in drug screening, facilitating the development of new anticancer agents as well as improvement of chemotherapy efficiency and its adaptation for personal treatment.

  19. Efficacy of photodynamic therapy against larvae of Aedes aegypti: confocal microscopy and fluorescence-lifetime imaging

    Science.gov (United States)

    de Souza, L. M.; Pratavieira, S.; Inada, N. M.; Kurachi, C.; Corbi, J.; Guimarães, F. E. G.; Bagnato, V. S.

    2014-03-01

    Recently a few demonstration on the use of Photodynamic Reaction as possibility to eliminate larvae that transmit diseases for men has been successfully demonstrated. This promising tool cannot be vastly used due to many problems, including the lake of investigation concerning the mechanisms of larvae killing as well as security concerning the use of photosensitizers in open environment. In this study, we investigate some of the mechanisms in which porphyrin (Photogem) is incorporated on the Aedes aegypti larvae previously to illumination and killing. Larvae at second instar were exposed to the photosensitizer and after 30 minutes imaged by a confocal fluorescence microscope. It was observed the presence of photosensitizer in the gut and at the digestive tract of the larva. Fluorescence-Lifetime Imaging showed greater photosensitizer concentration in the intestinal wall of the samples, which produces a strong decrease of the Photogem fluorescence lifetime. For Photodynamic Therapy exposition to different light doses and concentrations of porphyrin were employed. Three different light sources (LED, Fluorescent lamp, Sun light) also were tested. Sun light and fluorescent lamp shows close to 100% of mortality after 24 hrs. of illumination. These results indicate the potential use of photodynamic effect against the LARVAE of Aedes aegypti.

  20. Fluorescence lifetime of emitters with broad homogeneous linewidths modified in opal photonic crystals

    DEFF Research Database (Denmark)

    Nikolaev, Ivan S.; Lodahl, Peter; Vos, Willem L.

    2008-01-01

    We have investigated the dynamics of spontaneous emission from dye molecules embedded in opal photonic crystals. Fluorescence lifetimes of Rhodamine 6G (R6G) dye were measured as a function of both optical frequency and crystal lattice parameter of the polystyrene opals. Due to the broad...

  1. Fluorescence lifetime measurement with confocal endomicroscopy for direct analysis of tissue biochemistry in vivo

    Directory of Open Access Journals (Sweden)

    Youngjae Won

    2016-08-01

    Full Text Available Confocal endomicroscopy is a powerful tool for in vivo real-time imaging at cellular resolution inside a living body without tissue resection. Microscopic fluorescence lifetime measurement can provide information about localized biochemical conditions such as pH and the concentrations of oxygen and calcium. We hypothesized that combining these techniques could assist accurate cancer discrimination by providing both biochemical and morphological information. We designed a dual-mode experimental setup for confocal endomicroscopic imaging and fluorescence lifetime measurement and applied it to a mouse xenograft model of activated human pancreatic cancer generated by subcutaneous injection of AsPC-1 tumor cells. Using this method with pH-sensitive sodium fluorescein injection, we demonstrated discrimination between normal and cancerous tissues in a living mouse. With further development, this method may be useful for clinical cancer detection.

  2. Using multiphoton fluorescence lifetime imaging to characterize liver damage and fluorescein disposition in liver in vivo

    Science.gov (United States)

    Thorling, Camilla A.; Studier, Hauke; Crawford, Darrell; Roberts, Michael S.

    2016-03-01

    Liver disease is the fifth most common cause of death and unlike many other major causes of mortality, liver disease rates are increasing rather than decreasing. There is no ideal measurement of liver disease and although biopsies are the gold standard, this only allows for a spot examination and cannot follow dynamic processes of the liver. Intravital imaging has the potential to extract detailed information over a larger sampling area continuously. The aim of this project was to investigate whether multiphoton and fluorescence lifetime imaging microscopy could detect early liver damage and to assess whether it could detect changes in metabolism of fluorescein in normal and diseased livers. Four experimental groups were used in this study: 1) control; 2) ischemia reperfusion injury; 3) steatosis and 4) steatosis with ischemia reperfusion injury. Results showed that multiphoton microscopy could visualize morphological changes such as decreased fluorescence of endogenous fluorophores and the presence of lipid droplets, characteristic of steatosis. Fluorescence lifetime imaging microscopy showed increase in NADPH in steatosis with and without ischemia reperfusion injury and could detect changes in metabolism of fluorescein to fluorescein monoglurcuronide, which was impaired in steatosis with ischemia reperfusion injury. These results concluded that the combination of multiphoton microscopy and fluorescence lifetime imaging is a promising method of assessing early stage liver damage and that it can be used to study changes in drug metabolism in the liver as an indication of liver disease and has the potential to replace the traditional static liver biopsy currently used.

  3. GPU accelerated real-time confocal fluorescence lifetime imaging microscopy (FLIM) based on the analog mean-delay (AMD) method

    Science.gov (United States)

    Kim, Byungyeon; Park, Byungjun; Lee, Seungrag; Won, Youngjae

    2016-01-01

    We demonstrated GPU accelerated real-time confocal fluorescence lifetime imaging microscopy (FLIM) based on the analog mean-delay (AMD) method. Our algorithm was verified for various fluorescence lifetimes and photon numbers. The GPU processing time was faster than the physical scanning time for images up to 800 × 800, and more than 149 times faster than a single core CPU. The frame rate of our system was demonstrated to be 13 fps for a 200 × 200 pixel image when observing maize vascular tissue. This system can be utilized for observing dynamic biological reactions, medical diagnosis, and real-time industrial inspection. PMID:28018724

  4. A fusion-spliced near-field optical fiber probe using photonic crystal fiber for nanoscale thermometry based on fluorescence-lifetime measurement of quantum dots.

    Science.gov (United States)

    Fujii, Takuro; Taguchi, Yoshihiro; Saiki, Toshiharu; Nagasaka, Yuji

    2011-01-01

    We have developed a novel nanoscale temperature-measurement method using fluorescence in the near-field called fluorescence near-field optics thermal nanoscopy (Fluor-NOTN). Fluor-NOTN enables the temperature distributions of nanoscale materials to be measured in vivo/in situ. The proposed method measures temperature by detecting the temperature dependent fluorescence lifetimes of Cd/Se quantum dots (QDs). For a high-sensitivity temperature measurement, the auto-fluorescence generated from a fiber probe should be reduced. In order to decrease the noise, we have fabricated a novel near-field optical-fiber probe by fusion-splicing a photonic crystal fiber (PCF) and a conventional single-mode fiber (SMF). The validity of the novel fiber probe was assessed experimentally by evaluating the auto-fluorescence spectra of the PCF. Due to the decrease of auto-fluorescence, a six- to ten-fold increase of S/N in the near-field fluorescence lifetime detection was achieved with the newly fabricated fusion-spliced near-field optical fiber probe. Additionally, the near-field fluorescence lifetime of the quantum dots was successfully measured by the fabricated fusion-spliced near-field optical fiber probe at room temperature, and was estimated to be 10.0 ns.

  5. Fluorescence lifetime FRET imaging of receptor-ligand complexes in tumor cells in vitro and in vivo

    Science.gov (United States)

    Rudkouskaya, Alena; Sinsuebphon, Nattawut; Intes, Xavier; Mazurkiewicz, Joseph E.; Barroso, Margarida

    2017-02-01

    To guide the development of targeted therapies with improved efficacy and accelerated clinical acceptance, novel imaging methodologies need to be established. Toward this goal, fluorescence lifetime Förster resonance energy transfer (FLIM-FRET) imaging assays capitalize on the ability of antibodies or protein ligands to bind dimerized membrane bound receptors to measure their target engagement levels in cancer cells. Conventional FLIM FRET microscopy has been widely applied at visible wavelengths to detect protein-protein interactions in vitro. However, operation at these wavelengths restricts imaging quality and ability to quantitate lifetime changes in in vivo small animal optical imaging due to high auto-fluorescence and light scattering. Here, we have analyzed the uptake of iron-bound transferrin (Tf) probes into human breast cancer cells using FLIM-FRET microscopy in the visible and near-infrared (NIR) range. The development of NIR FLIM FRET microscopy allows for the use of quantitative lifetime-based molecular assays to measure drug-target engagement levels at multiple scales: from in vitro microscopy to in vivo small animal optical imaging (macroscopy). This novel approach can be extended to other receptors, currently targeted in oncology. Hence, lifetime-based molecular imaging can find numerous applications in drug delivery and targeted therapy assessment and optimization.

  6. Carrier Lifetimes in Fluorescent 6H-SiC for LEDs Application

    DEFF Research Database (Denmark)

    Grivickas, Vytautas; Gulbinas, Karolis; Jokubavičius, Valdas

    Recently it was shown a new approach based on all-semiconductor material technology which is composed with a near ultra-violet GaN LED excitation source and fluorescent silicon carbide (f-6H-SiC) substrate which generates a visible broad spectral light by N and B dopants and an efficient donor...... to acceptor pair recombination [1,2]. This combination can achieve higher electric-light conversion efficiency and high color rendering in comparison with today’s used blue GaN LED based and phosphors. The devices are promising candidates for general lightning applications and may obtain stability...... under co-linear and orthogonal probe geometry was used to measure carrier lifetimes in the layers under variable injection conditions. Same results are shown in Fig. 1 exaggerating the fact that longer electron lifetime responsible for higher emission and n-type doping should prevail the p-type doping...

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

    Science.gov (United States)

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

    2018-05-01

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

  8. Determination of the D mesons lifetimes

    International Nuclear Information System (INIS)

    Josa Mutuberria, M.I.

    1990-01-01

    The results from the experiment NA27, performed in the North Area of the Super Proton Synchrotron at CERN are presented. The experimental set up was the small, high resolution, rapid cyling bubble chamber LEBC coupled with the European Hybrid Spectrometer (EHS). More than 2 millions pictures were taken, with 1015000 in teractions in hydrogen. The stastistical sensitivity of the experiment was 38.5 events/μb. A clean sample of 700 charm particle decays was obtained. Estimators with minimal systematic and statistical errors are developed for the determination of the lifetimes of short-lived particles whose individual momenta are unknown. These estimators make use of the measured decay lengths and the a priori known production characteristics. In this way, it is possible to include identified but not fully reconstructed charm decays in the sample to determine their lifetime. The properties of these estimators were extensively studied by means of Montecarlo simulation. The detection of the short-lived particles through the impact parameter of their charged decay products leads to additional complications which are taken into account. The biases and statistical errors inherent in using simpler approximate lifetime estimators are also discussed. These estimators are applied to determine the lifetime of the D o and D +- mesons using the charm data sample from NA27. (Author)

  9. CONSTRAINING THE LIFETIME AND OPENING ANGLE OF QUASARS USING FLUORESCENT Ly α EMISSION: THE CASE OF Q0420–388

    International Nuclear Information System (INIS)

    Borisova, Elena; Lilly, Simon J.; Cantalupo, Sebastiano; Prochaska, J. Xavier; Rakic, Olivera; Worseck, Gabor

    2016-01-01

    A toy model is developed to understand how the spatial distribution of fluorescent emitters in the vicinity of bright quasars could be affected by the geometry of the quasar bi-conical radiation field and by its lifetime. The model is then applied to the distribution of high-equivalent-width Ly α emitters (with rest-frame equivalent widths above 100 Å, threshold used in, e.g., Trainor and Steidel) identified in a deep narrow-band 36 × 36 arcmin 2 image centered on the luminous quasar Q0420–388. These emitters are found near the edge of the field and show some evidence of an azimuthal asymmetry on the sky of the type expected if the quasar is radiating in a bipolar cone. If these sources are being fluorescently illuminated by the quasar, the two most distant objects require a lifetime of at least 15 Myr for an opening angle of 60° or more, increasing to more than 40 Myr if the opening angle is reduced to a minimum of 30°. However, some other expected signatures of boosted fluorescence are not seen at the current survey limits, e.g., a fall off in Ly α brightness, or equivalent width, with distance. Furthermore, to have most of the Ly α emission of the two distant sources to be fluorescently boosted would require the quasar to have been significantly brighter in the past. This suggests that these particular sources may not be fluorescent, invalidating the above lifetime constraints. This would cast doubt on the use of this relatively low equivalent width threshold and thus also on the lifetime analysis in Trainor and Steidel.

  10. CONSTRAINING THE LIFETIME AND OPENING ANGLE OF QUASARS USING FLUORESCENT Ly α EMISSION: THE CASE OF Q0420–388

    Energy Technology Data Exchange (ETDEWEB)

    Borisova, Elena; Lilly, Simon J.; Cantalupo, Sebastiano [Institute for Astronomy, ETH Zurich, Zurich, CH-8093 (Switzerland); Prochaska, J. Xavier [UCO/Lick Observatory, UC Santa Cruz, Santa Cruz, CA 95064 (United States); Rakic, Olivera; Worseck, Gabor, E-mail: borisova@phys.ethz.ch [Max-Planck-Institut für Astronomie, Heidelberg, D-69117 (Germany)

    2016-10-20

    A toy model is developed to understand how the spatial distribution of fluorescent emitters in the vicinity of bright quasars could be affected by the geometry of the quasar bi-conical radiation field and by its lifetime. The model is then applied to the distribution of high-equivalent-width Ly α emitters (with rest-frame equivalent widths above 100 Å, threshold used in, e.g., Trainor and Steidel) identified in a deep narrow-band 36 × 36 arcmin{sup 2} image centered on the luminous quasar Q0420–388. These emitters are found near the edge of the field and show some evidence of an azimuthal asymmetry on the sky of the type expected if the quasar is radiating in a bipolar cone. If these sources are being fluorescently illuminated by the quasar, the two most distant objects require a lifetime of at least 15 Myr for an opening angle of 60° or more, increasing to more than 40 Myr if the opening angle is reduced to a minimum of 30°. However, some other expected signatures of boosted fluorescence are not seen at the current survey limits, e.g., a fall off in Ly α brightness, or equivalent width, with distance. Furthermore, to have most of the Ly α emission of the two distant sources to be fluorescently boosted would require the quasar to have been significantly brighter in the past. This suggests that these particular sources may not be fluorescent, invalidating the above lifetime constraints. This would cast doubt on the use of this relatively low equivalent width threshold and thus also on the lifetime analysis in Trainor and Steidel.

  11. The increase of NADH fluorescence lifetime is associated with the metabolic change during osteogenic differentiation of human mesenchymal stem cells (hMSCs)

    Science.gov (United States)

    Guo, Han Wen; Yu, Jia Sin; Hsu, Shu Han; Wei, Yau Huei; Lee, Oscar K.; Wang, Hsing Wen

    2011-03-01

    Fluorescence lifetime of NADH had been used as an optical marker for monitoring cellular metabolism. In our pervious studies, we have demonstrated that NADH lifetime of hMSCs increase gradually with time of osteogenic differentiation. In this study, we measured NADH lifetime of hMSCs from a different donor as well as the corresponding metabolic indices such as ATP level, oxygen consumption and lactate release. We also measure the quantity of Complex I, III, IV and V. The results show that during differentiation more oxygen consumed, higher ATP level expressed and less lactate released, and the increase of NADH lifetime was associated with ATP level. Higher expression of the total Complex protein was observed at 3 and 4 weeks after differentiation than controls. However, Complex I expression did not show significant correlation with the increase of NADH fluorescence lifetime. In summary, we demonstrated that the change of NADH lifetime was associated with the metabolic change during osteogenic differentiation of hMSCs. The increase of NADH lifetime was in part due to the increased Complex protein interaction in mitochondria after differentiation.

  12. Distribution of diffusion times determined by fluorescence (lifetime) correlation spectroscopy

    Czech Academy of Sciences Publication Activity Database

    Pánek, Jiří; Loukotová, Lenka; Hrubý, Martin; Štěpánek, Petr

    2018-01-01

    Roč. 51, č. 8 (2018), s. 2796-2804 ISSN 0024-9297 R&D Projects: GA MŠk(CZ) LO1507 Institutional support: RVO:61389013 Keywords : polymer solution * fluorescence correlation spectroscopy * diffusion time distribution Subject RIV: CD - Macromolecular Chemistry OBOR OECD: Polymer science Impact factor: 5.835, year: 2016

  13. The modifier effects of chymotrypsin and trypsin enzymes on fluorescence lifetime distribution of "N-(1-pyrenyl)maleimide-bovine serum albumin" complex

    Science.gov (United States)

    Özyiğit, İbrahim Ethem; Karakuş, Emine; Pekcan, Önder

    2016-02-01

    Chymotrypsin and trypsin are the well known proteolytic enzymes, both of which are synthesized in the pancreas as their precursors - the inactive forms; chymotrypsinogen and trypsinogen - and then are released into the duodenum to cut proteins into smaller peptides. In this paper, the effects of activities of chymotrypsin and trypsin enzymes on fluorescence lifetime distributions of the substrat bovine serum albumin (BSA) modified with N-(1-pyrenyl)maleimide (PM) were examined. In the labeling study of BSA with PM, it is aimed to attach PM to the single free thiol (Cys34) and to all the free amine groups in accessible positions in order to produce excimers of pyrene planes of the possible highest amount to form the lifetime distributions in the widest range, that may show specifically distinguishing changes resulting from the activities of the proteases. The time resolved spectrofluorometer was used to monitor fluorescence decays, which were analyzed by using the exponential series method (ESM) to obtain the changes of lifetime distributions. After the exposure of the synthesized substrat PM-BSA to the enzymes, the fluorescence lifetime distributions exhibited different structures which were attributed to the different activities of the proteases.

  14. Laser induced fluorescence lifetime characterization of Bacillus endospore species using time correlated single photon counting analysis with the multi-exponential fit method

    Science.gov (United States)

    Smith, Clint; Edwards, Jarrod; Fisher, Andmorgan

    2010-04-01

    Rapid detection of biological material is critical for determining presence/absence of bacterial endospores within various investigative programs. Even more critical is that if select material tests positive for bacillus endospores then tests should provide data at the species level. Optical detection of microbial endospore formers such as Bacillus sp. can be heavy, cumbersome, and may only identify at the genus level. Data provided from this study will aid in characterization needed by future detection systems for further rapid breakdown analysis to gain insight into a more positive signature collection of Bacillus sp. Literature has shown that fluorescence spectroscopy of endospores could be statistically separated from other vegetative genera, but could not be separated among one another. Results of this study showed endospore species separation is possible using laser-induce fluorescence with lifetime decay analysis for Bacillus endospores. Lifetime decays of B. subtilis, B. megaterium, B. coagulans, and B. anthracis Sterne strain were investigated. Using the Multi-Exponential fit method data showed three distinct lifetimes for each species within the following ranges, 0.2-1.3 ns; 2.5-7.0 ns; 7.5-15.0 ns, when laser induced at 307 nm. The four endospore species were individually separated using principle component analysis (95% CI).

  15. FUNDUS AUTOFLUORESCENCE LIFETIMES AND CENTRAL SEROUS CHORIORETINOPATHY.

    Science.gov (United States)

    Dysli, Chantal; Berger, Lieselotte; Wolf, Sebastian; Zinkernagel, Martin S

    2017-11-01

    To quantify retinal fluorescence lifetimes in patients with central serous chorioretinopathy (CSC) and to identify disease specific lifetime characteristics over the course of disease. Forty-seven participants were included in this study. Patients with central serous chorioretinopathy were imaged with fundus photography, fundus autofluorescence, optical coherence tomography, and fluorescence lifetime imaging ophthalmoscopy (FLIO) and compared with age-matched controls. Retinal autofluorescence was excited using a 473-nm blue laser light and emitted fluorescence light was detected in 2 distinct wavelengths channels (498-560 nm and 560-720 nm). Clinical features, mean retinal autofluorescence lifetimes, autofluorescence intensity, and corresponding optical coherence tomography (OCT) images were further analyzed. Thirty-five central serous chorioretinopathy patients with a mean visual acuity of 78 ETDRS letters (range, 50-90; mean Snellen equivalent: 20/32) and 12 age-matched controls were included. In the acute stage of central serous chorioretinopathy, retinal fluorescence lifetimes were shortened by 15% and 17% in the respective wavelength channels. Multiple linear regression analysis showed that fluorescence lifetimes were significantly influenced by the disease duration (P autofluorescence lifetimes, particularly in eyes with retinal pigment epithelial atrophy, were associated with poor visual acuity. This study establishes that autofluorescence lifetime changes occurring in central serous chorioretinopathy exhibit explicit patterns which can be used to estimate perturbations of the outer retinal layers with a high degree of statistical significance.

  16. Determination of absolute Ba densities during dimming operation of fluorescent lamps by laser-induced fluorescence measurements

    International Nuclear Information System (INIS)

    Hadrath, S; Beck, M; Garner, R C; Lieder, G; Ehlbeck, J

    2007-01-01

    Investigations of fluorescent lamps (FL) are often focused on the electrodes, since the lifetime of the lamps is typically limited by the electrode lifetime and durability. During steady state operation, the work function lowering emitter material, in particular, barium, is lost. Greater barium losses occur under dimming conditions, in which reduced discharge currents lead to increased cathode falls, the result of the otherwise diminished heating of the electrode by the bombarding plasma ions. In this work the barium density near the electrodes of (FL), operating in high frequency dimming mode is investigated using the high-sensitivity method of laser-induced fluorescence. From these measurements we infer barium loss for a range of discharge currents and auxiliary coil heating currents. We show that the Ba loss can very easily be reduced by moderate auxiliary coil heating

  17. The modifier effects of chymotrypsin and trypsin enzymes on fluorescence lifetime distribution of "N-(1-pyrenyl)maleimide-bovine serum albumin" complex.

    Science.gov (United States)

    Özyiğit, İbrahim Ethem; Karakuş, Emine; Pekcan, Önder

    2016-02-05

    Chymotrypsin and trypsin are the well known proteolytic enzymes, both of which are synthesized in the pancreas as their precursors - the inactive forms; chymotrypsinogen and trypsinogen - and then are released into the duodenum to cut proteins into smaller peptides. In this paper, the effects of activities of chymotrypsin and trypsin enzymes on fluorescence lifetime distributions of the substrat bovine serum albumin (BSA) modified with N-(1-pyrenyl)maleimide (PM) were examined. In the labeling study of BSA with PM, it is aimed to attach PM to the single free thiol (Cys34) and to all the free amine groups in accessible positions in order to produce excimers of pyrene planes of the possible highest amount to form the lifetime distributions in the widest range, that may show specifically distinguishing changes resulting from the activities of the proteases. The time resolved spectrofluorometer was used to monitor fluorescence decays, which were analyzed by using the exponential series method (ESM) to obtain the changes of lifetime distributions. After the exposure of the synthesized substrat PM-BSA to the enzymes, the fluorescence lifetime distributions exhibited different structures which were attributed to the different activities of the proteases. Copyright © 2015 Elsevier B.V. All rights reserved.

  18. Polar plot representation of time-resolved fluorescence.

    Science.gov (United States)

    Eichorst, John Paul; Wen Teng, Kai; Clegg, Robert M

    2014-01-01

    Measuring changes in a molecule's fluorescence emission is a common technique to study complex biological systems such as cells and tissues. Although the steady-state fluorescence intensity is frequently used, measuring the average amount of time that a molecule spends in the excited state (the fluorescence lifetime) reveals more detailed information about its local environment. The lifetime is measured in the time domain by detecting directly the decay of fluorescence following excitation by short pulse of light. The lifetime can also be measured in the frequency domain by recording the phase and amplitude of oscillation in the emitted fluorescence of the sample in response to repetitively modulated excitation light. In either the time or frequency domain, the analysis of data to extract lifetimes can be computationally intensive. For example, a variety of iterative fitting algorithms already exist to determine lifetimes from samples that contain multiple fluorescing species. However, recently a method of analysis referred to as the polar plot (or phasor plot) is a graphical tool that projects the time-dependent features of the sample's fluorescence in either the time or frequency domain into the Cartesian plane to characterize the sample's lifetime. The coordinate transformations of the polar plot require only the raw data, and hence, there are no uncertainties from extensive corrections or time-consuming fitting in this analysis. In this chapter, the history and mathematical background of the polar plot will be presented along with examples that highlight how it can be used in both cuvette-based and imaging applications.

  19. Fluorescence lifetime imaging of microviscosity changes during ER autophagy in live cells

    Science.gov (United States)

    He, Ying; Samanta, Soham; Gong, Wanjun; Liu, Wufan; Pan, Wenhui; Yang, Zhigang; Qu, Junle

    2018-02-01

    Unfolded or misfolded protein accumulation inside Endoplasmic Reticulum (ER) will cause ER stress and subsequently will activate cellular autophagy to release ER stress, which would ultimately result in microviscosity changes. However, even though, it is highly significant to gain a quantitative assessment of microviscosity changes during ER autophagy to study ER stress and autophagy behaviors related diseases, it has rarely been reported yet. In this work, we have reported a BODIPY based fluorescent molecular rotor that can covalently bind with vicinal dithiols containing nascent proteins in ER and hence can result in ER stress through the inhibition of the folding of nascent proteins. The change in local viscosity, caused by the release of the stress in cells through autophagy, was quantified by the probe using fluorescence lifetime imaging. This work basically demonstrates the possibility of introducing synthetic chemical probe as a promising tool to diagnose ER-viscosity-related diseases.

  20. Fluorescence uranium determination

    International Nuclear Information System (INIS)

    Fernandez Cellini, R.; Crus Castillo, F. de la; Barrera Pinero, R.

    1960-01-01

    An equipment for analysis of uranium by fluorescence was developed in order to determine it at such a low concentration that it can not be determined by the most sensible analytical methods. this new fluorimeter was adapted to measure the fluorescence emitted by the phosphorus sodium fluoride-sodium carbonate-potasium carbonate-uranyl, being excited by ultraviolet light of 3,650 A the intensity of the light emitted was measure with a photomultiplicator RCA 5819 and the adequate electronic equipment. (Author) 19 refs

  1. Determination of the effective radiative lifetimes of the 6 3P1 atomic mercury level in low-pressure mercury discharges

    International Nuclear Information System (INIS)

    van de Weijer, P.; Cremers, R.M.M.

    1984-01-01

    Experiments are described in which low-pressure mercury, mercury-argon and mercury-krypton discharges were irradiated with a dye laser pulse at 365.5 nm, thus exciting mercury atoms from the metastable 6 3 P 2 level to the 6 3 D 2 level. The 6 3 D 2 level decays radiatively to the 6 P levels. By recording the time dependence of the overpopulation in the 6 3 P 1 and the 6 1 P 1 level at the fluorescence signals at 254 nm and 185 nm, respectively, the effective radiative lifetime of these levels were determined. The effective radiative lifetime of the 6 3 P 1 level was measured in the k 0 R regime 0.1-500. The 6 1 P 1 lifetime was determined for the following discharge conditions: tube diameter 10-36 mm, mercury density 7.10 18 -2.10 21 m -3 , and noble gas pressure 0, 130, 400 Pa

  2. In vivo imaging of cerebral energy metabolism with two-photon fluorescence lifetime microscopy of NADH.

    Science.gov (United States)

    Yaseen, Mohammad A; Sakadžić, Sava; Wu, Weicheng; Becker, Wolfgang; Kasischke, Karl A; Boas, David A

    2013-02-01

    Minimally invasive, specific measurement of cellular energy metabolism is crucial for understanding cerebral pathophysiology. Here, we present high-resolution, in vivo observations of autofluorescence lifetime as a biomarker of cerebral energy metabolism in exposed rat cortices. We describe a customized two-photon imaging system with time correlated single photon counting detection and specialized software for modeling multiple-component fits of fluorescence decay and monitoring their transient behaviors. In vivo cerebral NADH fluorescence suggests the presence of four distinct components, which respond differently to brief periods of anoxia and likely indicate different enzymatic formulations. Individual components show potential as indicators of specific molecular pathways involved in oxidative metabolism.

  3. Multimodal optical coherence tomography and fluorescence lifetime imaging with interleaved excitation sources for simultaneous endogenous and exogenous fluorescence.

    Science.gov (United States)

    Shrestha, Sebina; Serafino, Michael J; Rico-Jimenez, Jesus; Park, Jesung; Chen, Xi; Zhaorigetu, Siqin; Walton, Brian L; Jo, Javier A; Applegate, Brian E

    2016-09-01

    Multimodal imaging probes a variety of tissue properties in a single image acquisition by merging complimentary imaging technologies. Exploiting synergies amongst the data, algorithms can be developed that lead to better tissue characterization than could be accomplished by the constituent imaging modalities taken alone. The combination of optical coherence tomography (OCT) with fluorescence lifetime imaging microscopy (FLIM) provides access to detailed tissue morphology and local biochemistry. The optical system described here merges 1310 nm swept-source OCT with time-domain FLIM having excitation at 355 and 532 nm. The pulses from 355 and 532 nm lasers have been interleaved to enable simultaneous acquisition of endogenous and exogenous fluorescence signals, respectively. The multimodal imaging system was validated using tissue phantoms. Nonspecific tagging with Alexa Flour 532 in a Watanbe rabbit aorta and active tagging of the LOX-1 receptor in human coronary artery, demonstrate the capacity of the system for simultaneous acquisition of OCT, endogenous FLIM, and exogenous FLIM in tissues.

  4. Gentamicin differentially alters cellular metabolism of cochlear hair cells as revealed by NAD(P)H fluorescence lifetime imaging

    Science.gov (United States)

    Zholudeva, Lyandysha V.; Ward, Kristina G.; Nichols, Michael G.; Smith, Heather Jensen

    2015-05-01

    Aminoglycoside antibiotics are implicated as culprits of hearing loss in more than 120,000 individuals annually. Research has shown that the sensory cells, but not supporting cells, of the cochlea are readily damaged and/or lost after use of such antibiotics. High-frequency outer hair cells (OHCs) show a greater sensitivity to antibiotics than high- and low-frequency inner hair cells (IHCs). We hypothesize that variations in mitochondrial metabolism account for differences in susceptibility. Fluorescence lifetime microscopy was used to quantify changes in NAD(P)H in sensory and supporting cells from explanted murine cochleae exposed to mitochondrial uncouplers, inhibitors, and an ototoxic antibiotic, gentamicin (GM). Changes in metabolic state resulted in a redistribution of NAD(P)H between subcellular fluorescence lifetime pools. Supporting cells had a significantly longer lifetime than sensory cells. Pretreatment with GM increased NAD(P)H intensity in high-frequency sensory cells, as well as the NAD(P)H lifetime within IHCs. GM specifically increased NAD(P)H concentration in high-frequency OHCs, but not in IHCs or pillar cells. Variations in NAD(P)H intensity in response to mitochondrial toxins and GM were greatest in high-frequency OHCs. These results demonstrate that GM rapidly alters mitochondrial metabolism, differentially modulates cell metabolism, and provides evidence that GM-induced changes in metabolism are significant and greatest in high-frequency OHCs.

  5. Preparation and properties of Nd{sup 3+}:SrAlF{sub 5} nanocrystals embedded fluorophosphate transparent glass-ceramic with long fluorescence lifetime

    Energy Technology Data Exchange (ETDEWEB)

    Zheng, Ruilin; Wang, Jinlong; Zhang, Liaolin; Liu, Chunxiao; Wei, Wei [Nanjing University of Posts and Telecommunications, School of Optoelectronic Engineering, Nanjing (China)

    2016-07-15

    Nd{sup 3+}:SrAlF{sub 5} nanocrystals embedded fluorophosphate glass-ceramics were prepared by the melt quenching and subsequent thermal treatment method. The formation of SrAlF{sub 5} nanocrystals in the glass was confirmed by X-ray diffraction and scanning electron microscope. The fluorescence intensity and lifetime of the glass-ceramics increased with the increase of size of nanocrystals. Importantly, by controlling growth of nanocrystals, an obvious enhancement of lifetime (725 μs) emerged in the glass-ceramics heat-treated at 510 C and the transmittance can reach to 72.2 % at 1049 nm. The enhanced fluorescence intensity and lifetime were ascribed to the comfortable local environment to the Nd{sup 3+} ion and scattering of the nanoparticle embedded into the glass matrix. (orig.)

  6. Lifetime-based optical sensor for high-level pCO2 detection employing fluorescence resonance energy transfer

    International Nuclear Information System (INIS)

    Bueltzingsloewen, Christoph von; McEvoy, Aisling K.; McDonagh, Colette; MacCraith, Brian D.

    2003-01-01

    An optical sensor for the measurement of high levels of carbon dioxide in gas phase has been developed. It is based on fluorescence resonance energy transfer (FRET) between a long-lifetime ruthenium polypyridyl complex and the pH-active disazo dye Sudan III. The donor luminophore and the acceptor dye are both immobilised in a hydrophobic silica sol-gel/ethyl cellulose hybrid matrix material. Tetraoctylammonium hydroxide (TOA-OH) is used as an internal buffering system. Fluorescence lifetime is measured in the frequency domain, using low-cost phase modulation measurement technology. The use of Sudan III as an acceptor dye has enabled the sensor to have a dynamic range up to 100% carbon dioxide. The sensor displays 11.2 deg. phase shift between the limit of detection (LOD) of 0.06 and 100% CO 2 with a resolution of better than 2%. The encapsulation in the silica/polymer hybrid material has provided the sensor with good mechanical and chemical stability. The effect of molecular oxygen, humidity and temperature on the sensor performance was studied in detail

  7. Measurement of pH micro-heterogeneity in natural cheese matrices by fluorescence lifetime imaging

    Directory of Open Access Journals (Sweden)

    Zuzana eBurdikova

    2015-03-01

    Full Text Available Cheese, a product of microbial fermentation may be defined as a protein matrix entrapping fat, moisture, minerals and solutes as well as dispersed bacterial colonies. The growth and physiology of bacterial cells in these colonies may be influenced by the microenvironment around the colony, or alternatively the cells within the colony may modify the microenvironment (e.g. pH, redox potential due to their metabolic activity. While cheese pH may be measured at macro level there remains a significant knowledge gap relating to the degree of micro-heterogeneity of pH within the cheese matrix and its relationship with microbial, enzymatic and physiochemical parameters and ultimately with cheese quality, consistency and ripening patterns. The pH of cheese samples was monitored both at macroscopic scale and at microscopic scale, using a non-destructive microscopic technique employing C-SNARF-4 and Oregon Green 488 fluorescent probes. The objectives of this work were to evaluate the suitability of these dyes for microscale pH measurements in natural cheese matrices and to enhance the sensitivity and extend the useful pH range of these probes using fluorescence lifetime imaging (FLIM. In particular, fluorescence lifetime of Oregon Green 488 proved to be sensitive probe to map pH micro heterogeneity within cheese matrices. Good agreement was observed between macroscopic scale pH measurement by FLIM and by traditional pH methods, but in addition considerable localized microheterogeneity in pH was evident within the curd matrix with pH range between 4.0 and 5.5. This technique provides significant potential to further investigate the relationship between cheese matrix physico-chemistry and bacterial metabolism during cheese manufacture and ripening.

  8. Measurement of pH micro-heterogeneity in natural cheese matrices by fluorescence lifetime imaging.

    Science.gov (United States)

    Burdikova, Zuzana; Svindrych, Zdenek; Pala, Jan; Hickey, Cian D; Wilkinson, Martin G; Panek, Jiri; Auty, Mark A E; Periasamy, Ammasi; Sheehan, Jeremiah J

    2015-01-01

    Cheese, a product of microbial fermentation may be defined as a protein matrix entrapping fat, moisture, minerals and solutes as well as dispersed bacterial colonies. The growth and physiology of bacterial cells in these colonies may be influenced by the microenvironment around the colony, or alternatively the cells within the colony may modify the microenvironment (e.g., pH, redox potential) due to their metabolic activity. While cheese pH may be measured at macro level there remains a significant knowledge gap relating to the degree of micro-heterogeneity of pH within the cheese matrix and its relationship with microbial, enzymatic and physiochemical parameters and ultimately with cheese quality, consistency and ripening patterns. The pH of cheese samples was monitored both at macroscopic scale and at microscopic scale, using a non-destructive microscopic technique employing C-SNARF-4 and Oregon Green 488 fluorescent probes. The objectives of this work were to evaluate the suitability of these dyes for microscale pH measurements in natural cheese matrices and to enhance the sensitivity and extend the useful pH range of these probes using fluorescence lifetime imaging (FLIM). In particular, fluorescence lifetime of Oregon Green 488 proved to be sensitive probe to map pH micro heterogeneity within cheese matrices. Good agreement was observed between macroscopic scale pH measurement by FLIM and by traditional pH methods, but in addition considerable localized microheterogeneity in pH was evident within the curd matrix with pH range between 4.0 and 5.5. This technique provides significant potential to further investigate the relationship between cheese matrix physico-chemistry and bacterial metabolism during cheese manufacture and ripening.

  9. Segmented frequency-domain fluorescence lifetime measurements: minimizing the effects of photobleaching within a multi-component system.

    Science.gov (United States)

    Marwani, Hadi M; Lowry, Mark; Keating, Patrick; Warner, Isiah M; Cook, Robert L

    2007-11-01

    This study introduces a newly developed frequency segmentation and recombination method for frequency-domain fluorescence lifetime measurements to address the effects of changing fractional contributions over time and minimize the effects of photobleaching within multi-component systems. Frequency segmentation and recombination experiments were evaluated using a two component system consisting of fluorescein and rhodamine B. Comparison of experimental data collected in traditional and segmented fashion with simulated data, generated using different changing fractional contributions, demonstrated the validity of the technique. Frequency segmentation and recombination was also applied to a more complex system consisting of pyrene with Suwannee River fulvic acid reference and was shown to improve recovered lifetimes and fractional intensity contributions. It was observed that photobleaching in both systems led to errors in recovered lifetimes which can complicate the interpretation of lifetime results. Results showed clear evidence that the frequency segmentation and recombination method reduced errors resulting from a changing fractional contribution in a multi-component system, and allowed photobleaching issues to be addressed by commercially available instrumentation.

  10. Comparitive study of fluorescence lifetime quenching of rhodamine 6G by MoS2 and Au-MoS2

    Science.gov (United States)

    Shakya, Jyoti; Kasana, Parath; Mohanty, T.

    2018-04-01

    Time resolved fluorescence study of Rhodamine 6G (R6G) in the presence of Molybdenum disulfide (MoS2) nanosheets and gold doped MoS2 (Au-MoS2) have been carried out and discussed. We have analyzed the fluorescence decay curves of R6G and it is observed that Au-MoS2 is a better fluorescence lifetime quencher as compare to MoS2 nanosheets. Also, the energy transfer efficiency and energy transfer rate from R6G to MoS2 and Au-MoS2 has been calculated and found higher for Au-MoS2.

  11. Exciton-polaron quenching in organic thin-film transistors studied by fluorescence lifetime imaging microscopy

    DEFF Research Database (Denmark)

    Jensen, Per Baunegaard With; Leißner, Till; Osadnik, Andreas

    Organic semiconductors show great potential in electronic and optical applications. However, a major challenge is the degradation of the semiconductor materials that cause a reduction in device performance. Here, we present our investigations of Organic Thin Film Transistors (OTFT) based...... that correlates with the local charge density indicates a pronounced exciton quenching by the injected charges. Subsequent FLIM measurements on previously biased OTFT devices show a general decrease in fluorescence lifetime suggesting degradation of the organic semiconductor. This is correlated with the results...

  12. Mercury effects on Thalassiosira weissflogii: Applications of two-photon excitation chlorophyll fluorescence lifetime imaging and flow cytometry

    International Nuclear Information System (INIS)

    Wu Yun; Zeng Yan; Qu, Jianan Y.; Wang Wenxiong

    2012-01-01

    The toxic effects of inorganic mercury [Hg(II)] and methylmercury (MeHg) on the photosynthesis and population growth in a marine diatom Thalassiosira weissflogii were investigated using two methods: two-photon excitation fluorescence lifetime imaging (FLIM) and flow cytometry (FCM). For photosynthesis, Hg(II) exposure increased the average chlorophyll fluorescence lifetime, whereas such increment was not found under MeHg stress. This may be caused by the inhibitory effect of Hg(II) instead of MeHg on the electron transport chain. For population growth, modeled specific growth rate data showed that the reduction in population growth by Hg(II) mainly resulted from an increased number of injured cells, while the live cells divided at the normal rates. However, MeHg inhibitory effects on population growth were contributed by the reduced division rates of all cells. Furthermore, the cell images and the FCM data reflected the morphological changes of diatom cells under Hg(II)/MeHg exposure vividly and quantitatively. Our results demonstrated that the toxigenicity mechanisms between Hg(II) and MeHg were different in the algal cells.

  13. Fluorescence lifetime components reveal kinetic intermediate states upon equilibrium denaturation of carbonic anhydrase II.

    Science.gov (United States)

    Nemtseva, Elena V; Lashchuk, Olesya O; Gerasimova, Marina A; Melnik, Tatiana N; Nagibina, Galina S; Melnik, Bogdan S

    2017-12-21

    In most cases, intermediate states of multistage folding proteins are not 'visible' under equilibrium conditions but are revealed in kinetic experiments. Time-resolved fluorescence spectroscopy was used in equilibrium denaturation studies. The technique allows for detecting changes in the conformation and environment of tryptophan residues in different structural elements of carbonic anhydrase II which in its turn has made it possible to study the intermediate states of carbonic anhydrase II under equilibrium conditions. The results of equilibrium and kinetic experiments using wild-type bovine carbonic anhydrase II and its mutant form with the substitution of leucine for alanine at position 139 (L139A) were compared. The obtained lifetime components of intrinsic tryptophan fluorescence allowed for revealing that, the same as in kinetic experiments, under equilibrium conditions the unfolding of carbonic anhydrase II ensues through formation of intermediate states.

  14. Design, construction, and validation of a rotary multifunctional intravascular diagnostic catheter combining multispectral fluorescence lifetime imaging and intravascular ultrasound.

    Science.gov (United States)

    Bec, Julien; Xie, Hongtao; Yankelevich, Diego R; Zhou, Feifei; Sun, Yang; Ghata, Narugopal; Aldredge, Ralph; Marcu, Laura

    2012-10-01

    We report the development and validation of an intravascular rotary catheter for bimodal interrogation of arterial pathologies. This is based on a point-spectroscopy scanning time-resolved fluorescence spectroscopy technique enabling reconstruction of fluorescence lifetime images (FLIm) and providing information on arterial intima composition and intravascular ultrasound (IVUS) providing information on arterial wall morphology. The catheter design allows for independent rotation of the ultrasonic and optical channels within an 8 Fr outer diameter catheter sheath and integrates a low volume flushing channel for blood removal in the optical pathways. In the current configuration, the two channels consist of (a) a standard 3 Fr IVUS catheter with single element transducer (40 MHz) and (b) a side-viewing fiber optic (400 μm core). Experiments conducted in tissue phantoms showed the ability of the catheter to operate in an intraluminal setting and to generate coregistered FLIm and IVUS in one pull-back scan. Current results demonstrate the feasibility of the catheter for simultaneous bimodal interrogation of arterial lumen and for generation of robust fluorescence lifetime data under IVUS guidance. These results facilitate further development of a FLIm-IVUS technique for intravascular diagnosis of atherosclerotic cardiovascular diseases including vulnerable plaques.

  15. Effect of pharmacologically induced retinal degeneration on retinal autofluorescence lifetimes in mice.

    Science.gov (United States)

    Dysli, Chantal; Dysli, Muriel; Zinkernagel, Martin S; Enzmann, Volker

    2016-12-01

    Fluorescence lifetime imaging ophthalmoscopy (FLIO) was used to investigate retinal autofluorescence lifetimes in mouse models of pharmacologically induced retinal degeneration over time. Sodium iodate (NaIO 3 , 35 mg/kg intravenously) was used to induce retinal pigment epithelium (RPE) degeneration with subsequent loss of photoreceptors (PR) whereas N-methyl-N-nitrosourea (MNU, 45 mg/kg intraperitoneally) was employed for degeneration of the photoreceptor cell layer alone. All mice were measured at day 3, 7, 14, and 28 after the respective injection of NaIO 3 , MNU or NaCl (control). Fluorescence lifetime imaging was performed using a fluorescence lifetime imaging ophthalmoscope (Heidelberg Engineering, Heidelberg, Germany). Fluorescence was excited at 473 nm and fluorescence lifetimes were measured in a short and a long spectral channel (498-560 nm and 560-720 nm). Corresponding optical coherence tomography (OCT) images were consecutively acquired and histology was performed at the end of the experiments. Segmentation of OCT images and histology verified the cell type-specific degeneration process over time. Retinal autofluorescence lifetimes increased from day 3 to day 28 in mice after NaIO 3 treatment. Finally, at day 28, fluorescence lifetimes were prolonged by 8% in the short and 61% in the long spectral channel compared to control animals (p = 0.21 and p = 0.004, respectively). In mice after MNU treatment, the mean retinal autofluorescence lifetimes were already decreased at day 3 and retinal lifetimes were finally shortened by 27% in the short and 51% in the long spectral channel at day 28 (p = 0.0028). In conclusion, degeneration of the RPE with subsequent photoreceptor degeneration by NaIO 3 lead to longer mean fluorescence lifetimes of the retina compared to control mice, whereas during specific degeneration of the photoreceptor layer induced by MNU shorter lifetimes were measured. Therefore, short retinal fluorescence lifetimes may originate

  16. Characterization of a pulsed x-ray source for fluorescent lifetime measurements

    International Nuclear Information System (INIS)

    Blankespoor, S.C.; Derenzo, S.E.; Moses, W.W.; Rossington, C.S.; Ito, M.; Oba, K.

    1994-01-01

    To search for new, fast, inorganic scintillators, the authors have developed a bench-top pulsed x-ray source for determining fluorescent lifetimes and wavelengths of compounds in crystal or powdered form. This source uses a light-excited x-ray tube which produces x-rays when light from a laser diode strikes its photocathode. The x-ray tube has a tungsten anode, a beryllium exit window, a 30 kV maximum tube bias, and a 50 μA maximum average cathode current. The laser produces 3 x 10 7 photons at 650 nm per ∼100 ps pulse, with up to 10 7 pulses/sec. The time spread for the laser diode, x-ray tube, and a microchannel plate photomultiplier tube is less than 120 ps fwhm. The mean x-ray energy at tube biases of 20, 25, and 30 kV is 9.4, 10.3, and 11.1 keV, respectively. The authors measured 140, 230, and 330 x-ray photons per laser diode pulse per steradian, at tube biases of 20, 25, and 30 kV, respectively. Background x-rays due to dark current occur at a rate of 1 x 10 6 and 3 x 10 6 photons/sec/steradian at biases of 25 and 30 kV, respectively. Data characterizing the x-ray output with an aluminum filter in the x-ray beam are also presented

  17. High-sensitivity determination of Zn(II) and Cu(II) in vitro by fluorescence polarization

    Science.gov (United States)

    Thompson, Richard B.; Maliwal, Badri P.; Feliccia, Vincent; Fierke, Carol A.

    1998-04-01

    Recent work has suggested that free Cu(II) may play a role in syndromes such as Crohn's and Wilson's diseases, as well as being a pollutant toxic at low levels to shellfish and sheep. Similarly, Zn(II) has been implicated in some neural damage in the brain resulting from epilepsy and ischemia. Several high sensitivity methods exist for determining these ions in solution, including GFAAS, ICP-MS, ICP-ES, and electrochemical techniques. However, these techniques are generally slow and costly, require pretreatment of the sample, require complex instruments and skilled personnel, and are incapable of imaging at the cellular and subcellular level. To address these shortcomings we developed fluorescence polarization (anisotropy) biosensing methods for these ions which are very sensitivity, highly selective, require simple instrumentation and little pretreatment, and are inexpensive. Thus free Cu(II) or Zn(II) can be determined at picomolar levels by changes in fluorescence polarization, lifetime, or wavelength ratio using these methods; these techniques may be adapted to microscopy.

  18. Automated detection of breast cancer in resected specimens with fluorescence lifetime imaging

    Science.gov (United States)

    Phipps, Jennifer E.; Gorpas, Dimitris; Unger, Jakob; Darrow, Morgan; Bold, Richard J.; Marcu, Laura

    2018-01-01

    Re-excision rates for breast cancer lumpectomy procedures are currently nearly 25% due to surgeons relying on inaccurate or incomplete methods of evaluating specimen margins. The objective of this study was to determine if cancer could be automatically detected in breast specimens from mastectomy and lumpectomy procedures by a classification algorithm that incorporated parameters derived from fluorescence lifetime imaging (FLIm). This study generated a database of co-registered histologic sections and FLIm data from breast cancer specimens (N  =  20) and a support vector machine (SVM) classification algorithm able to automatically detect cancerous, fibrous, and adipose breast tissue. Classification accuracies were greater than 97% for automated detection of cancerous, fibrous, and adipose tissue from breast cancer specimens. The classification worked equally well for specimens scanned by hand or with a mechanical stage, demonstrating that the system could be used during surgery or on excised specimens. The ability of this technique to simply discriminate between cancerous and normal breast tissue, in particular to distinguish fibrous breast tissue from tumor, which is notoriously challenging for optical techniques, leads to the conclusion that FLIm has great potential to assess breast cancer margins. Identification of positive margins before waiting for complete histologic analysis could significantly reduce breast cancer re-excision rates.

  19. Effect of calcinations temperature on the luminescence intensity and fluorescent lifetime of Tb3+-doped hydroxyapatite (Tb-HA nanocrystallines

    Directory of Open Access Journals (Sweden)

    Hairong Yin

    2017-06-01

    Full Text Available Hydroxyapatite luminescent nanocrystallines doped with 6 mol.% Tb3+ (Tb-HA were prepared via chemical deposition method and calcined at different temperature, and the effects of calcinations temperature on the luminescence intensity and fluorescent lifetime were studied. TEM image of Tb-HA revealed that the shape of nanocrystallines changed from needle-like to short rod-like and sphere-like with the increase of calcinations temperature; while the particles sizes decreased from 190 nm to 110 nm. The crystallinity degree increased. The typical emission peaks attributed to Tb3+ ions were observed in emission spectra of 6 mol.% Tb-HA under 378 nm excitation. The luminescent intensity of Tb-HA, which showed the fluorescence quenching, firstly enhanced and then decreased at 700 °C; while the fluorescent lifetime increased firstly and then decreased after 600 °C. Furthermore, the ratio of intensity between 545 nm and 490 nm corresponding to electric-dipole and magnetic-dipole transition (IR: IO increases firstly and then decreases, which revealed that the proportion of substitute type and site of Ca2+ ions by Tb3+ ions were helpful to realize the substitute process and functional structure design.

  20. Magnetic field and temperature dependence of the fluorescence lifetime of Cr sup(3+) in GdA103

    International Nuclear Information System (INIS)

    Helman, J.S.; Caride, A.O.; Basso, H.C.; Terrile, M.C.; Carvalho, R.A.

    1991-01-01

    The fluorescence lifetime of Cr sup(3+) in GdA10 sub(3) was measured in the range 1.8 - 4.2 K in magnetic fields up to 6 T. The results show a remarkable dependence of the transition probabilities on magnetic order. A model based on the exchange interaction between Cr sup(3+) in highly excited states and the Gd sup(3+) ions is proposed. (author)

  1. Differentiation of ocular fundus fluorophores by fluorescence lifetime imaging using multiple excitation and emission wavelengths

    Science.gov (United States)

    Hammer, M.; Schweitzer, D.; Schenke, S.; Becker, W.; Bergmann, A.

    2006-10-01

    Ocular fundus autofluorescence imaging has been introduced into clinical diagnostics recently. It is in use for the observation of the age pigment lipofuscin, a precursor of age - related macular degeneration (AMD). But other fluorophores may be of interest too: The redox pair FAD - FADH II provides information on the retinal energy metabolism, advanced glycation end products (AGE) indicate protein glycation associated with pathologic processes in diabetes as well as AMD, and alterations in the fluorescence of collagen and elastin in connective tissue give us the opportunity to observe fibrosis by fluorescence imaging. This, however, needs techniques able to differentiate particular fluorophores despite limited permissible ocular exposure as well as excitation wavelength (limited by the transmission of the human ocular lens to >400 nm). We present an ophthalmic laser scanning system (SLO), equipped with picosecond laser diodes (FWHM 100 ps, 446 nm or 468 nm respectively) and time correlated single photon counting (TCSPC) in two emission bands (500 - 560 nm and 560 - 700 nm). The decays were fitted by a bi-exponential model. Fluorescence spectra were measured by a fluorescence spectrometer fluorolog. Upon excitation at 446 nm, the fluorescence of AGE, FAD, and lipofuscin were found to peak at 503 nm, 525 nm, and 600 nm respectively. Accordingly, the statistical distribution of the fluorescence decay times was found to depend on the different excitation wavelengths and emission bands used. The use of multiple excitation and emission wavelengths in conjunction with fluorescence lifetime imaging allows us to discriminate between intrinsic fluorophores of the ocular fundus. Taken together with our knowledge on the anatomical structure of the fundus, these findings suggest an association of the short, middle and long fluorescence decay time to the retinal pigment epithelium, the retina, and connective tissue respectively.

  2. Second-harmonic generation and fluorescence lifetime imaging microscopy through a rodent mammary imaging window

    Science.gov (United States)

    Young, Pamela A.; Nazir, Muhammad; Szulczewski, Michael J.; Keely, Patricia J.; Eliceiri, Kevin W.

    2012-03-01

    Tumor-Associated Collagen Signatures (TACS) have been identified that manifest in specific ways during breast tumor progression and that correspond to patient outcome. There are also compelling metabolic changes associated with carcinoma invasion and progression. We have characterized the difference in the autofluorescent properties of metabolic co-factors, NADH and FAD, between normal and carcinoma breast cell lines. Also, we have shown in vitro that increased collagen density alters metabolic genes which are associated with glycolysis and leads to a more invasive phenotype. Establishing the relationship between collagen density, cellular metabolism, and metastasis in physiologically relevant cancer models is crucial for developing cancer therapies. To study cellular metabolism with respect to collagen density in vivo, we use multiphoton fluorescence excitation microscopy (MPM) in conjunction with a rodent mammary imaging window implanted in defined mouse cancer models. These models are ideal for the study of collagen changes in vivo, allowing determination of corresponding metabolic changes in breast cancer invasion and progression. To measure cellular metabolism, we collect fluorescence lifetime (FLIM) signatures of NADH and FAD, which are known to change based on the microenvironment of the cells. Additionally, MPM systems are capable of collecting second harmonic generation (SHG) signals which are a nonlinear optical property of collagen. Therefore, MPM, SHG, and FLIM are powerful tools with great potential for characterizing key features of breast carcinoma in vivo. Below we present the current efforts of our collaborative group to develop intravital approaches based on these imaging techniques to look at defined mouse mammary models.

  3. Demonstration of the lack of cytotoxicity of unmodified and folic acid modified graphene oxide quantum dots, and their application to fluorescence lifetime imaging of HaCaT cells.

    Science.gov (United States)

    Goreham, Renee V; Schroeder, Kathryn L; Holmes, Amy; Bradley, Siobhan J; Nann, Thomas

    2018-01-24

    The authors describe the synthesis of water-soluble and fluorescent graphene oxide quantum dots via acid exfoliation of graphite nanoparticles. The resultant graphene oxide quantum dots (GoQDs) were then modified with folic acid. Folic acid receptors are overexpressed in cancer cells and hence can bind to functionalized graphene oxide quantum dots. On excitation at 305 nm, the GoQDs display green fluorescence with a peak wavelength at ~520 nm. The modified GoQDs are non-toxic to macrophage cells even after prolonged exposure and high concentrations. Fluorescence lifetime imaging and multiphoton microscopy was used (in combination) to image HeCaT cells exposed to GoQDs, resulting in a superior method for bioimaging. Graphical abstract Schematic representation of graphene oxide quantum dots, folic acid modified graphene oxide quantum dots (red), and the use of fluorescence lifetime to discriminate against green auto-fluorescence of HeCaT cells.

  4. 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.

  5. Nanodiamond arrays on glass for quantification and fluorescence characterisation.

    Science.gov (United States)

    Heffernan, Ashleigh H; Greentree, Andrew D; Gibson, Brant C

    2017-08-23

    Quantifying the variation in emission properties of fluorescent nanodiamonds is important for developing their wide-ranging applicability. Directed self-assembly techniques show promise for positioning nanodiamonds precisely enabling such quantification. Here we show an approach for depositing nanodiamonds in pre-determined arrays which are used to gather statistical information about fluorescent lifetimes. The arrays were created via a layer of photoresist patterned with grids of apertures using electron beam lithography and then drop-cast with nanodiamonds. Electron microscopy revealed a 90% average deposition yield across 3,376 populated array sites, with an average of 20 nanodiamonds per site. Confocal microscopy, optimised for nitrogen vacancy fluorescence collection, revealed a broad distribution of fluorescent lifetimes in agreement with literature. This method for statistically quantifying fluorescent nanoparticles provides a step towards fabrication of hybrid photonic devices for applications from quantum cryptography to sensing.

  6. Plasmonic enhancement of ultraviolet fluorescence

    Science.gov (United States)

    Jiao, Xiaojin

    Plasmonics relates to the interaction between electromagnetic radiation and conduction electrons at metallic interfaces or in metallic nanostructures. Surface plasmons are collective electron oscillations at a metal surface, which can be manipulated by shape, texture and material composition. Plasmonic applications cover a broad spectrum from visible to near infrared, including biosensing, nanolithography, spectroscopy, optoelectronics, photovoltaics and so on. However, there remains a gap in this activity in the ultraviolet (UV, research. Motivating factors in the study of UV Plasmonics are the direct access to biomolecular resonances and native fluorescence, resonant Raman scattering interactions, and the potential for exerting control over photochemical reactions. This dissertation aims to fill in the gap of Plasmonics in the UV with efforts of design, fabrication and characterization of aluminium (Al) and magnesium (Mg) nanostructures for the application of label-free bimolecular detection via native UV fluorescence. The first contribution of this dissertation addresses the design of Al nanostructures in the context of UV fluorescence enhancement. A design method that combines analytical analysis with numerical simulation has been developed. Performance of three canonical plasmonic structures---the dipole antenna, bullseye nanoaperture and nanoaperture array---has been compared. The optimal geometrical parameters have been determined. A novel design of a compound bullseye structure has been proposed and numerically analyzed for the purpose of compensating for the large Stokes shift typical of UV fluorescence. Second, UV lifetime modification of diffusing molecules by Al nanoapertures has been experimentally demonstrated for the first time. Lifetime reductions of ~3.5x have been observed for the high quantum yield (QY) laser dye p-terphenyl in a 60 nm diameter aperture with 50 nm undercut. Furthermore, quantum-yield-dependence of lifetime reduction has been

  7. Autofluorescence Lifetimes in Geographic Atrophy in Patients With Age-Related Macular Degeneration.

    Science.gov (United States)

    Dysli, Chantal; Wolf, Sebastian; Zinkernagel, Martin S

    2016-05-01

    To investigate fluorescence lifetime characteristics in patients with geographic atrophy (GA) in eyes with age-related macular degeneration and to correlate the measurements with clinical data and optical coherence tomography (OCT) findings. Patients with GA were imaged with a fluorescence lifetime imaging ophthalmoscope. Retinal autofluorescence lifetimes were measured in a short and a long spectral channel (498-560 nm and 560-720 nm). Mean retinal fluorescence lifetimes were analyzed within GA and the surrounding retina, and data were correlated with best corrected visual acuity and OCT measurements. Fluorescence lifetime maps of 41 eyes of 41 patients (80 ± 7 years) with GA were analyzed. Mean lifetimes within areas of atrophy were prolonged by 624 ± 276 ps (+152%) in the short spectral channel and 418 ± 186 ps (+83%) in the long spectral channel compared to the surrounding tissue. Autofluorescence lifetime abnormalities in GA occurred with particular patterns, similar to those seen in fundus autofluorescence intensity images. Within the fovea short mean autofluorescence lifetimes were observed, presumably representing macular pigment. Short lifetimes were preserved even in the absence of foveal sparing but were decreased in patients with advanced retinal atrophy in OCT. Short lifetimes in the fovea correlated with better best corrected visual acuity in both spectral channels. This study established that autofluorescence lifetime changes in GA present with explicit patterns. We hypothesize that the short lifetimes seen within the atrophy may be used to estimate damage induced by atrophy and to monitor disease progression in the context of natural history or interventional therapeutic studies.

  8. Determination of the average lifetime of bottom hadrons

    Energy Technology Data Exchange (ETDEWEB)

    Althoff, M; Braunschweig, W; Kirschfink, F J; Martyn, H U; Rosskamp, P; Schmitz, D; Siebke, H; Wallraff, W [Technische Hochschule Aachen (Germany, F.R.). Lehrstuhl fuer Experimentalphysik 1A und 1. Physikalisches Inst.; Eisenmann, J; Fischer, H M

    1984-12-27

    We have determined the average lifetime of hadrons containing b quarks produced in e/sup +/e/sup -/ annihilation to be tausub(B)=1.83 x 10/sup -12/ s. Our method uses charged decay products from both non-leptonic and semileptonic decay modes.

  9. Determination of the average lifetime of bottom hadrons

    Energy Technology Data Exchange (ETDEWEB)

    Althoff, M; Braunschweig, W; Kirschfink, F J; Martyn, H U; Rosskamp, P; Schmitz, D; Siebke, H; Wallraff, W; Eisenmann, J; Fischer, H M

    1984-12-27

    We have determined the average lifetime of hadrons containing b quarks produced in e e annihilation to be tausub(B)=1.83x10 S s. Our method uses charged decay products from both non-leptonic and semileptonic decay modes. (orig./HSI).

  10. 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

  11. Pulp tissue in sex determination: A fluorescent microscopic study

    Science.gov (United States)

    Nayar, Amit; Singh, Harkanwal Preet; Leekha, Swati

    2014-01-01

    Aims: To determine and compare the reliability of pulp tissue in determination of sex and to analyze whether caries have any effect on fluorescent body test. Materials and Methods: This study was carried on 50 maxillary and mandibular teeth (25 male teeth and 25 female teeth), which were indicated for extraction. The teeth are categorized into 5 groups, 10 each (5 from males and 5 from females) on the basis of caries progression. The pulp cells are stained with quinacrine hydrochloride and observed with fluorescent microscope for fluorescent body. Gender is determined by identification of Y chromosome fluorescence in dental pulp. Results: Fluorescent bodies were found to be more in sound teeth in males as the caries increase the mean percentage of fluorescent bodies observed decreases in males. We also observed the fluorescent spots in females, and the value of the spot increases in female as the caries progresses, thereby giving false positive results in females. Conclusion: Sex determination by fluorescent staining of the Y chromosome is a reliable technique in teeth with healthy pulps or caries with enamel or up to half way of dentin. Teeth with caries involving pulp cannot be used for sex determination. PMID:25125912

  12. Optimal Fluorescence Waveband Determination for Detecting Defective Cherry Tomatoes Using a Fluorescence Excitation-Emission Matrix

    Directory of Open Access Journals (Sweden)

    In-Suck Baek

    2014-11-01

    Full Text Available A multi-spectral fluorescence imaging technique was used to detect defective cherry tomatoes. The fluorescence excitation and emission matrix was used to measure for defects, sound surface and stem areas to determine the optimal fluorescence excitation and emission wavelengths for discrimination. Two-way ANOVA revealed the optimal excitation wavelength for detecting defect areas was 410 nm. Principal component analysis (PCA was applied to the fluorescence emission spectra of all regions at 410 nm excitation to determine the emission wavelengths for defect detection. The major emission wavelengths were 688 nm and 506 nm for the detection. Fluorescence images combined with the determined emission wavebands demonstrated the feasibility of detecting defective cherry tomatoes with >98% accuracy. Multi-spectral fluorescence imaging has potential utility in non-destructive quality sorting of cherry tomatoes.

  13. Sensitive determination of enoxacin in pharmaceutical formulations by its quench effect on the fluorescence of glutathione-capped CdTe quantum dots.

    Science.gov (United States)

    Yang, Qiong; Tan, Xuanping; Yang, Jidong

    2016-02-01

    A sensitive and simple method for the determination of enoxacin (ENX) was developed based on the fluorescence quenching effect of ENX for glutathione (GSH)-capped CdTe quantum dots (QDs). Under optimum conditions, a good linear relationship was obtained from 4.333 × 10(-9)  mol⋅L(-1) to 1.4 × 10(-5)  mol⋅L(-1) with a correlation coefficient (R) of 0.9987, and the detection limit (3σ/K) was 1.313 × 10(-9)  mol⋅L(-1). The corresponding mechanism has been proposed on the basis of electron transfer supported by ultraviolet-visible (UV) light absorption, fluorescence spectroscopy, and the measurement of fluorescence lifetime. The method has been applied to the determination of ENX in pharmaceutical formulations (enoxacin gluconate injections and commercial tablets) with satisfactory results. The proposed method manifested several advantages such as high sensitivity, short analysis time, low cost and ease of operation. Copyright © 2015 John Wiley & Sons, Ltd.

  14. 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.

  15. Steady state and time-resolved fluorescence spectroscopy of quinine sulfate dication bound to sodium dodecylsulfate micelles: Fluorescent complex formation

    Energy Technology Data Exchange (ETDEWEB)

    Joshi, Sunita; Pant, Debi D., E-mail: ddpant@pilani.bits-pilani.ac.in

    2014-01-15

    Interaction of quinine sulfate dication (QSD) with anionic, sodium dodecylsulphate (SDS) surfactant has been studied at different premicellar, micellar and postmicellar concentrations in aqueous phase using steady state, time-resolved fluorescence and fluorescence anisotropy techniques. At premicellar concentrations of SDS, the decrease in absorbance, appearance of an extra fluorescence band at lower wavelengths and tri-exponential decay behavior of fluorescence, are attributed to complex formation between QSD molecules and surfactant monomers. At postmicellar concentrations the red shift in fluorescence spectrum, increase in quantum yield and increase in fluorescence lifetimes are attributed to incorporation of solute molecules to micelles. At lower concentrations of SDS, a large shift in fluorescence is observed on excitation at the red edge of absorption spectrum and this is explained in terms of distribution of ion pairs of different energies in the ground state and the observed fluorescence lifetime behavior corroborates with this model. The temporal fluorescence anisotropy decay of QSD in SDS micelles allowed determination of restriction on the motion of the fluorophore. All the different techniques used in this study reveal that the photophysics of QSD is very sensitive to the microenvironments of SDS micelles and QSD molecules reside at the water-micelle interface. -- Highlights: • Probe molecule is very sensitive to microenvironment of micelles. • Highly fluorescent ion-pair formation has been observed. • Modulated photophysics of probe molecule in micellar solutions has been observed. • Probe molecules strongly bind with micelles and reside at probe–micelle interface.

  16. Fluorescence Decay Dynamics of Ethidium Bromide in Polymers

    International Nuclear Information System (INIS)

    Jee, Ah Young; Min Yung

    2010-01-01

    The fluorescence lifetimes of EB in five polymers covering LDPE, HDPE, PC, PS, and PAA were measured by picosecond time-correlated single photon counting. The lifetime change of EB has been previously described by hydrogen bonding ability. In this work, we have observed that the lifetime of EB depends strongly on the Young's modulus of medium. Thus, it is possible that the fluorescence decay dynamics of EB could be influenced by medium rigidity rather than hydrogen bonding ability in polymer. The medium influence on the fluorescence decay dynamics of ethidium bromide (EB) has been investigated in various environments. For example, Ohmstead and Kearns related the fluorescence lifetime of EB to the excited-state proton transfer process. In addition, they reported that the solvent viscosity plays a minor role in the excited state decay process of EB. Chirico et al. measured the fluorescence decay of EB as 1.7 ns in water and 6.5 ns in ethanol and concluded that hydrogen bonding ability is a key factor for the nonradiative relaxation. Pal et al. measured the fluorescence decay time of EB in acetone, acetonitrile, and their mixtures. They observed that the fluorescence decay processes were independent on the solvent polarity. These results show that the EB lifetime does not depend much on polarity or viscosity, but is mainly influenced by hydrogen bonding. Overall, EB is one of most widely used dyes for probing DNA. When EB is intercalated into the helical structure of DNA, a large increase in the fluorescence lifetime has been observed in comparison with water environment, and the fluorescence enhancement was attributed to the blocking of the excited-state proton transfer

  17. Fluorescent determination of neptunium in plutonium

    International Nuclear Information System (INIS)

    Alexandruk, V.M.; Babaev, A.S.; Dem'yanova, T.A.; Stepanov, A.V.

    1991-01-01

    This paper describes a new procedure for direct determination of Neptunium in Plutonium using laser induced time resolved fluorescence method. The procedure based on measurement of fluorescence intensity of Neptunium followed its concentration in effective layer of pellet of calcium fluoride. Detection limit of determination of Neptunium is 2 10 -12 g. At the level of Neptunium content in Plutonium more than 5 ppm relative standard deviation is equal 0.08-0.12. For carrying out of single measurement it is necessary neither more nor less 5 mkg Plutonium

  18. Confocal fluorescence techniques in industrial application

    Science.gov (United States)

    Eggeling, Christian; Gall, Karsten; Palo, Kaupo; Kask, Peet; Brand, Leif

    2003-06-01

    The FCS+plus family of evaluation tools for confocal fluorescence spectroscopy, which was developed during recent years, offers a comprehensive view to a series of fluorescence properties. Originating in fluorescence correlation spectroscopy (FCS) and using similar experimental equipment, a system of signal processing methods such as fluorescence intensity distribution analysis (FIDA) was created to analyze in detail the fluctuation behavior of fluorescent particles within a small area of detection. Giving simultaneous access to molecular parameters like concentration, translational and rotational diffusion, molecular brightness, and multicolor coincidence, this portfolio was enhanced by more traditional techniques of fluorescence lifetime as well as time-resolved anisotropy determination. The cornerstones of the FCS+plus methodology will be shortly described. The inhibition of a phosphatase enzyme activity gives a comprehensive industrial application that demonstrates FCS+plus' versatility and its potential for pharmaceutical drug discovery.

  19. Direct Vpr-Vpr Interaction in Cells monitored by two Photon Fluorescence Correlation Spectroscopy and Fluorescence Lifetime Imaging

    Directory of Open Access Journals (Sweden)

    Mély Yves

    2008-09-01

    Full Text Available Abstract Background The human immunodeficiency virus type 1 (HIV-1 encodes several regulatory proteins, notably Vpr which influences the survival of the infected cells by causing a G2/M arrest and apoptosis. Such an important role of Vpr in HIV-1 disease progression has fuelled a large number of studies, from its 3D structure to the characterization of specific cellular partners. However, no direct imaging and quantification of Vpr-Vpr interaction in living cells has yet been reported. To address this issue, eGFP- and mCherry proteins were tagged by Vpr, expressed in HeLa cells and their interaction was studied by two photon fluorescence lifetime imaging microscopy and fluorescence correlation spectroscopy. Results Results show that Vpr forms homo-oligomers at or close to the nuclear envelope. Moreover, Vpr dimers and trimers were found in the cytoplasm and in the nucleus. Point mutations in the three α helices of Vpr drastically impaired Vpr oligomerization and localization at the nuclear envelope while point mutations outside the helical regions had no effect. Theoretical structures of Vpr mutants reveal that mutations within the α-helices could perturb the leucine zipper like motifs. The ΔQ44 mutation has the most drastic effect since it likely disrupts the second helix. Finally, all Vpr point mutants caused cell apoptosis suggesting that Vpr-mediated apoptosis functions independently from Vpr oligomerization. Conclusion We report that Vpr oligomerization in HeLa cells relies on the hydrophobic core formed by the three α helices. This oligomerization is required for Vpr localization at the nuclear envelope but not for Vpr-mediated apoptosis.

  20. 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

  1. Precision lifetime measurements

    International Nuclear Information System (INIS)

    Tanner, C.E.

    1994-01-01

    Precision measurements of atomic lifetimes provide important information necessary for testing atomic theory. The authors employ resonant laser excitation of a fast atomic beam to measure excited state lifetimes by observing the decay-in-flight of the emitted fluorescence. A similar technique was used by Gaupp, et al., who reported measurements with precisions of less than 0.2%. Their program includes lifetime measurements of the low lying p states in alkali and alkali like systems. Motivation for this work comes from a need to test the atomic many-body-perturbation theory (MBPT) that is necessary for interpretation of parity nonconservation experiments in atomic cesium. The authors have measured the cesium 6p 2 P 1/2 and 6p 2 P 3/2 state lifetimes to be 34.934±0.094 ns and 30.499±0.070 ns respectively. With minor changes to the apparatus, they have extended their measurements to include the lithium 2p 2 P 1/2 and 2p 2 P 3/2 states

  2. Phosphorescence and delayed fluorescence properties of fluorone dyes in bio-related films

    International Nuclear Information System (INIS)

    Penzkofer, A.; Tyagi, A.; Slyusareva, E.; Sizykh, A.

    2010-01-01

    Graphical abstract: The spectral and temporal phosphorescence and delayed fluorescence behaviour of five fluorescein dyes in gelatine, starch, and chitosan is studied and basic parameters are determined. Research highlights: → Phosphorescence quantum yields of fluorone dyes in bio-related films are measured at room temperature. → Delayed fluorescence quantum yields of fluorone dyes in bio-related films are measured at room temperature. → Phosphorescence lifetimes of fluorone dyes in bio-related films are measured at room temperature. → Delayed fluorescence lifetimes of fluorone dyes in bio-related films are measured at room temperature. → General theory of short-pulse excited phosphorescence and delayed fluorescence is presented and relevant parameters are extracted. - Abstract: The phosphorescence and delayed fluorescence behaviour of the fluorone dyes disodium fluorescein (FL, uranine), 4,5-dibromofluorescein (DBF), eosin Y (EO), erythrosine B (ER), and rose bengal (RB) in bio-films of gelatine, starch, and chitosan at room temperature is studied. Phosphorescence and delayed fluorescence quantum yields and lifetimes were measured. The singlet-triplet dynamics is described and applied to the fluorone dyes for parameter extraction. For uranine films at room temperature no phosphorescence could be resolved. The efficiency of singlet-triplet intersystem crossing increased in the order φ ISC (DBF) ISC (EO) ISC (ER) ISC (RB) due to the heavy atom effect on spin-orbit coupling. The phosphorescence quantum yields increased in the order φ P (DBF) P (EO) P (RB) P (ER). The phosphorescence lifetimes followed the order τ P (DBF) > τ P (EO) > τ P (ER) > τ P (RB).

  3. Determination of uranium in seawater by fluorescence spectrometry

    International Nuclear Information System (INIS)

    Kawashima, Toshi; Kawakubo, Senkichi; Minegishi, Hisako.

    1984-01-01

    A Fluorescence spectrometry for the determination of uranium in seawater has been developed. Anion exchange separation of uranium from seawater followed by preparation of NaF-carbonate cake and by spectrometry for ultraviolet ray excited fluorescence of uranium on the fluoride host provide the trace determinaton of uranium at the subnano gram level. Anion exchange behavior, excitation-emission behavior of the uranium on the host and effects of foreign ions to the fluorescence have been presented. Appling the method to 1 ml of seawater 3 ppb of uranium has been determined. (author)

  4. Distinct roles of the photosystem II protein PsbS and zeaxanthin in the regulation of light harvesting in plants revealed by fluorescence lifetime snapshots.

    Science.gov (United States)

    Sylak-Glassman, Emily J; Malnoë, Alizée; De Re, Eleonora; Brooks, Matthew D; Fischer, Alexandra Lee; Niyogi, Krishna K; Fleming, Graham R

    2014-12-09

    The photosystem II (PSII) protein PsbS and the enzyme violaxanthin deepoxidase (VDE) are known to influence the dynamics of energy-dependent quenching (qE), the component of nonphotochemical quenching (NPQ) that allows plants to respond to fast fluctuations in light intensity. Although the absence of PsbS and VDE has been shown to change the amount of quenching, there have not been any measurements that can detect whether the presence of these proteins alters the type of quenching that occurs. The chlorophyll fluorescence lifetime probes the excited-state chlorophyll relaxation dynamics and can be used to determine the amount of quenching as well as whether two different genotypes with the same amount of NPQ have similar dynamics of excited-state chlorophyll relaxation. We measured the fluorescence lifetimes on whole leaves of Arabidopsis thaliana throughout the induction and relaxation of NPQ for wild type and the qE mutants, npq4, which lacks PsbS; npq1, which lacks VDE and cannot convert violaxanthin to zeaxanthin; and npq1 npq4, which lacks both VDE and PsbS. These measurements show that although PsbS changes the amount of quenching and the rate at which quenching turns on, it does not affect the relaxation dynamics of excited chlorophyll during quenching. In addition, the data suggest that PsbS responds not only to ΔpH but also to the Δψ across the thylakoid membrane. In contrast, the presence of VDE, which is necessary for the accumulation of zeaxanthin, affects the excited-state chlorophyll relaxation dynamics.

  5. Time-resolved fluorescence spectroscopy

    International Nuclear Information System (INIS)

    Gustavsson, Thomas; Mialocq, Jean-Claude

    2007-01-01

    This article addresses the evolution in time of light emitted by a molecular system after a brief photo-excitation. The authors first describe fluorescence from a photo-physical point of view and discuss the characterization of the excited state. Then, they explain some basic notions related to fluorescence characterization (lifetime and decays, quantum efficiency, so on). They present the different experimental methods and techniques currently used to study time-resolved fluorescence. They discuss basic notions of time resolution and spectral reconstruction. They briefly present some conventional methods: intensified Ccd cameras, photo-multipliers and photodiodes associated with a fast oscilloscope, and phase modulation. Other methods and techniques are more precisely presented: time-correlated single photon counting (principle, examples, and fluorescence lifetime imagery), streak camera (principle, examples), and optical methods like the Kerr optical effect (principle and examples) and fluorescence up-conversion (principle and theoretical considerations, examples of application)

  6. Time resolved fluorescence of cow and goat milk powder

    Science.gov (United States)

    Brandao, Mariana P.; de Carvalho dos Anjos, Virgílio; Bell., Maria José V.

    2017-01-01

    Milk powder is an international dairy commodity. Goat and cow milk powders are significant sources of nutrients and the investigation of the authenticity and classification of milk powder is particularly important. The use of time-resolved fluorescence techniques to distinguish chemical composition and structure modifications could assist develop a portable and non-destructive methodology to perform milk powder classification and determine composition. This study goal is to differentiate milk powder samples from cows and goats using fluorescence lifetimes. The samples were excited at 315 nm and the fluorescence intensity decay registered at 468 nm. We observed fluorescence lifetimes of 1.5 ± 0.3, 6.4 ± 0.4 and 18.7 ± 2.5 ns for goat milk powder; and 1.7 ± 0.3, 6.9 ± 0.2 and 29.9 ± 1.6 ns for cow's milk powder. We discriminate goat and cow powder milk by analysis of variance using Fisher's method. In addition, we employed quadratic discriminant analysis to differentiate the milk samples with accuracy of 100%. Our results suggest that time-resolved fluorescence can provide a new method to the analysis of powder milk and its composition.

  7. Anthracene Fluorescence Quenching by a Tetrakis (Ketocarboxamide Cavitand

    Directory of Open Access Journals (Sweden)

    Tibor Zoltan Janosi

    2014-01-01

    Full Text Available Quenching of both fluorescence lifetime and fluorescence intensity of anthracene was investigated in the presence of a newly derived tetrakis (ketocarboxamide cavitand at various concentrations. Time-correlated single photon counting method was applied for the lifetime measurements. A clear correlation between the fluorescence lifetime of anthracene as a function of cavitand concentration in dimethylformamide solution was observed. The bimolecular collisional quenching constant was derived from the decrease of lifetime. Fluorescence intensity was measured in the emission wavelength region around 400 nm as a result of excitation at 280 nm. Effective quenching was observed in the presence of the cavitand. The obtained Stern-Volmer plot displayed upward curvature. The results did not follow even extended Stern-Volmer behavior, often used to describe deviations from static bimolecular quenching. To explain our results we adopted the Smoluchowski model and obtained a reasonable estimate for the molecular radius of the cavitand in solution.

  8. Multimodal fluorescence imaging spectroscopy

    NARCIS (Netherlands)

    Stopel, Martijn H W; Blum, Christian; Subramaniam, Vinod; Engelborghs, Yves; Visser, Anthonie J.W.G.

    2014-01-01

    Multimodal fluorescence imaging is a versatile method that has a wide application range from biological studies to materials science. Typical observables in multimodal fluorescence imaging are intensity, lifetime, excitation, and emission spectra which are recorded at chosen locations at the sample.

  9. Quantitative time domain analysis of lifetime-based Förster resonant energy transfer measurements with fluorescent proteins: Static random isotropic fluorophore orientation distributions

    DEFF Research Database (Denmark)

    Alexandrov, Yuriy; Nikolic, Dino Solar; Dunsby, Christopher

    2018-01-01

    Förster resonant energy transfer (FRET) measurements are widely used to obtain information about molecular interactions and conformations through the dependence of FRET efficiency on the proximity of donor and acceptor fluorophores. Fluorescence lifetime measurements can provide quantitative...... into new software for fitting donor emission decay profiles. Calculated FRET parameters, including molar population fractions, are compared for the analysis of simulated and experimental FRET data under the assumption of static and dynamic fluorophores and the intermediate regimes between fully dynamic...... analysis of FRET efficiency and interacting population fraction. Many FRET experiments exploit the highly specific labelling of genetically expressed fluorescent proteins, applicable in live cells and organisms. Unfortunately, the typical assumption of fast randomization of fluorophore orientations...

  10. Emerging biomedical applications of time-resolved fluorescence spectroscopy

    Science.gov (United States)

    Lakowicz, Joseph R.; Szmacinski, Henryk; Koen, Peter A.

    1994-07-01

    Time-resolved fluorescence spectroscopy is presently regarded as a research tool in biochemistry, biophysics, and chemical physics. Advances in laser technology, the development of long-wavelength probes, and the use of lifetime-based methods are resulting in the rapid migration of time-resolved fluorescence to the clinical chemistry lab, to the patient's bedside, to flow cytometers, to the doctor's office, and even to home health care. Additionally, time-resolved imaging is now a reality in fluorescence microscopy, and will provide chemical imaging of a variety of intracellular analytes and/or cellular phenomena. In this overview paper we attempt to describe some of the opportunities available using chemical sensing based on fluorescence lifetimes, and to predict those applications of lifetime-based sensing which are most likely in the near future.

  11. Two-photon excited fluorescence microscopy application for ex vivo investigation of ocular fundus samples

    Science.gov (United States)

    Peters, Sven; Hammer, Martin; Schweitzer, Dietrich

    2011-07-01

    Two-photon excited fluorescence (TPEF) imaging of ocular tissue has recently become a promising tool in ophthalmology for diagnostic and research purposes. The feasibility and the advantages of TPEF imaging, namely deeper tissue penetration and improved high-resolution imaging of microstructures, have been demonstrated lately using human ocular samples. The autofluorescence properties of endogenous fluorophores in ocular fundus tissue are well known from spectrophotometric analysis. But fluorophores, especially when it comes to fluorescence lifetime, typically display a dependence of their fluorescence properties on local environmental parameters. Hence, a more detailed investigation of ocular fundus autofluorescence ideally in vivo is of utmost interest. The aim of this study is to determine space-resolved the stationary and time-resolved fluorescence properties of endogenous fluorophores in ex vivo porcine ocular fundus samples by means of two-photon excited fluorescence spectrum and lifetime imaging microscopy (FSIM/FLIM). By our first results, we characterized the autofluorescence of individual anatomical structures of porcine retina samples excited at 760 nm. The fluorescence properties of almost all investigated retinal layers are relatively homogenous. But as previously unknown, ganglion cell bodies show a significantly shorter fluorescence lifetime compared to the adjacent mueller cells. Since all retinal layers exhibit bi-exponential autofluorescence decays, we were able to achieve a more precise characterization of fluorescence properties of endogenous fluorophores compared to a present in vivo FLIM approach by confocal scanning laser ophthalmoscope (cSLO).

  12. Fluorescence kinetics of Trp-Trp dipeptide and its derivatives in water via ultrafast fluorescence spectroscopy.

    Science.gov (United States)

    Jia, Menghui; Yi, Hua; Chang, Mengfang; Cao, Xiaodan; Li, Lei; Zhou, Zhongneng; Pan, Haifeng; Chen, Yan; Zhang, Sanjun; Xu, Jianhua

    2015-08-01

    Ultrafast fluorescence dynamics of Tryptophan-Tryptophan (Trp-Trp/Trp2) dipeptide and its derivatives in water have been investigated using a picosecond resolved time correlated single photon counting (TCSPC) apparatus together with a femtosecond resolved upconversion spectrophotofluorometer. The fluorescence decay profiles at multiple wavelengths were fitted by a global analysis technique. Nanosecond fluorescence kinetics of Trp2, N-tert-butyl carbonyl oxygen-N'-aldehyde group-l-tryptophan-l-tryptophan (NBTrp2), l-tryptophan-l-tryptophan methyl ester (Trp2Me), and N-acetyl-l-tryptophan-l-tryptophan methyl ester (NATrp2Me) exhibit multi-exponential decays with the average lifetimes of 1.99, 3.04, 0.72 and 1.22ns, respectively. Due to the intramolecular interaction between two Trp residues, the "water relaxation" lifetime was observed around 4ps, and it is noticed that Trp2 and its derivatives also exhibit a new decay with a lifetime of ∼100ps, while single-Trp fluorescence decay in dipeptides/proteins shows 20-30ps. The intramolecular interaction lifetime constants of Trp2, NBTrp2, Trp2Me and NATrp2Me were then calculated to be 3.64, 0.93, 11.52 and 2.40ns, respectively. Candidate mechanisms (including heterogeneity, solvent relaxation, quasi static self-quenching or ET/PT quenching) have been discussed. Copyright © 2015. Published by Elsevier B.V.

  13. Determining of the Optimal Device Lifetime using Mathematical Renewal Models

    Directory of Open Access Journals (Sweden)

    Knežo Dušan

    2016-05-01

    Full Text Available Paper deals with the operations and equipment of the machine in the process of organizing production. During operation machines require maintenance and repairs, while in case of failure or machine wears it is necessary to replace them with new ones. For the process of replacement of old machines with new ones the term renewal is used. Qualitative aspects of the renewal process observe renewal theory, which is mainly based on the theory of probability and mathematical statistics. Devices lifetimes are closely related to the renewal of the devices. Presented article is focused on mathematical deduction of mathematical renewal models and determining optimal lifetime of the devices from the aspect of expenditures on renewal process.

  14. Simultaneous determination of glycols based on fluorescence anisotropy

    International Nuclear Information System (INIS)

    Garcia Sanchez, F.; Navas Diaz, A.; Lopez Guerrero, M.M.

    2007-01-01

    Simultaneous determination of non-fluorescent glycols in mixtures without separation or chemical transformation steps is described. Two methods based in the measure of fluorescence anisotropy of a probe such as fluorescein dissolved in the analyte or analyte mixtures are described. In the first method, the anisotropy spectra of pure and mixtures of analytes are used to quantitative determination (if the fluorophor concentration is in a range where fluorescence intensity is proportional to concentration). In the second method, a calibration curve anisotropy-concentration based on the application of the Perrin equation is established. The methods presented here are capable of directly resolving binary mixtures of non-fluorescent glycols on the basis of differences on the fluorescence anisotropy of a fluorescence tracer. Best analytical performances were obtained by application of the method based on Perrin equation. This method is simple, rapid and allows the determination of mixtures of glycols with reasonable accuracy and precision. Detection limits are limited by the quantum yield and anisotropy values of the tracer in the solvents. Recovery values are related to the differences in anisotropy values of the tracer in the pure solvents. Mixtures of glycerine/ethylene glycol (GL/EG), ethylene glycol/1,2-propane diol (EG/1,2-PPD) and polyethylene glycol 400/1,2-propane diol (PEG 400/1,2-PPD) were analysed and recovery values are within 95-120% in the Perrin method. Relative standard deviation are in the range 1.3-2.9% and detection limits in the range 3.9-8.9%

  15. In vivo multiphoton and fluorescence lifetime imaging microscopy of the healthy and cholestatic liver

    Science.gov (United States)

    Kuznetsova, Daria S.; Dudenkova, Varvara V.; Rodimova, Svetlana A.; Bobrov, Nikolai V.; Zagainov, Vladimir E.; Zagaynova, Elena V.

    2018-02-01

    A cholestatic liver disease presents one of the most common liver diseases and can potentially progress to cirrhosis or even cholangiocarcinoma. Conventional techniques are insufficient to precisely describe the complex internal structure, heterogeneous cell populations and the dynamics of biological processes of the liver. Currently, the methods of multiphoton and fluorescence lifetime imaging microscopy are actively introducing to biomedical research. Those methods are extremely informative and non-destructive that allows studying of a large number of processes occurring inside cells and tissues, analyzing molecular cellular composition, as well as evaluating the state of connective tissue fibers due to their ability to generate a second optical harmonic. Multiphoton and FLIM microscopy do not need additional staining of samples or the incorporation of any markers to study metabolism, lipid composition, microstructure analysis, evaluation of fibrous structures. These parameters have pronounced changes in hepatocytes of liver with common pathological diseases. Thereby in this study we investigated metabolic changes in the healthy and cholestatic liver based on the fluorescence of the metabolic co-factors NAD(P)H and FAD by multiphoton microscopy combined with FLIM. To estimate the contribution of energy metabolism and lipogenesis in the observed changes of the metabolic profile, a separate analysis of NADH and NADPH was presented. The data can be used to develop new criteria for the identification of hepatic pathology at the level of hepatocyte changes directed to personalized medicine in the future.

  16. QUANTIFYING THE SHORT LIFETIME WITH TCSPC-FLIM: FIRST MOMENT VERSUS FITTING METHODS

    Directory of Open Access Journals (Sweden)

    LINGLING XU

    2013-10-01

    Full Text Available Combing the time-correlated single photon counting (TCSPC with fluorescence lifetime imaging microscopy (FLIM provides promising opportunities in revealing important information on the microenvironment of cells and tissues, but the applications are thus far mainly limited by the accuracy and precision of the TCSPC-FLIM technique. Here we present a comprehensive investigation on the performance of two data analysis methods, the first moment (M1 method and the conventional least squares (Fitting method, in quantifying fluorescence lifetime. We found that the M1 method is more superior than the Fitting method when the lifetime is short (70 ~ 400 ps or the signal intensity is weak (<103 photons.

  17. Photoluminescence decay lifetime measurements of hemicyanine derivatives of different alkyl chain lengths

    International Nuclear Information System (INIS)

    Shim, Taekyu; Lee, Myounghee; Kim, Sungho; Sung, Jaeho; Rhee, Bum Ku; Kim, Doseok; Kim, Hyunsung; Yoon, Kyung Byung

    2004-01-01

    The fluorescence upconversion setup for the detection of photoluminescence (PL) decay lifetime with subpicosecond time resolution was constructed, and the photoluminescence phenomena of several hemicyanine dyes with alkyl chains of different chain lengths tethered to the N atom of the pyridine moiety (HC-n, n=6, 15, 22) in methanol were investigated. The average decay lifetimes of the solutions determined from the measured data by multi-order exponential decay curve fitting were ∼27 ps at the PL peak wavelength. It was found that the PL decay properties did not depend on the alkyl chain length in the molecule, implying that the twist of the alkylpyridinium ring of the molecule is not possible as a nonfluorescing relaxation pathway. The time-dependent PL spectra constructed from the PL lifetime data showed the dynamic Stokes shift of ∼1000 cm -1

  18. 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.

  19. Phosphorescence quantum yield determination with time-gated fluorimeter and Tb(III)-acetylacetonate as luminescence reference

    Energy Technology Data Exchange (ETDEWEB)

    Penzkofer, A., E-mail: alfons.penzkofer@physik.uni-regensburg.de [Fakultät für Physik, Universität Regensburg, Universitätsstrasse 31, D-93053 Regensburg (Germany)

    2013-03-29

    Highlights: ► Procedure for absolute phosphorescence quantum yield measurement is described. ► Experimental setup for absolute luminescence quantum yield standard calibration. ► Tb(acac){sub 3} proposed as phosphorescence quantum yield reference standard. ► Luminescence quantum yield of Tb(acac){sub 3} in cyclohexane measured. ► Luminescence lifetime of Tb(acac){sub 3} in cyclohexane measured. - Abstract: Phosphorescence quantum yield measurements of fluorescent and phosphorescent samples require the use of time-gated fluorimeters in order to discriminate against the fluorescence contribution. As reference standard a non-fluorescent luminescent compound is needed for absolute phosphorescence quantum yield determination. For this purpose the luminescence behavior of the rare earth chelate terbium(III)-acetylacetonate (Tb(acac){sub 3}) was studied (determination of luminescence quantum yield and luminescence lifetime). The luminescence quantum yield of Tb(acac){sub 3} was determined by using an external light source and operating the fluorimeter in chemo/bioluminescence mode with a fluorescent dye (rhodamine 6G in methanol) as reference standard. A procedure is developed for absolute luminescence (phosphorescence) quantum yield determination of samples under investigation with a time-gated fluorimeter using a non-fluorescent luminescent compound of known luminescence quantum yield and luminescence lifetime.

  20. Metabolic Mapping of Breast Cancer with Multiphoton Spectral and Lifetime Imaging

    Science.gov (United States)

    2007-03-01

    2002. Spectrally resolved fluorescence lifetime imaging microscopy. Appl. Spec- trosc. 56 :155-166. 38. Becker, W., A. Bergmann, E. Haustein , Z...photon fluores- cence lifetime imaging microscopy of macrophage-mediated antigen processing. J. Microsc. 185 :339-353. 45. Lin, H.J., P. Herman , and

  1. Determination of lifetimes and nonadiabatic correlations from measured dipole polarizabilities

    International Nuclear Information System (INIS)

    Curtis, Lorenzo J

    2007-01-01

    In atomic systems for which the total oscillator strength of excitations from the ground state is dominated by the transition to the lowest resonance level, the f-sum rule provides a bracketing inequality connecting the lifetime τ of that level to the dipole polarizability α d . This relationship has been used previously to deduce α d from τ. It is shown here that improved spectroscopic accuracies now permit this procedure to be inverted, with τ deduced from a value for α d obtained spectroscopically using the core polarization model. A similar quantitative relationship exists connecting the nonadiabatic correlation factor β to τ, and thus also to α d . The method is applied to a recent measurement of α d for Kr 6+ to obtain the values τ(4s4p 1 P 1 ) 0.096 ± 0.003 ns and β(Kr 6+ ) = 1.71 ± 0.03a 5 0 . It is shown that the use of this method to make precision lifetime determinations for a small number of ions in an isoelectronic sequence permits the exploitation of observed semiempirical regularities to specify the lifetimes of all ions in that sequence

  2. Evaluation of Fluorescent Analogs of Deoxycytidine for Monitoring DNA Transitions from Duplex to Functional Structures

    Directory of Open Access Journals (Sweden)

    Yogini P. Bhavsar

    2011-01-01

    Full Text Available Topological variants of single-strand DNA (ssDNA structures, referred to as “functional DNA,” have been detected in regulatory regions of many genes and are thought to affect gene expression. Two fluorescent analogs of deoxycytidine, Pyrrolo-dC (PdC and 1,3-diaza-2-oxophenoxazine (tC∘, can be incorporated into DNA. Here, we describe spectroscopic studies of both analogs to determine fluorescent properties that report on structural transitions from double-strand DNA (dsDNA to ssDNA, a common pathway in the transition to functional DNA structures. We obtained fluorescence-detected circular dichroism (FDCD spectra, steady-state fluorescence spectra, and fluorescence lifetimes of the fluorophores in DNA. Our results show that PdC is advantageous in fluorescence lifetime studies because of a distinct ~2 ns change between paired and unpaired bases. However, tC∘ is a better probe for FDCD experiments that report on the helical structure of DNA surrounding the fluorophore. Both fluorophores provide complementary data to measure DNA structural transitions.

  3. 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

  4. Using the fluorescence of DBO to study the aggregation of asphaltenes

    Energy Technology Data Exchange (ETDEWEB)

    Yang, Zixin; Bohne, Cornelia [Department of Chemistry, University of Victoria (Canada)], email: xyang@uvic.ca

    2010-07-01

    Asphaltene, operationally defined as the fraction of bitumen that is insoluble in heptane but soluble in toluene, is the least characterized component of crude oil. It can aggregate at low concentrations, causing problems in the reservoir and during transport and processing. Fluorescence techniques have been employed to characterize asphaltenes, e.g. by adding external probes. DBO (2,3-diazabicyclo [2.2.2]oct-2-ene), a fluorescence probe molecule with a long fluorescence lifetime, was made sensitive to the presence of aliphatic C-H bonds. DBO was used as an external fluorescent probe to characterize the aggregation of Athabasca asphaltene. The lifetime of DBO was measured using single photon counting. Preliminary lifetime measurements show that AA-5 quenches the emission of DBO, leading to a shortening of the DBO lifetime. The abrupt decrease in lifetime may be related to the interaction of DBO with the AA-5 aggregate; further studies are being performed to test this hypothesis. In conclusion, DBO interacts with asphaltene components and has the potential for being used as a probe to study the asphaltene aggregation.

  5. Fluorescence properties of europium and samarium. beta. -diketonates and their use in fluorometry

    Energy Technology Data Exchange (ETDEWEB)

    Huang, H; Hiraki, K; Nishikawa, Y [Kinki Univ., Higashi-Osaka, Osaka (Japan). Faculty of Science and Technology

    1981-01-01

    Several europium and samarium ..beta..-diketonates (tta, ntfa, bfa) complexed with 1, 10-phenanthroline, or with trioctylphosphine oxide (topo) were synthesized. The fluorescence properties of these compounds in benzene or hexane have been studied. Absorption and fluorescence spectra, fluorescence quantum yield, fluorescence sensitivity index (F.S.I.), and fluorescence lifetime were measured. From the measurement of fluorescence lifetime of the ..beta..-diketonates, the velocity of radiative process (k sub(f)/phi sub(f)) has almost the same value for benzene and hexane solvent. The red fluorescence (Em. max. : 619 nm) of Eu(III) in these chelates is attributed to transitions from /sup 5/D/sub 0/ ..-->.. /sup 7/F/sub 2/ levels of this ion, and the three-band spectrum (Em. max. : 569 nm, 606 nm, 650 nm) indicates the transitions from the /sup 4/G sub(5/2) ..-->.. /sup 6/H sub(5/2), /sup 4/G sub(5/2) ..-->.. /sup 6/H sub(7/2), and /sup 4/G sub(5/2) ..-->.. /sup 6/H sub(9/2) levels of Sm(III), respectively. These spectra are not changed by any solvents and ligands. From the results, the fluorescence of the ..beta..-diketonates in organic solvent has been attributed to m* ..-->.. m luminescence transition. The complexes of Eu(III) and Sm(III) show radiative transition within orbitals, composed exclusively of 4f orbitals of rare earth ions (m* ..-->.. m radiative transition). Fluorinated ligands show better sensitivity than unfluorinated ligands, and the best sensitivity is obtained with TTA-phen system, and/or TTA-topo system for the spectrofluorometric determination of the two metals. In the case of Eu determination, 619 nm emission wavelength is used (the determinable range : 0.2 -- 10 ppb Eu), and in the case of Sm determination, 650 nm emission wavelength is adopted (the determinable range : 0.1 -- 1 ppm Sm), because of much higher sensitivity than the other two peaks (569, 606 nm) without interference from europium complex.

  6. Determination of surface recombination velocity and bulk lifetime in detector grade silicon and germanium crystals

    International Nuclear Information System (INIS)

    Derhacobian, N.; Fine, P.; Walton, J.T.; Wong, Y.K.; Rossington, C.S.; Luke, P.N.

    1993-10-01

    Utility of a noncontact photoconductive decay (PCD) technique is demonstrated in measuring bulk lifetime, τ B , and surface recombination velocity, S, in detector grade silicon and germanium crystals. We show that the simple analytical equations which relate the observed effective lifetimes in PCD transients to τ B and S have a limited range of applicability. The noncontact PCD technique is used to determine the effect of several surface treatments on the observed effective lifetimes in Si and Ge. A degradation of the effective lifetime in Si is reported as result of the growth of a thin layer of native oxide at room temperature under atmospheric conditions

  7. Non-Euclidean phasor analysis for quantification of oxidative stress in ex vivo human skin exposed to sun filters using fluorescence lifetime imaging microscopy

    Science.gov (United States)

    Osseiran, Sam; Roider, Elisabeth M.; Wang, Hequn; Suita, Yusuke; Murphy, Michael; Fisher, David E.; Evans, Conor L.

    2017-12-01

    Chemical sun filters are commonly used as active ingredients in sunscreens due to their efficient absorption of ultraviolet (UV) radiation. Yet, it is known that these compounds can photochemically react with UV light and generate reactive oxygen species and oxidative stress in vitro, though this has yet to be validated in vivo. One label-free approach to probe oxidative stress is to measure and compare the relative endogenous fluorescence generated by cellular coenzymes nicotinamide adenine dinucleotides and flavin adenine dinucleotides. However, chemical sun filters are fluorescent, with emissive properties that contaminate endogenous fluorescent signals. To accurately distinguish the source of fluorescence in ex vivo skin samples treated with chemical sun filters, fluorescence lifetime imaging microscopy data were processed on a pixel-by-pixel basis using a non-Euclidean separation algorithm based on Mahalanobis distance and validated on simulated data. Applying this method, ex vivo samples exhibited a small oxidative shift when exposed to sun filters alone, though this shift was much smaller than that imparted by UV irradiation. Given the need for investigative tools to further study the clinical impact of chemical sun filters in patients, the reported methodology may be applied to visualize chemical sun filters and measure oxidative stress in patients' skin.

  8. Fluorescence spectroscopy and confocal microscopy of the mycotoxin citrinin in condensed phase and hydrogel films.

    Science.gov (United States)

    Lauer, Milena H; Gehlen, Marcelo H; de Jesus, Karen; Berlinck, Roberto G S

    2014-05-01

    The emission spectra, quantum yields and fluorescence lifetimes of citrinin in organic solvents and hydrogel films have been determined. Citrinin shows complex fluorescence decays due to the presence of two tautomers in solution and interconversion from excited-state double proton transfer (ESDPT) process. The fluorescence decay times associated with the two tautomers have values near 1 and 5 ns depending on the medium. In hydrogel films of agarose and alginate, fluorescence imaging showed that citrinin is not homogeneously dispersed and highly emissive micrometer spots may be formed. Fluorescence spectrum and decay analysis are used to recognize the presence of citrinin in hydrogel films using confocal fluorescence microscopy and spectroscopy.

  9. Fluorescence resonance energy transfer imaging of CFP/YFP labeled NDH in cyanobacterium cell

    International Nuclear Information System (INIS)

    Ji Dongmei; Lv Wei; Huang Zhengxi; Xia Andong; Xu Min; Ma Weimin; Mi Hualing; Ogawa Teruo

    2007-01-01

    The laser confocal scanning microscopy combined with time-correlated single photon counting imaging technique to obtain fluorescence intensity and fluorescence lifetime images for fluorescence resonance energy transfer measurement is reported. Both the fluorescence lifetime imaging microscopy (FLIM) and intensity images show inhomogeneous cyan fluorescent protein and yellow fluorescent protein (CFP /YFP) expression or inhomogeneous energy transfer between CFP and YFP over whole cell. The results presented in this work show that FLIM could be a potential method to reveal the structure-function behavior of NAD(P)H dehydrogenase complexes in living cell

  10. Sensitive and selective detection of adenine using fluorescent ZnS nanoparticles

    International Nuclear Information System (INIS)

    Meerabai Devi, L; Negi, Devendra P S

    2011-01-01

    We have used fluorescent ZnS nanoparticles as a probe for the determination of adenine. A typical 2 x 10 -7 M concentration of adenine quenches 39.3% of the ZnS fluorescence. The decrease in ZnS fluorescence as a function of adenine concentration was found to be linear in the concentration range 5 x 10 -9 -2 x 10 -7 M. The limit of detection (LOD) of adenine by this method is 3 nM. Among the DNA bases, only adenine quenched the fluorescence of ZnS nanoparticles in the submicromolar concentration range, thus adding selectivity to the method. The amino group of adenine was important in determining the quenching efficiency. Steady-state fluorescence experiments suggest that one molecule of adenine is sufficient to quench the emission arising from a cluster of ZnS consisting of about 20 molecules. Time-resolved fluorescence measurements indicate that the adenine molecules block the sites on the surface of ZnS responsible for emission with the longest lifetime component. This method may be applied for the determination of adenine in biological samples since the measurements have been carried out at pH 7.

  11. 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.

  12. Determination of ATP as a fluorescence probe with europium(III)-doxycycline.

    Science.gov (United States)

    Hou, Faju; Wang, Xiaolei; Jiang, Chongqiu

    2005-03-01

    A new spectrofluorimetric method has been developed for the determination of adenosine disodium triphosphate (ATP). We studied the interactions between the doxycycline (DC)-Eu3+ complex and adenosine disodium triphosphate (ATP) by using UV-visible absorption and fluorescence spectra. Using doxycycline (DC)-Eu3+ as a fluorescence probe, under the optimum conditions, ATP could remarkably enhance the fluorescence intensity of the DC-Eu3+ complex at lambda = 612 nm. The enhanced fluorescence intensity of the Eu3+ ion was in proportion to the concentration of ATP. The optimum conditions for the determination of ATP were also investigated. The linear ranges for ATP were 1.00 x 10(-7) - 2.00 x 10(-6) mol L(-1) with detection limits of 4.07 x 10(-8) mol L(-1). This method is simple, practical and relatively free of interference from coexisting substances, and can be successfully applied to the determination of ATP in samples. The mechanism of fluorescence enhancement between the doxycycline (DC)-Eu3+ complex and ATP was also studied.

  13. Lifetime measurements

    International Nuclear Information System (INIS)

    Poletti, A.R.

    1976-01-01

    Recent developments in experimental methods of measuring the lifetimes of excited nuclear states is reviewed in three main areas. (a) Doppler Shift Attenuation Measurements (DSAM) Times: 10 -14 - 10 -11 sec.; (b) Recoil Distance Measurements (RDM) Times: 10 -9 - 10 -12 sec.; (c) Direct Electronic Timing Times: down to 10 -10 sec.; A measurement of an excited state lifetime can answer a large number of different questions. Two examples are discussed: (a) The determination of the lifetime of an isomeric transition in 93 Tc and its use in determining an upper limit for the magnitude of the parity non-conserving matrix element - /Hsub(PN)/17/2 + >. (b) The dependence of the strength of M2 transitions on isospin in nuclei in the 1dsub(3/2) -1fsub(7/2) region. (author)

  14. Lifetime measurement of the 8s level in francium

    International Nuclear Information System (INIS)

    Gomez, E.; Sprouse, G.D.; Orozco, L.A.; Galvan, A. Perez

    2005-01-01

    We measure the lifetime of the 8s level of 210 Fr atoms on a magneto-optically trapped sample with time-correlated single-photon counting. The 7P 1/2 state serves as the resonant intermediate level for two-step excitation of the 8s level completed with a 1.3-μm laser. Analysis of the fluorescence decay through the 7P 3/2 level gives 53.30±0.44 ns for the 8s level lifetime

  15. From morphology to biochemical state - intravital multiphoton fluorescence lifetime imaging of inflamed human skin

    Science.gov (United States)

    Huck, Volker; Gorzelanny, Christian; Thomas, Kai; Getova, Valentina; Niemeyer, Verena; Zens, Katharina; Unnerstall, Tim R.; Feger, Julia S.; Fallah, Mohammad A.; Metze, Dieter; Ständer, Sonja; Luger, Thomas A.; Koenig, Karsten; Mess, Christian; Schneider, Stefan W.

    2016-03-01

    The application of multiphoton microscopy in the field of biomedical research and advanced diagnostics promises unique insights into the pathophysiology of inflammatory skin diseases. In the present study, we combined multiphoton-based intravital tomography (MPT) and fluorescence lifetime imaging (MPT-FLIM) within the scope of a clinical trial of atopic dermatitis with the aim of providing personalised data on the aetiopathology of inflammation in a non-invasive manner at patients’ bedsides. These ‘optical biopsies’ generated via MPT were morphologically analysed and aligned with classical skin histology. Because of its subcellular resolution, MPT provided evidence of a redistribution of mitochondria in keratinocytes, indicating an altered cellular metabolism. Two independent morphometric algorithms reliably showed an even distribution in healthy skin and a perinuclear accumulation in inflamed skin. Moreover, using MPT-FLIM, detection of the onset and progression of inflammatory processes could be achieved. In conclusion, the change in the distribution of mitochondria upon inflammation and the verification of an altered cellular metabolism facilitate a better understanding of inflammatory skin diseases and may permit early diagnosis and therapy.

  16. 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.

  17. Smart lanthanide coordination polymer fluorescence probe for mercury(II) determination

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Baoxia [Henan Key Laboratory Cultivation Base of Nanobiological Analytical Chemistry, College of Chemistry and Chemical Engineering, Shangqiu Normal University, Shangqiu 476000 (China); Huang, Yankai [Henan Key Laboratory Cultivation Base of Nanobiological Analytical Chemistry, College of Chemistry and Chemical Engineering, Shangqiu Normal University, Shangqiu 476000 (China); College of Chemistry and Molecular Engineering, Zhengzhou University, Zhengzhou 450001 (China); Zhu, Xu; Hao, Yuanqiang [Henan Key Laboratory Cultivation Base of Nanobiological Analytical Chemistry, College of Chemistry and Chemical Engineering, Shangqiu Normal University, Shangqiu 476000 (China); Ding, Yujie [College of Biochemical Engineering, Anhui Polytechnic University, Wuhu 241000 (China); Wei, Wei; Wang, Qi [Henan Key Laboratory Cultivation Base of Nanobiological Analytical Chemistry, College of Chemistry and Chemical Engineering, Shangqiu Normal University, Shangqiu 476000 (China); Qu, Peng, E-mail: qupeng0212@163.com [Henan Key Laboratory Cultivation Base of Nanobiological Analytical Chemistry, College of Chemistry and Chemical Engineering, Shangqiu Normal University, Shangqiu 476000 (China); College of Chemistry and Molecular Engineering, Zhengzhou University, Zhengzhou 450001 (China); Xu, Maotian, E-mail: xumaotian@sqnc.edu.cn [Henan Key Laboratory Cultivation Base of Nanobiological Analytical Chemistry, College of Chemistry and Chemical Engineering, Shangqiu Normal University, Shangqiu 476000 (China); College of Chemistry and Molecular Engineering, Zhengzhou University, Zhengzhou 450001 (China)

    2016-03-17

    Lanthanide coordination polymers (LCPs) have recently emerged as attractive biosensor materials due to their flexible components, high tailorable properties and unique luminescence features. In this work, we designed a smart LCP probe of Tb-CIP/AMP {(CIP, ciprofloxacin) (AMP, adenosine monophosphate)} for Hg{sup 2+} detection by using lanthanide ions as metal nodes, CIP as ligand molecule, and AMP as bridging linker and recognition unit. Tb-CIP/AMP emits strong green luminescence due to the inclusion of AMP, which withdraws the coordinated water molecules and shields Tb{sup 3+} from the quenching effect of O–H vibration in water molecules. The subsequent addition of Hg{sup 2+} into Tb-CIP/AMP can strongly quench the fluorescence because of the specific coordination interaction between AMP and Hg{sup 2+}. As a kind of Hg{sup 2+} nanosensor, the probe exhibited excellent selectivity for Hg{sup 2+} and high sensitivity with detection limit of 0.16 nM. In addition, the probe has long fluorescence lifetime up to millisecond and has been applied to detect Hg{sup 2+} in drinking water and human urine samples with satisfactory results. We envision that our strategy, in the future, could be extended to the designation of other LCP-based hypersensitive time-gated luminescence assays in biological media and biomedical imaging. - Highlights: • Lanthanide coordination polymer of Tb-CIP/AMP was synthesized via a simple self-assembly process. • AMP was employed as a bifunctional molecule for both fluorescence sensitization and target recognition. • Hypersensitive detection of Hg{sup 2+} was achieved based on time-resolved spectroscopy.

  18. Smart lanthanide coordination polymer fluorescence probe for mercury(II) determination

    International Nuclear Information System (INIS)

    Liu, Baoxia; Huang, Yankai; Zhu, Xu; Hao, Yuanqiang; Ding, Yujie; Wei, Wei; Wang, Qi; Qu, Peng; Xu, Maotian

    2016-01-01

    Lanthanide coordination polymers (LCPs) have recently emerged as attractive biosensor materials due to their flexible components, high tailorable properties and unique luminescence features. In this work, we designed a smart LCP probe of Tb-CIP/AMP {(CIP, ciprofloxacin) (AMP, adenosine monophosphate)} for Hg"2"+ detection by using lanthanide ions as metal nodes, CIP as ligand molecule, and AMP as bridging linker and recognition unit. Tb-CIP/AMP emits strong green luminescence due to the inclusion of AMP, which withdraws the coordinated water molecules and shields Tb"3"+ from the quenching effect of O–H vibration in water molecules. The subsequent addition of Hg"2"+ into Tb-CIP/AMP can strongly quench the fluorescence because of the specific coordination interaction between AMP and Hg"2"+. As a kind of Hg"2"+ nanosensor, the probe exhibited excellent selectivity for Hg"2"+ and high sensitivity with detection limit of 0.16 nM. In addition, the probe has long fluorescence lifetime up to millisecond and has been applied to detect Hg"2"+ in drinking water and human urine samples with satisfactory results. We envision that our strategy, in the future, could be extended to the designation of other LCP-based hypersensitive time-gated luminescence assays in biological media and biomedical imaging. - Highlights: • Lanthanide coordination polymer of Tb-CIP/AMP was synthesized via a simple self-assembly process. • AMP was employed as a bifunctional molecule for both fluorescence sensitization and target recognition. • Hypersensitive detection of Hg"2"+ was achieved based on time-resolved spectroscopy.

  19. Double-gated spectral snapshots for biomolecular fluorescence

    International Nuclear Information System (INIS)

    Nakamura, Ryosuke; Hamada, Norio; Ichida, Hideki; Tokunaga, Fumio; Kanematsu, Yasuo

    2007-01-01

    A versatile method to take femtosecond spectral snapshots of fluorescence has been developed based on a double gating technique in the combination of an optical Kerr gate and an image intensifier as an electrically driven gate set in front of a charge-coupled device detector. The application of a conventional optical-Kerr-gate method is limited to molecules with the short fluorescence lifetime up to a few hundred picoseconds, because long-lifetime fluorescence itself behaves as a source of the background signal due to insufficiency of the extinction ratio of polarizers employed for the Kerr gate. By using the image intensifier with the gate time of 200 ps, we have successfully suppressed the background signal and overcome the application limit of optical-Kerr-gate method. The system performance has been demonstrated by measuring time-resolved fluorescence spectra for laser dye solution and the riboflavin solution as a typical sample of biomolecule

  20. Characterization of porcine eyes based on autofluorescence lifetime imaging

    Science.gov (United States)

    Batista, Ana; Breunig, Hans Georg; Uchugonova, Aisada; Morgado, António Miguel; König, Karsten

    2015-03-01

    Multiphoton microscopy is a non-invasive imaging technique with ideal characteristics for biological applications. In this study, we propose to characterize three major structures of the porcine eye, the cornea, crystalline lens, and retina using two-photon excitation fluorescence lifetime imaging microscopy (2PE-FLIM). Samples were imaged using a laser-scanning microscope, consisting of a broadband sub-15 femtosecond (fs) near-infrared laser. Signal detection was performed using a 16-channel photomultiplier tube (PMT) detector (PML-16PMT). Therefore, spectral analysis of the fluorescence lifetime data was possible. To ensure a correct spectral analysis of the autofluorescence lifetime data, the spectra of the individual endogenous fluorophores were acquired with the 16-channel PMT and with a spectrometer. All experiments were performed within 12h of the porcine eye enucleation. We were able to image the cornea, crystalline lens, and retina at multiple depths. Discrimination of each structure based on their autofluorescence intensity and lifetimes was possible. Furthermore, discrimination between different layers of the same structure was also possible. To the best of our knowledge, this was the first time that 2PE-FLIM was used for porcine lens imaging and layer discrimination. With this study we further demonstrated the feasibility of 2PE-FLIM to image and differentiate three of the main components of the eye and its potential as an ophthalmologic technique.

  1. Solvent isotope effect on the fluorescence of azoalkanes

    International Nuclear Information System (INIS)

    Mirbach, M.J.; Mirbach, M.F.; Cherry, W.R.; Turro, N.J.; Engel, P.

    1977-01-01

    A study of fluorescence quantum yields and fluorescence lifetimes of two cyclic azoalkanes reveal a striking dependence of phisub(F) and tausub(F) on solvent and on isotopic substitution (OH → OD). A mechanism involving specific deactivation of the fluorescent state from a hydrogen bonded complex is proposed to rationalize the data. (orig./HK) [de

  2. 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.

  3. TR-LIF LIFETIME MEASUREMENTS AND HFR+CPOL CALCULATIONS OF RADIATIVE PARAMETERS IN VANADIUM ATOM (V I)

    International Nuclear Information System (INIS)

    Wang, Q.; Jiang, L. Y.; Shang, X.; Tian, Y. S.; Dai, Z. W.; Quinet, P.; Palmeri, P.; Zhang, W.

    2014-01-01

    Radiative lifetimes of 79 levels belonging to the 3d 3 4s4p, 3d 4 4p, 3d 3 4s5p, 3d 4 5p, and 3d 3 4s4d configurations of V I with energy from 26,604.807 to 46,862.786 cm –1 have been measured using time-resolved laser-induced fluorescence (TR-LIF) spectroscopy in laser-produced plasma. The lifetime values reported in this paper are in the range of 3.3-494 ns, and the uncertainties of these measurements are within ±10%. A good agreement was obtained with previous data. HFR+CPOL calculations have been performed and used to combine the calculated branching fractions with the available experimental lifetimes to determine semi-empirical transition probabilities for 784 V I transitions

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

    Science.gov (United States)

    DeRose, Paul C; Resch-Genger, Ute

    2010-03-01

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

  5. Absorbance and fluorescence studies on porphyrin Nanostructures ...

    African Journals Online (AJOL)

    The aim of this work was to study some photophysical properties of PNR for application as light harvester in dye sensitized solar cells. These properties included absorbance, fluorescence, and fluorescence quantum yield and lifetime. The results of Transmission Electron Microscope (TEM) images showed the formation of ...

  6. Quantitative frequency-domain fluorescence spectroscopy in tissues and tissue-like media

    Science.gov (United States)

    Cerussi, Albert Edward

    1999-09-01

    In the never-ending quest for improved medical technology at lower cost, modern near-infrared optical spectroscopy offers the possibility of inexpensive technology for quantitative and non-invasive diagnoses. Hemoglobin is the dominant chromophore in the 700-900 nm spectral region and as such it allows for the optical assessment of hemoglobin concentration and tissue oxygenation by absorption spectroscopy. However, there are many other important physiologically relevant compounds or physiological states that cannot be effectively sensed via optical methods because of poor optical contrast. In such cases, contrast enhancements are required. Fluorescence spectroscopy is an attractive component of optical tissue spectroscopy. Exogenous fluorophores, as well as some endogenous ones, may furnish the desperately needed sensitivity and specificity that is lacking in near-infrared optical tissue spectroscopy. The main focus of this thesis was to investigate the generation and propagation of fluorescence photons inside tissues and tissue-like media (i.e., scattering dominated media). The standard concepts of fluorescence spectroscopy have been incorporated into a diffusion-based picture that is sometimes referred to as photon migration. The novelty of this work lies in the successful quantitative recovery of fluorescence lifetimes, absolute fluorescence quantum yields, fluorophore concentrations, emission spectra, and both scattering and absorption coefficients at the emission wavelength from a tissue-like medium. All of these parameters are sensitive to the fluorophore local environment and hence are indicators of the tissue's physiological state. One application demonstrating the capabilities of frequency-domain lifetime spectroscopy in tissue-like media is a study of the binding of ethidium bromide to bovine leukocytes in fresh milk. Ethidium bromide is a fluorescent dye that is commonly used to label DNA, and hence visualize chromosomes in cells. The lifetime of

  7. Nuclear lifetime measurement

    International Nuclear Information System (INIS)

    Guillaume, Georges

    Three direct techniques of lifetime measurement are emphasized: electronic methods and two methods based on the Doppler effect (the recoil distance methods or RDM, the Doppler shift attenuation methods or DSAM). Said direct methods are concerned with the direct measurement of the radioactive decay constants of nuclear excited states. They allow lifetimes of nucleus bound states whose deexcitations occur by electromagnetic transitions, to be determined. Other methods for measuring lifetimes are also examined: microwave techniques and those involving the blocking effect in crystals (direct methods) and also various indirect methods of obtaining lifetimes (γ resonance scattering, capture reactions, inelastic electron and nucleus scattering, and Coulomb deexcitation) [fr

  8. 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.

  9. Investigation of the inclusion behavior between p-sulfoniccalix[8]arene and norfloxacin by fluorescence spectroscopy

    International Nuclear Information System (INIS)

    Wang Xueying; Luo Chuannan; Lv Zhen; Lu Fuguang

    2011-01-01

    The host-guest complexation between p-sulfoniccalix[8]arene (SC 8 A) and norfloxacin (NFLX) in aqueous solution was investigated by fluorescence spectroscopy. Strong fluorescence intensity of the NFLX aqueous solution alone and obvious fluorescence quenching of NFLX solution in the presence of SC 8 A were observed. The fluorescence lifetimes of NFLX and SC 8 A-NFLX inclusion complex were determined and the effect of temperature on SC 8 A-NFLX inclusion complex was studied. The static quenching of the inclusion was obtained, that is the SC 8 A can form a nonfluorescent ground-state inclusion complex with NFLX. As the results show, the combined ratio (n) was 1:1 and association constant K was 1.17x10 5 L/mol. Based on the experimental results, the mechanism of the inclusion complex was explored. The space matching, electrostatic force and hydrogen bond play important effects in the inclusion process. Subsequently, the addition of bovine serum albumin (BSA) solution led to the recovery of fluorescence intensity. It is indicated that BSA can liberate the NFLX into the solution by destructing the SC 8 A-NFLX inclusion complex. Hence SC 8 A may be used for controlled-release drug delivery in the pharmaceutical industry. - Highlights: → Fluorescence lifetimes of NFLX and SC8A-NFLX inclusion complex were determined. → Mechanism of the SC8A-NFLX inclusion complex was explored. → It is proved that SC8A can form a nonfluorescent ground-state inclusion complex with NFLX.

  10. Determination of paraquat in water samples using a sensitive fluorescent probe titration method.

    Science.gov (United States)

    Yao, Feihu; Liu, Hailong; Wang, Guangquan; Du, Liming; Yin, Xiaofen; Fu, Yunlong

    2013-06-01

    Paraquat (PQ), a nonselective herbicide, is non-fluorescent in aqueous solutions. Thus, its determination through direct fluorescent methods is not feasible. The supramolecular inclusion interaction of PQ with cucurbit[7]uril was studied by a fluorescent probe titration method. Significant quenching of the fluorescence intensity of the cucurbit[7]uril-coptisine fluorescent probe was observed with the addition of PQ. A new fluorescent probe titration method with high selectivity and sensitivity at the ng/mL level was developed to determine PQ in aqueous solutions with good precision and accuracy based on the significant quenching of the supramolecular complex fluorescence intensity. The proposed method was successfully used in the determination of PQ in lake water, tap water, well water, and ditch water in an agricultural area, with recoveries of 96.73% to 105.77%. The fluorescence quenching values (deltaF) showed a good linear relationship with PQ concentrations from 1.0 x 10(-8) to 1.2 x 10(-5) mol/L with a detection limit of 3.35 x 10(-9) mol/L. In addition, the interaction models of the supramolecular complexes formed between the host and the guest were established using theoretical calculations. The interaction mechanism between the cucurbit[7]uril and PQ was confirmed by 1H NMR spectroscopy.

  11. Multiphoton autofluorescence lifetime imaging of induced pluripotent stem cells

    Science.gov (United States)

    Uchugonova, Aisada

    2017-06-01

    The multiphoton fluorescence lifetime imaging tomograph MPTflex with its flexible 360-deg scan head, articulated arm, and tunable femtosecond laser source was employed to study induced pluripotent stem cell (iPS) cultures. Autofluorescence (AF) lifetime imaging was performed with 250-ps temporal resolution and submicron spatial resolution using time-correlated single-photon counting. The two-photon excited AF was based on the metabolic coenzymes NAD(P)H and flavin adenine dinucleotide/flavoproteins. iPS cells generated from mouse embryonic fibroblasts (MEFs) and cocultured with growth-arrested MEFs as feeder cells have been studied. Significant differences on AF lifetime signatures were identified between iPS and feeder cells as well as between their differentiating counterparts.

  12. Combination of confocal principle and aperture stop separation improves suppression of crystalline lens fluorescence in an eye model.

    Science.gov (United States)

    Klemm, Matthias; Blum, Johannes; Link, Dietmar; Hammer, Martin; Haueisen, Jens; Schweitzer, Dietrich

    2016-09-01

    Fluorescence lifetime imaging ophthalmoscopy (FLIO) is a new technique to detect changes in the human retina. The autofluorescence decay over time, generated by endogenous fluorophores, is measured in vivo. The strong autofluorescence of the crystalline lens, however, superimposes the intensity decay of the retina fluorescence, as the confocal principle is not able to suppress it sufficiently. Thus, the crystalline lens autofluorescence causes artifacts in the retinal fluorescence lifetimes determined from the intensity decays. Here, we present a new technique to suppress the autofluorescence of the crystalline lens by introducing an annular stop into the detection light path, which we call Schweitzer's principle. The efficacy of annular stops with an outer diameter of 7 mm and inner diameters of 1 to 5 mm are analyzed in an experimental setup using a model eye based on fluorescent dyes. Compared to the confocal principle, Schweitzer's principle with an inner diameter of 3 mm is able to reduce the simulated crystalline lens fluorescence to 4%, while 42% of the simulated retina fluorescence is preserved. Thus, we recommend the implementation of Schweitzer's principle in scanning laser ophthalmoscopes used for fundus autofluorescence measurements, especially the FLIO device, for improved image quality.

  13. Positronium Yields in Liquids Determined by Lifetime and Angular Correlation Measurements

    DEFF Research Database (Denmark)

    Mogensen, O. E.; Jacobsen, F. M.

    1982-01-01

    hydrocarbons), 3.2 (average value for 8 aromatic hydrocarbons), 2.6 (average value for 5 alcohols). Values of this ratio for various other liquids are also given. The results for the mixtures show how I'3 and I'1, vary as the Ps formation is inhibited (CCl4 mixtures) or enhanced C6F6 mixtures). The most......Positron lifetime and angular correlation spectra were measured for 36 pure liquids, CCl4 mixtures with hexane and diethylether, and C6F6 mixtures with hexane. Apparent ortho-Ps yields, I'3, were determined as the intensity of the long-lived component in the lifetime spectra, while the apparent...... para-Ps yields, I'3, were obtained as the intensity of the narrowest gaussian in a three-gaussian fit to the angular correlation spectra. The ratio I'3/I1, expected to be 3, was found to be instead 2.3 (average value for 3 ethers), 2.5 (average value for 10 linear, branched, and cyclic aliphatic...

  14. Synchronous fluorescence based biosensor for albumin determination by cooperative binding of fluorescence probe in a supra-biomolecular host-protein assembly.

    Science.gov (United States)

    Patra, Digambara

    2010-01-15

    A synchronous fluorescence probe based biosensor for estimation of albumin with high sensitivity and selectivity was developed. Unlike conventional fluorescence emission or excitation spectral measurements, synchronous fluorescence measurement offered exclusively a new synchronous fluorescence peak in the shorter wavelength range upon binding of chrysene with protein making it an easy identification tool for albumin determination. The cooperative binding of a fluorescence probe, chrysene, in a supramolecular host-protein assembly during various albumin assessments was investigated. The presence of supramolecular host molecules such as beta-cyclodextrin, curucurbit[6]uril or curucurbit[7]uril have little influence on sensitivity or limit of detection during albumin determination but reduced dramatically interference from various coexisting metal ion quenchers/enhancers. Using the present method the limit of detection for BSA and gamma-Globulin was found to be 0.005 microM which is more sensitive than reported values. Copyright 2009 Elsevier B.V. All rights reserved.

  15. A sensitive fluorescence quenching method for determination of bismuth with tiron

    Energy Technology Data Exchange (ETDEWEB)

    Taher, Mohammad Ali; Rahimi, Mina [Department of Chemistry, Shahid Bahonar University of Kerman, Kerman (Iran, Islamic Republic of); Fazelirad, Hamid, E-mail: hamidfazelirad@gmail.com [Department of Chemistry, Shahid Bahonar University of Kerman, Kerman (Iran, Islamic Republic of); Department of Chemistry, Science and Research Branch, Islamic Azad University, Yazd (Iran, Islamic Republic of); Young Researchers Society, Shahid Bahonar University of Kerman, P.O. Box 76175-133, Kerman (Iran, Islamic Republic of)

    2014-01-15

    We describe a fluorescence quenching method for determination of bismuth with tiron. The method is based on the reaction of tiron by bismuth(III) in acidic media. The influence of variables such as the pH, type of buffer, tiron concentration, reaction time and temperature were investigated. Under optimized conditions, the fluorescence quenching extent is proportional to the concentration of bismuth for Bi–tiron system at the range 0.13–2.09 μg mL{sup −1} and the detection limit is 0.05 μg mL{sup −1}. The proposed sensor presented good repeatability, evaluated in terms of relative standard deviation (R.S.D.=±0.498%) for 11 replicates. This sensitive, rapid and accurate method has been successfully applied to the determination of trace bismuth(III) in water and hair samples and certified reference materials. -- Highlights: • No previous paper report on use of fluorescence quenching for determination of Bi. • Fluorescence quenching of trion is a sensitive method for determination of Bi(III). • Under the optimum conditions the detection limit is very low (0.05 μg mL{sup −1}). • The procedure is simple and safe and has high tolerance limit to interferences.

  16. Laser induced fluorescence and phosphorescence of matrix isolated glyoxal - Evidence for exciplex formation in the A 1Au and a 3Au states

    Science.gov (United States)

    Van Ijzendoorn, L. J.; Baas, F.; Koernig, S.; Greenberg, J. M.; Allamandola, L. J.

    1986-01-01

    Laser-induced fluorescence and phosphorescence as well as infrared and visible absorption spectra of glyoxal in Ar, N2, and CO matrices are presented and analyzed. Glyoxal in its first excited electronic state is shown to form an exciplex with its nearest neighbors in all three matrices, and transitions normally forbidden dominate the emission spectra. The spectral characteristics of these complexes are similar to those of the Ar-glyoxal complex found in supersonic beam experiments. Due to the matrix cage effect, no vibrational predissociation is observed. The phosphorescence lifetime is determined and an upper limit is given for the fluorescence lifetime. This, in combination with the relative intensities of fluorescence and phosphorescence, can be used to place limits on the quantum yields of the various relaxation processes.

  17. From morphology to clinical pathophysiology: multiphoton fluorescence lifetime imaging at patients' bedside

    Science.gov (United States)

    Mess, Christian; Zens, Katharina; Gorzelanny, Christian; Metze, Dieter; Luger, Thomas A.; König, Karsten; Schneider, Stefan W.; Huck, Volker

    2017-02-01

    Application of multiphoton microscopy in the field of biomedical research and advanced diagnostics promises unique insights into the pathophysiology of skin diseases. By means of multiphoton excitation, endogenous biomolecules like NADH, collagen or elastin show autofluorescence or second harmonic generation. Thus, these molecules provide information about the subcellular morphology, epidermal architecture and physiological condition of the skin. To gain a deeper understanding of the linkage between cellular structure and physiological processes, non-invasive multiphotonbased intravital tomography (MPT) and fluorescence lifetime imaging (FLIM) were combined within the scopes of inflammatory skin, chronic wounds and drug delivery in clinical application. The optical biopsies generated via MPT were morphologically analyzed and aligned with classical skin histology. Because of its subcellular resolution, MPT provided evidence of a redistribution of mitochondria in keratinocytes, indicating an altered cellular metabolism. Independent morphometric algorithms reliably showed a perinuclear accumulation in lesional skin in contrast to an even distribution in healthy skin. Confirmatively, MPT-FLIM showed an obvious metabolic shift in lesions. Moreover, detection of the onset and progression of inflammatory processes could be achieved. The feasibility of primary in vivo tracking of applied therapeutic agents further broadened our scope: We examined the permeation and subsequent distribution of agents directly visualized in patientś skin in short-term repetitive measurements. Furthermore, we performed MPT-FLIM follow-up investigations in the long-term course of therapy. Therefore, clinical MPT-FLIM application offers new insights into the pathophysiology and the individual therapeutic course of skin diseases, facilitating a better understanding of the processes of inflammation and wound healing.

  18. Observation of rotating nuclear molecules and determination of their lifetimes

    Energy Technology Data Exchange (ETDEWEB)

    Comas, V.; Heinz, S.; Ackermann, D.; Heredia, J.; Hessberger, F.P.; Khuyagbaatar, J.; Kindler, B.; Lommel, B.; Mann, R. [GSI Helmholtzzentrum fuer Schwerionenforschung GmbH, Darmstadt (Germany); Hofmann, S. [GSI Helmholtzzentrum fuer Schwerionenforschung GmbH, Darmstadt (Germany); Goethe-Universitaet Frankfurt, Institut fuer Physik, Frankfurt (Germany)

    2012-12-15

    Long-living rotating nuclear molecules (or ''dinuclear systems'') have been observed at the velocity filter SHIP at GSI in reactions of {sup 64}Ni + {sup 207}Pb at Coulomb barrier energies. The rotation was directly revealed by the velocity spectra of deep inelastic target-like transfer products which are formed during the lifetime of the nuclear molecule and emitted after its breakup. The corresponding rotation angles were about 180 degree pointing to long nuclear interaction times or lifetimes of the system, respectively. We deduced the lifetimes from the lines in the velocity spectra originating from two different rotation angles. Further, the unambiguous correlation of a certain transfer product with its individual velocity spectrum allowed us to study the lifetimes as a function of the number of transferred protons. (orig.)

  19. Smart optical probes for near-infrared fluorescence imaging of Alzheimer's disease pathology

    International Nuclear Information System (INIS)

    Raymond, Scott B.; Bacskai, Brian J.; Skoch, Jesse; Hills, Ivory D.; Swager, Timothy M.; Nesterov, Evgueni E.

    2008-01-01

    Near-infrared fluorescent probes for amyloid-beta (Aβ) are an exciting option for molecular imaging in Alzheimer's disease research and may translate to clinical diagnostics. However, Aβ-targeted optical probes often suffer from poor specificity and slow clearance from the brain. We are designing smart optical probes that emit characteristic fluorescence signal only when bound to Aβ. We synthesized a family of dyes and tested Aβ-binding sensitivity with fluorescence spectroscopy and tissue-staining. Select compounds exhibited Aβ-dependent changes in fluorescence quantum yield, lifetime, and emission spectra that may be imaged microscopically or in vivo using new lifetime and spectral fluorescence imaging techniques. Smart optical probes that turn on when bound to Aβ will improve amyloid detection and may enable quantitative molecular imaging in vivo. (orig.)

  20. Theoretical and experimental investigation of atomic radiative lifetimes and hyperfine structures

    International Nuclear Information System (INIS)

    Joensson, Per.

    1992-01-01

    Atomic radiative lifetimes and hyperfine structures as well as other properties, such as total energy and specific mass shift, have been studied theoretically and experimentally. Computer programs to calculate hyperfine structure constants from non-relativistic multiconfiguration Hartree-Fock (MCHF) and relativistic multi-configuration Dirac-Fock (MCDF) wavefunctions have been written. Using these programs large-scale calculations of hyperfine structures in lithium and sodium have been performed. It is shown, that the MCHF method is able to predict hyperfine structures to an accuracy of a few per mille in lithium, whereas for the more complex hyperfine structures to an accuracy of a few per mille in lithium, whereas for the more complex sodium atom an accuracy of a few per cent is obtainable. For lithium convergence of the total energy, ionization energy, specific mass shift and hyperfine parameters has been studied with the MCHF method. Radiative lifetimes and hyperfine structures of excited states in sodium and silver have been experimentally determined using time-resolved laser spectroscopy. By recording the fluorescence light decay curves following VUV excitation, the radiative lifetimes and hyperfine structures of the 7p 2 P states in silver were measured. The delayed-coincidence technique has been used to make very accurate measurements of the radiative lifetimes and hyperfine structures of the lowest P states in sodium and silver

  1. Design of peptide substrates for nanosecond time-resolved fluorescence assays of proteases: 2,3-diazabicyclo[2.2.2]oct-2-ene as a noninvasive fluorophore.

    Science.gov (United States)

    Hennig, Andreas; Florea, Mara; Roth, Doris; Enderle, Thilo; Nau, Werner M

    2007-01-15

    Fluorescence protease assays were investigated with peptide substrates containing a 2,3-diazabicyclo[2.2.2]oct-2-ene-labeled asparagine (Dbo) as a fluorescent amino acid. The special characteristic of the fluorophore Dbo is its exceedingly long fluorescence lifetime (ca. 300 ns in water under air), which allows the use of nanosecond time-resolved fluorescence (Nano-TRF) detection to efficiently suppress shorter-lived background emission. In addition, the natural amino acids tryptophan and tyrosine can be employed as intramolecular fluorescence quenchers, which facilitates substrate design. Fourteen synthetic peptide substrates (composed of 2-19 amino acids) and five enzymes (trypsin, pepsin, carboxypeptidase A, leucine aminopeptidase, and chymotrypsin) were investigated and, in all 28 examined combinations, enzymatic activity was detected by monitoring the increase in steady state fluorescence with time and determining the reaction rates as kcat/Km values, which ranged from 0.2 to 80x10(6) M-1 min-1. The results suggest an excellent compatibility of the very small and hydrophilic fluorescent probe Dbo with solid-phase peptide synthesis and the investigated proteases. For all 14 peptides the fluorescence lifetimes before and after enzymatic cleavage were measured and Nano-TRF measurements were performed in 384-well microplates. The fluorescence lifetimes of the different peptides provide the basis for the rational design of Dbo-based fluorescent substrates for protease assays. Measurements in Nano-TRF mode revealed, in addition to efficient suppression of background fluorescence, an increased differentiation between cleaved and uncleaved substrate. The Dbo-based assays can be adapted for high-throughput screening.

  2. Characterization of a new series of fluorescent probes for imaging membrane order.

    Directory of Open Access Journals (Sweden)

    Joanna M Kwiatek

    Full Text Available Visualization and quantification of lipid order is an important tool in membrane biophysics and cell biology, but the availability of environmentally sensitive fluorescent membrane probes is limited. Here, we present the characterization of the novel fluorescent dyes PY3304, PY3174 and PY3184, whose fluorescence properties are sensitive to membrane lipid order. In artificial bilayers, the fluorescence emission spectra are red-shifted between the liquid-ordered and liquid-disordered phases. Using ratiometric imaging we demonstrate that the degree of membrane order can be quantitatively determined in artificial liposomes as well as live cells and intact, live zebrafish embryos. Finally, we show that the fluorescence lifetime of the dyes is also dependent on bilayer order. These probes expand the current palate of lipid order-sensing fluorophores affording greater flexibility in the excitation/emission wavelengths and possibly new opportunities in membrane biology.

  3. Positron annihilation lifetime spectroscopy source correction determination: A simulation study

    Energy Technology Data Exchange (ETDEWEB)

    Kanda, Gurmeet S.; Keeble, David J., E-mail: d.j.keeble@dundee.ac.uk

    2016-02-01

    Positron annihilation lifetime spectroscopy (PALS) can provide sensitive detection and identification of vacancy-related point defects in materials. These measurements are normally performed using a positron source supported, and enclosed by, a thin foil. Annihilation events from this source arrangement must be quantified and are normally subtracted from the spectrum before analysis of the material lifetime components proceeds. Here simulated PALS spectra reproducing source correction evaluation experiments have been systematically fitted and analysed using the packages PALSfit and MELT. Simulations were performed assuming a single lifetime material, and for a material with two lifetime components. Source correction terms representing a directly deposited source and various foil supported sources were added. It is shown that in principle these source terms can be extracted from suitably designed experiments, but that fitting a number of independent, nominally identical, spectra is recommended.

  4. Technological determinants of the lifetime well-being in the 21th century

    Directory of Open Access Journals (Sweden)

    Zhironkin Sergey

    2016-01-01

    Full Text Available The paper deals with technological determinants of the lifetime social well-being, as conditions of maintaining the level and style of consumption, social status throughout the whole life. Despite the high importance of the problem of the social well-being and the relevance of its solution for the development of modern society, the issues of its correlation with the technological level of production and the development of information technologies are still poorly researched. As the problem of the life-time wellbeing take on special significance, it is important to analyze its conditions, related not only to social benefits, but also to the scientific and technological progress. For the future innovation and the digital stage of social well-being is its network well-being. It is based on the usage of the Internet for professional activity and personal self-fulfillment during the whole conscious life. Network well-being also reflects the convergence of information and network, industrial, financial, marketing, and social technologies. The formation of the life-time network well-being requires from the Government some measures of social adjustment, fiscal policy and the regulation of employment to encourage Internet Education and Internet employment, enhancing the prestige of working in high-tech industries.

  5. Fluorescence detection of single molecules using pulsed near-field optical excitation and time correlated photon counting

    International Nuclear Information System (INIS)

    Ambrose, W.P.; Goodwin, P.M.; Martin, J.C.; Keller, R.A.

    1994-01-01

    Pulsed excitation, time correlated single photon counting and time gated detection are used in near-field optical microscopy to enhance fluorescence images and measure the fluorescence lifetimes of single molecules of Rhodamine 6G on silica surfaces. Time gated detection is used to reject prompt scattered background and to improve the image signal to noise ratio. The excited state lifetime of a single Rhodamine 6G molecule is found to depend on the position of the near-field probe. We attribute the lifetime variations to spontaneous emission rate alterations by the fluorescence reflected from and quenching by the aluminum coated probe

  6. [The intraoperative determination of intestinal vitality with a fluorescent indicator].

    Science.gov (United States)

    Ivanov, A; Terziev, I

    1997-01-01

    Intestinal obstruction due to strangulation is induced in dogs under experimental conditions, with intestinal wall vitality assessment done on the ground of standard clinical criteria, using fluorescence dye and UV rays, as well as histological study. Sensitivity, specificity and prognostic value of each of the methods employed are determined. The fluorescence method advantages are recorded, and the prospects of its clinical implementation are estimated.

  7. Study on the fluorescence characteristics of carbon dots

    Science.gov (United States)

    Mao, Xiao-Jiao; Zheng, Hu-Zhi; Long, Yi-Juan; Du, Juan; Hao, Jian-Yu; Wang, Ling-Ling; Zhou, Dong-Bo

    2010-02-01

    Herein, we prepared water-soluble fluorescent carbon dots with diameter about 1.5 nm from cheap commercial lampblack. These fluorescent carbon nanoparticles are stable toward photobleaching and stable in water for more than half a year without fluorescence decrease. In order to improve its fluorescence properties, we passivated these nanoparticles with bisamino-terminated polyethylene glycol (PEG 1500N). Therefore, both fluorescence quantum yield and lifetime increased after this progress. In addition, the passivated carbon dots were more inert to solvent than the bare one and showed different responses to pH change.

  8. Using chlorophyll fluorescence to determine stress in Eucalyptus ...

    African Journals Online (AJOL)

    Using chlorophyll fluorescence to determine stress in Eucalyptus grandis seedlings: scientific paper. ... Southern Forests: a Journal of Forest Science ... factors affect the functioning of the photosynthetic system, the status of the photosynthetic apparatus is a good indicator of the plant in terms of stress and stress adaptation.

  9. Dual fluorescence of single LH2 antenna nanorings

    International Nuclear Information System (INIS)

    Freiberg, A.; Raetsep, M.; Timpmann, K.; Trinkunas, G.

    2004-01-01

    A dual nature of fluorescence from LH2 pigment-protein complexes, which is a part of the light harvesting system of purple bacteria, is confirmed by fluorescence-lifetime dependence on recording wavelength and spectrally selective spectroscopy. An analysis based on the Holstein molecular crystal model, modified by allowing diagonal disorder, suggests coexistence of large- and small-radius self-trapped excitons, which serve as the origin of the dual fluorescence

  10. A new approach to determine accurately minority-carrier lifetime

    International Nuclear Information System (INIS)

    Idali Oumhand, M.; Mir, Y.; Zazoui, M.

    2009-01-01

    Electron or proton irradiations introduce recombination centers, which tend to affect solar cell parameters by reducing the minority-carrier lifetime (MCLT). Because this MCLT plays a fundamental role in the performance degradation of solar cells, in this work we present a new approach that allows us to get accurate values of MCLT. The relationship between MCLT in p-region and n-region both before and after irradiation has been determined by the new method. The validity and accuracy of this approach are justified by the fact that the degradation parameters that fit the experimental data are the same for both short-circuit current and the open-circuit voltages. This method is applied to the p + /n-InGaP solar cell under 1 MeV electron irradiation

  11. Improving fluorescence diagnosis of cancer by SLIM

    Science.gov (United States)

    Rück, Angelika; Dolp, Frank; Kinzler, Ingrid; Hauser, Carmen; Scalfi-Happ, Claudia

    2006-02-01

    Although during the last years, significant progress was made in cancer diagnosis, using either intrinsic or specially designed fluorophores, still problems exist, due to difficulties in spectral separation of highly overlapping probes or in lack of specificity. Many of the problems could be circumvented by focusing on time-resolved methods. In combination with spectral resolved detection (spectral fluorescence lifetime imaging, SLIM) highly sophisticated fluorescence lifetime imaging can be performed which might improve specificity of cell diagnosis. To record lifetime images (τ-mapping) with spectral resolution a setup was realized consisting of a laser scanning microscope equipped with a 16 channel array for time-correlated single photon counting (TCSPC) and a spectrograph in front of the array. A Ti:Saphir laser can be used for excitation or alternatively ps diode lasers. With this system the time- and spectral-resolved fluorescence characteristics of different fluorophores were investigated in solution and in cell culture. As an example, not only the mitochondria staining dye rhodamine 123 could be easily distinguished from DAPI, which intercalates into nucleic acids, but also different binding sites of DAPI. This was proved by the appearance of different lifetime components within different spectral channels. Another example is Photofrin, a photosensitizer which is approved for bladder cancer and for palliative lung and esophageal cancer in 20 countries, including the United States, Canada and many European countries. Photofrin is a complex mixture of different monomeric and aggregated porphyrins. The phototoxic efficiency during photodynamic therapy (PDT) seems to be correlated with the relative amounts of monomers and aggregates. With SLIM different lifetimes could be attributed to various, spectrally highly overlapping compounds. In addition, a detailed analysis of the autofluorescence by SLIM could explain changes of mitochondrial metabolism during

  12. Automatic segmentation of fluorescence lifetime microscopy images of cells using multiresolution community detection--a first study.

    Science.gov (United States)

    Hu, D; Sarder, P; Ronhovde, P; Orthaus, S; Achilefu, S; Nussinov, Z

    2014-01-01

    Inspired by a multiresolution community detection based network segmentation method, we suggest an automatic method for segmenting fluorescence lifetime (FLT) imaging microscopy (FLIM) images of cells in a first pilot investigation on two selected images. The image processing problem is framed as identifying segments with respective average FLTs against the background in FLIM images. The proposed method segments a FLIM image for a given resolution of the network defined using image pixels as the nodes and similarity between the FLTs of the pixels as the edges. In the resulting segmentation, low network resolution leads to larger segments, and high network resolution leads to smaller segments. Furthermore, using the proposed method, the mean-square error in estimating the FLT segments in a FLIM image was found to consistently decrease with increasing resolution of the corresponding network. The multiresolution community detection method appeared to perform better than a popular spectral clustering-based method in performing FLIM image segmentation. At high resolution, the spectral segmentation method introduced noisy segments in its output, and it was unable to achieve a consistent decrease in mean-square error with increasing resolution. © 2013 The Authors Journal of Microscopy © 2013 Royal Microscopical Society.

  13. Riboflavin enhanced fluorescence of highly reduced graphene oxide

    Science.gov (United States)

    Iliut, Maria; Gabudean, Ana-Maria; Leordean, Cosmin; Simon, Timea; Teodorescu, Cristian-Mihail; Astilean, Simion

    2013-10-01

    The improvement of graphene derivates' fluorescence properties is a challenging topic and very few ways were reported up to now. In this Letter we propose an easy method to enhance the fluorescence of highly reduced graphene oxide (rGO) through non-covalent binding to a molecular fluorophore, namely the riboflavin (Rb). While the fluorescence of Rb is quenched, the Rb - decorated rGO exhibits strong blue fluorescence and significantly increased fluorescence lifetime, as compared to its pristine form. The data reported here represent a promising start towards tailoring the optical properties of rGOs, having utmost importance in optical applications.

  14. The fluorescence intensities ratio is not a reliable parameter for evaluation of protein unfolding transitions.

    Science.gov (United States)

    Žoldák, Gabriel; Jancura, Daniel; Sedlák, Erik

    2017-06-01

    Monitoring the fluorescence of proteins, particularly the fluorescence of intrinsic tryptophan residues, is a popular method often used in the analysis of unfolding transitions (induced by temperature, chemical denaturant, and pH) in proteins. The tryptophan fluorescence provides several suitable parameters, such as steady-state fluorescence intensity, apparent quantum yield, mean fluorescence lifetime, position of emission maximum that are often utilized for the observation of the conformational/unfolding transitions of proteins. In addition, the fluorescence intensities ratio at different wavelengths (usually at 330 nm and 350 nm) is becoming an increasingly popular parameter for the evaluation of thermal transitions. We show that, under certain conditions, the use of this parameter for the analysis of unfolding transitions leads to the incorrect determination of thermodynamic parameters characterizing unfolding transitions in proteins (e.g., melting temperature) and, hence, can compromise the hit identification during high-throughput drug screening campaigns. © 2017 The Protein Society.

  15. Fluorescence lifetime correlation spectroscopy combined with lifetime tuning: New perspectives in supported phospholipid bilayer research

    Czech Academy of Sciences Publication Activity Database

    Benda, Aleš; Fagulová, Veronika; Deyneka, Alexander; Enderlain, J.; Hof, Martin

    2006-01-01

    Roč. 22, č. 23 (2006), s. 9580-9585 ISSN 0743-7463 R&D Projects: GA ČR GA203/05/2308; GA MŠk LC06063 Institutional research plan: CEZ:AV0Z40400503; CEZ:AV0Z10100522 Keywords : spectroscopy * fluorescence * FLCS Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 3.902, year: 2006

  16. X-ray Free Electron Laser Determination of Crystal Structures of Dark and Light States of a Reversibly Photoswitching Fluorescent Protein at Room Temperature

    Directory of Open Access Journals (Sweden)

    Christopher D. M. Hutchison

    2017-09-01

    Full Text Available The photochromic fluorescent protein Skylan-NS (Nonlinear Structured illumination variant mEos3.1H62L is a reversibly photoswitchable fluorescent protein which has an unilluminated/ground state with an anionic and cis chromophore conformation and high fluorescence quantum yield. Photo-conversion with illumination at 515 nm generates a meta-stable intermediate with neutral trans-chromophore structure that has a 4 h lifetime. We present X-ray crystal structures of the cis (on state at 1.9 Angstrom resolution and the trans (off state at a limiting resolution of 1.55 Angstrom from serial femtosecond crystallography experiments conducted at SPring-8 Angstrom Compact Free Electron Laser (SACLA at 7.0 keV and 10.5 keV, and at Linac Coherent Light Source (LCLS at 9.5 keV. We present a comparison of the data reduction and structure determination statistics for the two facilities which differ in flux, beam characteristics and detector technologies. Furthermore, a comparison of droplet on demand, grease injection and Gas Dynamic Virtual Nozzle (GDVN injection shows no significant differences in limiting resolution. The photoconversion of the on- to the off-state includes both internal and surface exposed protein structural changes, occurring in regions that lack crystal contacts in the orthorhombic crystal form.

  17. Determination of adenosine disodium triphosphate using prulifloxacin-terbium(III) as a fluorescence probe by spectrofluorimetry

    International Nuclear Information System (INIS)

    Yu Fengshan; Li Lin; Chen Fang

    2008-01-01

    A new spectrofluorimetric method is developed for determination of adenosine disodium triphosphate (ATP). The interactions between prulifloxacin (PUFX)-Tb 3+ complex and adenosine disodium triphosphate has been studied by using UV-vis absorption and fluorescence spectra. Using prulifloxacin-Tb 3+ as a fluorescence probe, under the optimum conditions, ATP can remarkably enhance the fluorescence intensity of the prulifloxacin-Tb 3+ complex at λ = 545 nm and the enhanced fluorescence intensity is in proportion to the concentration of ATP. Optimum conditions for the determination of ATP were also investigated. The dynamic range for the determination of ATP is 4.0 x 10 -7 to 2.0 x 10 -5 mol L -1 , and the detection limit (3 σ/k) is 1.7 x 10 -8 mol L -1 . This method is simple, practical and relatively free interference from coexisting substances and can be successfully applied to determination of ATP in real pharmaceutical samples. The mechanism of fluorescence enhancement of prulifloxacin-Tb 3+ complex by ATP was also discussed

  18. ZnSe quantum dots based fluorescence quenching method for determination of paeoniflorin

    International Nuclear Information System (INIS)

    Chen, Zhi; Chen, Jiayi; Liang, Qiaowen; Wu, Dudu; Zeng, Yuaner; Jiang, Bin

    2014-01-01

    Water soluble ZnSe quantum dots (QDs) modified by mercaptoacetic acid (MAA) were used to determinate paeoniflorin in aqueous solutions by the fluorescence spectroscopic technique. The results showed that the fluorescence of the modified ZnSe QDs could be quenched by paeoniflorin effectively in physiological buffer solution. The optimum fluorescence intensity was found to be at incubation time 10 min, pH 7.0 and temperature 25 °C. Under the optimal conditions, the detection limit of paeoniflorin was 7.30×10 −7 mol L −1 . Moreover, the quenching mechanism was discussed to be a static quenching procedure, which was proved by quenching rate constant K q (1.02×10 13 L mol −1 s −1 ). -- Highlights: • The fluorescence intensity of ZnSe QDs could be quenched by paeoniflorin. • Foreign substance showed insignificant effect for determination of paeoniflorin. • The quenching mechanism was discussed to be a static quenching procedure

  19. ZnSe quantum dots based fluorescence quenching method for determination of paeoniflorin

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Zhi [Center of Analysis, Guangdong Medical College, Dongguan 523808 (China); School of Chinese Herbal Medicine, Guangzhou University of Chinese Medicine, Guangzhou 510006 (China); Chen, Jiayi; Liang, Qiaowen [School of Chinese Herbal Medicine, Guangzhou University of Chinese Medicine, Guangzhou 510006 (China); Wu, Dudu [Center of Analysis, Guangdong Medical College, Dongguan 523808 (China); Zeng, Yuaner, E-mail: zengyuaner@126.com [School of Chinese Herbal Medicine, Guangzhou University of Chinese Medicine, Guangzhou 510006 (China); Jiang, Bin, E-mail: gzjiangbin@hotmail.com [School of Chinese Herbal Medicine, Guangzhou University of Chinese Medicine, Guangzhou 510006 (China)

    2014-01-15

    Water soluble ZnSe quantum dots (QDs) modified by mercaptoacetic acid (MAA) were used to determinate paeoniflorin in aqueous solutions by the fluorescence spectroscopic technique. The results showed that the fluorescence of the modified ZnSe QDs could be quenched by paeoniflorin effectively in physiological buffer solution. The optimum fluorescence intensity was found to be at incubation time 10 min, pH 7.0 and temperature 25 °C. Under the optimal conditions, the detection limit of paeoniflorin was 7.30×10{sup −7} mol L{sup −1}. Moreover, the quenching mechanism was discussed to be a static quenching procedure, which was proved by quenching rate constant K{sub q} (1.02×10{sup 13} L mol{sup −1} s{sup −1}). -- Highlights: • The fluorescence intensity of ZnSe QDs could be quenched by paeoniflorin. • Foreign substance showed insignificant effect for determination of paeoniflorin. • The quenching mechanism was discussed to be a static quenching procedure.

  20. Fluorescence lifetime studies of MeV erbium implanted silica glass

    International Nuclear Information System (INIS)

    Lidgard, A.; Polman, A.; Jacobsen, D.C.; Blonder, G.E.; Kistler, R.; Poate, J.M.; Becker, P.C.

    1991-01-01

    MeV erbium ion implantation into various SiO 2 glasses has been studied with the aim of incorporating the rare-earth dopant as an optically active ion in the silica network. The lifetime of the excited state ranges from 1.6 to 12.8 ms, depending on base material and implantation fluence. These results have positive implications for silica-based integrated optical technology. (Author)

  1. Fluorescence lifetime studies of MeV erbium implanted silica glass

    Energy Technology Data Exchange (ETDEWEB)

    Lidgard, A.; Polman, A.; Jacobsen, D.C.; Blonder, G.E.; Kistler, R.; Poate, J.M.; Becker, P.C. (AT and T Bell Labs., Murray Hill, NJ (USA))

    1991-05-23

    MeV erbium ion implantation into various SiO{sub 2} glasses has been studied with the aim of incorporating the rare-earth dopant as an optically active ion in the silica network. The lifetime of the excited state ranges from 1.6 to 12.8 ms, depending on base material and implantation fluence. These results have positive implications for silica-based integrated optical technology. (Author).

  2. Absorption and fluorescence spectroscopic characterization of BLUF domain of AppA from Rhodobacter sphaeroides

    Science.gov (United States)

    Zirak, P.; Penzkofer, A.; Schiereis, T.; Hegemann, P.; Jung, A.; Schlichting, I.

    2005-08-01

    The BLUF domain of the transcriptional anti-repressor protein AppA from the non-sulfur anoxyphototrophic purple bacterium Rhodobacter sphaeroides was characterized by absorption and emission spectroscopy. The BLUF domain constructs AppA 148 (consisting of amino-acid residues 1-148) and AppA 126 (amino-acid residues 1-126) are investigated. The cofactor of the investigated domains is found to consist of a mixture of the flavins riboflavin, FMN, and FAD. The dark-adapted domains exist in two different active receptor conformations (receptor states) with different sub-nanosecond fluorescence lifetimes (BLUF r,f and BLUF r,sl) and a small non-interacting conformation (BLUF nc). The active receptor conformations are transformed to putative signalling states (BLUF s,f and BLUF s,sl) of low fluorescence efficiency and picosecond fluorescence lifetime by blue-light excitation (light-adapted domains). In the dark at room temperature both signalling states recover back to the initial receptor states with a time constant of about 17 min. A quantum yield of signalling state formation of about 25% was determined by intensity dependent transmission measurements. A photo-cycle scheme is presented including photo-induced charge transfer complex formation, charge recombination, and protein binding pocket reorganisation.

  3. Absorption and fluorescence spectroscopic characterization of BLUF domain of AppA from Rhodobacter sphaeroides

    International Nuclear Information System (INIS)

    Zirak, P.; Penzkofer, A.; Schiereis, T.; Hegemann, P.; Jung, A.; Schlichting, I.

    2005-01-01

    The BLUF domain of the transcriptional anti-repressor protein AppA from the non-sulfur anoxyphototrophic purple bacterium Rhodobacter sphaeroides was characterized by absorption and emission spectroscopy. The BLUF domain constructs AppA 148 (consisting of amino-acid residues 1-148) and AppA 126 (amino-acid residues 1-126) are investigated. The cofactor of the investigated domains is found to consist of a mixture of the flavins riboflavin, FMN, and FAD. The dark-adapted domains exist in two different active receptor conformations (receptor states) with different sub-nanosecond fluorescence lifetimes (BLUF r,f and BLUF r,sl ) and a small non-interacting conformation (BLUF nc ). The active receptor conformations are transformed to putative signalling states (BLUF s,f and BLUF s,sl ) of low fluorescence efficiency and picosecond fluorescence lifetime by blue-light excitation (light-adapted domains). In the dark at room temperature both signalling states recover back to the initial receptor states with a time constant of about 17 min. A quantum yield of signalling state formation of about 25% was determined by intensity dependent transmission measurements. A photo-cycle scheme is presented including photo-induced charge transfer complex formation, charge recombination, and protein binding pocket reorganisation

  4. Combination of confocal principle and aperture stop separation improves suppression of crystalline lens fluorescence in an eye model

    OpenAIRE

    Klemm, Matthias; Blum, Johannes; Link, Dietmar; Hammer, Martin; Haueisen, Jens; Schweitzer, Dietrich

    2016-01-01

    Fluorescence lifetime imaging ophthalmoscopy (FLIO) is a new technique to detect changes in the human retina. The autofluorescence decay over time, generated by endogenous fluorophores, is measured in vivo. The strong autofluorescence of the crystalline lens, however, superimposes the intensity decay of the retina fluorescence, as the confocal principle is not able to suppress it sufficiently. Thus, the crystalline lens autofluorescence causes artifacts in the retinal fluorescence lifetimes d...

  5. Filter replacement lifetime prediction

    Science.gov (United States)

    Hamann, Hendrik F.; Klein, Levente I.; Manzer, Dennis G.; Marianno, Fernando J.

    2017-10-25

    Methods and systems for predicting a filter lifetime include building a filter effectiveness history based on contaminant sensor information associated with a filter; determining a rate of filter consumption with a processor based on the filter effectiveness history; and determining a remaining filter lifetime based on the determined rate of filter consumption. Methods and systems for increasing filter economy include measuring contaminants in an internal and an external environment; determining a cost of a corrosion rate increase if unfiltered external air intake is increased for cooling; determining a cost of increased air pressure to filter external air; and if the cost of filtering external air exceeds the cost of the corrosion rate increase, increasing an intake of unfiltered external air.

  6. Detection of radiation-induced brain necrosis in live rats using label-free time-resolved fluorescence spectroscopy (TRFS) (Conference Presentation)

    Science.gov (United States)

    Hartl, Brad A.; Ma, Htet S. W.; Sridharan, Shamira; Hansen, Katherine; Klich, Melanie; Perks, Julian; Kent, Michael; Kim, Kyoungmi; Fragoso, Ruben; Marcu, Laura

    2017-02-01

    Differentiating radiation-induced necrosis from recurrent tumor in the brain remains a significant challenge to the neurosurgeon. Clinical imaging modalities are not able to reliably discriminate the two tissue types, making biopsy location selection and surgical management difficult. Label-free fluorescence lifetime techniques have previously been shown to be able to delineate human brain tumor from healthy tissues. Thus, fluorescence lifetime techniques represent a potential means to discriminate the two tissues in real-time during surgery. This study aims to characterize the endogenous fluorescence lifetime signatures from radiation induced brain necrosis in a tumor-free rat model. Fischer rats received a single fraction of 60 Gy of radiation to the right hemisphere using a linear accelerator. Animals underwent a terminal live surgery after gross necrosis had developed, as verified with MRI. During surgery, healthy and necrotic brain tissue was measured with a fiber optic needle connected to a multispectral fluorescence lifetime system. Measurements of the necrotic tissue showed a 48% decrease in intensity and 20% increase in lifetimes relative to healthy tissue. Using a support vector machine classifier and leave-one-out validation technique, the necrotic tissue was correctly classified with 94% sensitivity and 97% specificity. Spectral contribution analysis also confirmed that the primary source of fluorescence contrast lies within the redox and bound-unbound population shifts of nicotinamide adenine dinucleotide. A clinical trial is presently underway to measure these tissue types in humans. These results show for the first time that radiation-induced necrotic tissue in the brain contains significantly different metabolic signatures that are detectable with label-free fluorescence lifetime techniques.

  7. Gated Detection Measurements of Phosphorescence Lifetimes

    Directory of Open Access Journals (Sweden)

    Yordan Kostov

    2004-10-01

    Full Text Available A low-cost, gated system for measurements of phosphorescence lifetimes is presented. An extensive description of the system operating principles and metrological characteristics is given. Remarkably, the system operates without optical filtering of the LED excitation source. A description of a practical system is also given and its performance is discussed. Because the device effectively suppresses high-level background fluorescence and scattered light, it is expected to find wide-spread application in bioprocess, environmental and biomedical fields.

  8. The reactivities of furocoumarin excited states with DNA in solution. A laser flash photolysis and fluorescence study.

    Science.gov (United States)

    Beaumont, P C; Parsons, B J; Navaratnam, S; Phillips, G O; Allen, J C

    1980-07-29

    The effect of DNA on both the fluorescence emission spectra and yields and lifetimes of the triplet stae of psoralen and 8-methoxypsoralen in aqueous solution has been determined. The changes in the fluorescence spectra are similar in nature for both of these furocoumarins and are attributed to binding of the drug to DNA. The yield of the 8-methoxypsoralen triplet state when bound to DNA was found to be similar, if not identical, to that measured in the absence of DNA. This contrasts sharply with data obtained for psoralen from which it is concluded that either the yield of bound psoralen triplet states is very low, if not zero, or that the lifetime of such species is less than 50 ns. The relevance of this data to the molecular basis of skin photosensitisation by furocoumarins is discussed.

  9. A New Theoretical Approach to Single-Molecule Fluorescence Optical Studies of RNA Dynamics

    International Nuclear Information System (INIS)

    Zhao Xinghai; Shan Guangcun; Bao Shuying

    2011-01-01

    Single-molecule fluorescence spectroscopy in condensed phases has many important chemical and biological applications. The single-molecule fluorescence measurements contain information about conformational dynamics on a vast range of time scales. Based on the data analysis protocols methodology proposed by X. Sunney Xie, the theoretical study here mainly focuses on the single-molecule studies of single RNA with interconversions among different conformational states, to with a single FRET pair attached. We obtain analytical expressions for fluorescence lifetime correlation functions that relate changes in fluorescence lifetime to the distance-dependent FRET mechanism within the context of the Smoluchowski diffusion model. The present work establishes useful guideline for the single-molecule studies of biomolecules to reveal the complicated folding dynamics of single RNA molecules at nanometer scale.

  10. Determination of several families of phytochemicals in different pre-cooked convenience vegetables: effect of lifetime and cooking.

    Science.gov (United States)

    Alarcón-Flores, M Isabel; Hernández-Sánchez, Francisco; Romero-González, Roberto; Plaza-Bolaños, Patricia; Martínez Vidal, J Luis; Garrido Frenich, Antonia

    2014-11-01

    Phytochemicals content, including several families such as phenolic acids, isoflavones, flavones, flavonols, isothiocyanates, and glucosinolates, was determined in pre-cooked convenience vegetables by ultra high performance liquid chromatography coupled to triple quadrupole tandem mass spectrometry (UHPLC-QqQ-MS/MS). It was observed that there is not a common behavior of the individual concentration of phytochemicals during the lifetime and cooking of the matrix, and compounds change their concentration without a specific trend. It was observed that neither lifetime nor cooking process have significant effects on the total content of phytochemicals except in broccoli, although some changes in the individual content of the target compounds were observed, suggesting that interconversion processes could be performed during the lifetime and/or cooking process of the product.

  11. Facile and Scalable Preparation of Fluorescent Carbon Dots for Multifunctional Applications

    Directory of Open Access Journals (Sweden)

    Dan Wang

    2017-06-01

    Full Text Available The synthesis of fluorescent nanomaterials has received considerable attention due to the great potential of these materials for a wide range of applications, from chemical sensing through bioimaging to optoelectronics. Herein, we report a facile and scalable approach to prepare fluorescent carbon dots (FCDs via a one-pot reaction of citric acid with ethylenediamine at 150 °C under ambient air pressure. The resultant FCDs possess an optical bandgap of 3.4 eV and exhibit strong excitation-wavelength-independent blue emission (λEm = 450 nm under either one- or two-photon excitation. Owing to their low cytotoxicity and long fluorescence lifetime, these FCDs were successfully used as internalized fluorescent probes in human cancer cell lines (HeLa cells for two-photon excited imaging of cells by fluorescence lifetime imaging microscopy with a high-contrast resolution. They were also homogenously mixed with commercial inks and used to draw fluorescent patterns on normal papers and on many other substrates (e.g., certain flexible plastic films, textiles, and clothes. Thus, these nanomaterials are promising for use in solid-state fluorescent sensing, security labeling, and wearable optoelectronics.

  12. Time-Resolved Fluorescence of Water-Soluble Pyridinium Salt: Sensitive Detection of the Conformational Changes of Bovine Serum Albumin.

    Science.gov (United States)

    Li, Lei; Yi, Hua; Jia, Menghui; Chang, Mengfang; Zhou, Zhongneng; Zhang, Sanjun; Pan, Haifeng; Chen, Yan; Chen, Jinquan; Xu, Jianhua

    2016-06-20

    In this paper, we report a pyridinium salt "turn-on" fluorescent probe, 4-[2-(4-Dimethylamino-phenyl)-vinyl]-1-methylpyridinium iodide (p-DASPMI), and applied its time-resolved fluorescence (TRF) to monitor the protein conformational changes. Both the fluorescence lifetime and quantum yield (QY) of p-DASPMI were increased about two orders of magnitude after binding to the protein bovine serum albumin (BSA). The free p-DASPMI in solution presents an ultrashort fluorescence lifetime (12.4 ps), thus it does not interfere the detection of bound p-DASPMI which has nanosecond fluorescence lifetime. Decay-associated spectra (DAS) show that p-DASPMI molecules bind to subdomains IIA and IIIA of BSA. The TRF decay profiles of p-DASPMI can be described by multi-exponential decay function ([Formula: see text]), and the obtained parameters, such as lifetimes ([Formula: see text]), fractional amplitudes ([Formula: see text]), and fractional intensities ([Formula: see text]), may be used to deduce the conformational changes of BSA. The pH and Cu 2+ induced conformational changes of BSA were investigated through the TRF of p-DASPMI. The results show that the p-DASPMI is a candidate fluorescent probe in studying the conformational changes of proteins through TRF spectroscopy and microscopy in the visible range. © The Author(s) 2016.

  13. 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.

  14. Nanostructure induced changes in lifetime and enhanced second-harmonic response of organic-plasmonic hybrids

    Energy Technology Data Exchange (ETDEWEB)

    Leißner, Till [NanoSYD, Mads Clausen Institute, University of Southern Denmark, Alsion 2, 6400 Sønderborg (Denmark); Department of Biochemistry and Molecular Biology, University of Southern Denmark, Campusvej 55, 5230 Odense (Denmark); Kostiučenko, Oksana; Rubahn, Horst-Günter; Fiutowski, Jacek, E-mail: fiutowski@mci.sdu.dk [NanoSYD, Mads Clausen Institute, University of Southern Denmark, Alsion 2, 6400 Sønderborg (Denmark); Brewer, Jonathan R. [Department of Biochemistry and Molecular Biology, University of Southern Denmark, Campusvej 55, 5230 Odense (Denmark)

    2015-12-21

    In this letter we show that the optical response of organic nanofibers, grown from functionalized para-quaterphenylene molecules, can be controlled by forming organic-plasmonic hybrid systems. The interaction between nanofibers and supporting regular arrays of nanostructures leads to a strongly enhanced second harmonic response. At the same time, the fluorescence lifetime of the nanofibers is reduced from 0.32 ns for unstructured gold films to 0.22 ns for gold nanosquare arrays, demonstrating efficient organic–plasmonic interaction. To study the origin of these effects, we applied two-photon laser scanning microscopy and fluorescence lifetime imaging microscopy. These findings provide an effective approach for plasmon-enhanced second-harmonic generation at the nanoscale, which is attractive for nanophotonic circuitry.

  15. Characterization of standard reference material 2944, Bi-ion-doped glass, spectral correction standard for red fluorescence

    International Nuclear Information System (INIS)

    DeRose, Paul C.; Smith, Melody V.; Anderson, Jeffrey R.; Kramer, Gary W.

    2013-01-01

    Standard Reference Material (SRM) 2944 is a cuvette-shaped, Bi-ion-doped glass, recommended for optimal use for relative spectral correction of emission from 590 nm to 805 nm and day-to-day performance verification of steady-state fluorescence spectrometers. Properties of this standard that influence its effective use or contribute to the uncertainty in its certified emission spectrum were explored here. These properties include its photostability, absorbance, dissolution rate in water, anisotropy and temperature coefficient of fluorescence intensity. The expanded uncertainties (k=2) in the certified spectrum are about 4% around the nominal peak maximum at 704 nm and increase to about 6% at the wings, using an excitation wavelength of 515 nm. -- Highlights: ► The fluorescence emission spectrum of SRM 2944 was determined for spectral correction. ► This Bi-ion-doped glass has been certified in the fluorescence region from 530 nm to 830 nm. ► Fluorescence properties of the glass were determined, e.g., anisotropy, lifetime. ► SRM 2944 is photostable under common visible lamp excitation, when UV light is not present

  16. New spectrofluorometer with pulsed intensified photodiodes array for direct trace determination of actinides and lanthanides in solutions

    International Nuclear Information System (INIS)

    Decambox, P.; Kirsch, B.; Mauchien, P.; Moulin, C.

    1989-01-01

    Actinides and lanthanides in solution at very low level are determined by Time-Resolved Laser-Induced Spectrofluorometry (TRLIS) with pulsed intensified photodiodes array detection. Temporal resolution allows discrimination against short lifetime fluorescence and measurement of fluorescence lifetime in various matrices. The use of laser source leads to excitation selectivity and high sensitivity. Multichannel detection allows to cover the entire wavelength range of interest which leads to emission selectivity and rapidity. These different advantages are present in the newly commercialized spectrofluorometer FLUO 2001 together with specially analytical adapted software. The apparatus and performances obtained for several actinides and lanthanides are presented

  17. Significant improvement of accuracy and precision in the determination of trace rare earths by fluorescence analysis

    International Nuclear Information System (INIS)

    Ozawa, L.; Hersh, H.N.

    1976-01-01

    Most of the rare earths in yttrium, gadolinium and lanthanum oxides emit characteristic fluorescent line spectra under irradiation with photons, electrons and x rays. The sensitivity and selectivity of the rare earth fluorescences are high enough to determine the trace amounts (0.01 to 100 ppM) of rare earths. The absolute fluorescent intensities of solids, however, are markedly affected by the synthesis procedure, level of contamination and crystal perfection, resulting in poor accuracy and low precision for the method (larger than 50 percent error). Special care in preparation of the samples is required to obtain good accuracy and precision. It is found that the accuracy and precision for the determination of trace (less than 10 ppM) rare earths by fluorescence analysis improved significantly, while still maintaining the sensitivity, when the determination is made by comparing the ratio of the fluorescent intensities of the trace rare earths to that of a deliberately added rare earth as reference. The variation in the absolute fluorescent intensity remains, but is compensated for by measuring the fluorescent line intensity ratio. Consequently, the determination of trace rare earths (with less than 3 percent error) is easily made by a photoluminescence technique in which the rare earths are excited directly by photons. Accuracy is still maintained when the absolute fluorescent intensity is reduced by 50 percent through contamination by Ni, Fe, Mn or Pb (about 100 ppM). Determination accuracy is also improved for fluorescence analysis by electron excitation and x-ray excitation. For some rare earths, however, accuracy by these techniques is reduced because indirect excitation mechanisms are involved. The excitation mechanisms and the interferences between rare earths are also reported

  18. Natural Radionuclides and Isotopic Signatures for Determining Carbonaceous Aerosol Sources, Aerosol Lifetimes, and Washout Processes

    International Nuclear Information System (INIS)

    Gaffney, Jeffrey

    2012-01-01

    This is the final technical report. The project description is as follows: to determine the role of aerosol radiative forcing on climate, the processes that control their atmospheric concentrations must be understood, and aerosol sources need to be determined for mitigation. Measurements of naturally occurring radionuclides and stable isotopic signatures allow the sources, removal and transport processes, as well as atmospheric lifetimes of fine carbonaceous aerosols, to be evaluated.

  19. Natural Radionuclides and Isotopic Signatures for Determining Carbonaceous Aerosol Sources, Aerosol Lifetimes, and Washout Processes

    Energy Technology Data Exchange (ETDEWEB)

    Gaffney, Jeffrey [Univ. of Arkansas, Little Rock, AR (United States)

    2012-12-12

    This is the final technical report. The project description is as follows: to determine the role of aerosol radiative forcing on climate, the processes that control their atmospheric concentrations must be understood, and aerosol sources need to be determined for mitigation. Measurements of naturally occurring radionuclides and stable isotopic signatures allow the sources, removal and transport processes, as well as atmospheric lifetimes of fine carbonaceous aerosols, to be evaluated.

  20. Rotationally cooled laser induced fluorescence determination of polycyclic aromatic hydrocarbons

    International Nuclear Information System (INIS)

    Warren, J.A.; Hayes, J.M.; Small, G.J.

    1982-01-01

    In recent years the development of new highly selective and sensitive methods for the characterization and determination of polycyclic aromatic hydrocarbons (PAHs) and their derivatives in complex mixtures has received considerable attention. High selectivity is associated here with the ability to distinguish between substitutional isomers of PAHs. Attainment of this selectivity with capillary column-gas chromatography-mass spectrometry for complex mixtures is very difficult and time-consuming. Alternative approaches are, therefore, required. Given that the majority of PAHs fluoresce with reasonable quantum yields and that high sensitivities are afforded by fluorescence detection, the possibility of developing high-resolution fluorescence based techniques is attractive. This is all the more so if the technique's selectivity does not rely on physical separation, e.g., chromatography. In this paper discussion is limited to such techniques

  1. Synthesis of water-soluble, ring-substituted squaraine dyes and their evaluation as fluorescent probes and labels

    Energy Technology Data Exchange (ETDEWEB)

    Tatarets, Anatoliy L. [SSI ' Institute for Single Crystals' of the National Academy of Sciences of Ukraine, 60 Lenin Ave., Kharkov 61001 (Ukraine); Fedyunyayeva, Irina A. [SSI ' Institute for Single Crystals' of the National Academy of Sciences of Ukraine, 60 Lenin Ave., Kharkov 61001 (Ukraine); Dyubko, Tatyana S. [SSI ' Institute for Single Crystals' of the National Academy of Sciences of Ukraine, 60 Lenin Ave., Kharkov 61001 (Ukraine); Povrozin, Yevgeniy A. [SSI ' Institute for Single Crystals' of the National Academy of Sciences of Ukraine, 60 Lenin Ave., Kharkov 61001 (Ukraine); Doroshenko, Andrey O. [Institute of Chemistry, V.N. Karazin National University, 4 Svobody Sq., Kharkov 61077 (Ukraine); Terpetschnig, Ewald A. [SETA BioMedicals, LLC, 2014 Silver Ct East, Urbana, IL 61801 (United States) and ISS, Inc., 1602 Newton Drive, Champaign, IL 61822 (United States)]. E-mail: ewaldte@juno.com; Patsenker, Leonid D. [SSI ' Institute for Single Crystals' of the National Academy of Sciences of Ukraine, 60 Lenin Ave., Kharkov 61001 (Ukraine); SETA BioMedicals, LLC, 2014 Silver Ct East, Urbana, IL 61801 (United States)

    2006-06-16

    A series of ring-substituted squaraines absorbing and emitting in the red and NIR spectral region was synthesized and their spectral and photophysical properties (quantum yields, fluorescence lifetimes) and photostabilities were measured and compared to Cy5, a commonly used fluorescent label. The absorption maxima in aqueous media were found to be between 628 and 667 nm and the emission maxima are between 642 and 685 nm. Squaraine dyes exhibit high extinction coefficients (163,000-265,000 M{sup -1} cm{sup -1}) and lower quantum yields (2-7%) in aqueous buffer but high quantum yields (up to 45%) and long fluorescence lifetimes (up to 3.3 ns) in presence of BSA. Dicyanomethylene- and thio-substituted squaraines exhibit an additional absorption around 400 nm with extinction coefficients between 21,500 and 44,500 M{sup -1} cm{sup -1}. These dyes are excitable not only with red but also with blue diode lasers or light emitting diodes. Due to the favourable spectral and photophysical properties these dyes can be used as fluorescent probes and labels for intensity- and fluorescence lifetime-based biomedical applications.

  2. Synthesis of water-soluble, ring-substituted squaraine dyes and their evaluation as fluorescent probes and labels

    International Nuclear Information System (INIS)

    Tatarets, Anatoliy L.; Fedyunyayeva, Irina A.; Dyubko, Tatyana S.; Povrozin, Yevgeniy A.; Doroshenko, Andrey O.; Terpetschnig, Ewald A.; Patsenker, Leonid D.

    2006-01-01

    A series of ring-substituted squaraines absorbing and emitting in the red and NIR spectral region was synthesized and their spectral and photophysical properties (quantum yields, fluorescence lifetimes) and photostabilities were measured and compared to Cy5, a commonly used fluorescent label. The absorption maxima in aqueous media were found to be between 628 and 667 nm and the emission maxima are between 642 and 685 nm. Squaraine dyes exhibit high extinction coefficients (163,000-265,000 M -1 cm -1 ) and lower quantum yields (2-7%) in aqueous buffer but high quantum yields (up to 45%) and long fluorescence lifetimes (up to 3.3 ns) in presence of BSA. Dicyanomethylene- and thio-substituted squaraines exhibit an additional absorption around 400 nm with extinction coefficients between 21,500 and 44,500 M -1 cm -1 . These dyes are excitable not only with red but also with blue diode lasers or light emitting diodes. Due to the favourable spectral and photophysical properties these dyes can be used as fluorescent probes and labels for intensity- and fluorescence lifetime-based biomedical applications

  3. Automatic Segmentation of Fluorescence Lifetime Microscopy Images of Cells Using Multi-Resolution Community Detection -A First Study

    Science.gov (United States)

    Hu, Dandan; Sarder, Pinaki; Ronhovde, Peter; Orthaus, Sandra; Achilefu, Samuel; Nussinov, Zohar

    2014-01-01

    Inspired by a multi-resolution community detection (MCD) based network segmentation method, we suggest an automatic method for segmenting fluorescence lifetime (FLT) imaging microscopy (FLIM) images of cells in a first pilot investigation on two selected images. The image processing problem is framed as identifying segments with respective average FLTs against the background in FLIM images. The proposed method segments a FLIM image for a given resolution of the network defined using image pixels as the nodes and similarity between the FLTs of the pixels as the edges. In the resulting segmentation, low network resolution leads to larger segments, and high network resolution leads to smaller segments. Further, using the proposed method, the mean-square error (MSE) in estimating the FLT segments in a FLIM image was found to consistently decrease with increasing resolution of the corresponding network. The MCD method appeared to perform better than a popular spectral clustering based method in performing FLIM image segmentation. At high resolution, the spectral segmentation method introduced noisy segments in its output, and it was unable to achieve a consistent decrease in MSE with increasing resolution. PMID:24251410

  4. X-ray fluorescence analysis for trace element determination in foodstuff chemistry

    International Nuclear Information System (INIS)

    Wildanger, W.

    The physical fundamentals of X-ray fluorescence analysis are given and the routine spectrometers described. The basic principles are given of analytical methods used in qualitative and quantitative fluorescence analyses. Examples are given of the use of the method in a number of fields and the possibility and usefulness is discussed for the determination of trace elements in foodstuffs. The preparation of samples, preliminary concentration of components and calibration methods are discussed. (M.K.)

  5. Quantum dots (QDs) based fluorescence probe for the sensitive determination of kaempferol

    Science.gov (United States)

    Tan, Xuanping; Liu, Shaopu; Shen, Yizhong; He, Youqiu; Yang, Jidong

    2014-12-01

    In this work, using the quenching of fluorescence of thioglycollic acid (TGA)-capped CdTe quantum dots (QDs), a novel method for the determination of kaempferol (KAE) has been developed. Under optimum conditions, a linear calibration plot of the quenched fluorescence intensity at 552 nm against the concentration of KAE was observed in the range of 4-44 μg mL-1 with a detection limit (3σ/K) of 0.79 μg mL-1. In addition, the detailed reaction mechanism has also been proposed on the basis of electron transfer supported by ultraviolet-visible (UV-vis) absorption and fluorescence (FL) spectroscopy. The method has been applied for the determination of KAE in pharmaceutical preparations with satisfactory results. The proposed method manifested several advantages such as high sensitivity, short analysis time, low cost and ease of operation.

  6. Determination of antioxidant content in biodiesel by fluorescence spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Magalhaes, Keurison F.; Caires, Anderson R.L. [Universidade Federal da Grande Dourados, MS (Brazil). Grupo de Optica Aplicada; Oliveira, Samuel L. [Universidade Federal de Mato Grosso do Sul (UFMS), MS (Brazil). Grupo de Optica e Fotonica

    2011-07-01

    Full text. Biodiesel is an alternative fuel composed by mono-alkyl esters obtained from vegetable oils or animal fats. Due to its chemical structure, biodiesel is highly susceptible to oxidation which leads to formation of insoluble gums and sediments that can block the filter system of fuel injection. Biodiesel made from vegetable oils typically has a small amount of natural antioxidants so that it is necessary to add synthetic antioxidants to enhance its stability and retain their properties for a longer period. The main antioxidants are synthetic phenolic compounds such as butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT) and tert-butyl hydroquinone (TBHQ) as well as natural antioxidants as tocopherols. The fluorescence spectroscopy has been applied for determination of phenolic compounds in oils. Here, a method based on fluorescence is proposed to quantify the BHA and TBHQ antioxidant concentration in biodiesel produced from sunflower and soybean oils. Soybean and sunflower biodiesel were obtained by transesterification of fatty alcohol in the presence of NaOH as catalyst. The reactions were carried out in the molar ratio of 6:1 methanol/oil. After the production and purification, biodiesel samples were stored. Biodiesel samples with BHA and TBHQ concentrations from 1000 to 8000 ppm (m/m) were pre- pared. These samples were diluted in ethanol (95%) in order to measure the fluorescence spectra. Fluorescence and excitation spectra of the solutions were recorded at room temperature using a spectrofluorimeter. The emission spectra were obtained under excitation at about 310nm and fluorescence in the 320-800nm range was evaluated. Biodiesel samples without BHA and TBHQ showed fluorescence band at about 420nm, which can be attributed to tocopherols inherent to the vegetable oils used in the biodiesel production. The addition of BHA and/or TBHQ is responsible for the appearance of a fluorescence band around 330nm. It was verified that the fluorescence

  7. Tryptophan and ATTO 590: mutual fluorescence quenching and exciplex formation.

    Science.gov (United States)

    Bhattacharjee, Ujjal; Beck, Christie; Winter, Arthur; Wells, Carson; Petrich, Jacob W

    2014-07-24

    Investigation of fluorescence quenching of probes, such as ATTO dyes, is becoming an increasingly important topic owing to the use of these dyes in super-resolution microscopies and in single-molecule studies. Photoinduced electron transfer is their most important nonradiative pathway. Because of the increasing frequency of the use of ATTO and related dyes to investigate biological systems, studies are presented for inter- and intramolecular quenching of ATTO 590 with tryptophan. In order to examine intramolecular quenching, an ATTO 590-tryptophan conjugate was synthesized. It was determined that tryptophan is efficiently quenching ATTO 590 fluorescence by excited-state charge transfer and two charge transfer complexes are forming. In addition, it was discovered that an exciplex (whose lifetime is 5.6 ns) can be formed between tryptophan and ATTO 590, and it is suggested that the possibility of such exciplex formation should be taken into account when protein fluorescence is monitored in a system tagged with ATTO dyes.

  8. [Determination of H2S in Rat Intestinal Perfusion Solution Based on Fluorescence Analysis].

    Science.gov (United States)

    Hou, Jun-feng; Li, Xin-xia; Shen, Xue-ru; Huojia, Miliban; Guan, Ming

    2015-08-01

    Under alkaline conditions, Fluorescein mercury has strong fluorescence, however, when it met S(2-), its fluorescence would quench, in view of the above, a fluorescence method for determination of H2S in biological samples was established. In the 0.1 mol · L(-1) NaOH dilution, when the concentration of fluorescein Mercury and Na2S was 5.0 × 10(-5) and 1.0 × 10(-5) mol · L(-1) respectively, the fluorescence intensity of system was determined at 522 nm. The results showed that, at the range of 4.0 × 10(-7)~2.0 × 10(-6) mol · L(-1), the concentration decreasing of H2S and fluorescence intensity had good linear relationship, r=0.9980, the RSD of precision test was 4.59% (n=7), the detection limit was 3.5 × 10(-8) mol · L(-1), the content of H2S in the sample were 1.01 × 10(-6) and 1.15 × 10(-6) mol · L(-1), and the recovery rate was 95.8%~101.0%, the method has the advantages of simple operation, high sensitivity, good selectivity, can accurately determine of H2S in intestinal perfused solution, and provides the basis for the determination of endogenous H2S.

  9. Determination of ππ scattering lengths from measurement of π+π- atom lifetime

    International Nuclear Information System (INIS)

    Adeva, B.; Afanasyev, L.; Benayoun, M.; Benelli, A.; Berka, Z.; Brekhovskikh, V.; Caragheorgheopol, G.; Cechak, T.; Chiba, M.; Chliapnikov, P.V.; Ciocarlan, C.; Constantinescu, S.; Costantini, S.; Curceanu, C.; Doskarova, P.; Dreossi, D.; Drijard, D.; Dudarev, A.; Ferro-Luzzi, M.; Fungueirino Pazos, J.L.

    2011-01-01

    The DIRAC experiment at CERN has achieved a sizeable production of π + π - atoms and has significantly improved the precision on its lifetime determination. From a sample of 21 227 atomic pairs, a 4% measurement of the S-wave ππ scattering length difference |a 0 -a 2 |=(0.2533 -0.0078 +0.0080 | stat +0.0078 -0.0073 | syst )M π + -1 has been attained, providing an important test of Chiral Perturbation Theory.

  10. [Fluorescence Determination of Trace Se with the Hydride-K13-Rhodamine 6G System].

    Science.gov (United States)

    Liang, Ai-hui; Li, Yuan; Huang, Shan-shan; Luo, Yang-he; Wen, Gui-qing; Jiang, Zhi-liang

    2015-05-01

    Se is a necessary trace element for human and animals, but the excess intake of Se caused poison. Thus, it is very important to determination of Se in foods and water. The target of this study is development of a new, sensitive and selective hydride generation-molecular fluorescence method for the determination of Se. In 0. 36 mol . L-1 sulfuric acid, NaBH4 as reducing agent, Se (IV) is reduced to H2 Se. Usin3-g I solution as absorption liquid3, I- is reduced to I- by H2Se. When adding rhodamine 6G, Rhodamine 6G and I3- form association particles, which lead to the fluorescence intensity decreased. When Se(IV) existing, Rhodamine 6G and I3- bind less, And the remaining amount of Rhodamine 6G increase. So the fluorescence intensity is enhanced. The analytical conditions were optimized, a 0. 36 ml . L-1 H2SO4, 21. 6.g . L-1 NaBH4, 23.3 µm . L-1 rhodamine 6G, and 50 µmol . L-1 KI3 were chosen for use. When the excitation wavelength is at 480nm, the Rayleigh scattering peak does not affect the fluorescence recording, and was selected for determination of Se. Under the selected conditions, Se(IV) concentration in the 0. 02~0. 60 µg . mL-1 range and the increase value of the fluorescence intensity (ΔF) at 562 nm linear relationship. The linear regression equation is ΔF562 nm =12. 6c + 20. 9. The detecton limit was 0.01 µ.g . L-1. The influence of coexistence substances on the hydride generatin-molecular fluorescence determination of 5. 07 X10(-6) mol . L-1 Se(IV) was considered in details. Results showed that this new fluorescence method is of high selectivity, that is, 0. 5 mmol. L-1 Ba2+, Ca2+, Zn2+ and Fe3+, 0. 25 mmol . L-1 . Mg2+, 0. 05 mmol . L-1 K+, 0. 2 mmol . L-1 Al3+, 0. 025 mmol . L-1 Te(VI) do not interfere with the determination. The influence of Hg2+, CD2+ and Cu2+ that precipitate with Se(IV), can be eliminated by addition of complex reagent. This hydride generation-molecular fluorescence method has been applied to determination of trace Se in water

  11. Fluorescent BODIPY Rotor: Viscometer for Cellular Organelles and Membrane-Mimicking Vesicles

    Science.gov (United States)

    Kimball, J.; Raut, S.; Fudala, R.; Doan, H.; Maliwal, B.; Sabnis, N.; Lacko, A.; Gryczynski, I.; Dzyuba, S.; Gryczynski, Z.

    2015-03-01

    Many cellular processes, such as mass and signal transport, metabolism and protein-protein interactions are governed in part by diffusion, and thus affected by their local microviscosity. Changes in this microviscosity has also been linked to various diseases, including atherosclerosis, Alzheimer's disease and diabetes. Therefore, directly measuring the heterogeneous viscosity of cellular constitutes can lead to greater understanding of these processes. To this effect, a novel homodiemeric BODIPY dye was evaluated as a fluorescent rotor probe for this application. A linear dependence on viscosity in the range of typical cellular microviscosity was established for steady-state and time-resolved properties of the dye. It was then embedded in vitro to membrane-mimicking lipid vesicles (DPPC, POPC, and POPC plus cholesterol) and results indicated it to be a viable sensor for lifetime-based determination of microviscosity. The BODIPY dye was lastly endocytosed by SKOV3 cells and Fluorescence Lifetime Imaging Microscopy (FLIM) was performed, successfully mapping the viscosity of internal cell components. This work was supported by the NIH Grant R01EB12003, the NSF Grant CBET-1264608, and the INFOR Grant from TCU.

  12. X-ray fluorescence spectroscopic determination of heavy metals and ...

    African Journals Online (AJOL)

    Purpose: To determine the heavy metal and trace element composition of the powdered aerial parts of Origanum sipyleum L. and its water extract. Methods: The heavy metal and trace elements content of the powdered plant material and 2 % aqueous extract were evaluated by x-ray fluorescence spectroscopy with silicon ...

  13. Fluorescence decay data analysis correcting for detector pulse pile-up at very high count rates

    Science.gov (United States)

    Patting, Matthias; Reisch, Paja; Sackrow, Marcus; Dowler, Rhys; Koenig, Marcelle; Wahl, Michael

    2018-03-01

    Using time-correlated single photon counting for the purpose of fluorescence lifetime measurements is usually limited in speed due to pile-up. With modern instrumentation, this limitation can be lifted significantly, but some artifacts due to frequent merging of closely spaced detector pulses (detector pulse pile-up) remain an issue to be addressed. We propose a data analysis method correcting for this type of artifact and the resulting systematic errors. It physically models the photon losses due to detector pulse pile-up and incorporates the loss in the decay fit model employed to obtain fluorescence lifetimes and relative amplitudes of the decay components. Comparison of results with and without this correction shows a significant reduction of systematic errors at count rates approaching the excitation rate. This allows quantitatively accurate fluorescence lifetime imaging at very high frame rates.

  14. Quantitative determination of localized tissue oxygen concentration in vivo by two-photon excitation phosphorescence lifetime measurements

    NARCIS (Netherlands)

    Mik, Egbert G.; van Leeuwen, Ton G.; Raat, Nicolaas J.; Ince, Can

    2004-01-01

    This study describes the use of two-photon excitation phosphorescence lifetime measurements for quantitative oxygen determination in vivo. Doubling the excitation wavelength of Pd-porphyrin from visible light to the infrared allows for deeper tissue penetration and a more precise and confined

  15. Determination of chlorine in coal by X-ray fluorescence spectrometry method

    Energy Technology Data Exchange (ETDEWEB)

    Marek, S.; Bojarska, K. [Central Mining Institute, Katowice (Poland). Dept. of Environmental Monitoring

    1997-12-31

    Determination of chlorine contents in coal is essential for both environmental protection and its technological use. The existing method of chlorine determination in coal are titration methods which have considerable errors particularly in the low concentration range. The elaborated method with the use of X-ray fluorescence spectrometry in a comparison to the other methods is much faster and has better precision and accuracy. The principle of the method lies in the measurement of X-ray fluorescence radiation intensity which is emitted by chlorine in a sample and its comparison with standards. The calibration of the elaborated XRF method is based on natural coals having various concentrations of chlorine within the whole range of its occurrence in Polish coals. Concentrations for the calibration purpose were obtained by the determination of chlorine contents in selected coals by atomic absorption spectrometry method. The procedure of sample preparation for direct X-ray measurements, instrumental measuring conditions and the way of calibration curve preparation are described in the paper. All X-ray measurements were done with a Phillips sequential X-ray fluorescence spectrometer. A double anode Cr-Au X-ray tube with maximum power 3000 MW was used as the excitation source. 5 figs., 4 tabs.

  16. Absorption and fluorescence spectroscopic characterization of BLUF domain of AppA from Rhodobacter sphaeroides

    Energy Technology Data Exchange (ETDEWEB)

    Zirak, P. [Institut II - Experimentelle und Angewandte Physik, Universitaet Regensburg, Universitaetstrasse 31, D-93053 Regensburg (Germany); Penzkofer, A. [Institut II - Experimentelle und Angewandte Physik, Universitaet Regensburg, Universitaetstrasse 31, D-93053 Regensburg (Germany)], E-mail: alfons.penzkofer@physik.uni-regensburg.de; Schiereis, T. [Institut fuer Biologie, Experimentelle Biophysik, Humboldt-Universitaet zu Berlin, Invalidenstrasse 42, D-10115 Berlin (Germany); Hegemann, P. [Institut fuer Biologie, Experimentelle Biophysik, Humboldt-Universitaet zu Berlin, Invalidenstrasse 42, D-10115 Berlin (Germany); Jung, A. [Max-Planck-Institut fuer medizinische Forschung, Abteilung Biomolekulare Mechanismen, Jahnstrasse 29, D-69120 Heidelberg (Germany); Schlichting, I. [Max-Planck-Institut fuer medizinische Forschung, Abteilung Biomolekulare Mechanismen, Jahnstrasse 29, D-69120 Heidelberg (Germany)

    2005-08-08

    The BLUF domain of the transcriptional anti-repressor protein AppA from the non-sulfur anoxyphototrophic purple bacterium Rhodobacter sphaeroides was characterized by absorption and emission spectroscopy. The BLUF domain constructs AppA{sub 148} (consisting of amino-acid residues 1-148) and AppA{sub 126} (amino-acid residues 1-126) are investigated. The cofactor of the investigated domains is found to consist of a mixture of the flavins riboflavin, FMN, and FAD. The dark-adapted domains exist in two different active receptor conformations (receptor states) with different sub-nanosecond fluorescence lifetimes (BLUF{sub r,f} and BLUF{sub r,sl}) and a small non-interacting conformation (BLUF{sub nc}). The active receptor conformations are transformed to putative signalling states (BLUF{sub s,f} and BLUF{sub s,sl}) of low fluorescence efficiency and picosecond fluorescence lifetime by blue-light excitation (light-adapted domains). In the dark at room temperature both signalling states recover back to the initial receptor states with a time constant of about 17 min. A quantum yield of signalling state formation of about 25% was determined by intensity dependent transmission measurements. A photo-cycle scheme is presented including photo-induced charge transfer complex formation, charge recombination, and protein binding pocket reorganisation.

  17. Sensitivity of in vivo X-ray fluorescence determination of skeletal lead stores

    International Nuclear Information System (INIS)

    Sokas, R.K.; Besarab, A.; McDiarmid, M.A.; Shapiro, I.M.; Bloch, P.

    1990-01-01

    Eighteen patients with known past occupational lead exposure underwent parenteral diagnostic chelation with ethylenediaminetetraacetic acid and x-ray fluorescent determination of in vivo skeletal lead stores at the distal styloid process of the ulna and at the temporal base bone using a cobalt 57 source and measuring lead Ka x-rays. X-ray fluorescent lead measurements in both locations correlated with results of diagnostic chelation. Using a post-chelation urinary excretion of greater than 600 micrograms lead/24 h as the definition of high-lead stores, sensitivity of x-ray fluorescence at the wrist and temple was 56% and 39%, respectively

  18. Validity of LIDAS (LIfetime Depression Assessment Self-report): a self-report online assessment of lifetime major depressive disorder.

    Science.gov (United States)

    Bot, M; Middeldorp, C M; de Geus, E J C; Lau, H M; Sinke, M; van Nieuwenhuizen, B; Smit, J H; Boomsma, D I; Penninx, B W J H

    2017-01-01

    There is a paucity of valid, brief instruments for the assessment of lifetime major depressive disorder (MDD) that can be used in, for example, large-scale genomics, imaging or biomarker studies on depression. We developed the LIfetime Depression Assessment Self-report (LIDAS), which assesses lifetime MDD diagnosis according to DSM criteria, and is largely based on the widely used Composite International Diagnostic Interview (CIDI). Here, we tested the feasibility and determined the sensitivity and specificity for measuring lifetime MDD with this new questionnaire, with a regular CIDI as reference. Sensitivity and specificity analyses of the online lifetime MDD questionnaire were performed in adults with (n = 177) and without (n = 87) lifetime MDD according to regular index CIDIs, selected from the Netherlands Study of Depression and Anxiety (NESDA) and Netherlands Twin Register (NTR). Feasibility was tested in an additional non-selective, population-based sample of NTR participants (n = 245). Of the 753 invited persons, 509 (68%) completed the LIDAS, of which 419 (82%) did this online. User-friendliness of the instrument was rated high. Median completion time was 6.2 min. Sensitivity and specificity for lifetime MDD were 85% [95% confidence interval (CI) 80-91%] and 80% (95% CI 72-89%), respectively. This LIDAS instrument gave a lifetime MDD prevalence of 20.8% in the population-based sample. Measuring lifetime MDD with an online instrument was feasible. Sensitivity and specificity were adequate. The instrument gave a prevalence of lifetime MDD in line with reported population prevalences. LIDAS is a promising tool for rapid determination of lifetime MDD status in large samples, such as needed for genomics studies.

  19. Fluorescent metal nanoshell and CK19 detection on single cell image

    International Nuclear Information System (INIS)

    Zhang, Jian; Fu, Yi; Li, Ge; Lakowicz, Joseph R.; Zhao, Richard Y.

    2011-01-01

    Highlights: → Novel metal nanoshell as fluorescence imaging agent. → Fluorescent mAb-metal complex with enhanced intensity and shortened lifetime. → Immuno-interactions of mAb-metal complexes with CK19 molecules on CNCAP and HeLa cell surfaces. → Isolation of conjugated mAb-metal complexes from cellular autofluorescence on cell image. -- Abstract: In this article, we report the synthesis strategy and optical properties of a novel type of fluorescence metal nanoshell when it was used as imaging agent for fluorescence cell imaging. The metal nanoshells were made with 40 nm silica cores and 10 nm silver shells. Unlike typical fluorescence metal nanoshells which contain the organic dyes in the cores, novel metal nanoshells were composed of Cy5-labelled monoclonal anti-CK19 antibodies (mAbs) on the external surfaces of shells. Optical measurements to the single nanoparticles showed that in comparison with the metal free labelled mAbs, the mAb-Ag complexes displayed significantly enhanced emission intensity and dramatically shortened lifetime due to near-field interactions of fluorophores with metal. These metal nanoshells were found to be able to immunoreact with target cytokeratin 19 (CK19) molecules on the surfaces of LNCAP and HeLa cells. Fluorescence cell images were recorded on a time-resolved confocal microscope. The emissions from the metal nanoprobes could be clearly isolated from the cellular autofluorescence backgrounds on the cell images as either individuals or small clusters due to their stronger emission intensities and shorter lifetimes. These emission signals could also be precisely counted on single cell images. The count number may provide an approach for quantifying the target molecules in the cells.

  20. Application of time-correlated single photon counting and stroboscopic detection methods with an evanescent-wave fibre-optic sensor for fluorescence-lifetime-based pH measurements

    International Nuclear Information System (INIS)

    Henning, Paul E; Geissinger, Peter

    2012-01-01

    Quasi-distributed optical fibre sensor arrays containing luminescent sensor molecules can be read out spatially resolved utilizing optical time-of-flight detection (OTOFD) methods, which employ pulsed laser interrogation of the luminosensors and time-resolved detection of the sensor signals. In many cases, sensing is based on a change in sensor luminescence intensity; however, sensing based on luminescence lifetime changes is preferable because it reduces the need for field calibration. Because in OTOFD detection is time-resolved, luminescence-lifetime information is already available through the signal pulses, although in practise applications were restricted to sensors with long luminescence lifetimes (hundreds of ns). To implement lifetime-based sensing in crossed-optical-fibre-sensor arrays for sensor molecules with lifetimes less than 10 ns, two time-domain methods, time-correlated single photon counting and stroboscopic detection, were used to record the pH-dependent emission of a fluorescein derivative covalently attached to a highly-porous polymer. A two-term nonexponential decay function yielded both a good fit for experimental lifetime data during reconvolution and a pH response that matches Henderson–Hasselbalch behaviour, yielding a sensor accuracy of 0.02 pH units. Moreover, strong agreement was obtained for the two lifetime determination methods and with intensity-based measurements taken previously. (paper)

  1. Radiative lifetime and collisional quenching of carbon monochloride (A 2Δ) in an alternating current glow discharge

    International Nuclear Information System (INIS)

    Gottscho, R.A.; Burton, R.H.; Davis, G.P.

    1982-01-01

    Glow discharges are widely employed in semiconductor processing but are relatively poorly understood owing to, in part, a lack of reliable, quantitative diagnostics. Laser-induced fluorescence promises to be a useful in situ, nonintrusive probe for species concentrations and gas-phase temperatures, but requires the determination of fluoresence yields (i.e., radiative vs nonradiative decay rates) as a function of the plasma state and molecular rotational quantum number. In this work, carbon tetrachloride plasmas, which are used in the dry etching of such materials as Al, Si, GaAs, and InP, are examined using the laser-induced fluorescence technique. The quantum yield phi of CCl A 2 Δ→X 2 Pi fluorescence is determined as a function of pressure, flow-rate, power, electrode temperature, and feedstock composition. Total pressure and addition of Cl 2 to the feedstock are found to be most important in reducing the quantum yield; other plasma parameters and addition of O 2 , He, Ar, or N 2 are found to be of secondary importance. The radiative lifetime of carbon monochloride CCl, A 2 Δ (v = 0) is found to be 105 +- 3 ns and to be independent of rotational quantum number up to J = 45.5. The weak dependence of CCl laser-induced fluorescence on most plasma variables makes it nearly ideal as a simple, direct, and quantitative temperature and concentration diagnostic

  2. Origins of fluorescence in evolved bacteriophytochromes.

    Science.gov (United States)

    Bhattacharya, Shyamosree; Auldridge, Michele E; Lehtivuori, Heli; Ihalainen, Janne A; Forest, Katrina T

    2014-11-14

    Use of fluorescent proteins to study in vivo processes in mammals requires near-infrared (NIR) biomarkers that exploit the ability of light in this range to penetrate tissue. Bacteriophytochromes (BphPs) are photoreceptors that couple absorbance of NIR light to photoisomerization, protein conformational changes, and signal transduction. BphPs have been engineered to form NIR fluorophores, including IFP1.4, Wi-Phy, and the iRFP series, initially by replacement of Asp-207 by His. This position was suggestive because its main chain carbonyl is within hydrogen-bonding distance to pyrrole ring nitrogens of the biliverdin chromophore, thus potentially functioning as a crucial transient proton sink during photoconversion. To explain the origin of fluorescence in these phytofluors, we solved the crystal structures of IFP1.4 and a comparison non-fluorescent monomeric phytochrome DrCBDmon. Met-186 and Val-288 in IFP1.4 are responsible for the formation of a tightly packed hydrophobic hub around the biliverdin D ring. Met-186 is also largely responsible for the blue-shifted IFP1.4 excitation maximum relative to the parent BphP. The structure of IFP1.4 revealed decreased structural heterogeneity and a contraction of two surface regions as direct consequences of side chain substitutions. Unexpectedly, IFP1.4 with Asp-207 reinstalled (IFPrev) has a higher fluorescence quantum yield (∼9%) than most NIR phytofluors published to date. In agreement, fluorescence lifetime measurements confirm the exceptionally long excited state lifetimes, up to 815 ps, in IFP1.4 and IFPrev. Our research helps delineate the origin of fluorescence in engineered BphPs and will facilitate the wide-spread adoption of phytofluors as biomarkers. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

  3. Laser fluorescence determination of radioactive waste cotton fabric in the exploration of uranium content

    International Nuclear Information System (INIS)

    Wang Jiangong

    2010-01-01

    In order to meet the dosage test the operational needs of the laser fluorescence determination of trace radioactive waste cotton fabric uranium research and exploration, to determine the sample ashing time, measured dosage of acidity and digestion and other technical parameters, gives the laser fluorescence determination of radioactive abandoned cotton fabric of trace uranium method. Method of high sensitivity, strong anti-interference, the detection limit of 0.025μg/g(Burning down dregs), relative standard deviation was 3.96%, the mean recovery 93.3%-103% for masks, gloves and other radioactive waste to the determination of trace uranium. (authors)

  4. Determination of sparfloxacin with CdSe/CdS quantum dots as fluorescent probes

    Energy Technology Data Exchange (ETDEWEB)

    Hou, Ming, E-mail: gxglzws@foxmail.com; Yan, Xiaoya; Xiong, Ling

    2015-01-15

    Water-soluble CdSe/CdS quantum dots (QDs) modified with thioglycolic acid (TGA) were synthesized. A novel method for determination of sparfloxacin (SPFX) has been developed based on quenching of the fluorescence of QDs at 556 nm wavelength. The optimum fluorescence intensity was found in 0.067 mol L{sup −1} KH{sub 2}PO{sub 4}–Na{sub 2}HPO{sub 4} buffer solution at pH 6.47 of 3.0×10{sup −5} mol L{sup −1} QDs. When the concentration of quantum dots is 3.0×10{sup −5} mol L{sup −1} the fluorescence quenching intensity of QDs is linearly proportional to the concentration of SPFX from 0.5 μg mL{sup −1} to 30 μg mL{sup −1}, with correlation coefficient R=0.9983. The detection limit for SPFX was 0.1391 μg mL{sup −1}. The method was used for determination of SPFX in tablets, and the results agreed with the claimed value. Trace amounts of SPFX in milk were also determined with the recovery of 95.3–106.8%. - Highlights: • Water-soluble CdSe/CdS quantum dots modified with thioglycolic acid were synthesized. • Determination of sparfloxacin was based on quenching of the fluorescence of QDs. • The detection limit for sparfloxacin was 0.1391 μg mL{sup −1}. • The method has been used successfully to determine SPFX in tablets and milk.

  5. X-ray fluorescence determination of cobalt in iron-manganese oceanic concretions

    International Nuclear Information System (INIS)

    Ivanenko, V.V.; Kustov, V.N.; Metelev, A.Yu.; Rakita, K.A.

    1989-01-01

    A method was developed for resolution of weak analytical lines for elements determined by radionuclide-excited X-ray fluorescence multi-element analysis. The method was used aboart for determining cobalt and some other commercially valuable elements in iron-manganese concretions of Pacific ocean 109 Cd was used as an ionizing radiation source

  6. Improving, characterizing and predicting the lifetime of organic photovoltaics

    DEFF Research Database (Denmark)

    Gevorgyan, Suren A.; Heckler, Ilona Maria; Bundgaard, Eva

    2017-01-01

    This review summarizes the recent progress in the stability and lifetime of organic photovoltaics (OPVs). In particular, recently proposed solutions to failure mechanisms in different layers of the device stack are discussed comprising both structural and chemical modifications. Upscaling...... characterization reported recently. Lifetime testing and determination is another challenge in the field of organic solar cells and the final sections of this review discuss the testing protocols as well as the generic marker for device lifetime and the methodology for comparing all the lifetime landmarks in one...... common diagram. These tools were used to determine the baselines for OPV lifetime tested under different ageing conditions. Finally, the current status of lifetime for organic solar cells is presented and predictions are made for progress in the near future....

  7. Fluorescent turn-on determination of the activity of peptidases using peptide templated gold nanoclusters

    International Nuclear Information System (INIS)

    Luo, Junjun; Wang, Liqiang; Zeng, Ke; Shen, Congcong; Qian, Pin; Yang, Minghui; Rasooly, Avraham; Qu, Fengli

    2016-01-01

    The fluorescence intensity of gold nanoclusters (AuNCs) is inversely related to the length of a peptide immobilized on its surface. This finding has been exploited to design a turn-on fluorescent method for the determination of the activity of peptidase. The β-site amyloid precursor protein-cleaving enzyme 1 (BACE1) was chosen as a model peptidase. BACE1 cleaves the peptide substrates on AuNCs, and the fluorescence intensity of the AuNCs (at exCitation/emission wavelengths of 320/405 nm) carrying the rest of the cleaved peptide is significantly higher than that of the AuNCs with uncleaved peptide. Transmission electron microscopy revealed a decrease in the size of the AuNCs which is assumed cause fluorescence enhancement. The assay was applied to the determination of BACE1 activity in spiked cell lysates, and recoveries were between 96.9 and 104.0 %. (author)

  8. High-resolution imaging of basal cell carcinoma: a comparison between multiphoton microscopy with fluorescence lifetime imaging and reflectance confocal microscopy.

    Science.gov (United States)

    Manfredini, Marco; Arginelli, Federica; Dunsby, Christopher; French, Paul; Talbot, Clifford; König, Karsten; Pellacani, Giovanni; Ponti, Giovanni; Seidenari, Stefania

    2013-02-01

    The aim of this study was to compare morphological aspects of basal cell carcinoma (BCC) as assessed by two different imaging methods: in vivo reflectance confocal microscopy (RCM) and multiphoton tomography with fluorescence lifetime imaging implementation (MPT-FLIM). The study comprised 16 BCCs for which a complete set of RCM and MPT-FLIM images were available. The presence of seven MPT-FLIM descriptors was evaluated. The presence of seven RCM equivalent parameters was scored in accordance to their extension. Chi-squared test with Fisher's exact test and Spearman's rank correlation coefficient were determined between MPT-FLIM scores and adjusted-RCM scores. MPT-FLIM and RCM descriptors of BCC were coupled to match the descriptors that define the same pathological structures. The comparison included: Streaming and Aligned elongated cells, Streaming with multiple directions and Double alignment, Palisading (RCM) and Palisading (MPT-FLIM), Typical tumor islands, and Cell islands surrounded by fibers, Dark silhouettes and Phantom islands, Plump bright cells and Melanophages, Vessels (RCM), and Vessels (MPT-FLIM). The parameters that were significantly correlated were Melanophages/Plump Bright Cells, Aligned elongated cells/Streaming, Double alignment/Streaming with multiple directions, and Palisading (MPT-FLIM)/Palisading (RCM). According to our data, both methods are suitable to image BCC's features. The concordance between MPT-FLIM and RCM is high, with some limitations due to the technical differences between the two devices. The hardest difficulty when comparing the images generated by the two imaging modalities is represented by their different field of view. © 2012 John Wiley & Sons A/S.

  9. Measurement of the D/sup 0/ lifetime

    Energy Technology Data Exchange (ETDEWEB)

    Althoff, M; Braunschweig, W; Gerhards, R; Kirschfink, F J; Martyn, H U; Rosskamp, R; Wallraff, W; Bock, B; Eisenmann, J; Fischer, H M

    1986-10-01

    We have determined the D/sup 0/ lifetime from reconstructed vertices of D/sup 0/ mesons produced in e/sup +/e/sup -/ annihilations at an average center of mass energy of 42.2 GeV. From fifteen events the D/sup 0/ lifetime was determined to be (4.3+-1.7+-0.8)x10/sup -13/s.

  10. Laser diagnostics in combustion. Elastic scattering and picosecond laser-induced fluorescence

    Energy Technology Data Exchange (ETDEWEB)

    Ossler, Frederik

    1999-05-01

    .01 and 10 bar. The effective lifetimes generally decrease with temperature and increase with pressure and in different ways for the two substances which gives implications of their use to combustion related studies. The fluorescence emission from gas phase aromatic hydrocarbons has been studied at elevated temperatures and at atmospheric pressure for different concentrations of oxygen (0-10%). Picosecond laser radiation at 266 nm was used for the excitation. The lifetimes for most of the substances decreased between one and two orders of magnitude when the temperature was increased between 400 and 1200 K. The dependence of the lifetimes on temperature and oxygen concentration indicate that the relaxation dynamics is determined by sequences and the density of vibrational states. The results give also some implications to combustion related diagnostics 170 refs, 30 figs, 2 tabs

  11. Lifetime-vibrational interference effects in resonantly excited x-ray emission spectra of CO

    Energy Technology Data Exchange (ETDEWEB)

    Skytt, P.; Glans, P.; Gunnelin, K. [Uppsala Univ. (Sweden)] [and others

    1997-04-01

    The parity selection rule for resonant X-ray emission as demonstrated for O{sub 2} and N{sub 2} can be seen as an effect of interference between coherently excited degenerate localized core states. One system where the core state degeneracy is not exact but somewhat lifted was previously studied at ALS, namely the resonant X-ray emission of amino-substituted benzene (aniline). It was shown that the X-ray fluorescence spectrum resulting from excitation of the C1s at the site of the {open_quotes}aminocarbon{close_quotes} could be described in a picture separating the excitation and the emission processes, whereas the spectrum corresponding to the quasi-degenerate carbons could not. Thus, in this case it was necessary to take interference effects between the quasi-degenerate intermediate core excited states into account in order to obtain agreement between calculations and experiment. The different vibrational levels of core excited states in molecules have energy splittings which are of the same order of magnitude as the natural lifetime broadening of core excitations in the soft X-ray range. Therefore, lifetime-vibrational interference effects are likely to appear and influence the band shapes in resonant X-ray emission spectra. Lifetime-vibrational interference has been studied in non-resonant X-ray emission, and in Auger spectra. In this report the authors discuss results of selectively excited soft X-ray fluorescence spectra of molecules, where they focus on lifetime-interference effects appearing in the band shapes.

  12. Determination of ammonium on an integrated microchip with LED-induced fluorescence detection.

    Science.gov (United States)

    Xue, Shuhua; Uchiyama, Katsumi; Li, Hai-Fang

    2012-01-01

    A simply fabricated microfluidic device integrated with a fluorescence detection system has been developed for on-line determination of ammonium in aqueous samples. A 365-nm light-emitting diode (LED) as an excitation source and a minor band pass filter were mounted into a polydimethylsiloxane (PDMS)-based microchip for the purpose of miniaturization of the entire analytical system. The ammonium sample reacted with o-phthaldialdehyde (OPA) on-chip with sodium sulfite as reducing reagent to produce a fluorescent isoindole derivative, which can emit fluorescence signal at about 425 nm when excited at 365 nm. Effects of pH, flow rate of solutions, concentrations of OPA-reagent, phosphate and sulfite salt were investigated. The calibration curve of ammonium in the range of 0.018-1.8 microg/mL showed a good linear relationship with R2 = 0.9985, and the detection limit was (S/N = 3) 3.6 x 10(-4) microg/mL. The relative standard deviation was 2.8% (n = 11) by calculating at 0.18 microg/mL ammonium for repeated detection. The system was applied to determine the ammonium concentration in rain and river waters, even extent to other analytes fluorescence detection by the presented device.

  13. Spectrofluorimetric determination of trace amount of coenzyme II using ciprofloxacin-terbium complex as a fluorescent probe

    International Nuclear Information System (INIS)

    Bian Weiwei; Wang Yusheng; Zhu Xiaojing; Jiang Chongqiu

    2006-01-01

    A new spectrofluorimetric method was developed for the determination of trace amount of nicotinamide adenine dinucleotide phosphate (NADP). Using terbium ion (Tb 3+ )-ciprofloxacin (CIP) complex as a fluorescent probe, in the buffer solution of pH=9.00, NADP can remarkably enhance the fluorescence intensity of the Tb 3+ -CIP complex at λ=545nm and the enhanced fluorescence intensity of Tb 3+ ion is in proportion to the concentration of NADP. Optimum conditions for the determination of NADP were also investigated. The dynamic range for the determination of NADP is 4.9x10 -7 -3.7x10 -6 molL -1 with detection limit of 1.3x10 -7 molL -1 . This method is simple, practical and relatively free interference from coexisting substances and can be successfully applied to determination of NADP in synthetic water samples. Moreover, the enhancement mechanisms of the fluorescence intensity in the Tb 3+ -CIP system and the Tb 3+ -CIP-NADP system have been also discussed

  14. The fluorescence behaviour of methyl and phenyl salicylate

    Science.gov (United States)

    Ford, D.; Thistlethwaite, P. J.; Woolfe, G. J.

    1980-01-01

    Fluorcsccnce lifetimes tor the 450 nm emission of methyl and phenyl salicylate in various solvents have been measured. Qucnching studics on the 340 nm fluorescence of these molecules point to the existence of three distinct ground state conformers.

  15. Measurement of the D/sup 0/ lifetime

    Energy Technology Data Exchange (ETDEWEB)

    Althoff, M; Braunschweig, W; Gerhards, R; Kirschfink, F J; Martyn, H U; Rosskamp, P; Wallraff, W; Bock, B; Eisenmann, J; Fischer, H M

    1986-04-01

    We have determined the D/sup 0/ lifetime from reconstructed vertices of D/sup 0/ mesons produced in e/sup +/e/sup -/ annihilations at an average center of mass energy of 42.2 GeV. From fifteen events the D/sup 0/ lifetime was determined to be (4.3/sup +2.0//sub -1.4/ +- 0.8) . 10/sup -13/s.

  16. A reagentless enzymatic fluorescent biosensor for glucose based on upconverting glasses, as excitation source, and chemically modified glucose oxidase.

    Science.gov (United States)

    Del Barrio, Melisa; Cases, Rafael; Cebolla, Vicente; Hirsch, Thomas; de Marcos, Susana; Wilhelm, Stefan; Galbán, Javier

    2016-11-01

    Upon near-infrared excitation Tm(3+)+Yb(3+) doped fluorohafnate glasses present upconversion properties and emit visible light. This property permits to use these glasses (UCG) as excitation sources for fluorescent optical biosensors. Taking this into account, in this work a fluorescent biosensor for glucose determination is designed and evaluated. The biosensor combines the UCG and the fluorescence of the enzyme glucose oxidase chemically modified with a fluorescein derivative (GOx-FS), whose intensity is modified during the enzymatic reaction with glucose. Optical parameters have been optimized and a mathematical model describing the behavior of the analytical signal is suggested. Working in FIA mode, the biosensor responds to glucose concentrations up to, at least, 15mM with a limit of detection of 1.9mM. The biosensor has a minimum lifetime of 9 days and has been applied to glucose determination in drinks. The applicability of the sensor was tested by glucose determination in two fruit juices. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. Fluorescent Nanodiamonds in Biomedical Applications.

    Science.gov (United States)

    Mitura, Katarzyna Anna; Włodarczyk, Elżbieta

    2018-04-18

    Nanoparticles have an extended surface and a large surface area, which is the ratio of the size of the surfacearea to the volume. A functionalized surface can give rise to more modifications and therefore allows this nanomaterial to have new properties. Fluorescent molecules contain fluorophore, which is capable of being excited via the absorption of light energy at a specific wavelength and subsequently emitting radiation energy of a longer wavelength. A chemically modified surface of nanodiamond (ND; by carboxylation) demonstrated biocompatibility with DNA, cytochrome C, and antigens. In turn, fluorescent nanodiamonds (FNDs) belong to a group of new nanomaterials. Their surface can be modified by joining functional groups such as carboxyl, hydroxyl, or amino, after which they can be employed as a fluorescence agent. Their fluorescent properties result from defects in the crystal lattice. FNDs reach dimensions of 4-100 nm, have attributes such as photostability, long fluorescence lifetimes (10 ns), and fluorescence emission between 600 and 700 nm. They are also nontoxic, chemically inert, biocompatible, and environmentally harmless. The main purpose of this article was to present the medical applications of various types of modified NDs.

  18. A new front-face optical cell for measuring weak fluorescent emissions with time resolution in the picosecond time scale.

    Science.gov (United States)

    Gryczynski, Z; Bucci, E

    1993-11-01

    Recent developments of ultrafast fluorimeters allow measuring time-resolved fluorescence on the picosecond time scale. This implies one is able to monitor lifetimes and anisotropy decays of highly quenched systems and of systems that contain fluorophores having lifetimes in the subnanosecond range; both systems that emit weak signals. The combination of weak signals and very short lifetimes makes the measurements prone to distortions which are negligible in standard fluorescence experiments. To cope with these difficulties, we have designed a new optical cell for front-face optics which offers to the excitation beam a horizontal free liquid surface in the absence of interactions with optical windows. The new cell has been tested with probes of known lifetimes and anisotropies. It proved very useful in detecting tryptophan fluorescence in hemoglobin. If only diluted samples are available, which cannot be used in front-face optics, regular square geometry can still be utilized by inserting light absorbers into a cuvette of 1 cm path length.

  19. Time-resolved laser-induced fluorescence system

    Science.gov (United States)

    Bautista, F. J.; De la Rosa, J.; Gallegos, F. J.

    2006-02-01

    Fluorescence methods are being used increasingly in the measurement of species concentrations in gases, liquids and solids. Laser induced fluorescence is spontaneous emission from atoms or molecules that have been excited by laser radiation. Here we present a time resolved fluorescence instrument that consists of a 5 μJ Nitrogen laser (337.1 nm), a sample holder, a quartz optical fiber, a spectrometer, a PMT and a PC that allows the measurement of visible fluorescence spectra (350-750 nm). Time response of the system is approximately 5 ns. The instrument has been used in the measurement of colored bond paper, antifreeze, diesel, cochineal pigment and malignant tissues. The data acquisition was achieved through computer control of a digital oscilloscope (using General Purpose Interface Bus GPIB) and the spectrometer via serial (RS232). The instrument software provides a graphic interface that lets make some data acquisition tasks like finding fluorescence spectra, and fluorescence lifetimes. The software was developed using the Lab-View 6i graphic programming package and can be easily managed in order to add more functions to it.

  20. Comparison of fluorescence-enhancing reagents and optimization of laser fluorimetric technique for the determination of dissolved uranium

    International Nuclear Information System (INIS)

    Ceren Kuetahyali; Joaquin Cobos; Rondinella, V.V.

    2011-01-01

    Results from tests aimed at optimizing an instrumental procedure for the direct and fast determination of uranium in solution by laser fluorescence are presented. A comparison of sample fluorescence measured using different fluorescence enhancing reagents was performed: sodium pyrophosphate, orthophosphoric acid, sulphuric acid and a commercially available fluorescence enhancer were tested for the determination of uranium. From the experimental results, 0.01 M Na 4 P 2 O 7 x 10H 2 O showed the best performance. Effects of reagent pH, different matrices, different concentrations of dissolved Th, and sample volume were investigated. Applications of the improved procedure for the determination of uranium in samples arising from UO 2 -based high level nuclear waste dissolution studies are described. (author)

  1. The generalized centroid difference method for picosecond sensitive determination of lifetimes of nuclear excited states using large fast-timing arrays

    Energy Technology Data Exchange (ETDEWEB)

    Régis, J.-M., E-mail: regis@ikp.uni-koeln.de [Institut für Kernphysik der Universität zu Köln, Zülpicher Str. 77, 50937 Köln (Germany); Mach, H. [Departamento de Física Atómica y Nuclear, Universidad Complutense, 28040 Madrid (Spain); Simpson, G.S. [Laboratoire de Physique Subatomique et de Cosmologie Grenoble, 53, rue des Martyrs, 38026 Grenoble Cedex (France); Jolie, J.; Pascovici, G.; Saed-Samii, N.; Warr, N. [Institut für Kernphysik der Universität zu Köln, Zülpicher Str. 77, 50937 Köln (Germany); Bruce, A. [School of Computing, Engineering and Mathematics, University of Brighton, Lewes Road, Brighton BN2 4GJ (United Kingdom); Degenkolb, J. [Institut für Kernphysik der Universität zu Köln, Zülpicher Str. 77, 50937 Köln (Germany); Fraile, L.M. [Departamento de Física Atómica y Nuclear, Universidad Complutense, 28040 Madrid (Spain); Fransen, C. [Institut für Kernphysik der Universität zu Köln, Zülpicher Str. 77, 50937 Köln (Germany); Ghita, D.G. [Horia Hulubei National Institute for Physics and Nuclear Engineering, 77125 Bucharest (Romania); and others

    2013-10-21

    A novel method for direct electronic “fast-timing” lifetime measurements of nuclear excited states via γ–γ coincidences using an array equipped with N∈N equally shaped very fast high-resolution LaBr{sub 3}(Ce) scintillator detectors is presented. Analogous to the mirror symmetric centroid difference method, the generalized centroid difference method provides two independent “start” and “stop” time spectra obtained by a superposition of the N(N−1)γ–γ time difference spectra of the N detector fast-timing system. The two fast-timing array time spectra correspond to a forward and reverse gating of a specific γ–γ cascade. Provided that the energy response and the electronic time pick-off of the detectors are almost equal, a mean prompt response difference between start and stop events is calibrated and used as a single correction for lifetime determination. These combined fast-timing arrays mean γ–γ time-walk characteristics can be determined for 40keVlifetime determination limit of the method over the total dynamic range is mainly determined by the statistics. The setup of an N=4 detector fast-timing array delivered an absolute time resolving power of 3 ps for 10 000 γ–γ events per total fast timing array start and stop time spectrum. The new method is tested over the total dynamic range by the measurements of known picosecond lifetimes in standard γ-ray sources.

  2. Fluorescent chemosensor for pyridine based on N-doped carbon dots.

    Science.gov (United States)

    Campos, B B; Abellán, C; Zougagh, M; Jimenez-Jimenez, J; Rodríguez-Castellón, E; Esteves da Silva, J C G; Ríos, A; Algarra, M

    2015-11-15

    Fluorescent carbon dots (CDs) and its nitrogen doped (N-CDs) nanoparticles have been synthesized from lactose as precursor using a bottom-up hydrothermal methodology. The synthesized nanoparticles have been characterized by elemental analysis, FTIR, Raman, TEM, DLS, XPS, and steady-state and life-time fluorescence. The synthesized carbon nanoparticles, CDs and N-CDs, have a size at about 7.7±2.4 and 50±15nm, respectively, and quantum yields of 8% (CDs) and 11% (N-CDs). These techniques demonstrated the effectiveness of the synthesis procedure and the functionalization of the CDs surface with amine and amide groups in the presence of NH3 in aqueous media. The effect of excitation wavelength and pH on the luminescent properties was studied. Under the optimal conditions, the nitrogen doped nanoparticles can be used as pyridine sensor in aqueous media because they show an enhancement of its fluorescence with a good linear relationship. The analytical method is simple, reproducible and very sensitive for pyridine determination. Copyright © 2015 Elsevier Inc. All rights reserved.

  3. A dihydrazone based fluorescent probe for selective determination of Al{sup 3+} ions

    Energy Technology Data Exchange (ETDEWEB)

    Pratap Singh, Divya; Singh, Vinod P., E-mail: singvp@yahoo.co.in

    2014-11-15

    A highly selective fluorescent sensor N,N′-bis((2-hydroxynaphthalen-1-yl)methylene) oxalohydrazide (H{sub 2}ohn) for the determination of Al{sup 3+} ions was synthesized and characterized by different physico-chemical and spectroscopic techniques. The single crystal structure of H{sub 2}ohn receptor has also been reported. The H{sub 2}ohn shows an enhanced fluorescence in the presence of Al{sup 3+} ions in ethanol–water (2:3 v/v) solution. Other cations viz. Na{sup +}, K{sup +}, Mg{sup 2+}, Ca{sup 2+}, Mn{sup 2+}, Fe{sup 3+}, Co{sup 2+}, Ni{sup 2+}, Cu{sup 2+}, Zn{sup 2+}, Pb{sup 2+}, Cd{sup 2+} and Hg{sup 2+} show no appreciable change in fluorescence intensity. The binding mode of H{sub 2}ohn receptor with Al{sup 3+} was studied by UV–visible, fluorescence and {sup 1}H NMR titrations. The receptor acts as a dibasic hexadentate ligand and interacts with two Al{sup 3+} ions with a high binding constant K{sub B}=2.62×10{sup 11} M{sup −2}. The lowest detection limit for Al{sup 3+} complex of H{sub 2}ohn was determined to be 8.56×10{sup −10} M. The structures of H{sub 2}ohn and its Al(III) complex were also optimized by DFT calculations. - Highlights: • A dihydrazone based fluorescent sensor is synthesized. • Characterized by IR, NMR, UV−visible and mass spectral analysis. • The receptor serves as a selective fluorescent sensor for Al{sup 3+} over other cations. • The sensing property is monitored by UV−visible, fluorescence and NMR spectroscopy. • A high binding constant of the receptor with Al{sup 3+} is reported here.

  4. Characterization of antibody-chelator conjugates: Determination of chelator content by terbium fluorescence titration

    Energy Technology Data Exchange (ETDEWEB)

    Brandt, K.D.; Schnobrich, K.E.; Johnson, D.K. (Abbott Laboratories, Department 90M, Abbott Park, IL (United States))

    1991-01-01

    Fluorescence titrations were performed by adding varying mole ratios of terbium(III) to antibody conjugates formed by benzyl isothiocyanate derivatives of three different polyaminopolycarboxylate chelators (NTA, EDTA, and DTPA) and the results compared to values for average chelator content obtained by cobalt-57 binding assays. For two different murine monoclonal antibodies, the average chelator content obtained by terbium fluorescence titration correlated closely with that measured by the cobalt-57 binding assay. It is concluded that lanthanide fluorescence titrations provide a useful alternative to radiometal binding assays for the determination of chelator content in protein-chelator conjugates.

  5. Characterization of antibody-chelator conjugates: Determination of chelator content by terbium fluorescence titration

    International Nuclear Information System (INIS)

    Brandt, K.D.; Schnobrich, K.E.; Johnson, D.K.

    1991-01-01

    Fluorescence titrations were performed by adding varying mole ratios of terbium(III) to antibody conjugates formed by benzyl isothiocyanate derivatives of three different polyaminopolycarboxylate chelators (NTA, EDTA, and DTPA) and the results compared to values for average chelator content obtained by cobalt-57 binding assays. For two different murine monoclonal antibodies, the average chelator content obtained by terbium fluorescence titration correlated closely with that measured by the cobalt-57 binding assay. It is concluded that lanthanide fluorescence titrations provide a useful alternative to radiometal binding assays for the determination of chelator content in protein-chelator conjugates

  6. ''LIFETIME'': a computer program for analyzing Doppler-shift recoil-distance nuclear lifetime data

    International Nuclear Information System (INIS)

    Wells, J.C.; Fewell, M.P.; Johnson, N.R.

    1985-10-01

    The program LIFETIME is designed to extract lifetimes of nuclear levels from Doppler-shift recoil-distance experiments by performing a least-square fit to the experimental data (shifted and unshifted photopeak intensities and branching ratios). Initial populations of levels and transition rates between levels are treated as variable parameters. In terms of these parameters the population of each level as a function of time is determined by the Bateman equations, and the shifted and unshifted intensities are calculated. 19 refs., 5 figs

  7. Fluorescence uranium determination; Determinacion de uranio por fluorescencia. I. Proyecto de equipo y comprobacion

    Energy Technology Data Exchange (ETDEWEB)

    Fernandez Cellini, R; Crus Castillo, F. de la; Barrera Pinero, R

    1960-07-01

    An equipment for analysis of uranium by fluorescence was developed in order to determine it at such a low concentration that it can not be determined by the most sensible analytical methods. this new fluorimeter was adapted to measure the fluorescence emitted by the phosphorus sodium fluoride-sodium carbonate-potasium carbonate-uranyl, being excited by ultraviolet light of 3,650 A the intensity of the light emitted was measure with a photomultiplicator RCA 5819 and the adequate electronic equipment. (Author) 19 refs.

  8. Determination of the average lifetime of b-baryons

    International Nuclear Information System (INIS)

    Abreu, P.; Adam, W.

    1996-01-01

    The average lifetime of b-baryons has been studied using 3.10 6 hadronic Z 0 decays collected by the DELPHI detector at LEP. Three methods have been used, based on the measurement of different observables: the proper decay time distribution of 206 vertices reconstructed with a Λ, a lepton and an oppositely charged pion; the impact parameter distribution of 441 muons with high transverse momentum accompanied by a Λ in the same jet; and the proper decay time distribution of 125 Λ c -lepton decay vertices with the Λ c exclusively reconstructed through its pKπ, pK 0 and Λ3π decay modes. The combined result is: τ(b-baryon)=(1.254 +0.121 -0.109 (stat) ±0.04(syst) +0.03 -0.05 (syst)) ps where the first systematic error is due to experimental uncertainties and the second to the uncertainties in the modelling of the b-baryon production and semi-leptonic decay. Including the measurement recently published by DELPHI based on a sample of proton-muon vertices, the average b-baryon lifetime is: τ(b-baryon)=(1.255 +0.115 -0.102 (stat) ±0.05) ps. (orig.)

  9. Fluorescein Derivatives in Intravital Fluorescence Imaging

    Directory of Open Access Journals (Sweden)

    Michael S. Roberts

    2013-08-01

    Full Text Available Intravital fluorescence microscopy enables the direct imaging of fluorophores in vivo and advanced techniques such as fluorescence lifetime imaging (FLIM enable the simultaneous detection of multiple fluorophores. Consequently, it is now possible to record distribution and metabolism of a chemical in vivo and to optimise the delivery of fluorophores in vivo. Recent clinical applications with fluorescein and other intravital fluorescent stains have occurred in neurosurgery, dermatology [including photodynamic therapy (PDT] and endomicroscopy. Potential uses have been identified in periodontal disease, skin graft and cancer surgery. Animal studies have demonstrated that diseased tissue can be specifically stained with fluorophore conjugates. This review focuses on the fluorescein derived fluorophores in common clinical use and provides examples of novel applications from studies in tissue samples.

  10. Determination of uranium in geologic materials by laser-excited fluorescence

    International Nuclear Information System (INIS)

    McHugh, J.B.

    1982-01-01

    A laser-excited fluorescence method is described for the determination of trace amounts of uranium in rocks and soils. The limit of detection is less than 1 ppm, and the relative standard deviation ranges from 2.6 to 12.5%. The method was evaluated by using known geological reference samples

  11. The use of fluorescent intrabodies to detect endogenous gankyrin in living cancer cells

    International Nuclear Information System (INIS)

    Rinaldi, Anne-Sophie; Freund, Guillaume; Desplancq, Dominique; Sibler, Annie-Paule; Baltzinger, Mireille; Rochel, Natacha; Mély, Yves; Didier, Pascal; Weiss, Etienne

    2013-01-01

    Expression of antibody fragments in mammalian cells (intrabodies) is used to probe the target protein or interfere with its biological function. We previously described the in vitro characterisation of a single-chain Fv (scFv) antibody fragment (F5) isolated from an intrabody library that binds to the oncoprotein gankyrin (GK) in solution. Here, we have isolated several other scFvs that interact with GK in the presence of F5 and tested whether they allow, when fused to fluorescent proteins, to detect by FRET endogenous GK in living cells. The binding of pairs of scFvs to GK was analysed by gel filtration and the ability of each scFv to mediate nuclear import/export of GK was determined. Binding between scFv-EGFP and RFP-labelled GK in living cells was detected by fluorescence lifetime imaging microscopy (FLIM). After co-transfection of two scFvs fused to EGFP and RFP, respectively, which form a tri-molecular complex with GK in vitro, FRET signal was measured. This system allowed us to observe that GK is monomeric and distributed throughout the cytoplasm and nucleus of several cancer cell lines. Our results show that pairs of fluorescently labelled intrabodies can be monitored by FLIM–FRET microscopy and that this technique allows the detection of lowly expressed endogenous proteins in single living cells. Highlights: ► Endogenous GK in living cells was targeted with pairs of fluorescently-tagged scFvs. ► Tri-molecular complexes containing two scFvs and one molecule GK were formed. ► GK was detected using fluorescence lifetime-based FRET imaging. ► GK is monomeric and homogeneously distributed in several cancer cell lines. ► This technique may have many applications in live-cell imaging of endogenous proteins

  12. The use of fluorescent intrabodies to detect endogenous gankyrin in living cancer cells

    Energy Technology Data Exchange (ETDEWEB)

    Rinaldi, Anne-Sophie; Freund, Guillaume; Desplancq, Dominique; Sibler, Annie-Paule; Baltzinger, Mireille [Ecole Supérieure de Biotechnologie de Strasbourg, UMR 7242, CNRS/Université de Strasbourg, boulevard Sébastien Brant, 67412 Illkirch (France); Rochel, Natacha [Institut de Génétique et de Biologie Moléculaire et Cellulaire, UMR 7104, CNRS/INSERM/Université de Strasbourg, rue Laurent Fries, 67404 Illkirch (France); Mély, Yves; Didier, Pascal [Faculté de Pharmacie, UMR 7213, CNRS/Université de Strasbourg, route du Rhin, 67401 Illkirch (France); Weiss, Etienne, E-mail: eweiss@unistra.fr [Ecole Supérieure de Biotechnologie de Strasbourg, UMR 7242, CNRS/Université de Strasbourg, boulevard Sébastien Brant, 67412 Illkirch (France)

    2013-04-01

    Expression of antibody fragments in mammalian cells (intrabodies) is used to probe the target protein or interfere with its biological function. We previously described the in vitro characterisation of a single-chain Fv (scFv) antibody fragment (F5) isolated from an intrabody library that binds to the oncoprotein gankyrin (GK) in solution. Here, we have isolated several other scFvs that interact with GK in the presence of F5 and tested whether they allow, when fused to fluorescent proteins, to detect by FRET endogenous GK in living cells. The binding of pairs of scFvs to GK was analysed by gel filtration and the ability of each scFv to mediate nuclear import/export of GK was determined. Binding between scFv-EGFP and RFP-labelled GK in living cells was detected by fluorescence lifetime imaging microscopy (FLIM). After co-transfection of two scFvs fused to EGFP and RFP, respectively, which form a tri-molecular complex with GK in vitro, FRET signal was measured. This system allowed us to observe that GK is monomeric and distributed throughout the cytoplasm and nucleus of several cancer cell lines. Our results show that pairs of fluorescently labelled intrabodies can be monitored by FLIM–FRET microscopy and that this technique allows the detection of lowly expressed endogenous proteins in single living cells. Highlights: ► Endogenous GK in living cells was targeted with pairs of fluorescently-tagged scFvs. ► Tri-molecular complexes containing two scFvs and one molecule GK were formed. ► GK was detected using fluorescence lifetime-based FRET imaging. ► GK is monomeric and homogeneously distributed in several cancer cell lines. ► This technique may have many applications in live-cell imaging of endogenous proteins.

  13. Response of Substituted Indoleacetic Acids in the Indolo-alpha-pyrone Fluorescence Determination

    DEFF Research Database (Denmark)

    Engvild, Kjeld Christensen; Böttger, M.; Kaiser, P.

    1978-01-01

    .-pyrones. Other halogenated indoleacetic acids show between zero and 60% of the fluorescence of IAA. Apparently the concentration of IAA cannot be determined in crude extracts in the presence of 4-chloro- or 5-hydroxy-indoleacetic acid, because separate determinations of each of these compounds are not possible...

  14. Modeling and optimization of membrane lifetime in dead-end ultra filtration

    NARCIS (Netherlands)

    Zondervan, E.; Roffel, B.

    2008-01-01

    In this paper, a membrane lifetime model is developed and experimentally validated. The lifetime model is based on the Weibull probability density function. The lifetime model can be used to determine an unambiguous characteristic membrane lifetime. Experimental results showed that membrane lifetime

  15. Precision determination of the lifetime of the neutral pion

    CERN Multimedia

    2002-01-01

    The experiment aims at a determination of the lifetime of the neutral pion with an accuracy of 1\\%, as against at present 5\\% for the best measurement by the indirect, Primakoff method, and 18\\% by a direct measurement. \\\\\\\\2 gold foils with different separations are traversed by the extracted proton beam to the High Intensity Area. Positive particles of 160 GeV/c momentum from this target are transported by the H6 beam to the North Area. The electron component is separated from heavier particles by synchrotron losses in the bending magnets, and is further identified by a lead glass Cerenkov counter, in coincidence with scintillation counters (Fig. 1).\\\\\\\\ The electron flux from the double foil target depends on the distance between the foils, since for small separations the neutral pions will have to decay before electrons can be materialised by the decay $\\gamma$ rays.\\\\\\\\ The average momentum of neutral pions from a 400 GeV pN reaction, giving a 160 GeV/c electron, is about 250 GeV/c, and the corresponding...

  16. A ratiometric nanoprobe based on silver nanoclusters and carbon dots for the fluorescent detection of biothiols

    Science.gov (United States)

    Zhang, Shuming; Lin, Bixia; Yu, Ying; Cao, Yujuan; Guo, Manli; Shui, Lingling

    2018-04-01

    Ratiometric fluorescent probes could eliminate the influence from experimental factors and improve the detection accuracy. In this article, a ratiometric nanoprobe was constructed based on silver nanoclusters (AgNCs) with nitrogen-doped carbon dots (NCDs) and used for the detection of biothiols. The fluorescence peak of AgNCs was observed at 650 nm with excitation wavelength at 370 nm. In order to construct the ratiometric fluorescent probe, NCDs with the excitation and emission wavelengths at 370 nm and 450 nm were selected. After adding AgNCs, the fluorescence of NCDs was quenched. The mechanism of the fluorescence quenching was studied by fluorescence, UV-Vis absorption and the fluorescence lifetime spectra. The results indicated that the quenching could be ascribed to the inner filter effect (IFE). With the addition of biothiols, the fluorescence of AgNCs at 650 nm decreased due to the breakdown of AgNCs, and the fluorescence of NCDs at 450 nm recovered accordingly. Thus, the relationship between the ratio of the fluorescence intensities (I450/I650) and biothiol concentration was used to establish the determination method for biothiols. Cysteine (Cys) was taken as the model of biothiols, and the working curve for Cys was I450/I650 = 0.60CCys - 1.86 (CCys: μmol/L) with the detection limit of 0.14 μmol/L (S/N = 3). Then, the method was used for the detection of Cys in human urine and serum samples with satisfactory accuracy and recovery ratios. Furthermore, the probe could be applied for the visual semi-quantitative determination of Cys by naked eyes.

  17. Critical tonicity determination of sperm using fluorescent staining and flow cytometry

    Energy Technology Data Exchange (ETDEWEB)

    Noiles, E.E.; Ruffing, N.A.; Kleinhans, F.W.; Mark, L.A.; Watson, P.F.; Critser, J.K. (Methodist Hospital, Indianapolis, IN (USA)); Horstman, L. (Purdue Univ., Lafayette, IN (USA). School of Veterinary Medicine); Mazur, P. (Oak Ridge National Lab., TN (USA))

    1990-01-01

    The use of cryopreserved, rather than fresh, mammalian semen for artificial insemination confers several important medical and/or economic advantages. However, current methods for cryopreservation of both human and bovine spermatozoa result in approximately only a 50% survival rate with thawing, obviously reducing the fertilizing capacity of the semen. A primary consideration during the cooling process is to avoid intracellular ice crystal formation with its lethal consequences to the cell. Current techniques achieve this by controlling the cooling rate. Computation of the time necessary for this dehydration, and hence, the cooling rate, is dependent upon knowledge of the water permeability coefficient (L{sub {rho}}) and its activation energy. The fluorophore, 6-carboxyfluoroscein diacetate (CFDA), which is nonfluorescent, readily crosses the intact plasma membrane. Intracellular esterases hydrolyze CFDA to 6-carboxyfluoroscein, a fluorescent, membrane-impermeable fluorophore. Consequently, spermatozoa with intact plasma membranes fluoresce bright green (Garner et. al., 1986), but those with disrupted membranes do not. Therefore, the purpose of this study was to use loss of CFDA fluorescence to determine the osmolality at which 50% of the spermatozoa will swell and lyse (critical tonicity, CT). These data will then be used to determine the L{sub {rho}} and its activation energy for sperm, thus increasing the knowledge available in cellular cryopreservation. 15 refs., 3 figs.

  18. Monte Carlo modeling of time-resolved fluorescence for depth-selective interrogation of layered tissue.

    Science.gov (United States)

    Pfefer, T Joshua; Wang, Quanzeng; Drezek, Rebekah A

    2011-11-01

    Computational approaches for simulation of light-tissue interactions have provided extensive insight into biophotonic procedures for diagnosis and therapy. However, few studies have addressed simulation of time-resolved fluorescence (TRF) in tissue and none have combined Monte Carlo simulations with standard TRF processing algorithms to elucidate approaches for cancer detection in layered biological tissue. In this study, we investigate how illumination-collection parameters (e.g., collection angle and source-detector separation) influence the ability to measure fluorophore lifetime and tissue layer thickness. Decay curves are simulated with a Monte Carlo TRF light propagation model. Multi-exponential iterative deconvolution is used to determine lifetimes and fractional signal contributions. The ability to detect changes in mucosal thickness is optimized by probes that selectively interrogate regions superficial to the mucosal-submucosal boundary. Optimal accuracy in simultaneous determination of lifetimes in both layers is achieved when each layer contributes 40-60% of the signal. These results indicate that depth-selective approaches to TRF have the potential to enhance disease detection in layered biological tissue and that modeling can play an important role in probe design optimization. Published by Elsevier Ireland Ltd.

  19. X-Ray fluorescence determination of the mobile forms of toxic elements in meadow chernozems

    International Nuclear Information System (INIS)

    Belikov, K.N.; Blank, A.B.; Shevtsov, N.I.

    1997-01-01

    An X-ray fluorescence method for determining mobile forms of Mn, V, Cr, Co, Ni, Zn, and Pb in meadow chernozems was developed. It is based on the extraction of analytes with an acetate-ammonia buffer solution and the evaporation of the obtained filtrate with carbon powder followed by the X-ray fluorescence analysis of the dry residue. The effect of concomitants on the analytical signals of elements under determination was examined. It was suggested to spike samples with barium in order to decrease analytical errors. (author)

  20. Lifetime of B hadrons from CDF

    International Nuclear Information System (INIS)

    Miao, Ting.

    1996-08-01

    A review of the lifetimes of B hadrons measured by the CDF collaboration at Fermilab is presented. The data corresponds to 110 pb -1 of p anti p collisions at √s = 1.8 TeV. The inclusive B hadron lifetime is measured using a high statistics sample of B → J/ΨΧ decays. Species specific lifetimes of the B + , B 0 , B 0 s , and Λ 0 b are determined using both fully reconstructed decays and partially reconstructed decays consisting of a lepton associated with a charm hadron

  1. Computer-assisted sperm morphometry fluorescence-based analysis has potential to determine progeny sex

    Directory of Open Access Journals (Sweden)

    Pilar Santolaria

    2016-01-01

    Full Text Available This study was designed to determine the ability of computer-assisted sperm morphometry analysis (CASA-Morph with fluorescence to discriminate between spermatozoa carrying different sex chromosomes from the nuclear morphometrics generated and different statistical procedures in the bovine species. The study was divided into two experiments. The first was to study the morphometric differences between X- and Y-chromosome-bearing spermatozoa (SX and SY, respectively. Spermatozoa from eight bulls were processed to assess simultaneously the sex chromosome by FISH and sperm morphometry by fluorescence-based CASA-Morph. SX cells were larger than SY cells on average (P < 0.001 although with important differences between bulls. A simultaneous evaluation of all the measured features by discriminant analysis revealed that nuclear area and average fluorescence intensity were the variables selected by stepwise discriminant function analysis as the best discriminators between SX and SY. In the second experiment, the sperm nuclear morphometric results from CASA-Morph in nonsexed (mixed SX and SY and sexed (SX semen samples from four bulls were compared. FISH allowed a successful classification of spermatozoa according to their sex chromosome content. X-sexed spermatozoa displayed a larger size and fluorescence intensity than nonsexed spermatozoa (P < 0.05. We conclude that the CASA-Morph fluorescence-based method has the potential to find differences between X- and Y-chromosome-bearing spermatozoa in bovine species although more studies are needed to increase the precision of sex determination by this technique.

  2. Quantitative determination of uranium in organic solution by X-ray fluorescence

    International Nuclear Information System (INIS)

    Leyt, D.V. de; Colangelo, C.H.

    1987-01-01

    An X-ray fluorescent method for the determination of uranium in tributilphosphate-kerosene-nitriacid solution has been developed. Chemical properties of the matrix elements were studied in order to select a convenient procedure to determine samples and standards on the same way. The method avoids the destruction of the organic material and has proved to be very useful for the fast control of uranium concentration. (Author) [es

  3. Fluorescence enhancement by Au nanostructures: nanoshells and nanorods.

    Science.gov (United States)

    Bardhan, Rizia; Grady, Nathaniel K; Cole, Joseph R; Joshi, Amit; Halas, Naomi J

    2009-03-24

    Metallic nanoparticles influence the quantum yield and lifetime of adjacent fluorophores in a manner dependent on the properties of the nanostructure. Here we directly compare the fluorescence enhancement of the near-infrared fluorophore IR800 by Au nanoshells (NSs) and Au nanorods (NRs), where human serum albumin (HSA) serves as a spacer layer between the nanoparticle and the fluorophore. Our measurements reveal that the quantum yield of IR800 is enhanced from approximately 7% as an isolated fluorophore to 86% in a NSs-HSA-IR800 complex and 74% in a NRs-HSA-IR800 complex. This dramatic increase in fluorescence shows tremendous potential for contrast enhancement in fluorescence-based bioimaging.

  4. 5f state interaction with inner coordination sphere ligands: einsteinium 3+ ion fluorescence in aqueous and organic phases

    International Nuclear Information System (INIS)

    Beitz, J.V.; Wester, D.W.; Williams, C.W.

    1983-01-01

    The interaction between 5f electron states of einsteinium 3+ ion and coordinated ligands in solution has been probed using laser-induced fluorescence. Aquo einsteinium 3+ ion was observed to fluoresce from its first excited J = 5 state in a broad-band peaking at 9260 wavenumbers. The observed fluorescence lifetimes were 1.05 microseconds and 2.78 microseconds in H 2 O and D 2 O (99+ % D atom), respectively. The non-radiative decay rates derived from the lifetime data are compared with previously reported data for Cm, Sm, Eu, Tb, and Dy aquo 3+ ions. The 5f actinide states exhibit substantially greater non-radiative decay rates than do lanthanide 4f states of similar energy gap. This provides evidence that actinide 5f electrons interact more strongly with their inner coordination sphere than do lanthanide ion 4f electrons. The fluorescence lifetime of einsteinium 3+ ion complexed with 1 formal di(2-ethylhexyl)orthophosphoric acid in h-heptane was 2.34 microseconds. 3 figures, 1 table

  5. Mapping Charge Carrier Density in Organic Thin-Film Transistors by Time-Resolved Photoluminescence Lifetime Studies

    DEFF Research Database (Denmark)

    Leißner, Till; Jensen, Per Baunegaard With; Liu, Yiming

    2017-01-01

    The device performance of organic transistors is strongly influenced by the charge carrier distribution. A range of factors effect this distribution, including injection barriers at the metal-semiconductor interface, the morphology of the organic film, and charge traps at the dielectric/organic...... interface or at grain boundaries. In our comprehensive experimental and analytical work we demonstrate a method to characterize the charge carrier density in organic thin-film transistors using time-resolved photoluminescence spectroscopy. We developed a numerical model that describes the electrical...... and optical responses consistently. We determined the densities of free and trapped holes at the interface between the organic layer and the SiO2 gate dielectric by comparison to electrical measurements. Furthermore by applying fluorescence lifetime imaging microscopy we determine the local charge carrier...

  6. Laser-time resolved fluorimetric determination of trace of boron in U3O8

    International Nuclear Information System (INIS)

    Xu Yongyuan; Wang Yulong; Wang Qin

    1988-01-01

    In this work, a laser-time resolved fluorimetric determinatin of trace of boron in U 3 O 8 had been developed. The boron complex with dibenzoyl methane (DBM) in a suitable medium is excited by a small nitrogen laser and emits the delay fluorescence with lifetime of 2 ms which is much longer than that of the fluorescence of uranium. Since the fluorescence of uranium doesn't interfere with determination of boron in the time resolved fluorimetric method boron need not be separated from uranium in advance. Thus the determination is very rapid and simple. The limit of determination is 0.02 ngB/ml. When 10 mgU is taken, 0.01 ppm of boron in uranium can be determined. Several samples of U 3 O 8 with boron content from 0.04 to 0.5 ppm have been determined by using this method. The results of determination have been accordant with other methods

  7. Fluorescent sensing and determination of mercury (II) ions in water ...

    African Journals Online (AJOL)

    In this study we report on a fluorescent sensing probe based on a naphthyl azo dye modified dibenzo-18-crown-6-ether (DB18C6) for the detection and determination of mercury (II) ions in water. The probe showed high sensitivity and selectivity towards the mercury (II) ion among various alkali, alkaline earth, and transition ...

  8. Blood selenium content determination by X-ray fluorescence

    International Nuclear Information System (INIS)

    Mainardi, R.T.

    1987-01-01

    The presence of some elements in small amounts (traces) in the human body is of foremost importance for the prevention and treatment of several diseases. It has been recently shown that traces of selenium in blood are closely related to the occurrence of miotonic distrophy, a muscular disease that is affecting a significant percentage of the population. This work describes a simple procedure to determine selenium in human blood serum by energy dispersive X-ray fluorescence analysis. Final quantification is achieved through the addition of titanium as an internal standard. (Author) [es

  9. Determination of coal ash content by the combined x-ray fluorescence and scattering spectrum

    Science.gov (United States)

    Mikhailov, I. F.; Baturin, A. A.; Mikhailov, A. I.; Borisova, S. S.; Fomina, L. P.

    2018-02-01

    An alternative method is proposed for the determination of the inorganic constituent mass fraction (ash) in solid fuel by the ratio of Compton and Rayleigh X-ray scattering peaks IC/IR subject to the iron fluorescence intensity. An original X-ray optical scheme with a Ti/Mo (or Sc/Cu) double-layer secondary radiator allows registration of the combined fluorescence-and-scattering spectrum at the specified scattering angle. An algorithm for linear calibration of the Compton-to-Rayleigh IC/IR ratio is proposed which uses standard samples with two certified characteristics: mass fractions of ash (Ad) and iron oxide (WFe2O3). Ash mass fractions have been determined for coals of different deposits in the wide range of Ad from 9.4% to 52.7% mass and WFe2O3 from 0.3% to 4.95% mass. Due to the high penetrability of the probing radiation with energy E > 17 keV, the sample preparation procedure is rather simplified in comparison with the traditional method of Ad determination by the sum of fluorescence intensities of all constituent elements.

  10. Determination of {pi}{pi} scattering lengths from measurement of {pi}{sup +{pi}-} atom lifetime

    Energy Technology Data Exchange (ETDEWEB)

    Adeva, B. [Santiago de Compostela University (Spain); Afanasyev, L. [JINR Dubna (Russian Federation); Benayoun, M. [LPNHE des Universites Paris VI/VII, IN2P3-CNRS (France); Benelli, A. [Zurich University (Switzerland); Berka, Z. [Czech Technical University in Prague, Prague (Czech Republic); Brekhovskikh, V. [IHEP Protvino (Russian Federation); Caragheorgheopol, G. [IFIN-HH, National Institute for Physics and Nuclear Engineering, Bucharest (Romania); Cechak, T. [Czech Technical University in Prague, Prague (Czech Republic); Chiba, M. [Tokyo Metropolitan University (Japan); Chliapnikov, P.V. [IHEP Protvino (Russian Federation); Ciocarlan, C.; Constantinescu, S. [IFIN-HH, National Institute for Physics and Nuclear Engineering, Bucharest (Romania); Costantini, S. [Basel University (Switzerland); Curceanu, C. [IFIN-HH, National Institute for Physics and Nuclear Engineering, Bucharest (Romania); Doskarova, P. [Czech Technical University in Prague, Prague (Czech Republic); Dreossi, D. [INFN, Sezione di Trieste and Trieste University, Trieste (Italy); Drijard, D., E-mail: Daniel.Drijard@cern.ch [CERN, Geneva (Switzerland); Dudarev, A. [JINR Dubna (Russian Federation); Ferro-Luzzi, M. [CERN, Geneva (Switzerland); Fungueirino Pazos, J.L. [Santiago de Compostela University (Spain)

    2011-10-05

    The DIRAC experiment at CERN has achieved a sizeable production of {pi}{sup +{pi}-} atoms and has significantly improved the precision on its lifetime determination. From a sample of 21 227 atomic pairs, a 4% measurement of the S-wave {pi}{pi} scattering length difference |a{sub 0}-a{sub 2}|=(0.2533{sub -0.0078}{sup +0.0080}|{sub stat}{sup +0.0078}{sub -0.0073}|{sub syst})M{sub {pi}}{sup +-1} has been attained, providing an important test of Chiral Perturbation Theory.

  11. Imaging molecular interactions in cells by dynamic and static fluorescence anisotropy (rFLIM and emFRET)

    NARCIS (Netherlands)

    Lidke, D.S.; Nagy, P.; Barisas, B.G.; Heintzmann, R.; Post, Janine Nicole; Lidke, K.A.; Clayton, A.H.A.; Arndt-jovin, D.J.; Jovin, T.M.

    2003-01-01

    We report the implementation and exploitation of fluorescence polarization measurements, in the form of anisotropy fluorescence lifetime imaging microscopy (rFLIM) and energy migration Förster resonance energy transfer (emFRET) modalities, for wide-field, confocal laser-scanning microscopy and flow

  12. Fast and accurate automated cell boundary determination for fluorescence microscopy

    Science.gov (United States)

    Arce, Stephen Hugo; Wu, Pei-Hsun; Tseng, Yiider

    2013-07-01

    Detailed measurement of cell phenotype information from digital fluorescence images has the potential to greatly advance biomedicine in various disciplines such as patient diagnostics or drug screening. Yet, the complexity of cell conformations presents a major barrier preventing effective determination of cell boundaries, and introduces measurement error that propagates throughout subsequent assessment of cellular parameters and statistical analysis. State-of-the-art image segmentation techniques that require user-interaction, prolonged computation time and specialized training cannot adequately provide the support for high content platforms, which often sacrifice resolution to foster the speedy collection of massive amounts of cellular data. This work introduces a strategy that allows us to rapidly obtain accurate cell boundaries from digital fluorescent images in an automated format. Hence, this new method has broad applicability to promote biotechnology.

  13. Autoindicating optical properties of laccase as the base of an optical biosensor film for phenol determination

    Energy Technology Data Exchange (ETDEWEB)

    Sanz, J.; Marcos, S. de; Galban, J. [University of Zaragoza, Analytical Biosensors Group (GBA), Analytical Chemistry Department, Faculty of Sciences, Zaragoza (Spain)

    2012-08-15

    In the context of sustainable analytical chemistry, phenol has been determined through its enzymatic reaction with laccase. The method has been studied and optimized through the autoindicating optical properties of laccase both by intrinsic molecular absorption and fluorescence. The method shows a linear range from 9.79.10{sup -6} to 7.50.10{sup -4} M with a relative standard deviation of 1.07 %. The molecular absorption methodology has been implemented in a polyacrylamide film for the design of an autoindicating optical sensor. In order to increase the lifetime of the sensor, the reversibility study of the enzymatic reaction has proposed, as a novelty, the regeneration of laccase with an oxidase-type enzyme (glucose oxidase). The lifetime of the sensor film has improved from 15 to 30 measurements. The reaction mechanism has also been studied and confirmed by fluorescence and molecular absorption. The method leads to the determination of phenol in environmental samples. (orig.)

  14. Angular distributions as lifetime probes

    Energy Technology Data Exchange (ETDEWEB)

    Dror, Jeff Asaf; Grossman, Yuval [Department of Physics, LEPP, Cornell University,Ithaca, NY 14853 (United States)

    2014-06-27

    If new TeV scale particles are discovered, it will be important to determine their width. There is, however, a problematic region, where the width is too small to be determined directly, and too large to generate a secondary vertex. For a collection of colored, spin polarized particles, hadronization depolarizes the particles prior to their decay. The amount of depolarization can be used to probe the lifetime in the problematic region. In this paper we apply this method to a realistic scenario of a top-like particle that can be produced at the LHC. We study how depolarization affects the angular distributions of the decay products and derive an equation for the distributions that is sensitive to the lifetime.

  15. Simultaneous determination of Magnolol and Honokiol by amino acid ionic liquid synchronous fluorescence spectrometry

    Science.gov (United States)

    Liu, Wei; Zhu, Xiashi

    2018-05-01

    A novel method based on amino acid ionic liquids (AAILs) as an additive synchronous fluorescence spectrometry is proposed for simultaneous determination of magnolol (MN) and honokiol (HN) in traditional Chinese medicine Houpu. The overlapping fluorescence spectrum of MN and HN could be completely separated in the AAILs medium. Experiment parameters (the type and concentration of AAILs, pH values and temperature) were discussed. The detection limits of MN and HN reached 1.46 ng/mL, 0.92 ng/mL and the recovery rates ranged from 98.6%-100.7%, 99.7%-100.6%, respectively. This methods was successfully employed for simultaneously determination of MN and HN in real samples. No significant differences could be found in the results of this method and the pharmacopoeia of People's Republic of China 2015 (Ch.P.2015). The experiment mechanisms were discussed by the Gaussian simulation and fluorescence quantum yield.

  16. An X-ray fluorescence method for the determination of metals thicknesses

    International Nuclear Information System (INIS)

    Vazquez, Cristina; Leyt, D.V. de; Riveros, J.A.

    1987-01-01

    An absolute method for the determination of the thickness of a metal film deposited on a metallic substrate is described. The method is based on the measurement of fluorescent intensity ratios for two lines from the substrate element. Additionally, the proposed method can be employed to determine the density of the deposited material or the incident angle of primary radiation and take off angle, if the metal film thickness is known. (Author) [es

  17. Long term optical stability of fluorescent solar concentrator plates

    NARCIS (Netherlands)

    Slooff, L.H.; Bakker, N.J.; Sommeling, P.M.; Büchtemann, A.; Wedel, A.; Sark, W.G.J.H.M. van

    2014-01-01

    Fluorescent solar concentrators offer an alternative approach for low-cost photovoltaic energy conversion. For successful application, not only the power conversion efficiency and cost are important, but also lifetime or stability of the devices. As today’s concentrator is made of polymer sheets

  18. Long-term optical stability of fluorescent solar concentrator plates

    NARCIS (Netherlands)

    Slooff, Lenneke H.; Bakker, Nicolaas J.; Sommeling, Paul M.; Büchtemann, Andreas; Wedel, Armin; Van Sark, Wilfried G J H M

    2014-01-01

    Fluorescent solar concentrators offer an alternative approach for low-cost photovoltaic energy conversion. For successful application, not only the power conversion efficiency and cost are important, but also lifetime or stability of the devices. As today's concentrator is made of polymer sheets

  19. Fluorescence lifetime spectroscopy: potential for in-vivo estimation of skin fluorophores changes after low power laser treatment

    Science.gov (United States)

    Ferulova, Inesa; Lihachev, Alexey; Spigulis, Janis

    2013-11-01

    The impact of visible cwlaser irradiation on skin autofluorescence lifetimes was investigated in spectral range from 450 nm to 600 nm. Skin optical provocations were performed during 1 min by 405 nm low power cw laser with power density up to 20 mW/cm2. Autofluorescence lifetimes were measured before and immediately after the optical provocation.

  20. Measurement of the $\\Omega_{c}^{0}$ lifetime

    CERN Document Server

    Adamovich, M.I.; Alexandrov, Yu.A.; Barberis, D.; Beck, M.; Berat, C.; Beusch, W.; Boss, M.; Brons, S.; Bruckner, W.; Buenerd, M.; Buscher, C.; Charignon, F.; Chauvin, J.; Chudakov, E.A.; Dropmann, F.; Engelfried, J.; Faller, F.; Fournier, A.; Gerasimov, S.; Godbersen, M.; Grafstrom, P.; Haller, T.; Heidrich, M.; Hurst, R.B.; Konigsmann, Kay; Konorov, I.; Martens, K.; Martin, P.; Masciocchi, S.; Michaels, R.; Muller, U.; Newsom, C.; Paul, S.; Povh, B.; Ren, Z.; Rey-Campagnolle, M.; Rosner, G.; Rossi, L.; Rudolph, H.; Schmitt, L.; Siebert, H.W.; Simon, A.; Smith, V.J.; Thilmann, O.; Trombini, A.; Vesin, E.; Volkemer, B.; Vorwalter, K.; Walcher, T.; Walder, G.; Werding, R.; Wittmann, E.; Zavertyaev, M.V.

    1995-01-01

    We present the measurement of the lifetime of the Omega_c we have performed using three independent data samples from two different decay modes. Using a Sigma- beam of 340 GeV/c we have obtained clean signals for the Omega_c decaying into Xi- K- pi+ pi+ and Omega- pi+ pi- pi+, avoiding topological cuts normally used in charm analysis. The short but measurable lifetime of the Omega_c is demonstrated by a clear enhancement of the signals at short but finite decay lengths. Using a continuous maximum likelihood method we determined the lifetime to be tau(Omega_c) = 55 +13-11(stat) +18-23(syst) fs. This makes the Omega_c the shortest living weakly decaying particle observed so far. The short value of the lifetime confirms the predicted pattern of the charmed baryon lifetimes and demonstrates that the strong interaction plays a vital role in the lifetimes of charmed hadrons.

  1. Excited-state lifetime measurement of silicon vacancy centers in diamond by single-photon frequency upconversion

    Science.gov (United States)

    Rong, Youying; Ma, Jianhui; Chen, Lingxiao; Liu, Yan; Siyushev, Petr; Wu, Botao; Pan, Haifeng; Jelezko, Fedor; Wu, E.; Zeng, Heping

    2018-05-01

    We report a method with high time resolution to measure the excited-state lifetime of silicon vacancy centers in bulk diamond avoiding timing jitter from the single-photon detectors. Frequency upconversion of the fluorescence emitted from silicon vacancy centers was achieved from 738 nm to 436 nm via sum frequency generation with a short pump pulse. The excited-state lifetime can be obtained by measuring the intensity of upconverted light while the pump delay changes. As a probe, a pump laser with pulse duration of 11 ps provided a high temporal resolution of the measurement. The lifetime extracted from the pump–probe curve was 0.755 ns, which was comparable to the timing jitter of the single-photon detectors.

  2. Fluorescence quenching of newly synthesized biologically active coumarin derivative by aniline in binary solvent mixtures

    International Nuclear Information System (INIS)

    Evale, Basavaraj G.; Hanagodimath, S.M.

    2009-01-01

    The fluorescence quenching of newly synthesized coumarin (chromen-2-one) derivative, 4-(5-methyl-3-furan-2-yl-benzofuran-2-yl)-7-methyl-chromen-2-one (MFBMC) by aniline in different solvent mixtures of benzene and acetonitrile was determined at room temperature (296 K) by steady-state fluorescence measurements. The quenching is found to be appreciable and positive deviation from linearity was observed in the Stern-Volmer (S-V) plots in all the solvent mixtures. This could be explained by static and dynamic quenching models. The positive deviation in the S-V plot is interpreted in terms of ground-state complex formation model and sphere of action static quenching model. Various rate parameters for the fluorescence quenching process have been determined by using the modified Stern-Volmer equation. The sphere of action static quenching model agrees very well with experimental results. The dependence of Stern-Volmer constant K SV , on dielectric constant ε of the solvent mixture suggests that the fluorescence quenching is diffusion-limited. Further with the use of finite sink approximation model, it is concluded that these bimolecular quenching reactions are diffusion-limited. Using lifetime (τ o ) data, the distance parameter R' and mutual diffusion coefficient D are estimated independently.

  3. Preparation of Plasmonic Platforms of Silver Wires on Gold Mirrors and Their Application to Surface Enhanced Fluorescence

    Science.gov (United States)

    2015-01-01

    In this report we describe a preparation of silver wires (SWs) on gold mirrors and its application to surface enhanced fluorescence (SEF) using a new methodology. Silica protected gold mirrors were drop-coated with a solution of silver triangular nanoprisms. The triangular nanoprisms were slowly air-dried to get silver wires that self-assembled on the gold mirrors. Fluorescence enhancement was studied using methyl azadioxatriangulenium chloride (Me-ADOTA·Cl) dye in PVA spin-coated on a clean glass coverslip. New Plasmonic Platforms (PPs) were assembled by placing a mirror with SWs in contact with a glass coverslip spin-coated with a uniform Me-ADOTA·Cl film. It was shown that surface enhanced fluorescence is a real phenomenon, not just an enhancement of the fluorescence signal due to an accumulation of the fluorophore on rough nanostructure surfaces. The average fluorescence enhancement was found to be about 15-fold. The lifetime of Me-ADOTA·Cl dye was significantly reduced (∼4 times) in the presence of SWs. Moreover, fluorescence enhancement and lifetime did not show any dependence on the excitation light polarization. PMID:25296293

  4. Lifetimes of charm and beauty hadrons

    International Nuclear Information System (INIS)

    Bellini, G.; Dornan, P.J.

    1997-01-01

    Major breakthroughs have been achieved in the determination of the lifetimes of charm and beauty hadrons. Much larger data samples than previously have become available and new experimental devices and techniques have been developed and employed. The lifetimes of all weakly decaying singly charmed hadrons have been measured, some with an accuracy of a few percent. The difference in the shortest lifetime - τ(Ω c ) - and the longest one - τ(D + ) - is given by a factor of close to ten. The experimental status of beauty lifetimes, while less complete, has still reached a new level of quality and is now better than 5% for the commoner states. New theoretical tools, based mainly on heavy quark expansions, have been developed; they incorporate as well as transcend earlier phenomenological descriptions. The observed pattern in the charm lifetime ratios is reproduced in a semi-quantitative manner as well as could be expected; as far as the beauty lifetime ratios are concerned some problems may well be emerging. The maturity level achieved in the measurements bodes quite well for future challenges where reliable and efficient tracking of the decay vertices will be crucial. (orig.)

  5. 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.

  6. 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.

  7. Determination of copper in geological materials by X-ray fluorescence

    International Nuclear Information System (INIS)

    Roca, M.; Bayon, A.

    1981-01-01

    X-ray fluorescence has been applied to the determination of copper content of geological materials in the concentration range of 0.01 to % CuO. A molybdenum target tube Is used, samples being presented in finely-ground powder form. Various methods for the correction for background and Instrumental copper interferences have been considered. To correct for matrix effects different tube scattered primary radiations have been tested as references or internal standards. MoK(41 - (C) provides the most suitable results. The use of influence empirical coefficients for the effect of iron on copper and of mass absorption coefficients has also been considered. For samples with a high content of lead, several procedures to correct for I t s influence have been investigated. Comparison between data obtained by X-ray fluorescence and wet-chemical techniques indicated good agreement. (Author) 6 refs

  8. Rapid fluorometric determination of perfluorooctanoic acid by its quenching effect on the fluorescence of quantum dots

    International Nuclear Information System (INIS)

    Liu, Qi; Huang, Aizhen; Wang, Nan; Zheng, Guan; Zhu, Lihua

    2015-01-01

    Analysis of perfluorooctanoic acid (PFOA) usually requires a combination of high-performance liquid chromatography and mass spectrometry, which is expensive and time-consuming. In the present work, water-soluble CdS quantum dots (QDs) were employed to develop a simple and rapid fluorometric method for the determination of PFOA. Strongly fluorescent CdS QDs were prepared by using 3-mercaptopropionic acid (MPA) as a stabilizer. It was observed that PFOA strongly quenched the fluorescence emission of the MPA-CdS QDs because PFOA promotes the aggregation of MPA-CdS QDs through a fluorine–fluorine affinity interaction. Under optimum conditions, the fluorescence intensity of MPA-CdS QDs was observed to decrease linearly with an increase in the concentration of PFOA from 0.5 to 40 μmol L −1 , with a limit of detection of 0.3 μmol L −1 . This new method was successfully implemented for the analysis of PFOA-spiked textile samples, with recoveries ranging from 95% to 113%. - Highlights: • PFOA significantly quenched the fluorescence emission of quantum dots (QDs). • A rapid and simple fluorescence sensor was proposed for determining PFOA by QDs. • PFOA determination could be completed within approximately 10 min. • The developed method had a working range of 0.5 to 40 μmol L −1 and a detection limit of 0.3 μmol L −1

  9. In vivo time-gated fluorescence imaging with biodegradable luminescent porous silicon nanoparticles.

    Science.gov (United States)

    Gu, Luo; Hall, David J; Qin, Zhengtao; Anglin, Emily; Joo, Jinmyoung; Mooney, David J; Howell, Stephen B; Sailor, Michael J

    2013-01-01

    Fluorescence imaging is one of the most versatile and widely used visualization methods in biomedical research. However, tissue autofluorescence is a major obstacle confounding interpretation of in vivo fluorescence images. The unusually long emission lifetime (5-13 μs) of photoluminescent porous silicon nanoparticles can allow the time-gated imaging of tissues in vivo, completely eliminating shorter-lived (50-fold in vitro and by >20-fold in vivo when imaging porous silicon nanoparticles. Time-gated imaging of porous silicon nanoparticles accumulated in a human ovarian cancer xenograft following intravenous injection is demonstrated in a live mouse. The potential for multiplexing of images in the time domain by using separate porous silicon nanoparticles engineered with different excited state lifetimes is discussed.

  10. The photoluminescence of a fluorescent lamp: didactic experiments on the exponential decay

    Science.gov (United States)

    Onorato, Pasquale; Gratton, Luigi; Malgieri, Massimiliano; Oss, Stefano

    2017-01-01

    The lifetimes of the photoluminescent compounds contained in the coating of fluorescent compact lamps are usually measured using specialised instruments, including pulsed lasers and/or spectrofluorometers. Here we discuss how some low cost apparatuses, based on the use of either sensors for the educational lab or commercial digital photo cameras, can be employed to the same aim. The experiments do not require that luminescent phosphors are hazardously extracted from the compact fluorescent lamp, that also contains mercury. We obtain lifetime measurements for specific fluorescent elements of the bulb coating, in good agreement with the known values. We also address the physical mechanisms on which fluorescence lamps are based in a simplified way, suitable for undergraduate students; and we discuss in detail the physics of the lamp switch-off by analysing the time dependent spectrum, measured through a commercial fiber-optic spectrometer. Since the experiment is not hazardous in any way, requires a simple setup up with instruments which are commonly found in educational labs, and focuses on the typical features of the exponential decay, it is suitable for being performed in the undergraduate laboratory.

  11. Computer-assisted sperm morphometry fluorescence-based analysis has potential to determine progeny sex.

    Science.gov (United States)

    Santolaria, Pilar; Pauciullo, Alfredo; Silvestre, Miguel A; Vicente-Fiel, Sandra; Villanova, Leyre; Pinton, Alain; Viruel, Juan; Sales, Ester; Yániz, Jesús L

    2016-01-01

    This study was designed to determine the ability of computer-assisted sperm morphometry analysis (CASA-Morph) with fluorescence to discriminate between spermatozoa carrying different sex chromosomes from the nuclear morphometrics generated and different statistical procedures in the bovine species. The study was divided into two experiments. The first was to study the morphometric differences between X- and Y-chromosome-bearing spermatozoa (SX and SY, respectively). Spermatozoa from eight bulls were processed to assess simultaneously the sex chromosome by FISH and sperm morphometry by fluorescence-based CASA-Morph. SX cells were larger than SY cells on average (P potential to find differences between X- and Y-chromosome-bearing spermatozoa in bovine species although more studies are needed to increase the precision of sex determination by this technique.

  12. Lifetime of {sup 44}Ti as probe for supernova models

    Energy Technology Data Exchange (ETDEWEB)

    Goerres, J; Meissner, J; Schatz, H; Stech, E; Tischhauser, P; Wiescher, M [Univ. of Notre Dame, Notre Dame, IN (United States); Bazin, D; Harkewicz, R; Hellstroem, M; Sherrill, B; Steiner, M [Michigan State Univ., East Lansing, MI (United States); Boyd, R N [Ohio State Univ., Columbus, OH (United States); Buchmann, L [TRIUMF, Vancouver, BC (Canada); Hartmann, D H [Clemson Univ., Clemson, SC (United States); Hinnefeld, J D [Indiana Univ. South Bend, South Bend, IN (United States)

    1998-06-01

    The recent observation of {sup 44}Ti radioactivity in the supernova remnant Cassiopeia A with the Compton Gamma Ray Observatory allows the determination of the absolute amount of {sup 44}Ti. This provides a test for current supernova models. The main uncertainty is the lifetime of {sup 44}Ti. We report a new measurement of the lifetime of {sup 44}Ti applying a novel technique. A mixed radioactive beam containing {sup 44}Ti as well as {sup 22}Na was implanted and the resulting {gamma}-activity was measured. This allowed the determination of the lifetime of {sup 44}Ti relative to the lifetime of {sup 22}Na, {tau} = (87.0 {+-} 1.9) y. With this lifetime, the {sup 44}Ti abundance agrees with theoretical predictions within the remaining observational uncertainties. (orig.)

  13. Catalyst deterioration over the lifetime of small utility engines.

    Science.gov (United States)

    Doll, Nicholas J; Reisel, John R

    2007-10-01

    In this paper, the deterioration of catalysts in small, four-stroke, spark-ignition engines is described. The laboratory testing performed followed a proven test method that mimics the lifetime of a small air-cooled utility engine operating under normal field conditions. The engines used were single-cylinder, 6.5-hp, side-valve engines. These engines have a nominal 125-hr lifetime. The effectiveness of the catalysts was determined by testing exhaust emissions before and after the catalyst to determine the catalyst's efficiency. This was done several times during the lifetime of the engines to determine the deterioration in the performance of the catalysts at lowering pollutant emissions. Additional testing was performed on the catalysts to determine wear patterns, contamination, and recoverable activity. The results indicate that considerable catalyst deterioration is occurring over the lifetime of the engine. The results reveal that soot buildup, poisons, and active surface loss appear to be the contributing factors to the deterioration. These results were determined after analyzing the exhaust emissions data, scanning electron microscope results analysis, and the impact of regeneration attempts. An ANOVA statistical analysis was performed, and it was determined that the emissions are also impacted, to some degree, by time and the engine itself.

  14. Multiwavelength FLIM: new concept for fluorescence diagnosis

    Science.gov (United States)

    Rück, Angelika; Lorenz, S.; Hauser, Carmen; Mosch, S.; Kalinina, S.

    2012-03-01

    Fluorescence guided tumor resection is very well accepted in the case of bladder cancer and brain tumor, respectively. However, false positive results are one of the major problems, which will make the discrimination between tumor tissue and inflammation difficult. In contrast fluorescence lifetime imaging (FLIM) and especially spectral resolved FLIM (SLIM) can significantly improve the analysis. The fluorescence decay of a fluorophore in many cases does not show a simple monoexponential profile. A very complex situation arises, when more than one compound has to be analyzed. This could be the case when endogenous fluorophores of living cells and tissues have to be discriminated to identify oxidative metabolic changes. Other examples are PDT, when different photosensitizer metabolites are observed simultaneously. In those cases a considerable improvement could be achieved when time-resolved and spectral-resolved techniques are simultaneously incorporated. Within this presentation the principles of spectral and time-resolved fluorescence imaging will be discussed. Successful applications as autofluorescence and 5-ALA induced porphyrin fluorescence will be described in more detail.

  15. Study on fluorescence properties of carbogenic nanoparticles and their application for the determination of ferrous succinate

    Energy Technology Data Exchange (ETDEWEB)

    Sun Wen [Department of Analytical Chemistry, China Pharmaceutical University, Nanjing 210009 (China); Du Yingxiang, E-mail: du_yingxiang@126.co [Department of Analytical Chemistry, China Pharmaceutical University, Nanjing 210009 (China) and Key Laboratory of Drug Quality Control and Pharmacovigilance, Ministry of Education, China Pharmaceutical University, Nanjing 210009 (China) and Key Laboratory of Modern Chinese Medicines, Ministry of Education, China Pharmaceutical University, Nanjing 210009 (China); Wang Yunqing [Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai 264003 (China)

    2010-08-15

    A new type of fluorescent nanomaterial named carbogenic nanoparticles (NPs) has drawn considerable attention recently. In this study, we adopted a direct and simple synthetic method to produce the carbogenic NPs and investigated the fluorescence properties of the as-prepared carbogenic NPs in detail. It was found that the fluorescence of carbogenic NPs was stable with the variance of environmental conditions such as pH, temperature and UV irradiation. More interestingly, we found carbogenic NPs exhibited high selectivity and sensitivity towards ferric ions. Under optimum conditions, a good linear relationship could be obtained between the fluorescence intensity and concentration of ferric ions in the range of 5.0x10{sup -5}-5.0x10{sup -4} mol L{sup -1}, and the limit of detection is 11.2 {mu}mol L{sup -1}. Based on the fluorescence quenching of carbogenic NPs, a rapid and specific quantitative method was proposed for the determination of ferrous succinate. The content of ferrous succinate in commercial tablets determined by the present method was agreed with the spectrophotometric method results and the reproducibility and the recovery of the proposed method were satisfactory.

  16. Determination of the activity of telomerase in cancer cells by using BSA-protected gold nanoclusters as a fluorescent probe.

    Science.gov (United States)

    Xu, Yujuan; Zhang, Peng; Wang, Zhen; Lv, Shaoping; Ding, Caifeng

    2018-02-27

    Gold nanoclusters (AuNCs) protected with a bovine serum albumin (BSA) coating are known to emit red fluorescence (peaking at 650 nm) on photoexcitation with ultraviolet light (365 nm). On addition of Cu(II) ions, fluorescence is quenched because Cu(II) complexes certain amino acid units in the BSA chain. Fluorescence is, however, restored if pyrophosphate (PPi) is added because it will chelate Cu(II) and remove it from the BSA coating on the AuNCs. Because PPi is involved in the function of telomerase, the BSA@AuNCs loaded with Cu(II) can act as a fluorescent probe for determination of the activity of telomerase. A fluorescent assay was worked out for telomerase that is highly sensitive and has a wide linear range (10 nU to 10 fM per mL). The fluorescent probe was applied to the determination of telomerase activity in cervix carcinoma cells via imaging. It is shown that tumor cells can be well distinguished from normal cells by monitoring the differences in intracellular telomerase activity. Graphical abstract Gold nanoclusters (AuNCs) protected by bovine serum albumin (BSA) and displaying red photoluminescence were prepared as fluorescent probe for the determination of telomerase activity and used for imaging of cervix carcinoma (HeLa) cells.

  17. Determination of Fe and Zn in healing plants by radionuclide X-ray fluorescence analysis

    International Nuclear Information System (INIS)

    Harangozo, M.; Toelgyessy, J.; Tomecek, O.; Ruzicka, I.; Cejpek, K.

    1999-01-01

    Radionuclide X-ray fluorescence method was used for the determination of Fe and Zn in healing plants (Sage, Peppermint, Stinging, Common Agrimony, Milfoil, Ribwort, Tansy, White Dead-Nettle). 238 Pu exciting source and Si/Li semiconductor detector were used for the determination. (author)

  18. Proximal Sensing of Plant-Pathogen Interactions in Spring Barley with Three Fluorescence Techniques

    Directory of Open Access Journals (Sweden)

    Georg Leufen

    2014-06-01

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

  19. Lifetime measurements of hadrons containing heavy quarks

    International Nuclear Information System (INIS)

    Forden, G.E.

    1985-01-01

    Recent lifetime measurements of heavy particles at PETRA and PEP are reviewed. A comparison of the methods used is given. The world averages for the lifetimes of the D 0 and D +- mesons are found to be (tau/dub D/ 0 ) - 3.97 +/- 0.3 x 10 -13 sec and (tau/dub D +-/) = 8.6 +/- 0.7 x 10 -13 sec. This difference in lifetimes is discussed in light of recent information about exclusive decays. The world average for the lifetime of bottom hadrons is determined to be (tau/sub b/) = 11.0 +/- 1.5 x 10 -13 sec and new estimates for the b quark mixing elements, absolute value V/sub bu/ and absolute value V/sub bc/, are given

  20. [Sensitive Determination of Chondroitin Sulfate by Fluorescence Recovery of an Anionic Aluminum Phthalocyanine-Cationic Surfactant Ion-Association Complex Used as a Fluorescent Probe Emitting at Red Region].

    Science.gov (United States)

    Chen, Lin; Huang, Ping; Yang, Hui-qing; Deng, Ya-bin; Guo, Meng-lin; Li, Dong-hui

    2015-08-01

    Determination of chondroitin sulfate in the biomedical field has an important value. The conventional methods for the assay of chondroitin sulfate are still unsatisfactory in sensitivity, selectivity or simplicity. This work aimed at developing a novel method for sensitive and selective determination of chondroitin sulfate by fluorimetry. We found that some kinds of cationic surfactants have the ability to quench the fluorescence of tetrasulfonated aluminum phthalocyanine (AlS4Pc), a strongly fluorescent compound which emits at red region, with high efficiency. But, the fluorescence of the above-mentioned fluorescence quenching system recovered significantly when chondroitin sulfate (CS) exits. Tetradecyl dimethyl benzyl ammonium chloride(TDBAC) which was screened from all of the candidates of cationic surfactants was chosen as the quencher because it shows the most efficient quenching effect. It was found that the fluorescence of AlS4Pc was extremely quenched by TDBAC because of the formation of association complex between AlS4Pc and TDBAC. Fluorescence of the association complex recovered dramatically after the addition of chondroitin sulfate (CS) due to the ability of chondroitin sulfate to shift the association equilibrium of the association, leading to the release of AlS4Pc, thus resulting in an increase in the fluorescence of the reaction system. Based on this phenomenon, a novel method with simplicity, accuracy and sensitivity was developed for quantitative determination of CS. Factors including the reaction time, influencing factors and the effect of coexisting substances were investigated and discussed. Under optimum conditions the linear range of the calibration curve was 0.20~10.0 μg · mL(-1). The detection limit for CS was 0.070 μg · mL(-1). The method has been applied to the analysis of practical samples with satisfied results. This work expands the applications of AlS4Pc in biomedical area.

  1. Real-time monitoring of luminescent lifetime changes of PtOEP oxygen sensing film with LED/photodiode-based time-domain lifetime device.

    Science.gov (United States)

    Ji, Shaomin; Wu, Wanhua; Wu, Yubo; Zhao, Taiyang; Zhou, Fuke; Yang, Yubin; Zhang, Xin; Liang, Xiaofen; Wu, Wenting; Chi, Lina; Wang, Zhonggang; Zhao, Jianzhang

    2009-05-01

    A cost-effective LED/photodiode(PD)-based time-domain luminescent lifetime measuring device with rugged electronics and simplified algorithms was assembled and successfully used to characterize oxygen sensing films, by continuously monitoring phosphorescence lifetime changes of phosphorescent platinum octaethylporphyrin (PtOEP) in cardo poly(aryl ether ketone) polymer (IMPEK-C) vs. variation of the oxygen partial pressure in a gas mixture (O(2)/N(2)). The results determined by both phosphorescence lifetime and intensity monitoring were compared and the lifetime mode gave results which are in good agreement with the intensity mode. The lifetime-based linear Stern-Volmer plot indicates that the PtOEP molecules are nearly homogeneously distributed in the sensing film. The phosphorescent lifetime of the PtOEP film changes from 75 micros in neat N(2) to less than 2 micros in neat O(2). The sensing system (by combination of the PtOEP sensing film with the home-assembled lifetime device) gives a high lifetime-based O(2) sensing resolution, e.g. about 2 micros Torr(-1) for low O(2) concentration (below 3.5% O(2), V/V). This feasible lifetime device configuration is affordable to most sensor laboratories and the device may facilitate the study of O(2) sensing material with the continuous lifetime monitoring method.

  2. Determination of the dispersion coefficient, in rivers through radioactive and fluorescent tracers

    International Nuclear Information System (INIS)

    Almeida Rosso, T.C. de.

    1986-03-01

    The purpose of this work was to determine experimentally the dispersion coefficient in natural streams through the aplication of radioactive and fluorescent tracers. The technique used to obtain the experimental data was the simmultaneous injection of the radioactive tracer Bromim - 82 and the fluorescent ones, Amidorodamine G. extra and Uranine. A comparison among these different tracers is shown as well as the performance of Uranine in a poluented stream in the presence of suspended sediments. The site chosen for the experiments was the Piabanha River, between Pedro do Rio and Areal, located at Rio de Janeiro state, in a nearly 20 Km strech. (Author) [pt

  3. Rapid extraction and x-ray fluorescence determination of gold in rock

    International Nuclear Information System (INIS)

    Lobanov, F.I.; Logunova, S.A.; Popov, A.M.; Krasnopevtseva, E.V.

    1994-01-01

    The optimal conditions for gold extractional recovery by melts of aliphatic monocarboxylic acids mixtures (C ≥ 16) with additions of di-2-ethylhexyldithiophosphoric acid, di-n-oxtylsulfide and alkylaniline were determined. A rapid method of extraction and X-ray fluorescence detection of gold in the presence of considerable amounts of Fe, Cu, Pb, Cd, Zn and Bi was developed

  4. Rapid fluorometric determination of perfluorooctanoic acid by its quenching effect on the fluorescence of quantum dots

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Qi; Huang, Aizhen; Wang, Nan, E-mail: nwang@hust.edu.cn; Zheng, Guan; Zhu, Lihua

    2015-05-15

    Analysis of perfluorooctanoic acid (PFOA) usually requires a combination of high-performance liquid chromatography and mass spectrometry, which is expensive and time-consuming. In the present work, water-soluble CdS quantum dots (QDs) were employed to develop a simple and rapid fluorometric method for the determination of PFOA. Strongly fluorescent CdS QDs were prepared by using 3-mercaptopropionic acid (MPA) as a stabilizer. It was observed that PFOA strongly quenched the fluorescence emission of the MPA-CdS QDs because PFOA promotes the aggregation of MPA-CdS QDs through a fluorine–fluorine affinity interaction. Under optimum conditions, the fluorescence intensity of MPA-CdS QDs was observed to decrease linearly with an increase in the concentration of PFOA from 0.5 to 40 μmol L{sup −1}, with a limit of detection of 0.3 μmol L{sup −1}. This new method was successfully implemented for the analysis of PFOA-spiked textile samples, with recoveries ranging from 95% to 113%. - Highlights: • PFOA significantly quenched the fluorescence emission of quantum dots (QDs). • A rapid and simple fluorescence sensor was proposed for determining PFOA by QDs. • PFOA determination could be completed within approximately 10 min. • The developed method had a working range of 0.5 to 40 μmol L{sup −1} and a detection limit of 0.3 μmol L{sup −1}.

  5. High sensitive determination of zinc with novel water-soluble small molecular fluorescent sensor

    International Nuclear Information System (INIS)

    Weng Ying; Chen Zilin; Wang Fang; Xue Lin; Jiang Hua

    2009-01-01

    A high sensitive method of quantitative analysis for the determination of zinc in the nutrition supplements has been developed by using a novel water-soluble fluorescent sensor HQ3: (8-pyridylmethyloxy-2-methyl-quinoline). Under the optimized condition of 67 mM phosphate buffer, pH 7.4, and 5% (v/v) DMSO, the zinc concentration showed good linear relationship with fluorescence intensity in the range of 7.5 x 10 -8 to 2.5 x 10 -5 M with the detection limit of 1.5 x 10 -8 M. HQ3 exhibited high selectivity to zinc comparing with other metal ions except for cadmium. The developed analytical method was successfully used for determining the content of zinc in a real sample of zinc gluconate solution of Sanchine.

  6. Steady state and time resolved fluorescence studies of azadioxatriangulenium (ADOTA) fluorophore in silica and PVA thin films

    DEFF Research Database (Denmark)

    Chib, Rahul; Raut, Sangram; Shah, Sunil

    2015-01-01

    A cationic azadioxatriangulenium dye was entrapped in silica thin films obtained by the sol-gel process and in poly (vinyl) alcohol (PVA) thin films. Azadioxatriangulenium is a red emitting fluorophore with a long fluorescence lifetime of ∼20 ns. The fluorescent properties of azadioxatriangulenium...

  7. FastFLIM, the all-in-one engine for measuring photoluminescence lifetime of 100 picoseconds to 100 milliseconds

    Science.gov (United States)

    Sun, Yuansheng; Coskun, Ulas; Liao, Shih-Chu Jeff; Barbieri, Beniamino

    2018-02-01

    Photoluminescence (PL) refers to light emission initiated by any form of photon excitation. PL spectroscopy and microscopy imaging has been widely applied in material, chemical and life sciences. Measuring its lifetime yields a new dimension of the PL imaging and opens new opportunities for many PL applications. In solar cell research, quantification of the PL lifetime has become an important evaluation for the characteristics of the Perovskite thin film. Depending upon the PL process (fluorescence, phosphorescence, photon upconversion, etc.), the PL lifetimes to be measured can vary in a wide timescale range (e.g. from sub-nanoseconds to microseconds or even milliseconds) - it is challenging to cover this wide range of lifetime measurements by a single technique efficiently. Here, we present a novel digital frequency domain (DFD) technique named FastFLIM, capable of measuring the PL lifetime from 100 ps to 100 ms at the high data collection efficiency (up to 140-million counts per second). Other than the traditional nonlinear leastsquare fitting analysis, the raw data acquired by FastFLIM can be directly processed by the model-free phasor plots approach for instant and unbiased lifetime results, providing the ideal routine for the PL lifetime microscopy imaging.

  8. Disruption of the hydrogen bonding network determines the pH-induced non-fluorescent state of the fluorescent protein ZsYellow by protonation of Glu221.

    Science.gov (United States)

    Bae, Ji-Eun; Kim, In Jung; Nam, Ki Hyun

    2017-11-04

    Many fluorescent proteins (FPs) exhibit fluorescence quenching at a low pH. This pH-induced non-fluorescent state of an FP serves as a useful indicator of the cellular pH. ZsYellow is widely used as an optical marker in molecular biology, but its pH-induced non-fluorescent state has not been characterized. Here, we report the pH-dependent spectral properties of ZsYellow, which exhibited the pH-induced non-fluorescence state at a pH below 4.0. We determined the crystal structures of ZsYellow at pH 3.5 (non-fluorescence state) and 8.0 (fluorescence state), which revealed the cis-configuration of the chromophore without pH-induced isomerization. In the non-fluorescence state, Arg95, which is involved in stabilization of the exited state of the chromophore, was found to more loosely interact with the carbonyl oxygen atom of the chromophore when compared to the interaction at pH 8.0. In the fluorescence state, Glu221, which is involved in the hydrogen bonding network around the chromophore, stably interacted with Gln42 and His202. By contrast, in the non-fluorescence state, the protonated conserved Glu221 residue exhibited a large conformational change and was separated from His202 by 5.46 Å, resulting in breakdown of the hydrogen bond network. Our results provide insight into the critical role of the conserved Glu221 residue for generating the pH-induced non-fluorescent state. Copyright © 2017 Elsevier Inc. All rights reserved.

  9. Synthesis and characterization of titania-based monodisperse fluorescent europium nanoparticles for biolabeling

    International Nuclear Information System (INIS)

    Tan Mingqian; Wang Guilan; Ye Zhiqiang; Yuan Jingli

    2006-01-01

    Inorganic-organic hybrid titania-based nanoparticles covalently bound to a fluorescent Eu 3+ chelate of 4,4'-bis(1'',1'',1'',2'',2'',3'',3''-heptafluoro-4'',6''-hexanedion-6''-yl) chlorosulfo-o-terphenyl (BHHCT-Eu 3+ ) were synthesized by a sol-gel technique. A conjugate of BHHCT with 3-[2-(2-aminoethylamino) ethylamino]propyl-trimethoxysilane (APTS) was used as a precursor for the nanoparticle preparation and monodisperse nanoparticles consisting of titania network and silica sub-network covalently bound to the Eu 3+ chelate were prepared by the copolymerization of APTS-BHHCT conjugate, titanium tetraisopropoxide (TTIP) and free APTS in EuCl 3 water-alcohol solution. The effects of reaction conditions on size and fluorescence lifetime of the nanoparticles were investigated. The characterizations by transmission electron microscopy and fluorometric methods indicate that the nanoparticles are near spherical and strongly fluorescent having a fluorescence quantum yield of 11.6% and a long fluorescence lifetime of ∼0.4 ms. The direct-introduced amino groups on the nanoparticle's surface by using free APTS in nanoparticle preparation facilitated the biolabeling process of the nanoparticles. The nanoparticle-labeled streptavidin (SA) was prepared and used in a sandwich-type time-resolved fluoroimmunoassay (TR-FIA) of human prostate-specific antigen (PSA) by using a 96-well microtiter plate as the solid phase carrier. The method gives a detection limit of 66 pg/ml for the PSA assay

  10. Fluorescence spectral studies of Gum Arabic: Multi-emission of Gum Arabic in aqueous solution

    Energy Technology Data Exchange (ETDEWEB)

    Dhenadhayalan, Namasivayam, E-mail: ndhena@gmail.com [Department of Chemistry, National Taiwan University, Taipei, Taiwan (China); Mythily, Rajan, E-mail: rajanmythily@gmail.com [Department of Chemistry, Dwaraka Doss Goverdhan Doss Vaishnav College (Autonomous), 833, Gokul Bagh, E.V.R. Periyar Road, Arumbakkam, Chennai 600 106 (India); Kumaran, Rajendran, E-mail: kumaranwau@rediffmail.com [Department of Chemistry, Dwaraka Doss Goverdhan Doss Vaishnav College (Autonomous), 833, Gokul Bagh, E.V.R. Periyar Road, Arumbakkam, Chennai 600 106 (India)

    2014-11-15

    Gum Arabic (GA), a food hydrocolloid is a natural composite obtained from the stems and branches of Acacia Senegal and Acacia Seyal trees. GA structure is made up of highly branched arabinogalactan polysaccharides. Steady-state absorption, fluorescence, and time-resolved fluorescence spectral studies of acid hydrolyzed GA solutions were carried out at various pH conditions. The fluorescence in GA is predominantly attributed to the presence of tyrosine and phenylalanine amino acids. The presence of multi-emissive peaks at different pH condition is attributed to the exposure of the fluorescing amino acids to the aqueous phase, which contains several sugar units, hydrophilic and hydrophobic moieties. Time-resolved fluorescence studies of GA exhibits a multi-exponential decay with different fluorescence lifetime of varying amplitude which confirms that tyrosine is confined to a heterogeneous microenvironment. The existence of multi-emissive peaks with large variation in the fluorescence intensities were established by 3D emission contour spectral studies. The probable location of the fluorophore in a heterogeneous environment was further ascertained by constructing a time-resolved emission spectrum (TRES) and time-resolved area normalized emission spectrum (TRANES) plots. Fluorescence spectral technique is used as an analytical tool in understanding the photophysical properties of a water soluble complex food hydrocolloid containing an intrinsic fluorophore located in a multiple environment is illustrated. - Highlights: • The Manuscript deals with the steady state absorption, emission, fluorescence lifetime and time-resolved emission spectrum studies of Gum Arabic in aqueous medium at various pH conditions. • The fluorescence emanates from the tyrosine amino acid present in GA. • Change in pH results in marked variation in the fluorescence spectral properties of tyrosine. • Fluorescence spectral techniques are employed as a tool in establishing the

  11. Statistical analysis of lifetime determinations in the presence of large errors

    International Nuclear Information System (INIS)

    Yost, G.P.

    1984-01-01

    The lifetimes of the new particles are very short, and most of the experiments which measure decay times are subject to measurement errors which are not negligible compared with the decay times themselves. Bartlett has analyzed the problem of lifetime estimation if the error on each event is small or zero. For the case of non-negligible measurement errors, σsub(i), on each event, we are interested in a few basic questions: How well does maximum likelihood work. That is, (a) are the errors reasonable, (b) is the answer unbiased, and (c) are there other estimators with superior performance. We concentrate on the results of our Monte Carlo investigation for the case in which the experiment is sensitive over all times -infinity< xsub(i)< infinity

  12. Determination of rare-earth elements in rocks by isotope-excited X-ray fluorescence spectrometry

    DEFF Research Database (Denmark)

    Kunzendorf, Helmar; Wollenberg, H.A.

    1970-01-01

    Isotope-excited X-ray fluorescence spectrometry furnishes a rapid determination of rare-earth elements in unprepared rock samples. The samples are excited by 241Am γ-rays, generating X-ray spectra on a multichannel pulse-height analyser. Gaussian peaks of the Kα and Kβ X-ray energies are treated ......-ray spectrometric scan of a longitudinally sliced drill core showed a close correlation between rare-earth abundances and appropriate minerals.......Isotope-excited X-ray fluorescence spectrometry furnishes a rapid determination of rare-earth elements in unprepared rock samples. The samples are excited by 241Am γ-rays, generating X-ray spectra on a multichannel pulse-height analyser. Gaussian peaks of the Kα and Kβ X-ray energies are treated...

  13. Determination of As in tobacco by using electrochemical hydride generation at a Nafion® solid polymer electrolyte cell hyphenated with atomic fluorescence spectrometry

    Science.gov (United States)

    Yang, Qinghua; Gan, Wuer; Deng, Yun; Sun, Huihui

    2011-11-01

    In the present work, a novel solid polymer electrolyte hydride generation (SPE-HG) cell was developed. The home-made SPE-HG cell, mainly composed of three components (Nafion®117 membrane for separating and H + exchanging, a soft graphite felt cathode and a Ti mesh modified by Ir anode), was employed for detecting As by coupling to atomic fluorescence spectrometry (AFS). The H + generated by electrolysis of pure water in anode chamber transferred to cathode chamber through SPE, and immediately reacted with As 3 + to generate AsH 3. The relative mechanisms and operation conditions for hydride generation of As were investigated in detail. The developed cell employed water as an alternative of acid anolyte, with virtues of low-cost, more than 6 months lifetime and environment friendly compared with the conventional cell. Under the optimized conditions, the limit of determination of As 3 + for sample blank solution was 0.12 μg L - 1 , the RSD was 2.9% for 10 consecutive measurements of 5 μg L - 1 As 3 + standard solution. The accuracy of the method was verified by the determination of As in the reference Tea (GBW07605) and the developed method was successfully applied to determine trace amounts of As in tobacco samples with recovery from 97% to 103%.

  14. Porous glasses as a matrix for incorporation of photonic materials. Pore determination by positron annihilation lifetime spectroscopy

    Science.gov (United States)

    Reisfeld, Pore determination by positron annihilation lifetime spectroscopy R.; Saraidarov, T.; Jasinska, B.

    2004-07-01

    Porous glasses prepared by the sol-gel technique have a variety of applications when incorporated by photonic materials: tunable lasers, sensors, luminescence solar concentrators, semiconductor quantum dots, biological markers. The known methods of pore size determinations, the nitrogen adsorption and mercury porosimetry allow to determine the sizes of open pores. Positron annihilation lifetime spectroscopy (PALS) allows to determine pore sizes also of closed pores. As an example we have performed measurements of non-doped zirconia-silica-polyurethane (ZSUR) ormocer glasses and the same glasses doped with lead sulfide quantum dots. The pore radii range between 0.25-0.38 nm, total surface area 15.5-23.8 m 2/g.

  15. Study of lifetimes of fluorescence levels of tetravalent uranium in the incommensurate phase of thorium tetrabromide and tetrachloride

    International Nuclear Information System (INIS)

    Milicic, A.

    1989-01-01

    The lifetimes of radiative levels of tetravalent uranium in the incommensurate phase of thorium tetrahalides have been measured as a function of different parameters: site symmetry, temperature and concentration. The incommensurate phase of thorium tetrabromide and tetrachloride is characterized by a continuous distribution of site symmetries induced by a continuous and weak displacement of the halides around the thorium (uranium) ions. At low temperature, 4.2 K, the lifetime variation as a function of excited classes of symmetry is governed by the radiative process probability as well as the energy transfer between uranium ions in different sites. At higher temperature, a model based on a Boltzmann equilibrium between closed energy levels is able to reproduce the experimental lifetime variation as a function of the temperature, for a given class of symmetry. For the variation of lifetime as a function of uranium ion concentrations, at high dilution and in the case of U 4+ : ThBr 4 , there is a competition between the energy transfer and thermal population of excited states [fr

  16. Lifetimes of partial charge transfer exciplexes of 9-cyanophenanthrene and 9-cyanoanthracene

    OpenAIRE

    Dolotova, Elena; Dogadkin, Denis; Soboleva, Irina; Kuzmin, Michael; Nicolet, Olivier; Vauthey, Eric

    2003-01-01

    The fluorescence decays of several exciplexes with partial charge transfer have been investigated in solvents of various polarity. The measured lifetimes are found to be in reasonable agreement with the activation enthalpy and entropy of exciplex decay obtained earlier from the temperature dependence of the exciplex emission quantum yields. For exciplexes with 9-cyanophenanthrene substantial contribution of the higher local excited state into the exciplex electronic structure is found and bor...

  17. Nuclear lifetimes

    International Nuclear Information System (INIS)

    Caraca, J.M.G.

    1976-01-01

    The importance of the results obtained in experiments of measurement of lifetimes for a detailed knowledge of nuclear structure is referred. Direct methods of measurement of nuclear lifetimes are described, namely, electronic methods, recoil-distance method, doppler shift atenuation method and blocking-method. A brief reference is made to indirect methods for measurement of life-times

  18. Determination of trace aluminum by fluorescence quenching method based on catalysis of potassium chlorate oxidizing alizarin red

    Science.gov (United States)

    Shao-Qin, Lin; Xuan, Lin; Shi-Rong, Hu; Li-Qing, Zeng; Yan, Wang; Li, Chen; Jia-Ming, Liu; Long-Di, Li

    2005-11-01

    A new method for the determination of trace aluminum has been proposed. It is based on the fact that alizarin red can emit strong and stable fluorescence at 80 °C for 30 min and Al 3+ can effectively catalyze potassium chlorate oxidizing alizarin red to form non-fluorescence complex which cause the fluorescence quenching. The linear dynamic range of this method is 0.040-4.00 ng l -1 with a detection limit of 5.3 pg l -1. The regression equation can be expressed as Δ If = 8.731 + 21.73 c (ng l -1), with the correlation coefficient r = 0.9992 ( n = 6). This sensitive, rapid and accurate method has been applied to the determination of trace aluminum(III) in human hair and tea samples successfully. What is more, the mechanism of catalyzing potassium chlorate oxidizing alizarin red by the fluorescence quenching method is also discussed.

  19. Improved b lifetime measurement from MAC

    International Nuclear Information System (INIS)

    Ford, W.T.

    1984-03-01

    Two recent publications, from the MAC and Mark II collaborations, have reported the somewhat surprising result that the lifetime of particles made up of b quarks is in the 1 to 2 picosecond range, or somewhat longer than the lifetimes of charm particles. Although the charm decays are favored transitions while those of b particles depend upon off-diagonal elements of the weak flavor mixing matrix, the smallness of the b decay rates in face of the large available phase space indicates that the off-diagonal elements are indeed very small. The possibility for complete determination of the mixing matrix was brought significantly nearer by the availability of the lifetime information; what is needed now is to reduce the uncertainty of the measurements, which was about 33% for both experiments. We describe here an extension of the b lifetime study with the MAC detector, incorporating some new data and improvements in the analysis. 12 references

  20. Fluorescent carbon dots and nanodiamonds for biological imaging: preparation, application, pharmacokinetics and toxicity.

    Science.gov (United States)

    Liu, Jia-Hui; Yang, Sheng-Tao; Chen, Xin-Xin; Wang, Haifang

    2012-10-01

    The rapid advancement of nanotechnology has brought us some new types of fluorescent probes, which are indispensable for bioimaging in life sciences. Because of their innate biocompatibility, good resistance against photobleaching, long fluorescence lifetime and wide fluorescence spectral region, fluorescent carbon quantum dots (C-Dots) and nanosized diamonds (nanodiamonds, NDs) are gradually evolving into promising reagents for bioimaging. In this review, we summarize the recent achievements in fluorescent C-Dots and NDs with emphases on their preparation, properties, imaging application, pharmacokinetics and toxicity. Perspectives on further investigations and opportunities to develop C-Dots and NDs into the safer and more sensitive imaging probes for both living cells and animal models are discussed.

  1. Determination of selenium via the fluorescence quenching effect of selenium on hemoglobin-catalyzed peroxidative reaction.

    Science.gov (United States)

    Chen, Ya-Hong; Zhang, Ya-Nan; Tian, Feng-Shou

    2015-05-01

    A new method for the determination of selenium based on its fluorescence quenching on the hemoglobin-catalyzed reaction of H2 O2 and l-tyrosine has been established. The effect of pH, foreign ions and the optimization of variables on the determination of selenium was examined. The calibration curve was found to be linear between the fluorescence quenching (F0 /F) and the concentration of selenium within the range of 0.16-4.00 µg/mL. The detection limit was 1.96 ng/mL and the relative standard deviation was 3.14%. This method can be used for the determination of selenium in Se-enriched garlic bulbs with satisfactory results. Copyright © 2014 John Wiley & Sons, Ltd.

  2. Determination of Europium by fluorescence using pyrrolidinium based task specific ionic liquid

    International Nuclear Information System (INIS)

    Kumar, Satendra; Gayan, Siuli Maji; Sankaran, K.

    2012-01-01

    Determination of lanthanides using fluorescence is a major challenge in aqueous medium due to their poor molar absorptivities and low quantum yield. To overcome these problems, ligand sensitized fluorescence has been widely used for trace level detection of lanthanides in solutions. Recently, ionic liquid a non aqueous medium has been used to observe the fluorescence of lanthanides. In this work we have used pyrrolidinium based ionic liquid for the study of europium fluorescence. The ionic liquid (bmpyr)(BA) was prepared using the metathesis reaction involving 1-butyl,1-methyl pyrrolidinium chloride (bmpyr)(CI). (bmpyr)(Cl) crystals and silver benzoate were added according to their mole equivalents in methanol and stirred for 4 hrs. The filtrate was concentrated using a rotary evaporator, dried for 10 h at room temperature, and then for 40 h at 105℃under reduced pressure. This results in a white crystalline hygroscopic solid which was characterized by FT-lR spectroscopy. The yield of the product was ∼ 95%. Butyl-1, methyl pyrrolidinium bis(trifluoromethanesulfonyl)amide (bmpyr)(TF 2 N) is used for dissolve and dilute (bmpyr)(BA) and fluorescence and life time of Eu 3+ were obtained. For the concentration of Eu 3+ used in this study (2x10 -7 M), the optimum concentration of the sensitizing ionic liquid was found to be 10 -4 M. An enhancement factor of about 32000 was found in this ionic liquid compare to aqueous medium. Fluorescence life time of europium in this ionic liquid is 1100 μs which is ten times more than the life time of europium in aqueous medium (110 μs), indicating a reduction in the rates of non-radiative processes which was provided by the ionic liquid. This study leads to detection of europium in ppb level. (author)

  3. The neutron lifetime experiment PENeLOPE

    Energy Technology Data Exchange (ETDEWEB)

    Schreyer, Wolfgang [Technische Universitaet Muenchen (Germany); Collaboration: PENeLOPE-Collaboration

    2015-07-01

    The neutron lifetime τ{sub n}=880.3±1.1 s is an important parameter in the Standard Model of particle physics and in Big Bang cosmology. Several systematic corrections of previously published results reduced the PDG world average by several σ in the last years and call for a new experiment with complementary systematics. The experiment PENeLOPE, currently under construction at the Physik-Department of Technische Universitaet Muenchen, aims to determine the neutron lifetime with a precision of 0.1 s. It will trap ultra-cold neutrons in a magneto-gravitational trap using a large superconducting magnet and will measure their lifetime by both neutron counting and online proton detection. This presentation gives an overview over the latest developments of the experiment.

  4. Contactless Spectral-dependent Charge Carrier Lifetime Measurements in Silicon Photovoltaic Materials

    Science.gov (United States)

    Roller, John; Hamadani, Behrang; Dagenais, Mario

    Charge carrier lifetime measurements in bulk or unfinished photovoltaic (PV) materials allow for a more accurate estimate of power conversion efficiency in completed solar cells. In this work, carrier lifetimes in PV-grade silicon wafers are obtained by way of quasi-steady state photoconductance measurements. These measurements use a contactless RF system coupled with varying narrow spectrum input LEDs, ranging in wavelength from 460 nm to 1030 nm. Spectral dependent lifetime measurements allow for determination of bulk and surface properties of the material, including the intrinsic bulk lifetime and the surface recombination velocity. The effective lifetimes are fit to an analytical physics-based model to determine the desired parameters. Passivated and non-passivated samples are both studied and are shown to have good agreement with the theoretical model.

  5. The X-ray fluorescent method for determination of total sulphur in bituminous coals

    International Nuclear Information System (INIS)

    Widowska-Kusmierska, J.; Siess, K.

    1979-01-01

    The X-ray fluorescent technique for the determination of total sulphur covering concentrations from 0,1 to 10% has been applied for bituminous coals showing a great variability in qualitative and quantitative composition of mineral matter (ash). The described method is a quick one giving results during one hour. The obtained good accuracy of determinations gives prospects for wide industrial application. (author)

  6. Rapid determination of ampicillin in bovine milk by liquid chromatography with fluorescence detection

    Energy Technology Data Exchange (ETDEWEB)

    Ang, C.Y.W.; Luo, Wenhong [National Center for Toxicological Research, Jefferson, AR (United States)

    1997-01-01

    A rapid and sensitive liquid chromatographic (LC) method was developed for the determination of ampicillin residues in raw bovine milk, processed skim milk, and pasteurized, homogenized whole milk with vitamin D. Milk samples were deproteinized with trichloroacetic acid (TCA) and acetonictrile. After centrifugation, the clear supernatant was reacted with formaldehyde and TCA under heat. The major fluorescent derivative of ampicillin was then determined by reversed-phase LC with fluorescence detection. Average recoveries of ampicillin fortified at 5, 10, and 20 ppb (ng/mL) were all >85% with coefficients of variation <10%. Limits of detection ranged from 0.31 to 0.51 ppb and limits of quantitation, from 0.66 to 1.2 ppb. After appropriate validation, this method should be suitable for rapid analysis of milk for ampicillin residues at the tolerance level of 10 ppb. 16 refs., 4 figs., 3 tabs.

  7. A method to quantify FRET stoichiometry with phasor plot analysis and acceptor lifetime ingrowth.

    Science.gov (United States)

    Chen, WeiYue; Avezov, Edward; Schlachter, Simon C; Gielen, Fabrice; Laine, Romain F; Harding, Heather P; Hollfelder, Florian; Ron, David; Kaminski, Clemens F

    2015-03-10

    FRET is widely used for the study of protein-protein interactions in biological samples. However, it is difficult to quantify both the FRET efficiency (E) and the affinity (Kd) of the molecular interaction from intermolecular FRET signals in samples of unknown stoichiometry. Here, we present a method for the simultaneous quantification of the complete set of interaction parameters, including fractions of bound donors and acceptors, local protein concentrations, and dissociation constants, in each image pixel. The method makes use of fluorescence lifetime information from both donor and acceptor molecules and takes advantage of the linear properties of the phasor plot approach. We demonstrate the capability of our method in vitro in a microfluidic device and also in cells, via the determination of the binding affinity between tagged versions of glutathione and glutathione S-transferase, and via the determination of competitor concentration. The potential of the method is explored with simulations. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

  8. Time-resolved fluorescence microscopy (FLIM) as an analytical tool in skin nanomedicine.

    Science.gov (United States)

    Alexiev, Ulrike; Volz, Pierre; Boreham, Alexander; Brodwolf, Robert

    2017-07-01

    The emerging field of nanomedicine provides new approaches for the diagnosis and treatment of diseases, for symptom relief, and for monitoring of disease progression. Topical application of drug-loaded nanoparticles for the treatment of skin disorders is a promising strategy to overcome the stratum corneum, the upper layer of the skin, which represents an effective physical and biochemical barrier. The understanding of drug penetration into skin and enhanced penetration into skin facilitated by nanocarriers requires analytical tools that ideally allow to visualize the skin, its morphology, the drug carriers, drugs, their transport across the skin and possible interactions, as well as effects of the nanocarriers within the different skin layers. Here, we review some recent developments in the field of fluorescence microscopy, namely Fluorescence Lifetime Imaging Microscopy (FLIM)), for improved characterization of nanocarriers, their interactions and penetration into skin. In particular, FLIM allows for the discrimination of target molecules, e.g. fluorescently tagged nanocarriers, against the autofluorescent tissue background and, due to the environmental sensitivity of the fluorescence lifetime, also offers insights into the local environment of the nanoparticle and its interactions with other biomolecules. Thus, FLIM shows the potential to overcome several limits of intensity based microscopy. Copyright © 2017 Elsevier B.V. All rights reserved.

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

    Science.gov (United States)

    2015-01-01

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

  10. The measurement of subnanosecond nuclear lifetimes

    International Nuclear Information System (INIS)

    White, D.C.S.

    1974-01-01

    This research dealt with the measurement of subnanosecond nuclear lifetimes using the pulsed beam delayed-coincidence technique. Measurements were performed on isotopes in the f7/2 shell and specifically the isotopes of titanium and vanadium. Experimental investigations were also pursued in 59 Ni and 65 Zn. Several new lifetimes were determined and confirmation was obtained for some previous values which were measured with different techniques. More information was also obtained on certain levels where previous results are in disagreement. (author)

  11. ConA-based glucose sensing using the long-lifetime azadioxatriangulenium fluorophore

    Science.gov (United States)

    Cummins, Brian; Simpson, Jonathan; Gryczynski, Zygmunt; Sørensen, Thomas Just; Laursen, Bo W.; Graham, Duncan; Birch, David; Coté, Gerard

    2014-02-01

    Fluorescent glucose sensing technologies have been identified as possible alternatives to current continuous glucose monitoring approaches. We have recently introduced a new, smart fluorescent ligand to overcome the traditional problems of ConA-based glucose sensors. For this assay to be translated into a continuous glucose monitoring device where both components are free in solution, the molecular weight of the smart fluorescent ligand must be increased. We have identified ovalbumin as a naturally-occurring glycoprotein that could serve as the core-component of a 2nd generation smart fluorescent ligand. It has a single asparagine residue that is capable of displaying an N-linked glycan and a similar isoelectric point to ConA. Thus, binding between ConA and ovalbumin can potentially be monovalent and sugar specific. This work is the preliminary implementation of fluorescently-labeled ovalbumin in the ConA-based assay. We conjugate the red-emitting, long-lifetime azadioxatriangulenium (ADOTA+) dye to ovalbumin, as ADOTA have many advantageous properties to track the equilibrium binding of the assay. The ADOTA-labeled ovalbumin is paired with Alexa Fluor 647-labeled ConA to create a Förster Resonance Energy Transfer (FRET) assay that is glucose dependent. The assay responds across the physiologically relevant glucose range (0-500 mg/dL) with increasing intensity from the ADOTA-ovalbumin, showing that the strategy may allow for the translation of the smart fluorescent ligand concept into a continuous glucose monitoring device.

  12. Fluorescent silver nanoclusters for ultrasensitive determination of chromium(VI) in aqueous solution

    International Nuclear Information System (INIS)

    Zhang, Jian Rong; Zeng, Ai Lian; Luo, Hong Qun; Li, Nian Bing

    2016-01-01

    Highlights: • Fluorescent Ag nanoclusters were first applied to Cr(VI) detection. • The proposed method is simple, rapid, and environmentally friendly. • The sensor shows a wide linear range, low detection limit, and good selectivity. • The system can also be used for the indirect assay of total chromium and Cr(III). • The analyses in real water samples are satisfactory. - Abstract: In this work, a simple and sensitive Cr(VI) sensor is proposed based on fluorescent polyethyleneimine-stabilized Ag nanoclusters, which allows the determination over a wide concentration range of 0.1 nM–3.0 μM and with a detection limit as low as 0.04 nΜ and a good selectivity. The quenching mechanism was discussed in terms of the absorption and fluorescence spectra, suggesting that Cr(VI) is connected to Ag nanoclusters by hydrogen bond between the oxygen atom at the vertex of tetrahedron structure of Cr(VI) and the amino nitrogen of polyethyleneimine that surrounded Ag nanoclusters and electron transfer from Ag nanoclusters to highly electron-deficient Cr(VI) results in fluorescence quenching. Despite the failure to quench the fluorescence efficiently, Cr(III) can also be measured using the proposed Ag nanoclusters by being oxidized to Cr(VI) in alkaline solution (pH ∼9) containing H 2 O 2 . Therefore, our approach could be used to detect Cr(VI), Cr(III) and the total chromium level in aqueous solution. In addition, Cr(VI) analysis in real water samples were satisfactory, indicating this method could be practically promising for chromium measurements.

  13. Surface plasmon-enhanced molecular fluorescence induced by gold nanostructures

    International Nuclear Information System (INIS)

    Teng, Y.; Ueno, K.; Shi, X.; Aoyo, D.; Misawa, H.; Qiu, J.

    2012-01-01

    The authors report on surface plasmon-enhanced fluorescence of Eosin Y molecules induced by gold nanostructures. Al 2 O 3 films deposited by atomic layer deposition with sub-nanometer resolution were used as the spacer layer to control the distance between molecules and the gold surface. As the thickness of the Al 2 O 3 film increased, the fluorescence intensity first increased and then decreased. The highest enhancement factor is achieved with a 1 nm Al 2 O 3 film. However, the trend for the fluorescence lifetime is the opposite. It first decreased and then increased. The changes in the fluorescence quantum yield were also calculated. The yield shows a similar trend to the fluorescence intensity. The competition between the surface plasmon-induced increase in the radiative decay rate and the gold-induced fluorescence quenching is responsible for the observed phenomenon. In addition, this competition strongly depends on the thickness of the spacer layer between Eosin Y molecules and the gold surface. (Copyright copyright 2012 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

  14. In vivo detection of oral epithelial cancer using endogenous fluorescence lifetime imaging: a pilot human study (Conference Presentation)

    Science.gov (United States)

    Jo, Javier A.; Hwang, Dae Yon; Palma, Jorge; Cheng, Shuna; Cuenca, Rodrigo; Malik, Bilal; Jabbour, Joey; Cheng, Lisa; Wright, John; Maitland, Kristen

    2016-03-01

    Endogenous fluorescence lifetime imaging (FLIM) provides direct access to the concomitant functional and biochemical changes accompanying tissue transition from benign to precancerous and cancerous. Since FLIM can noninvasively measure different and complementary biomarkers of precancer and cancer, we hypothesize that it will aid in clinically detecting early oral epithelial cancer. Our group has recently demonstrated the detection of benign from premalignant and malignant lesions based on endogenous multispectral FLIM in the hamster cheek-pouch model. Encouraged by these positive preliminary results, we have developed a handheld endoscope capable of acquiring multispectral FLIM images in real time from the oral mucosa. This novel FLIM endoscope is being used for imaging clinically suspicious pre-malignant and malignant lesions from patients before undergoing tissue biopsy for histopathological diagnosis of oral epithelial cancer. Our preliminary results thus far are already suggesting the potential of endogenous FLIM for distinguishing a variety of benign lesions from advanced dysplasia and squamous cell carcinoma (SCC). To the best of out knowledge, this is the first in vivo human study aiming to demonstrate the ability to predict the true malignancy of clinically suspicious lesions using endogenous FLIM. If successful, the resulting clinical tool will allow noninvasive real-time detection of epithelial precancerous and cancerous lesions in the oral mucosa and could potentially be used to assist at every step involved on the clinical management of oral cancer patients, from early screening and diagnosis, to treatment and monitoring of recurrence.

  15. New fluorescence spectroscopic method for the simultaneous determination of alkaloids in aqueous extract of green coffee beans.

    Science.gov (United States)

    Yisak, Hagos; Redi-Abshiro, Mesfin; Chandravanshi, Bhagwan Singh

    2018-05-11

    There is no fluorescence spectroscopic method for the determination of trigonelline and theobromine in green coffee beans. Therefore, the objective of this study was to develop a new fluorescence spectroscopic method to determine the alkaloids simultaneously in the aqueous extract of green coffee beans. The calibration curves were linear in the range 2-6, 1-6, 1-5 mg/L for caffeine, theobromine and trigonelline, respectively, with R 2  ≥ 0.9987. The limit of detection and limit of quantification were 2, 6 and 7 µg/L and 40, 20 and 20 µg/L for caffeine, theobromine and trigonelline, respectively. Caffeine and trigonelline exhibited well separated fluorescence excitation spectra and therefore the two alkaloids were selectively quantified in the aqueous extract of green coffee. While theobromine showed overlapping fluorescence excitation spectra with caffeine and hence theobromine could not be determined in the aqueous extract of green coffee beans. The amount of caffeine and trigonelline in the three samples of green coffee beans were found to be 0.95-1.10 and 1.00-1.10% (w/w), respectively. The relative standard deviations (RSD ≤ 4%) of the method for the three compounds of interest were of very good. The accuracy of the developed analytical method was evaluated by spiking standard caffeine and trigonelline to green coffee beans and the average recoveries were 99 ± 2% for both the alkaloids. A fast, sensitive and reliable fluorescence method for the simultaneous determination of caffeine and trigonelline in the aqueous extract of green coffee beans was developed and validated. The developed method reflected an effective performance to the direct determination of the two alkaloids in the aqueous extract of green coffee beans.

  16. Fluorescence properties of 3-amino phenylboronic acid and its interaction with glucose and ZnS:Cu quantum dots.

    Science.gov (United States)

    Kur-Kowalska, Karolina; Przybyt, Małgorzata; Ziółczyk, Paulina; Sowiński, Przemysław; Miller, Ewa

    2014-08-14

    Preliminary results of a study of the interaction between 3-amino phenylboronic acid and glucose or ZnS:Cu quantum dots are presented in this paper. ZnS:Cu quantum dots with mercaptopropionic acid as a capping agent were obtained and characterized. Quenching of 3-amino phenylboronic acid fluorescence was studied by steady-state and timeresolved measurements. For fluorescence quenching with glucose the results of steady-state measurements fulfill Stern-Volmer equation. The quenching constants are increasing with growing pH. The decay of fluorescence is monoexponential with lifetime about 8.4 ns, which does not depend on pH and glucose concentration indicating static quenching. The quenching constant can be interpreted as apparent equilibrium constant of estrification of boronic group with diol. Quantum dots are also quenching 3-amino phenylboronic acid fluorescence. Fluorescence lifetime, in this case, is slightly decreasing with increasing concentration of quantum dots. The quenching constants are increasing slightly with pH's growth. Quenching mechanism of 3-amino phenylboronic acid fluorescence by quantum dots needs further experiments to be fully explained. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Fluorescent Inhibitors as Tools To Characterize Enzymes: Case Study of the Lipid Kinase Phosphatidylinositol 4-Kinase IIIβ (PI4KB).

    Science.gov (United States)

    Humpolickova, Jana; Mejdrová, Ivana; Matousova, Marika; Nencka, Radim; Boura, Evzen

    2017-01-12

    The lipid kinase phosphatidylinositol 4-kinase IIIβ (PI4KB) is an essential host factor for many positive-sense single-stranded RNA (+RNA) viruses including human pathogens hepatitis C virus (HCV), Severe acute respiratory syndrome (SARS), coxsackie viruses, and rhinoviruses. Inhibitors of PI4KB are considered to be potential broad-spectrum virostatics, and it is therefore critical to develop a biochemical understanding of the kinase. Here, we present highly potent and selective fluorescent inhibitors that we show to be useful chemical biology tools especially in determination of dissociation constants. Moreover, we show that the coumarin-labeled inhibitor can be used to image PI4KB in cells using fluorescence-lifetime imaging microscopy (FLIM) microscopy.

  18. Determination of the cathode fall voltage in fluorescent lamps by measurement of the operating voltage

    International Nuclear Information System (INIS)

    Hilscher, A.

    2002-01-01

    A new method for the determination of the cathode fall voltage of fluorescent lamps is shown. The cathode fall voltage can be determined by measurement of the lamp operating voltage at constant lamp wall temperature, constant discharge current and variation of the electrode heating current. Commercial lamps, which do not need to be specially prepared, can be used for the measurement. The results show good correlation to other measurements of the cathode fall voltage at various discharge currents by means of capacitive coupling. The measured values of the cathode fall voltage are used for determining the minimum, target and maximum setting of the sum of the squares of the pin currents of one electrode (the so-called SOS value) as a function of the discharge current in fluorescent lamp dimming. (author)

  19. Lifetime of the metastable 23S1 state in stored Li+ ions

    International Nuclear Information System (INIS)

    Knight, R.D.

    1979-04-01

    A laser-induced fluorescence technique combined with the observation of spontaneous magnetic dipole photons from the highly metastable 2 3 S 1 state of Li + was used to measure the radiative lifetime of this state. The ions are created by electron impact on a lithium atomic beam and are subsequently stored for periods of many seconds in an RF-quadrupole ion trap. A tunable dye laser excites the 2 3 S--2 3 P, transition at 5485A, and the intercombination electric dipole transition 2 3 P 1 --1 1 S 0 at 202A is observed. This process depletes the metastable population in a time tau/sub d/ 3 S 1 / and provides a measure of the total number of metastables. Comparison with the rate of 210A spontaneously emitted photons yields a measured value for the 2 3 S 1 radiative lifetime of tau/sub rad/ = 58.6 +- 12.9 sec, where the quoted error represents 95% confidence levels. The theoretical lifetime is tau/sub theory/ = 49.0 sec. The measured value includes data taken with both 6 Li + and 7 Li + isotopes and was corrected for the slightly different detector efficiencies at 202A and 210A. A careful study of nonradiative quenching of the metastable state was necessary to understand observed differences between tau/sub rad/ and tau/sub 3 S 1 /, the total metastable lifetime. Spatial density profiles of the ions within the trap, useful for determining the ion temperature, were obtained by scanning the laser beam horizontally across the ion trap while storing 2 3 P 1 -- 1 S 0 photon counts as a function of the laser beam's position. Agreement with a simple equilibrium model, including space charge effects, is satisfactory. A study of the optical pumping process is necessary to understand the laser-ion interaction, and observational and theoretical data are presented. 47 references

  20. Determining the ice-binding planes of antifreeze proteins by fluorescence-based ice plane affinity.

    Science.gov (United States)

    Basu, Koli; Garnham, Christopher P; Nishimiya, Yoshiyuki; Tsuda, Sakae; Braslavsky, Ido; Davies, Peter

    2014-01-15

    Antifreeze proteins (AFPs) are expressed in a variety of cold-hardy organisms to prevent or slow internal ice growth. AFPs bind to specific planes of ice through their ice-binding surfaces. Fluorescence-based ice plane affinity (FIPA) analysis is a modified technique used to determine the ice planes to which the AFPs bind. FIPA is based on the original ice-etching method for determining AFP-bound ice-planes. It produces clearer images in a shortened experimental time. In FIPA analysis, AFPs are fluorescently labeled with a chimeric tag or a covalent dye then slowly incorporated into a macroscopic single ice crystal, which has been preformed into a hemisphere and oriented to determine the a- and c-axes. The AFP-bound ice hemisphere is imaged under UV light to visualize AFP-bound planes using filters to block out nonspecific light. Fluorescent labeling of the AFPs allows real-time monitoring of AFP adsorption into ice. The labels have been found not to influence the planes to which AFPs bind. FIPA analysis also introduces the option to bind more than one differently tagged AFP on the same single ice crystal to help differentiate their binding planes. These applications of FIPA are helping to advance our understanding of how AFPs bind to ice to halt its growth and why many AFP-producing organisms express multiple AFP isoforms.

  1. Single-label kinase and phosphatase assays for tyrosine phosphorylation using nanosecond time-resolved fluorescence detection.

    Science.gov (United States)

    Sahoo, Harekrushna; Hennig, Andreas; Florea, Mara; Roth, Doris; Enderle, Thilo; Nau, Werner M

    2007-12-26

    The collision-induced fluorescence quenching of a 2,3-diazabicyclo[2.2.2]oct-2-ene-labeled asparagine (Dbo) by hydrogen atom abstraction from the tyrosine residue in peptide substrates was introduced as a single-labeling strategy to assay the activity of tyrosine kinases and phosphatases. The assays were tested for 12 different combinations of Dbo-labeled substrates and with the enzymes p60c-Src Src kinase, EGFR kinase, YOP protein tyrosine phosphatase, as well as acid and alkaline phosphatases, thereby demonstrating a broad application potential. The steady-state fluorescence changed by a factor of up to 7 in the course of the enzymatic reaction, which allowed for a sufficient sensitivity of continuous monitoring in steady-state experiments. The fluorescence lifetimes (and intensities) were found to be rather constant for the phosphotyrosine peptides (ca. 300 ns in aerated water), while those of the unphosphorylated peptides were as short as 40 ns (at pH 7) and 7 ns (at pH 13) as a result of intramolecular quenching. Owing to the exceptionally long fluorescence lifetime of Dbo, the assays were alternatively performed by using nanosecond time-resolved fluorescence (Nano-TRF) detection, which leads to an improved discrimination of background fluorescence and an increased sensitivity. The potential for inhibitor screening was demonstrated through the inhibition of acid and alkaline phosphatases by molybdate.

  2. Occupational risk and lifetime exposure

    International Nuclear Information System (INIS)

    Lapp, R.E.

    1991-01-01

    Any lowering of annual radiation limits for occupational exposure should be based on industry experience with lifetime doses and not on a worst case career exposure of 47 years. Two decades of experience show a lifetime accumulation of less than 1.5 rem for workers with measurable exposure. This is 5% of the normal lifetime exposure of Americans to natural and medical radiation. Any epidemiology of the US nuclear power workforce's two decade long exposure would have to focus on excess leukemia. Application of the Hiroshima and Nagasaki cancer mortality shows that too few leukemias would be expressed to permit a feasible epidemiology. Ionizing radiation appears to be a mild carcinogen as compared to physical and chemical agents presented in the occupational environment. A realistic factor in determining any change in occupational exposure limits for ionizing radiation should take into account the past performance of the licensee and potential health effects applicable to the workplace. Specifically, the lifetime exposure data for workers at nuclear power plants and naval shipyards should be considered. The nuclear industry and the US Navy have detailed data on the annual exposure of workers with a combined collective exposure approaching 1 million worker-rem. The lifetime dose for naval personnel and shipyard workers averages 1.1 rem J 1990. Shipyard workers have an annual dose of 0.28 rem per work-year and a mean exposure time of 4.4 years. The data apply to workers with measurable dose

  3. Photoluminescence behavior of riboflavin and lumiflavin in liquid solutions and solid films

    Energy Technology Data Exchange (ETDEWEB)

    Penzkofer, A., E-mail: alfons.penzkofer@physik.uni-regensburg.de [Fakultaet fuer Physik, Universitaet Regensburg, Universitaetsstrasse 31, D-93053 Regensburg (Germany)

    2012-05-25

    Highlights: Black-Right-Pointing-Pointer Delayed fluorescence and phosphorescence spectra of flavins in starch films measured. Black-Right-Pointing-Pointer Quantum yield of singlet-triplet intersystem determined with a new approach. Black-Right-Pointing-Pointer Theory developed for determination of luminescence quantum yields of films. Black-Right-Pointing-Pointer Delayed fluorescence and phosphorescence lifetimes of flavins in starch films measured. Black-Right-Pointing-Pointer Singlet and triplet relevant parameters of riboflavin and lumiflavin determined. - Abstract: The absorption and emission behavior of riboflavin and lumiflavin in water, tetrahydrofuran (THF), water-starch, THF-polystyrene, starch films, and polystyrene films was studied at room temperature. Absorption cross-section spectra, fluorescence quantum distributions, and fluorescence quantum yields were determined. For the starch films additionally phosphorescence and delayed fluorescence spectra as well as phosphorescence lifetimes and delayed fluorescence lifetimes were measured and their quantum yields of intersystem-crossing, intrinsic triplet-based phosphorescence quantum yields, T{sub 1}-S{sub 0} radiative lifetimes, and S{sub 0}-T{sub 1} absorption strengths were calculated. A method of absolute intrinsic luminescence quantum distribution and quantum yield determination for dye doped films on transparent plates with a fluorimeter is described.

  4. Reliability-based assessment of polyethylene pipe creep lifetime

    International Nuclear Information System (INIS)

    Khelif, Rabia; Chateauneuf, Alaa; Chaoui, Kamel

    2007-01-01

    Lifetime management of underground pipelines is mandatory for safe hydrocarbon transmission and distribution systems. The use of high-density polyethylene tubes subjected to internal pressure, external loading and environmental variations requires a reliability study in order to define the service limits and the optimal operating conditions. In service, the time-dependent phenomena, especially creep, take place during the pipe lifetime, leading to significant strength reduction. In this work, the reliability-based assessment of pipe lifetime models is carried out, in order to propose a probabilistic methodology for lifetime model selection and to determine the pipe safety levels as well as the most important parameters for pipeline reliability. This study is enhanced by parametric analysis on pipe configuration, gas pressure and operating temperature

  5. Reliability-based assessment of polyethylene pipe creep lifetime

    Energy Technology Data Exchange (ETDEWEB)

    Khelif, Rabia [LaMI-UBP and IFMA, Campus de Clermont-Fd, Les Cezeaux, BP 265, 63175 Aubiere Cedex (France); LR3MI, Departement de Genie Mecanique, Universite Badji Mokhtar, BP 12, Annaba 23000 (Algeria)], E-mail: rabia.khelif@ifma.fr; Chateauneuf, Alaa [LGC-University Blaise Pascal, Campus des Cezeaux, BP 206, 63174 Aubiere Cedex (France)], E-mail: alaa.chateauneuf@polytech.univ-bpclermont.fr; Chaoui, Kamel [LR3MI, Departement de Genie Mecanique, Universite Badji Mokhtar, BP 12, Annaba 23000 (Algeria)], E-mail: chaoui@univ-annaba.org

    2007-12-15

    Lifetime management of underground pipelines is mandatory for safe hydrocarbon transmission and distribution systems. The use of high-density polyethylene tubes subjected to internal pressure, external loading and environmental variations requires a reliability study in order to define the service limits and the optimal operating conditions. In service, the time-dependent phenomena, especially creep, take place during the pipe lifetime, leading to significant strength reduction. In this work, the reliability-based assessment of pipe lifetime models is carried out, in order to propose a probabilistic methodology for lifetime model selection and to determine the pipe safety levels as well as the most important parameters for pipeline reliability. This study is enhanced by parametric analysis on pipe configuration, gas pressure and operating temperature.

  6. Lifetime measurements

    International Nuclear Information System (INIS)

    Fossan, D.B.; Warburton, E.K.

    1974-01-01

    Lifetime measurements are discussed, concentrating on the electronic technique, the recoil distance method (RDM), and the Doppler shift attenuation method (DSAM). A brief review of several indirect timing techniques is given, and their specific advantages and applicability are considered. The relationship between lifetimes of nuclear states and the nuclear structure information obtained from them is examined. A short discussion of channeling and microwave methods of lifetime measurement is presented. (23 figures, 171 references) (U.S.)

  7. Determination of Ivermectin in Medicated Feeds by Liquid Chromatography with Fluorescence Detection

    Science.gov (United States)

    2013-01-01

    A labour- and time-effective analytical procedure for determination of ivermectin in medicated feed at recommended level of 2.0 mg kg−1 has been developed and validated. The analyte was extracted from grinded feed samples with acetonitrile and derivatisated with N-methylimidazole and trifluoracetic anhydride. The fluorescent derivatives were analysed by liquid chromatography method using C8 column. The isocratic conditions using acetonitrile, methanol, water, and tetrahydrofuran were applied. Fluorescence detection was performed at 365 nm (excitation) and 475 nm (emission) wavelengths. The total analysis time was 10 min. The validation results of the method (within-laboratory reproducibility 4.0% CV, mean recovery 100.1%) confirm the appropriate precision and accuracy of the developed method. PMID:24453835

  8. Selective detection of Co2+ by fluorescent nano probe: Diagnostic approach for analysis of environmental samples and biological activities

    Science.gov (United States)

    Mahajan, Prasad G.; Dige, Nilam C.; Desai, Netaji K.; Patil, Shivajirao R.; Kondalkar, Vijay V.; Hong, Seong-Karp; Lee, Ki Hwan

    2018-06-01

    Nowadays scientist over the world are engaging to put forth improved methods to detect metal ion in an aqueous medium based on fluorescence studies. A simple, selective and sensitive method was proposed for detection of Co2+ ion using fluorescent organic nanoparticles. We synthesized a fluorescent small molecule viz. 4,4‧-{benzene-1,4-diylbis-[(Z)methylylidenenitrilo]}dibenzoic acid (BMBA) to explore its suitability as sensor for Co2+ ion and biocompatibility in form of nanoparticles. Fluorescence nanoparticles (BMBANPs) prepared by simple reprecipitation method. Aggregation induced enhanced emission of BMBANPs exhibits the narrower particle size of 68 nm and sphere shape morphology. The selective fluorescence quenching was observed by addition of Co2+ and does not affected by presence of other coexisting ion solutions. The photo-physical properties, viz. UV-absorption, fluorescence emission, and lifetime measurements are in support of ligand-metal interaction followed by static fluorescence quenching phenomenon in emission of BMBANPs. Finally, we develop a simple analytical method for selective and sensitive determination of Co2+ ion in environmental samples. The cell culture E. coli, Bacillus sps., and M. tuberculosis H37RV strain in the vicinity of BMBANPs indicates virtuous anti-bacterial and anti-tuberculosis activity which is of additional novel application shown by prepared nanoparticles.

  9. Chlorine determination in (U, Pu)C fuel by total reflection X-ray fluorescence spectrometry

    International Nuclear Information System (INIS)

    Misra, Nand Lal; Dhara, Sangita; Mudher, Khush Dev Singh; Aggarwal, Suresh K.; Thakur, Uday Kumar; Shah, Dipti; Sawant, R.M.; Ramakumar, K.L.

    2007-01-01

    A Total Reflection X-ray Fluorescence (TXRF) method for the determination of chlorine in (U,Pu)C has been developed. The method involves calibration of the instrument with standard solutions and validation of TXRF determination of chlorine using synthetic standard solutions. Cl K α line excited with W L α source was used for TXRF determinations of chlorine. Chlorine present in trace amounts in (U,Pu)C samples was first separated by pyro hydrolysis. The evolved chlorine, in form of HCl, was collected in 5 mM NaOH solution. This solution was analyzed for chlorine by Total Reflection X-ray Fluorescence Spectrometry using cobalt as an internal standard. The TXRF detection limit of chlorine was found to be 3.6 pg with sample size of 30 μL. In order to assess the applicability of TXRF method for chlorine determinations in other nuclear materials, one U 3 O 8 trace element standard was also analyzed for chlorine in similar way. The precision of the method was found to be 25% (1 σ) at ng level in most of the cases. (author)

  10. Imaging Amyloid Tissues Stained with Luminescent Conjugated Oligothiophenes by Hyperspectral Confocal Microscopy and Fluorescence Lifetime Imaging.

    Science.gov (United States)

    Nyström, Sofie; Bäck, Marcus; Nilsson, K Peter R; Hammarström, Per

    2017-10-20

    Proteins that deposit as amyloid in tissues throughout the body can be the cause or consequence of a large number of diseases. Among these we find neurodegenerative diseases such as Alzheimer's and Parkinson's disease afflicting primarily the central nervous system, and systemic amyloidosis where serum amyloid A, transthyretin and IgG light chains deposit as amyloid in liver, carpal tunnel, spleen, kidney, heart, and other peripheral tissues. Amyloid has been known and studied for more than a century, often using amyloid specific dyes such as Congo red and Thioflavin T (ThT) or Thioflavin (ThS). In this paper, we present heptamer-formyl thiophene acetic acid (hFTAA) as an example of recently developed complements to these dyes called luminescent conjugated oligothiophenes (LCOs). hFTAA is easy to use and is compatible with co-staining in immunofluorescence or with other cellular markers. Extensive research has proven that hFTAA detects a wider range of disease associated protein aggregates than conventional amyloid dyes. In addition, hFTAA can also be applied for optical assignment of distinct aggregated morphotypes to allow studies of amyloid fibril polymorphism. While the imaging methodology applied is optional, we here demonstrate hyperspectral imaging (HIS), laser scanning confocal microscopy and fluorescence lifetime imaging (FLIM). These examples show some of the imaging techniques where LCOs can be used as tools to gain more detailed knowledge of the formation and structural properties of amyloids. An important limitation to the technique is, as for all conventional optical microscopy techniques, the requirement for microscopic size of aggregates to allow detection. Furthermore, the aggregate should comprise a repetitive β-sheet structure to allow for hFTAA binding. Excessive fixation and/or epitope exposure that modify the aggregate structure or conformation can render poor hFTAA binding and hence pose limitations to accurate imaging.

  11. Mobility-lifetime product in epitaxial GaAs X-ray detectors

    Energy Technology Data Exchange (ETDEWEB)

    Sun, G.C. [GESEC R and D, Universite Pierre et Marie Curie, Bat.11, 140 rue de Lourmel, 75015 Paris (France)]. E-mail: guocsun@ccr.jussieu.fr; Zazoui, M. [LPMC, Faculte des Sciences et Techniques-Mohammedia, B.P. 146 Bd Hassan II, Mohammedia, Maroc (Morocco); Talbi, N. [Faculte des Sciences, Universite de Gabes, Route de Medenine, 6029 Gabes (Tunisia); Khirouni, K. [Faculte des Sciences, Universite de Gabes, Route de Medenine, 6029 Gabes (Tunisia); Bourgoin, J.C. [GESEC R and D, Universite Pierre et Marie Curie, Bat.11, 140 rue de Lourmel, 75015 Paris (France)

    2007-04-01

    Self-supported thick (200-500 {mu}m), non-intentionally doped, epitaxial GaAs layers are good candidates for X-ray imaging for the following reasons. Their electronic properties are homogeneous over large areas, they can be grown at low cost, the technology to realize pixel detectors of various size is standard, the defect concentration is low and the fluorescence yield is small. Here, we characterize the defects present in the material and evaluate the mobility-lifetime product, using Deep Level Transient Spectroscopy combined with current-voltage and charge collection measurements.

  12. Interpretation of measurements of dynamic fluorescence of the eye

    Science.gov (United States)

    Schweitzer, Dietrich; Hammer, Martin; Jentsch, Susanne; Schenke, Stefan

    2007-09-01

    First pathological alterations occur at cellular level, most in metabolism. An indirect estimation of metabolic activity in cells is measurement of microcirculation. Measurements of tissue autofluorescence are potentially suited for direct investigation of cellular metabolism. Besides redox pairs of co-enzymes (NADH-NAD, FADH2-FAD) several other fluorophores are excited in tissue. In addition, a number of anatomical structures are simultaneously excited, when investigating the eye-ground. In this study, spectral and time resolved comparison was performed between purified substances, single ocular structures and in vivo measurements of the time-resolved autofluorescence at the human eye. In human eyes, the ageing pigment lipofuscin covers other fluorophores at the fundus in long - wave visible range. Applying lifetime measurements, weakly emitting fluorophores can be detected, when the lifetimes are different from the strongly emitting fluorophore. For this, the autofluorescence was excited at 468 nm and detected in two spectral ranges (500 nm-560 nm, 560 nm-700 nm). In tri-exponential fitting, the short lifetime corresponds to retinal pigment epithelium, the mean lifetime corresponds probably to neural retina and the long lifetime is caused by fluorescence of connective tissue.

  13. Radiative-lifetime measurements and calculations of odd-parity highly excited levels in Ba i

    International Nuclear Information System (INIS)

    Zhang Wei; Du Shan; Palmeri, Patrick; Quinet, Pascal; Biemont, Emile; Dai Zhenwen

    2010-01-01

    Natural radiative lifetime measurements have been performed for 70 odd-parity highly excited levels of neutral barium in the energy range from 308 15.512 to 417 59.93 cm -1 by a time-resolved laser-induced fluorescence technique in a laser-produced plasma. The lifetime values measured in this paper are in the range from 11.3 to 901 ns. They are compared with the published lifetimes of four levels. Two of them are in good agreement, whereas for the other two our measurements are slightly longer than the published data. The reasons for the discrepancies are discussed. Comparisons with theoretical results of the Hartree-Fock method with relativistic corrections illustrate the difficulties associated with the use of Cowan's codes for obtaining accurate branching fractions for transitions depopulating highly excited levels along the Rydberg series of heavy neutral elements. This work will be useful to extend the set of oscillator strengths available in Ba i.

  14. Some applications of x-ray fluorescence spectrography to the determination of uranium and thorium

    International Nuclear Information System (INIS)

    Jones, R.W.

    1959-04-01

    Several methods for the determination of uranium and thorium by X-ray fluorescence spectrography are described. In pure solutions the sensitivity for these elements is 5-10 ppm. For solutions containing gross concentrations of impurities, strontium is added as an internal standard. Precision and accuracy of the determinations are about 1% when working in the optimum concentration range. (author)

  15. Radiative lifetime and Lande-factor measurements of the Se I 4p35S 5S2 level using pulsed laser spectroscopy

    International Nuclear Information System (INIS)

    Zerne, R.

    1992-01-01

    This diploma project consists of spectroscopic examinations of atomic selenium. Natural selenium was thermally dissociated in a quarts resonance cell keeping the background pressure of selenium molecules low by differential heating. The 4p 3 5S 5 S 2 level was excited by frequency-tripled pulsed dye-laser radiation at 207 nm. From time-resolved recording of the fluorescence decay at 216 nm the natural radiative lifetime of the 5 S 2 level was determined to be 493(15) ns. Quantum-beat and optical double resonance measurements in an external magnetic field yielded g j = 2.0004(10) for the Lande factor

  16. Determinations of silicon and phosphorus in Pepperbush standard reference material by neutron activation and x-ray fluorescence methods

    International Nuclear Information System (INIS)

    Mizumoto, Yoshihiko; Nishio, Hirofumi; Hayashi, Takeshi; Kusakabe, Toshio; Iwata, Shiro.

    1987-01-01

    Silicon and phosphorus contents in Pepperbush standard reference material were determined by neutron activation and X-ray fluorescence methods. In neutron activation analysis, β-ray spectra of 32 P produced by 31 P(n,γ) 32 P reaction on Pepperbush and standard samples were measured by a low background β-ray spectrometer. In X-ray fluorescence analysis, the standard samples were prepared by mixing the Pepperbush powder with silicon dioxide and diammonium hydrogenphosphate. Characteristic X-rays from the samples were analyzed by a wavelength dispersive X-ray fluorescence spectrometer. From the β and X-ray intensities, silicon and phosphorus contents in Pepperbush were determined to be 1840 ± 80 and 1200 ± 50 μg g -1 , respectively. (author)

  17. Fluorescent derivatives of nucleotides. Metal ion interactions and pH dependency.

    Science.gov (United States)

    Vanderkooi, J M; Weiss, C J; Woodrow, G V

    1979-02-01

    The fluorescence parameters of ethenoadenosine derivatives are influenced by metal cations and pH, as summarized here. The pH profile of ethenoadenosine determined by fluorescence intensity gives a normal titration curve and is not affected by ionic strength. In contrast, the pH titration curves of etheno-ATP, etheno ADP, and etheno AMP depend upon ionic strength. At high ionic strength normal curves are obtained, whereas at low ionic strength anomalies are obtained; this suggests that the phosphates can interact with the ring, possibly by hydrogen binding to the ring nitrogens. The room temperature fluorescence of ethenoadenosine occurs from the base form, although excitation of either the acid or base forms can contribute to the emission. This result can be explained if the excited state pK is lower than the ground state pK, and if deprotonation occurs within the time scale of the excited state. At low pH values the fluorescence lifetime of the base form is dependent upon the buffer concentration, indicating that the reverse reaction, protonation, occurs. The affinity constants for the binding of metals to the ethenoadenosine phosphates resemble those for the corresponding adenosine phosphates. Ni(II) and Co(II) are more effective than Mn(II) in quenching the fluorescence of ethenoadenosine phosphates; this result is predicted by Förster's theory for energy transfer based upon the overlap between donor emission spectrum and acceptor absorption spectrum. The diamagnetic ions Mg(II), Ca(II), and Zn(II) do not appear to affect the fluorescence of the ethenoadenosine phosphates directly, but rather to affect the conformation of the molecule, thereby affecting the quantum yield.

  18. Fluorescent investigation of the interactions between N-(p-chlorophenyl)-N'-(1-naphthyl) thiourea and serum albumin: Synchronous fluorescence determination of serum albumin

    International Nuclear Information System (INIS)

    Cui Fengling; Wang Junli; Cui Yanrui; Li Jianping

    2006-01-01

    The interactions between N-(p-chlorophenyl)-N'-(1-naphthyl) thiourea and serum albumin were investigated by fluorescence spectroscopy and UV absorption spectrum under physiological conditions. The results of spectroscopic measurements suggested that N-(p-chlorophenyl)-N'-(1-naphthyl) thiourea should have a strong ability to quench the intrinsic fluorescence of both bovine serum albumin and human serum albumin through static quenching procedure, and the hydrophobic interaction was the predominant intermolecular force stabilizing the complex. Thermodynamic parameter enthalpy changes (ΔH) and entropy changes (ΔS) were calculated according to the Vant'Hoff equation. The binding distances between N-(p-chlorophenyl)-N'-(1-naphthyl) thiourea and the proteins were evaluated on the basis of the theory of Foester energy transfer. In addition, the effects of other ions on the binding constants of complexes were also discussed. Synchronous fluorescence technology was successfully applied to the determination of serum albumins added to the CPNT solution

  19. Continuous determination of gaseous ammonia in the ambient atmosphere using fluorescence derivatization

    Science.gov (United States)

    Abbas, Rana; Tanner, Roger L.

    A method for continuous determination of ambient ammonia levels employing o-phthalaldehyde fluorescence derivatization is described. A simplified Venturi scrubber and gas-liquid separator have been employed for reproducible measurements of ⩾ 0.1 ppb ambient ammonia with less than 2 min time resolution. The scrubbing efficiency of the ammonia gas collection system was determined to be 29 ± 1 %. During 4 d in August 1979 ambient ammonia levels at the Brookhaven National Laboratory site averaged about 1.5 ± 1.1 ppb during afternoon daylight hours.

  20. Fluorescence method for the determination of carcinogenic N-nitrosamines using N-(8-methoxy-5-quinolinesulfonyl)-aziridine

    International Nuclear Information System (INIS)

    Gracheva, I.N.; Kovel'man, I.R.; Pimenova, V.V.; Tochilkin, A.I.; Zhukova, G.F.

    1986-01-01

    The determination of carcinogenic N-nitrosamines (NA) in the environment and particularly in food products is, according to the authors, extremely important. The objective of the present work is the development of an easily accessible and highly sensitive method for the determination of NA. NA, isolated from food products, are denitrosated to their corresponding amines, which are then determined in the form of fluorescent derivatives by reaction with N-(8-methoxy-5-quinolinesulfonyl) aziridine. The validity of the results of NA analysis from food product was confirmed by gas-liquid chromatography using a thermoluminescent detector TEA-502. The limit of detection is 1x10 -10 mole of N-nitrosamine. The possibility of separating and determining the new fluorescent derivatives III(a-f) on silufol plates, which are easily obtained and widely used in analytical practice, was examined

  1. Heritability of lifetime ecstasy use.

    Science.gov (United States)

    Verweij, Karin J H; Treur, Jorien L; Vreeker, Annabel; Brunt, Tibor M; Willemsen, Gonneke; Boomsma, Dorret I; Vink, Jacqueline M

    2017-09-01

    Ecstasy is a widely used psychoactive drug that users often take because they experience positive effects such as increased euphoria, sociability, elevated mood, and heightened sensations. Ecstasy use is not harmless and several immediate and long term side effects have been identified. Lifetime ecstasy use is likely to be partly influenced by genetic factors, but no twin study has determined the heritability. Here, we apply a classical twin design to a large sample of twins and siblings to estimate the heritability of lifetime ecstasy use. The sample comprised 8500 twins and siblings aged between 18 and 45 years from 5402 families registered at the Netherlands Twin Registry. In 2013-2014 participants filled out a questionnaire including a question whether they had ever used ecstasy. We used the classical twin design to partition the individual differences in liability to ecstasy use into that due to genetic, shared environmental, and residual components. Overall, 10.4% of the sample had used ecstasy during their lifetime, with a somewhat higher prevalence in males than females. Twin modelling indicated that individual differences in liability to lifetime ecstasy use are for 74% due to genetic differences between individuals, whereas shared environmental and residual factors explain a small proportion of its liability (5% and 21%, respectively). Although heritability estimates appeared to be higher for females than males, this difference was not significant. Lifetime ecstasy use is a highly heritable trait, which indicates that some people are genetically more vulnerable to start using ecstasy than others. Copyright © 2017. Published by Elsevier B.V.

  2. An image fiber based fluorescent probe with associated signal processing scheme for biomedical diagnostics

    International Nuclear Information System (INIS)

    Vaishakh, M; Murukeshan, V M; Seah, L K

    2008-01-01

    A dual-modality image fiber based fluorescent probe that can be used for depth sensitive imaging and suppression of fluorescent emissions with nanosecond lifetime difference is proposed and illustrated in this paper. The system can give high optical sectioning and employs an algorithm for obtaining phase sensitive images. The system can find main application in in vivo biomedical diagnostics for detecting biochemical changes for distinguishing malignant tissue from healthy tissue

  3. Novel Fluorescent Microemulsion: Probing Properties, Investigating Mechanism, and Unveiling Potential Application.

    Science.gov (United States)

    Hou, Mengna; Dang, Leping; Liu, Tiankuo; Guo, Yun; Wang, Zhanzhong

    2017-08-09

    Nanoscale microemulsions have been utilized as delivery carriers for nutraceuticals and active biological drugs. Herein, we designed and synthesized a novel oil in water (O/W) fluorescent microemulsion based on isoamyl acetate, polyoxyethylene castor oil EL (CrEL), and water. The microemulsion emitted bright blue fluorescence, thus exhibiting its potential for active drug detection with label-free strategy. The microemulsion exhibited excitation-dependent emission and distinct red shift with longer excitation wavelengths. Lifetime and quantum yield of fluorescent microemulsion were 2.831 ns and 5.0%, respectively. An excellent fluorescent stability of the microemulsion was confirmed by altering pH, ionic strength, temperature, and time. Moreover, we proposed a probable mechanism of fluorochromic phenomenon, in connection with the aromatic ring structure of polyoxyethylene ether substituent in CrEL. Based on our findings, we concluded that this new fluorescent microemulsion is a promising drug carrier that can facilitate active drug detection with a label-free strategy. Although further research is required to understand the exact mechanism behind its fluorescence property, this work provided valuable guidance to develop new biosensors based on fluorescent microemulsion.

  4. Choice of excitation source for determination of rare earth elements with radioisotope excited X ray fluorescence

    International Nuclear Information System (INIS)

    Zhang Quanshi; Chang Yongfu

    2000-01-01

    The comparisons of two radioisotope source ( 241 Am and 238 Pu) which are the most available in the radioisotope excited X Ray Fluorescence (XRF) analysis technique and two characteristic X ray series (KX and LX) analyzed for the determination of the rare-earth (RE) elements were investigated in detail. According to the principle of emission and detection of X ray , the relative excitation efficiencies were calculated by the some fundamental physical parameters including the photoelectric mass attenuation coefficient, the fluorescent yield, the absorption jump factor, the emission probability of the detected fluorescent line with reference to other liens of the same series etc., The advantages and disadvantages of the two conditions are discussed. These results may determine the optimal excitation and detection conditions for different rare-earth elements. The experimental results with nine rare-earth elements (Ce, Nd, Sm, Tb, Tm, Ho, Er, Yb and Lu) are in agreement with the results of theoretical calculations

  5. Lifetime measurements in Crsub(I) by laser excitation from the metastable states

    International Nuclear Information System (INIS)

    Kwiatkowski, M.; Micali, G.; Werner, K.; Zimmermann, P.

    1981-01-01

    A combination of collisional and laser excitation was used to measure radiative lifetimes in Cr I. By a discharge an atomic beam of metastable atoms in the 3d 5 4sa 5 S, a 5 G, b 5 D, a 3 I, b 1 I and 3d 4 4s 2 a 5 D terms was produced. Spatially separated from the place of collisional excitation laser radiation selectively populated levels belonging to the 3d 5 4p z 5 P, y 5 P, u 5 F, u 5 D, x 3 I, y 1 I, 3d 5 5pz 5 G and 3d 4 4s4px 5 G terms. Time-resolved observation of the reemitted resonance fluorescence yielded the lifetimes of 28 levels. The values are compared with other experimental and theoretical results. (orig.)

  6. The rotational predissociation of HeH+ energy and lifetime measurements, ch. 3

    International Nuclear Information System (INIS)

    Locht, R.; Maas, J.G.; Asselt, N.P.F.B. van; Los, J.

    1976-01-01

    Relative lifetimes and energies above the dissociation limit have been determined for the rotational predissociation of several quasi-bound levels of the X'Σ + state of 4 HeH + . In particular, the lifetimes are very sensitive to the shape of the potential energy curve. These measurements are used to discriminate between two ab initio potential curves which differ by only 0.00004 a.u. (approximately 1 meV). Using the lifetime data, relative population factors were determined for the observed levels

  7. 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

  8. Prospects for a new cold neutron beam measurement of the neutron lifetime

    Energy Technology Data Exchange (ETDEWEB)

    Dewey, M., E-mail: mdewey@nist.go [National Institute of Standards and Technology, Gaithersburg, MD 20899 (United States); Coakley, K., E-mail: kevin.coakley@nist.go [National Institute of Standards and Technology, Boulder, CO 80305 (United States); Gilliam, D., E-mail: david.gilliam@nist.go [National Institute of Standards and Technology, Gaithersburg, MD 20899 (United States); Greene, G., E-mail: greenegl@ornl.go [Department of Physics, University of Tennessee, Knoxville, TN 37996 (United States); Physics Division, Oak Ridge National Lab, Building 6010, Oak Ridge, TN 37831 (United States); Laptev, A., E-mail: alaptev@nist.go [Los Alamos National Laboratory, Los Alamos, NM 87545 (United States); Nico, J., E-mail: jnico@nist.go [National Institute of Standards and Technology, Gaithersburg, MD 20899 (United States); Snow, W., E-mail: wsnow@indiana.ed [Indiana University/IUCF, Bloomington, IN 47408 (United States); Wietfeldt, F., E-mail: few@tulane.ed [Tulane University, New Orleans, LA 70118 (United States); Yue, A., E-mail: ayue@nist.go [Department of Physics, University of Tennessee, Knoxville, TN 37996 (United States)

    2009-12-11

    In the most accurate cold neutron beam determination of the neutron lifetime based on the absolute counting of decay protons, the largest uncertainty was attributed to the absolute determination of the capture flux of the cold neutron beam. Currently an experimental effort is underway at the National Institute of Standards and Technology (NIST) that will significantly reduce this contribution to the uncertainty in the lifetime determination. The next largest source of uncertainty is the determination of the absolute count rate of decay protons, which contributes to the experimental uncertainty approximately at the 1 s level. Experience with the recent neutron radiative decay experiment, which used the neutron lifetime apparatus, has provided valuable insights into ways to reduce other uncertainties. In addition, the cold neutron fluence rate at NIST is presently 1.5 times greater than in the 2003 measurement, and there is the prospect for a significantly higher rate with the new guide hall expansion. This paper discusses an approach for achieving a determination of the neutron lifetime with an accuracy of approximately 1 s.

  9. Enzyme entrapment in polyaniline films observed via fluorescence anisotropy and antiquenching

    Science.gov (United States)

    Nemzer, Louis R.; McCaffrey, Marisa; Epstein, Arthur J.

    2014-05-01

    The facile entrapment of oxidoreductase enzymes within polyaniline polymer films by inducing hydrophobic collapse using phosphate buffered saline (PBS) has been shown to be a cost-effective method for fabricating organic biosensors. Here, we use fluorescence anisotropy measurements to verify enzyme immobilization and subsequent electron donation to the polymer matrix, both prerequisites for an effective biosensor. Specifically, we measure a three order of magnitude decrease in the ratio of the fluorescence to rotational lifetimes. In addition, the observed fluorescence antiquenching supports the previously proposed model that the polymer chain assumes a severely coiled conformation when exposed to PBS. These results help to empirically reinforce the theoretical basis previously laid out for this biosensing platform.

  10. 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.

  11. The role of defects in fluorescent silicon carbide layers grown by sublimation epitaxy

    DEFF Research Database (Denmark)

    Schimmel, Saskia; Kaiser, Michl; Jokubavicius, Valdas

    2014-01-01

    Donor-acceptor co-doped SiC is a promising light converter for novel monolithic all-semiconductor white LEDs due to its broad-band donor-acceptor pair luminescence and potentially high internal quantum efficiency. Besides sufficiently high doping concentrations in an appropriate ratio yielding...... short radiative lifetimes, long nonradiative lifetimes are crucial for efficient light conversion. The impact of different types of defects is studied by characterizing fluorescent silicon carbide layers with regard to photoluminescence intensity, homogeneity and efficiency taking into account...

  12. Effects of short term changes in the blood glucose level on the autofluorescence lifetime of the human retina in healthy volunteers

    Science.gov (United States)

    Klemm, Matthias; Nagel, Edgar; Schweitzer, Dietrich; Schramm, Stefan; Haueisen, Jens

    2016-03-01

    Purpose: Fluorescence lifetime imaging ophthalmoscopy (FLIO) provides in vivo metabolic mapping of the ocular fundus. Changes in FLIO have been found in e.g. diabetes patients. The influence of short term metabolic changes caused by blood glucose level changes on is unknown. Aim of this work is the detection of short-term changes in fundus autofluorescence lifetime during an oral glucose tolerance test. Methods: FLIO was performed in 10 healthy volunteers (29+/-4 years, fasting for 12h) using a scanning laser ophthalmoscope (30° fundus, 34μm resolution, excitation with 473nm diode laser with 70 ps pulses at 80 MHz repetition rate, detection in two spectral channels 500-560nm (ch1) and 560-720nm (ch2) using the timecorrelated single photon counting method). The blood glucose level (BGL) was measured by an Accu-Chek® Aviva self-monitoring device. Before and after a glucose drink (300ml solution, containing 75g of glucose (Accu-Chek® Dextrose O.G.T.), BGL and FLIO were measured every 15min. The FLIMX software package was applied to compute the average fluorescence lifetime τ on the inner ring of the ETDRS grid using a modified 3-exponential approach. Results: The results are given as mean +/- standard deviation over all volunteers in ch1. Baseline measurement: BGL: 5.3+/-0.4 mmol/l, τ1: 49+/-6ps. A significant reduction (α=5% Wilcoxon rank-sum test) in τ1 is detected after 15min (BGL: 8.4+/-1.1 mmol/l, τ1: 44+/-5ps) and after 90min (BGL: 6.3+/-1.4 mmol/l, τ1: 41+/-5ps). Results of ch2 show smaller reductions in the fluorescence lifetimes over time.

  13. Hue-shifted monomeric variants of Clavularia cyan fluorescent protein: identification of the molecular determinants of color and applications in fluorescence imaging

    Directory of Open Access Journals (Sweden)

    Davidson Michael W

    2008-03-01

    mutagenesis study, we conclude that the two histidine residues in close proximity to the chromophore are approximately equal determinants of the blue-shifted fluorescence emission of mTFP1. With respect to live cell imaging applications, the mTFP1-derived mWasabi should be particularly useful in two-color imaging in conjunction with a Sapphire-type variant or as a fluorescence resonance energy transfer acceptor with a blue FP donor. In all fusions attempted, both mTFP1 and mWasabi give patterns of fluorescent localization indistinguishable from that of well-established avGFP variants.

  14. The determination, by x-ray-fluorescence spectrometry, of gold in activated charcoal

    International Nuclear Information System (INIS)

    Austen, C.E.

    1977-01-01

    A rapid method is described for the determination of gold in activated charcoal by X-ray-fluorescence spectrometry. Compensation for matrix effects is achieved by means of platinum that is added for use as an internal standard. Calibration is achieved by use of a series of synthetic standards that are made by the spiking of barren charcoal with gold and platinum. The limit of determination is about 8 p.p.m. of gold, and the relative standard deviation is 1,2 per cent at a concentration level of 2300 p.p.m

  15. Fluorescent carbon quantum dot hydrogels for direct determination of silver ions.

    Science.gov (United States)

    Cayuela, A; Soriano, M L; Kennedy, S R; Steed, J W; Valcárcel, M

    2016-05-01

    The paper reports for the first time the direct determination of silver ion (Ag(+)) using luminescent Carbon Quantum Dot hydrogels (CQDGs). Carbon Quantum Dots (CQDs) with different superficial moieties (passivate-CQDs with carboxylic groups, thiol-CQDs and amine-CQDs) were used to prepare hybrid gels using a low molecular weight hydrogelator (LMWG). The use of the gels results in considerable fluorescence enhancement and also markedly influences selectivity. The most selective CQDG system for Ag(+) ion detection proved to be those containing carboxylic groups onto their surface. The selectivity towards Ag(+) ions is possibly due to its flexible coordination sphere compared with other metal ions. This fluorescent sensing platform is based on the strong Ag-O interaction which can quench the photoluminescence of passivate-CQDs (p-CQDs) through charge transfer. The limit of detection (LOD) and quantification (LOQ) of the proposed method were 0.55 and 1.83µgmL(-1), respectively, being applied in river water samples. Copyright © 2016 Elsevier B.V. All rights reserved.

  16. Assessment of background gamma radiation and determination of excess lifetime cancer risk in Sabzevar City, Iran in 2014

    Directory of Open Access Journals (Sweden)

    Akbar Eslami

    2016-01-01

    Full Text Available Background: Background gamma radiation levels vary in different locations and depended on many factors such as radiation properties of soil, building materials as well as construction types which human lives on it. People are always exposed to ionizing radiation, which could badly influence their health. The aim of this study was to evaluate the background gamma-ray dose rate and the estimated annual effective dose equivalent and determination of excess lifetime cancer risk in Sabzevar City, Iran. Methods: The aim of this cross-sectional study was to determine the dose rate of background gamma radiation in outdoor an indoor areas, 26 stations were selected using the map of the Sabzevar City. The amount of gamma radiation was measured at 4 months (September to January in 2014 year. The dosimeter used in this study was a survey meter, that is designed for monitoring radiation of x, gamma and beta rays. Results: The obtained results show that there are significant differences between the indoor and outdoor exposures (P> 0.05. We did not observe significant differences between the time of sampling and sampling locations, (P<0.05. The minimum and maximum values of dose rate were found 66±20 nSvh-1 and 198±28 nSvh-1. The annual effective dose for Sabzevar residents was estimated to be 0.85 mSv and also the amount of excess lifetime cancer risk was estimated 3.39×10-3. Conclusion: According to the results, the excess lifetime cancer risk and the annual effective dose for the Sabzavar City residents due to the background gamma radiation was higher than the global average (0.5 mSv. The epidemiological studies have been proposed to evaluate the risk of chronic diseases associated with natural radiation exposure among residents.

  17. Preparation and preliminary characterization of crystallizing fluorescent derivatives of chicken egg white lysozyme

    Science.gov (United States)

    Sumida, John P.; Forsythe, Elizabeth L.; Pusey, Marc L.

    2001-11-01

    Fluorescence is one of the most versatile and powerful tools for the study of macromolecules. While most proteins are intrinsically fluorescent, working at crystallization concentrations require the use of covalently prepared derivatives added as tracers. This approach requires derivatives that do not markedly affect the crystal packing. We have prepared fluorescent derivatives of chicken egg white lysozyme with probes bound to one of two different sites on the protein molecule. Lucifer yellow and 5-(2-aminoethyl)aminonapthalene-1-sulfonic acid (EDANS) have been attached to the side chain carboxyl of Asp 101 using a carbodiimide coupling procedure. Asp 101 lies within the active site cleft, and it is believed that the probes are "buried" within that cleft. Lucifer yellow and EDANS probes with iodoacetamide reactive groups have been bound to His 15, located on the "back side" of the molecule relative to the active site. All the derivatives fluoresce in the solution and the crystalline states. Fluorescence characterization has focused on determination of binding effects on the probe quantum yield, lifetime, absorption and emission spectra, and quenching by added solutes. Quenching studies show that, as postulated, the Asp 101-bound probes are partially sheltered from the bulk solution by their location within the active site cleft. Probes bound to His 15 have quenching constants about equal to those for the free probes, indicating that this site is highly exposed to the bulk solution.

  18. Sensitization of uranium fluorescence using 2,6-pyridinedicarboxylic acid: Application for the determination of uranium in the presence of lanthanides

    International Nuclear Information System (INIS)

    Maji, S.; Viswanathan, K.S.

    2009-01-01

    The 2,6-pyridinedicarboxylic acid (PDA) has been shown to efficiently sensitize and enhance the fluorescence of uranium in aqueous medium. Interestingly, this ligand stabilizes the UO 2 2+ species, which without the ligand is known to be in a negligible concentration, in aqueous medium at pH 6. The ligand sensitized enhancement of UO 2 2+ fluorescence by PDA, provides an analytical tool for the determination of uranium at trace levels, in aqueous medium. Furthermore, PDA is also known to enhance the fluorescence of lanthanides; consequently, the simultaneous determination of uranium and lanthanides, using PDA as a fluorescence sensitizing agent, becomes a possibility, which has been demonstrated in this work. We have shown that the use of PDA yields detection limits of 2.2x10 -7 M for UO 2 2+ , 1x10 -8 M for Tb 3+ and 5x10 -9 M for Eu 3+ in the simultaneous determination of these analytes.

  19. Determinations of elements in pepperbush standard reference material by neutron activation and X-ray fluorescence analyses

    International Nuclear Information System (INIS)

    Mizumoto, Yoshihiko; Okada, Takayuki; Tatsumi, Toshiya; Kusakabe, Toshio; Katsurayama, Kousuke; Iwata, Shiro.

    1988-01-01

    Elemental contents in Pepperbush standard reference material have been determined by neutron activation and X-ray fluorescence analyses. The standard samples of orchard leaves, tomato leaves, pine needles and Kale are used for the experiment. In the neutron activation analysis, gamma-ray spectra of nuclei produced by (n,γ) reaction on Pepperbush and standard samples are measured with Ge detectors. In the X-ray fluorescence analysis, the samples are excited with X-rays from X-ray tube with rhodium anode, and the characteristic X-rays from samples are measured with a proportional counter or NaI(Tl) detector. From the gamma- and X-ray intensities, the elemental contents in Pepperbush are determined. As a result, the contents of seventeen elements, such as sodium, calcium, iron, etc., in Pepperbush are determined. (author)

  20. Determination of heavy metals in polar snow and ice by laser-excited atomic fluorescence spectrometry

    International Nuclear Information System (INIS)

    Bolshov, M.A.; Boutron, C.F.

    1994-01-01

    The new laser-excited atomic fluorescence spectrometry technique offers unrivalled sensitivity for the determination of trace metals in a wide variety of samples. This has allowed the direct determination of Pb, Cd and Bi in Antarctic and Greenland snow and ice down to the sub pg/g level. (authors). 11 refs., 2 figs

  1. Determination of nuclear spins of short-lived isotopes by laser induced fluorescence

    International Nuclear Information System (INIS)

    Buchinger, F.; Dabkiewicz, P.; Kremmling, H.; Kuehl, T.; Mueller, A.C.; Schuessler, H.A.

    1980-01-01

    The spins of several nuclear ground and isomeric states have been measured for a number of mercury isotopes. The fluorescent light from the 6s6p 3 P 1 state is observed at 2537 Angstroem after excitation with the frequency doubled output of a pulsed dye laser. Four different laser induced fluorescence techniques were tested for their applicability: double resonance, Hanle effect, time delayed integral Hanle beats, and time resolved quantum beats. The sensitivity and selectivity of these models are compared with emphasis on the determination of spins of nuclei far from beta-stability, where short half lives and low production yields restrict the number of available atoms. The experiments were carried out on-line with the ISOLDE isotope separator at CERN at densities as low as 10 6 atoms/cm 3 . Results for the very neutron deficient high spin mercury isomers with half lives of several seconds, but also for the ground states of the abundant low spin stable mercury isotopes, are given as examples. The test measurements determined the nuclear spins of the odd sup(185m-191m)Hg isomers to be I = 13/2. (orig.)

  2. Determination of low concentrations of thorium in geological materials by X-ray fluorescence

    International Nuclear Information System (INIS)

    Roca, M.; Bayon, A.

    1981-01-01

    An X-ray fluorescence method for the determination of thorium in geological samples down to 2 ppm ThO 2 has been developed. To achieve this determination limit an exposed area of the sample 42.5 mm in diameter is used, working with a molybdenum target tube operated at 90 kV and 30 m A. Corrections for background and line interference of the Rb Kα radiation have been carefully considered and empirical correction coefficients calculated. (Author) 3 refs

  3. Synchronous fluorescence determination of ciprofloxacin in the pharmaceutical formulation and human serum based on the perturbed luminescence of rare-earth ions

    Energy Technology Data Exchange (ETDEWEB)

    Tong Changlun, E-mail: cltong@zju.edu.c [Key Laboratory of Environmental Remediation and Ecological Health, Ministry of Education, College of Environmental and Resource Sciences, Zhejiang University, Hangzhou 310029 (China); Zhuo Xiajun; Guo Yun; Fang Yueheng [Key Laboratory of Environmental Remediation and Ecological Health, Ministry of Education, College of Environmental and Resource Sciences, Zhejiang University, Hangzhou 310029 (China)

    2010-11-15

    A simple, rapid and sensitive synchronous fluorescence method was developed for the determination of ciprofloxacin (CPFX) in the pharmaceutical formulation and human serum. The results show that when Y{sup 3+} is added into the CPFX solution, the characteristic fluorescence of Y{sup 3+} is not emitted whereas the fluorescence intensity of CPFX is significantly enhanced. The synchronous fluorescence technology is employed in this method to directly determine trace amount of CPFX in human serum. A linear relationship between the fluorescence intensity and the CPFX concentration is obtained in the range of 1.0x10{sup -9} {approx}5.0x10{sup -6} mol L{sup -1}. The limit of detection (LOD) of this method attains as low as 2.0x10{sup -10} mol L{sup -1} (S/N=3). The selectivity of this method is also very good. Common metal ions, rare-earth ions and some pharmaceuticals, which are usually used together with CPFX in the clinic, do not interfere with the determination of CPFX under general conditions.

  4. Autofluorescence lifetime imaging during transoral robotic surgery: a clinical validation study of tumor detection (Conference Presentation)

    Science.gov (United States)

    Lagarto, João. L.; Phipps, Jennifer E.; Unger, Jakob; Faller, Leta M.; Gorpas, Dimitris; Ma, Dinglong M.; Bec, Julien; Moore, Michael G.; Bewley, Arnaud F.; Yankelevich, Diego R.; Sorger, Jonathan M.; Farwell, Gregory D.; Marcu, Laura

    2017-02-01

    Autofluorescence lifetime spectroscopy is a promising non-invasive label-free tool for characterization of biological tissues and shows potential to report structural and biochemical alterations in tissue owing to pathological transformations. In particular, when combined with fiber-optic based instruments, autofluorescence lifetime measurements can enhance intraoperative diagnosis and provide guidance in surgical procedures. We investigate the potential of a fiber-optic based multi-spectral time-resolved fluorescence spectroscopy instrument to characterize the autofluorescence fingerprint associated with histologic, morphologic and metabolic changes in tissue that can provide real-time contrast between healthy and tumor regions in vivo and guide clinicians during resection of diseased areas during transoral robotic surgery. To provide immediate feedback to the surgeons, we employ tracking of an aiming beam that co-registers our point measurements with the robot camera images and allows visualization of the surgical area augmented with autofluorescence lifetime data in the surgeon's console in real-time. For each patient, autofluorescence lifetime measurements were acquired from normal, diseased and surgically altered tissue, both in vivo (pre- and post-resection) and ex vivo. Initial results indicate tumor and normal regions can be distinguished based on changes in lifetime parameters measured in vivo, when the tumor is located superficially. In particular, results show that autofluorescence lifetime of tumor is shorter than that of normal tissue (p robot assisted cancer removal interventions.

  5. Determination of acetylsalicylic acid and salicylic acid in foods, using HPLC with fluorescence detection.

    NARCIS (Netherlands)

    Venema, D.P.; Hollman, P.C.H.; Janssen, P.L.T.M.K.; Katan, M.B.

    1996-01-01

    We developed a specific and sensitive HPLC method with fluorescence detection for the determination of free acetylsalicylic acid, free salicylic acid, and free salicylic acid plus salicylic acid after alkaline hydrolysis (free-plus-bound) in foods. Acetylsalicylic acid was detected after postcolumn

  6. Time-resolved measurements of the external electric field effects on fluorescence in electron donor and acceptor pairs of N-ethylcarbazole and dimethyl terephthalate doped in a polymer film

    International Nuclear Information System (INIS)

    Iimori, Toshifumi; Yoshizawa, Tomokazu; Nakabayashi, Takakazu; Ohta, Nobuhiro

    2005-01-01

    Electric-field-induced change in fluorescence decay has been measured for electron donor and acceptor pairs of N-ethylcarbazole (ECZ) and dimethyl terephthalate (DMTP) doped in a polymer film. Field-induced change in lifetime of the fluorescence emitted from the locally excited state of ECZ clearly shows that the electron transfer from the excited state of ECZ to DMTP is enhanced by an external electric field ( F ). A comparison is made between the experimental results of the field effect on decay profile of the ECZ fluorescence and the simulated results. Time-resolved electrofluorescence spectra as well as the field-induced change in decay profile of exciplex fluorescence show that exciplex fluorescence is quenched by F at the early stage of time following photoexcitation, but enhanced by F at a later stage of time. Both the decrease in the initial population of the fluorescent exciplex and the lengthening of the exciplex fluorescence in lifetime are shown to be induced by F

  7. Time-resolved measurements of the external electric field effects on fluorescence in electron donor and acceptor pairs of N-ethylcarbazole and dimethyl terephthalate doped in a polymer film

    Energy Technology Data Exchange (ETDEWEB)

    Iimori, Toshifumi [Research Institute for Electronic Science (RIES), Hokkaido University, Sapporo 060-0812 (Japan); Graduate School of Environmental Earth Science, Hokkaido University, Sapporo 060-0810 (Japan); Yoshizawa, Tomokazu [Graduate School of Environmental Earth Science, Hokkaido University, Sapporo 060-0810 (Japan); Nakabayashi, Takakazu [Research Institute for Electronic Science (RIES), Hokkaido University, Sapporo 060-0812 (Japan); Graduate School of Environmental Earth Science, Hokkaido University, Sapporo 060-0810 (Japan); Ohta, Nobuhiro [Research Institute for Electronic Science (RIES), Hokkaido University, Sapporo 060-0812 (Japan); Graduate School of Environmental Earth Science, Hokkaido University, Sapporo 060-0810 (Japan)], E-mail: nohta@es.hokudai.ac.jp

    2005-12-07

    Electric-field-induced change in fluorescence decay has been measured for electron donor and acceptor pairs of N-ethylcarbazole (ECZ) and dimethyl terephthalate (DMTP) doped in a polymer film. Field-induced change in lifetime of the fluorescence emitted from the locally excited state of ECZ clearly shows that the electron transfer from the excited state of ECZ to DMTP is enhanced by an external electric field ( F ). A comparison is made between the experimental results of the field effect on decay profile of the ECZ fluorescence and the simulated results. Time-resolved electrofluorescence spectra as well as the field-induced change in decay profile of exciplex fluorescence show that exciplex fluorescence is quenched by F at the early stage of time following photoexcitation, but enhanced by F at a later stage of time. Both the decrease in the initial population of the fluorescent exciplex and the lengthening of the exciplex fluorescence in lifetime are shown to be induced by F.

  8. Fluorescent investigation of the interactions between N-(p-chlorophenyl)-N'-(1-naphthyl) thiourea and serum albumin: Synchronous fluorescence determination of serum albumin

    Energy Technology Data Exchange (ETDEWEB)

    Cui Fengling [School of Chemistry and Environmental Science, Key Laboratory for Environmental Pollution Control Technology of Henan Province, Henan Normal University, Xinxiang, Hennan 453007 (China)]. E-mail: fenglingcui@hotmail.com; Wang Junli [School of Chemistry and Environmental Science, Key Laboratory for Environmental Pollution Control Technology of Henan Province, Henan Normal University, Xinxiang, Hennan 453007 (China); Cui Yanrui [School of Chemistry and Environmental Science, Key Laboratory for Environmental Pollution Control Technology of Henan Province, Henan Normal University, Xinxiang, Hennan 453007 (China); Li Jianping [School of Chemistry and Environmental Science, Key Laboratory for Environmental Pollution Control Technology of Henan Province, Henan Normal University, Xinxiang, Hennan 453007 (China)

    2006-07-07

    The interactions between N-(p-chlorophenyl)-N'-(1-naphthyl) thiourea and serum albumin were investigated by fluorescence spectroscopy and UV absorption spectrum under physiological conditions. The results of spectroscopic measurements suggested that N-(p-chlorophenyl)-N'-(1-naphthyl) thiourea should have a strong ability to quench the intrinsic fluorescence of both bovine serum albumin and human serum albumin through static quenching procedure, and the hydrophobic interaction was the predominant intermolecular force stabilizing the complex. Thermodynamic parameter enthalpy changes ({delta}H) and entropy changes ({delta}S) were calculated according to the Vant'Hoff equation. The binding distances between N-(p-chlorophenyl)-N'-(1-naphthyl) thiourea and the proteins were evaluated on the basis of the theory of Foester energy transfer. In addition, the effects of other ions on the binding constants of complexes were also discussed. Synchronous fluorescence technology was successfully applied to the determination of serum albumins added to the CPNT solution.

  9. Single excitation-emission fluorescence spectrum (EEF) for determination of cetane improver in diesel fuel.

    Science.gov (United States)

    Insausti, Matías; Fernández Band, Beatriz S

    2015-04-05

    A highly sensitive spectrofluorimetric method has been developed for the determination of 2-ethylhexyl nitrate in diesel fuel. Usually, this compound is used as an additive in order to improve cetane number. The analytical method consists in building the chemometric model as a first step. Then, it is possible to quantify the analyte with only recording a single excitation-emission fluorescence spectrum (EEF), whose data are introduced in the chemometric model above mentioned. Another important characteristic of this method is that the fuel sample was used without any pre-treatment for EEF. This work provides an interest improvement to fluorescence techniques using the rapid and easily applicable EEF approach to analyze such complex matrices. Exploding EEF was the key to a successful determination, obtaining a detection limit of 0.00434% (v/v) and a limit of quantification of 0.01446% (v/v). Copyright © 2015 Elsevier B.V. All rights reserved.

  10. Positron annihilation lifetime spectroscopy study of Kapton thin foils

    Science.gov (United States)

    Kanda, G. S.; Ravelli, L.; Löwe, B.; Egger, W.; Keeble, D. J.

    2016-01-01

    Variable energy positron annihilation lifetime spectroscopy (VE-PALS) experiments on polyimide material Kapton are reported. Thin Kapton foils are widely used in a variety of mechanical, electronic applications. PALS provides a sensitive probe of vacancy-related defects in a wide range of materials, including open volume in polymers. Varying the positron implantation energy enables direct measurement of thin foils. Thin Kapton foils are also commonly used to enclose the positron source material in conventional PALS measurements performed with unmoderated radionuclide sources. The results of depth-profiled positron lifetime measurements on 7.6 μm and 25 μm Kapton foils are reported and determine a dominant 385(1) ps lifetime component. The absence of significant nanosecond lifetime component due to positronium formation is confirmed.

  11. Critical aggregates concentration of fatty esters present in biodiesel determined by turbidity and fluorescence.

    Science.gov (United States)

    Froehner, Sandro; Sánez, Juan; Dombroski, Luiz Fernando; Gracioto, Maria Paula

    2017-09-01

    Biodiesel for combustible engine is available as mixture of fossil diesel and fatty esters obtained by transesterification of vegetable oils. The use of biodiesel reduces the amount of SO x , mainly. However, it was already observed that biodiesel has a different behavior in environment in cases of accidental spill and groundwater contamination. It was noticed that the biodegradation of hydrocarbons (cyclic and aliphatic) in the presence of biodiesel are speeded, although the mechanism is still unclear. Considering the chemical structure of fatty esters, it was investigated the formation of aggregates in water solution by fatty esters present in commercial biodiesel. In Brazil, biodiesel is composed by 95% of fossil diesel and 5% of fatty esters mixture. In this work, fatty esters were treated as neutral surfactant, i.e., it was treated as a molecule with polar and non-polar part. Turbidity and fluorescence were used to determine the critical aggregates concentration (CAC). Water solutions containing fatty esters were examined exploiting changes in turbidity and fluorescence intensity of pyrene. Abrupt changes were attributed to aggregates formation, following the same behavior of traditional amphiphilic compounds. It was determined the CAC for ethyl palmitate, ethyl stearate, ethyl oleate, and ethyl linoleate. The values of CAC for fatty esters varied from 1.91 to 4.27 μmol/L, while CAC for the mixture of esters (biodiesel) was 2.01 for methyl esters and 1.19 for ethyl esters, both prepared using soybean oil. The aggregates formation was also determined by fluorescence measurements considering the changes in intensity of peaks I and III of pyrene. Pyrene senses the changes in environment polarity. The values found of CAC by fluorescence for individual ethyl esters varied from 1.85 to 3.21 μmol/L, while mixtures of ethyl esters was 2.23 and 2.07 μmol/L for mixture of methyl esters. The results clearly showed that fatty esters form aggregates and might be

  12. The determination, by x-ray-fluorescence spectrometry, of gold and uranium on resin

    International Nuclear Information System (INIS)

    Jacobs, J.J.; Balaes, A.M.E.

    1983-01-01

    The problems encountered in the determination of gold and uranium that are present simultaneously in a sample of resin were considered, and new background positions, as well as correction factors for background lift and partial spectral overlap, were determined for use in the X-ray-fluorescence measurement of these elements. The agreement between the results obtained by the use of the X-ray-fluorescence method and those obtained by atomic-absorption spectrophotometry were found to be satisfactory. The relative standard deviation in the former measurements is 0,005 at a concentration of 1000 p.p.m., and the working range of the calibrations is 15 to 1000 p.p.m. These limits can be extended by further dilution of the sample. The limits of determination are 6 p.p.m. in the briquette prepared from a sample of resin and 18 p.p.m. in the sample when the maximum mass of the sample is 3 g. The procedure is intended primarily for use in the rapid determination of gold and uranium when no analyses for other elements are required. The time taken for the analysis is 2 hours for 10 samples when 6 standards are used. A computer programme that was developed for the processing of the data is appended as part of a laboratory method

  13. Comparison of N-alkyl acridine orange dyes as fluorescence probes for the determination of cardiolipin

    Energy Technology Data Exchange (ETDEWEB)

    Kaewsuya, P.; Miller, J.D. [Department of Chemistry and Biochemistry, Miami University, Oxford, OH 45056 (United States); Danielson, N.D. [Department of Chemistry and Biochemistry, Miami University, Oxford, OH 45056 (United States)], E-mail: danielnd@muohio.edu; Sanjeevi, J.; James, P.F. [Department of Zoology, Miami University, Oxford, OH 45056 (United States)

    2008-09-26

    The phospholipid (PL), cardiolipin (CL), is found almost exclusively in the inner membrane of mitochondria and loss of CL is considered as an important indication of cell apoptosis. Previously, 10-N-nonyl acridine orange (NAO) has been used as a fluorescent probe for the visualization of CL in mitochondrial cell membranes and in solution. In this work for the determination of CL, we have synthesized two new fluorescent probes, n-tetradecyl acridine orange (C14-AO), and n-octadecyl acridine orange (C18-AO) by reacting acridine orange with the corresponding n-alkyl bromide. Using excitation and emission wavelengths at about 500 and 525 nm and varying the percentage of methanol in water as the solvent, no interaction between CL and the fluorescent probes at 75% is noted but a proportional quenching of the fluorescence signal by CL is observed at 50% or less for C14-AO and 60% or less for C18-AO. Binding efficiency of these fluorescent probes to CL is compared using dye concentrations of 5, 10, and 20 {mu}M. C18-AO shows a better sensitivity than C14-AO and NAO, respectively, but is less selective. For C14-AO, the detection limit and limit of quantitation are 0.07 and 0.21 {mu}M, respectively, which are better than those previously reported for NAO. One anionic PL, phosphatidic acid, shows some quenching interference to both the C14 and C18 dyes but only at concentrations above the working range for sample analysis. The CL in mitochondrial membrane samples is determined by standard addition using C14-AO. The level of CL in the outer mitochondrial membrane compared to the inner membrane is significantly increased due to the addition of cadmium chloride into the cells causing cell apoptosis.

  14. Determination of adenine based on the fluorescence recovery of the L-Tryptophan-Cu(2+) complex.

    Science.gov (United States)

    Duan, Ruilin; Li, Chunyan; Liu, Shaopu; Liu, Zhongfang; Li, Yuanfang; Yuan, Yusheng; Hu, Xiaoli

    2016-01-05

    A simple and sensitive method for determination of adenine was developed based on fluorescence quenching and recovery of L-Tryptophan (L-Trp). The fluorescence of L-Trp could efficiently quenched by copper ion compared with other common metal ions. Upon addition of adenine (Ade) in L-Trp-Cu(II) system, the fluorescence was reoccurred. Under the optimum conditions, the recovery fluorescence intensity was linearly correlated with the concentration of adenine in the range from 0.34 to 25.0μmolL(-1), with a correlation coefficient (R(2)) of 0.9994. The detection limit (3σ/k) was 0.046μmolL(-1), indicating that this method could applied to detect trace adenine. In this study, amino acids including L-Trp, D-Trp, L-Tyr, D-Tyr, L-Phe, D-Phe were investigated and only L-Trp could well chelated copper ion. Additionally, the mechanism of quench and recovery also were discussed and the method was successfully applied to detect the adenine in DNA with satisfactory results. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. Fluorescent Biosensor for Phosphate Determination Based on Immobilized Polyfluorene-Liposomal Nanoparticles Coupled with Alkaline Phosphatase.

    Science.gov (United States)

    Kahveci, Zehra; Martínez-Tomé, Maria José; Mallavia, Ricardo; Mateo, C Reyes

    2017-01-11

    This work describes the development of a novel fluorescent biosensor based on the inhibition of alkaline phosphatase (ALP). The biosensor is composed of the enzyme ALP and the conjugated cationic polyfluorene HTMA-PFP. The working principle of the biosensor is based on the fluorescence quenching of this polyelectrolyte by p-nitrophenol (PNP), a product of the hydrolysis reaction of p-nitrophenyl phosphate (PNPP) catalyzed by ALP. Because HTMA-PFP forms unstable aggregates in buffer, with low fluorescence efficiency, previous stabilization of the polyelectrolyte was required before the development of the biosensor. HTMA-PFP was stabilized through its interaction with lipid vesicles to obtain stable blue-emitting nanoparticles (NPs). Fluorescent NPs were characterized, and the ability to be quenched by PNP was evaluated. These nanoparticles were coupled to ALP and entrapped in a sol-gel matrix to produce a biosensor that can serve as a screening platform to identify ALP inhibitors. The components of the biosensor were examined before and after sol-gel entrapment, and the biosensor was optimized to allow the determination of phosphate ion in aqueous medium.

  16. Determination of non-ionic surfactants in technologic liquors and effluents by X-ray fluorescent spectrometry

    International Nuclear Information System (INIS)

    Dankowski, P.; Majda, J.

    1977-01-01

    The method has been worked out for determination of non-ionic surfactants in technological liquors and effluents, based on the X-ray fluorescent spectrometry with an initial concentration by means of the phosphomolybdic acid. The method is suitable for a quantitative determination of ethylene oxide adducts in a wide range of applicable concentrations, the trace-ones included. (M.Z.)

  17. Laser-induced fluorescence with an OPO system. Part II: direct determination of lead content in seawater by electrothermal atomization-laser-excited atomic fluorescence (ETA-LEAF).

    Science.gov (United States)

    Le Bihan, A; Lijour, Y; Giamarchi, P; Burel-Deschamps, L; Stephan, L

    2003-03-01

    Fluorescence was induced by coupling a laser with an optical parametric oscillator (OPO) to develop an analytical method for the direct determination of lead content, at ultra-trace level, in seawater by electrothermal atomization-laser-excited atomic fluorescence (ETA-LEAF). The optimization of atomization conditions, laser pulse energy, and mainly temporal parameters allowed us to reach a 3 fg detection limit (0.3 ng L(-1)) despite the low repetition rate of the device. The expected error on predicted concentrations of lead, at trace levels, in seawater was below 15%.

  18. Measurements of heavy quark and lepton lifetimes

    International Nuclear Information System (INIS)

    Jaros, J.A.

    1985-02-01

    The PEP/PETRA energy range has proved to be well-suited for the study of the lifetimes of hadrons containing the b and c quarks and the tau lepton for several reasons. First, these states comprise a large fraction of the total interaction rate in e + e - annihilation and can be cleanly identified. Second, the storage rings have operated at high luminosity and so produced these exotic states copiously. And finally, thanks to the interplay of the Fermi coupling strength, the quark and lepton masses, and the beam energy, the expected decay lengths are in the 1/2 mm range and so are comparatively easy to measure. This pleasant coincidence of cleanly identified and abundant signal with potentially large effects has made possible the first measurements of two fundamental weak couplings, tau → nu/sub tau/W and b → cW. These measurements have provided a sharp test of the standard model and allowed, for the first time, the full determination of the magnitudes of the quark mixing matrix. This paper reviews the lifetime studies made at PEP during the past year. It begins with a brief review of the three detectors, DELCO, MAC and MARK II, which have reported lifetime measurements. Next it discusses two new measurements of the tau lifetime, and briefly reviews a measurement of the D 0 lifetime. Finally, it turns to measurements of the B lifetime, which are discussed in some detail. 18 references, 14 figures, 1 table

  19. Lifetime and spin measurements in 40Ar

    International Nuclear Information System (INIS)

    Southon, J.

    1976-01-01

    Lifetimes of levels in 40 Ar populated by the 40 Ar(p,p') reaction have been measured using the Doppler shift attenuation method with a p-γ coincidence technique. A solid argon target was used. The lifetimes determined were (in psec.): 1461 keV level, 1.95 +- 0.15; 2121 keV, >25; 2524 keV, 0.53 +- 0.06; 2893 keV, 4.4 [+2.6,-1.3]; 3208 keV, 0.27. A comprehensive set of branching ratios was also derived and the spins and parities of the 3208 and 4481 keV states were determined to be 2 + and 1 +- respectively. Some of these results suggest that 2 particle -2 hole and 4 particle - 4 hole components are strongly mixed in the low-lying positive parity states in a manner similar to the 2 particle and 4 particle - 2 hole mixing that occurs in 42 Ca. An additional lifetime measurement for the recently discovered high spin state at 3464 keV was carried out using direct electronic timing. The level was excited by the 37 Cl(α,p) reaction and was found to have a lifetime of 1.00 +- 0.03 nsec, which taken together with other evidence indicates that its spin and parity are 6 + . The E2 transition strengths of the 40 Ar 6 + - 4 + - 2 + - 0 + cascade can be simply interpreted in terms of a weak coupling model. (author)

  20. Thorium determination by X-ray Fluorescence Spectrometry in simulated thorex process solutions

    International Nuclear Information System (INIS)

    Yamaura, M.; Matsuda, H.T.

    1989-01-01

    The X-ray fluorescence method for thorium determination in aqueous and organic (TBP-n-dodecane) solutions is described. The thin film-technique for sample preparation and a suitable internal standard have been used. Some parameters as analytical line, internal standard, filter paper, paper geometry, sample volume and measurement conditions were studied. Uranium, fission products, corrosion products and thorex reagent components were studied as interfering elements in the thorium analysis, as well as the matrix effect by using the thorex process simulated solutions the method to thorium determination in irradiated thorium solutions was applied. (M.J.C.) [pt

  1. COMPARISON OF TWO TEMPERATURE MEASUREMENT METHODS BY UPCONVERSION FLUORESCENCE SPECTRA OF ERBIUM-DOPED LEAD-FLUORIDE NANO-GLASS-CERAMICS

    Directory of Open Access Journals (Sweden)

    V. A. Aseev

    2015-05-01

    Full Text Available The study and compare of two temperature measurement methods is performed for the case of a lead-fluoride nano-glassceramics in the range from 317 to 423 K with a view to their application to temperature sensors. A method of temperature measurement by means of violet, green and red upconversion fluorescence spectra regression on latent structures and a method of temperature measurement by two fluorescence bands intensity ratio in green range are considered. It is shown that a four-dimensional space of latent structures is an optimum one in terms of temperature measurement accuracy. It made possible temperature determining with a relative error not larger than 0.15% at temperatures higher than 340 K by making use of fluorescence spectra training set with the step of 10 K. The method using two green bands fluorescence intensity ratio is inferior by the accuracy. Independence of pump power fluctuations is a significant advantage of the second method. To take advantage of the first method a stabilization of the pump power is necessary. The results of the work can be taken into account while developing optical temperature sensors with a better performance (in relation to accuracy and measurement range compared to existing ones which utilize temperature redistribution of fluorescence intensities in two closely-spaced bands or temperature dependence of fluorescence lifetime.

  2. Enhanced 3D fluorescence live cell imaging on nanoplasmonic substrate

    International Nuclear Information System (INIS)

    Gartia, Manas Ranjan; Hsiao, Austin; Logan Liu, G; Sivaguru, Mayandi; Chen Yi

    2011-01-01

    We have created a randomly distributed nanocone substrate on silicon coated with silver for surface-plasmon-enhanced fluorescence detection and 3D cell imaging. Optical characterization of the nanocone substrate showed it can support several plasmonic modes (in the 300-800 nm wavelength range) that can be coupled to a fluorophore on the surface of the substrate, which gives rise to the enhanced fluorescence. Spectral analysis suggests that a nanocone substrate can create more excitons and shorter lifetime in the model fluorophore Rhodamine 6G (R6G) due to plasmon resonance energy transfer from the nanocone substrate to the nearby fluorophore. We observed three-dimensional fluorescence enhancement on our substrate shown from the confocal fluorescence imaging of chinese hamster ovary (CHO) cells grown on the substrate. The fluorescence intensity from the fluorophores bound on the cell membrane was amplified more than 100-fold as compared to that on a glass substrate. We believe that strong scattering within the nanostructured area coupled with random scattering inside the cell resulted in the observed three-dimensional enhancement in fluorescence with higher photostability on the substrate surface.

  3. Enhanced 3D fluorescence live cell imaging on nanoplasmonic substrate

    Energy Technology Data Exchange (ETDEWEB)

    Gartia, Manas Ranjan [Department of Nuclear, Plasma and Radiological Engineering, University of Illinois, Urbana, IL 61801 (United States); Hsiao, Austin; Logan Liu, G [Department of Bioengineering, University of Illinois, Urbana, IL 61801 (United States); Sivaguru, Mayandi [Institute for Genomic Biology, University of Illinois, Urbana, IL 61801 (United States); Chen Yi, E-mail: loganliu@illinois.edu [Department of Electrical and Computer Engineering, University of Illinois, Urbana, IL 61801 (United States)

    2011-09-07

    We have created a randomly distributed nanocone substrate on silicon coated with silver for surface-plasmon-enhanced fluorescence detection and 3D cell imaging. Optical characterization of the nanocone substrate showed it can support several plasmonic modes (in the 300-800 nm wavelength range) that can be coupled to a fluorophore on the surface of the substrate, which gives rise to the enhanced fluorescence. Spectral analysis suggests that a nanocone substrate can create more excitons and shorter lifetime in the model fluorophore Rhodamine 6G (R6G) due to plasmon resonance energy transfer from the nanocone substrate to the nearby fluorophore. We observed three-dimensional fluorescence enhancement on our substrate shown from the confocal fluorescence imaging of chinese hamster ovary (CHO) cells grown on the substrate. The fluorescence intensity from the fluorophores bound on the cell membrane was amplified more than 100-fold as compared to that on a glass substrate. We believe that strong scattering within the nanostructured area coupled with random scattering inside the cell resulted in the observed three-dimensional enhancement in fluorescence with higher photostability on the substrate surface.

  4. Sensitization of uranium fluorescence using 2,6-pyridinedicarboxylic acid: Application for the determination of uranium in the presence of lanthanides

    Energy Technology Data Exchange (ETDEWEB)

    Maji, S. [Materials Chemistry Division, Chemistry Group, Indira Gandhi Centre for Atomic Research, Kalpakkam 603 102 (India); Viswanathan, K.S., E-mail: vish@igcar.gov.i [Materials Chemistry Division, Chemistry Group, Indira Gandhi Centre for Atomic Research, Kalpakkam 603 102 (India)

    2009-11-15

    The 2,6-pyridinedicarboxylic acid (PDA) has been shown to efficiently sensitize and enhance the fluorescence of uranium in aqueous medium. Interestingly, this ligand stabilizes the UO{sub 2}{sup 2+} species, which without the ligand is known to be in a negligible concentration, in aqueous medium at pH 6. The ligand sensitized enhancement of UO{sub 2}{sup 2+} fluorescence by PDA, provides an analytical tool for the determination of uranium at trace levels, in aqueous medium. Furthermore, PDA is also known to enhance the fluorescence of lanthanides; consequently, the simultaneous determination of uranium and lanthanides, using PDA as a fluorescence sensitizing agent, becomes a possibility, which has been demonstrated in this work. We have shown that the use of PDA yields detection limits of 2.2x10{sup -7} M for UO{sub 2}{sup 2+}, 1x10{sup -8} M for Tb{sup 3+} and 5x10{sup -9} M for Eu{sup 3+} in the simultaneous determination of these analytes.

  5. Micellar enhanced spectrofluorimetric methods: application to the determination of pyrene

    Energy Technology Data Exchange (ETDEWEB)

    Singh, H.; Hinze, W.L.

    1982-01-01

    The effects of cationic hexadecyltrimethylammonium chloride (CTAC), nonionic polyoxyethylene(9.5)p-1,1,3,3-tetramethylbutylphenol, Triton X-100 (TX-100), and anionic sodium dodecylsulfate (NaLS) surfactant micelles upon the spectrofluorimetric determination of pyrene is described. It was found that the intensity of the pyrene fluorescence is enhanced from 3 to 16 times in the presence of the micellar systems. Possible reasons for this micellar induced enhanced fluorescence are discussed. The spectral parameters, fluorescence lifetimes, quantum yields, lower detection limits, and analytical figures of merit for pyrene in CTAC, NaLS, TX-100, ethanol, and water are compared. The detection limit of pyrene in the presence of micelles (approx. 1.0 x 10/sup -10/ M) is about an order of magnitude lower than that possible in alcohol alone. A brief discussion on the predicted general applicability of this new technique in fluorimetric analysis is also given. 4 figures, 2 tables.

  6. Feasibility of the simultaneous determination of polycyclic aromatic hydrocarbons based on two-dimensional fluorescence correlation spectroscopy

    Science.gov (United States)

    Yang, Renjie; Dong, Guimei; Sun, Xueshan; Yang, Yanrong; Yu, Yaping; Liu, Haixue; Zhang, Weiyu

    2018-02-01

    A new approach for quantitative determination of polycyclic aromatic hydrocarbons (PAHs) in environment was proposed based on two-dimensional (2D) fluorescence correlation spectroscopy in conjunction with multivariate method. 40 mixture solutions of anthracene and pyrene were prepared in the laboratory. Excitation-emission matrix (EEM) fluorescence spectra of all samples were collected. And 2D fluorescence correlation spectra were calculated under the excitation perturbation. The N-way partial least squares (N-PLS) models were developed based on 2D fluorescence correlation spectra, showing a root mean square error of calibration (RMSEC) of 3.50 μg L- 1 and root mean square error of prediction (RMSEP) of 4.42 μg L- 1 for anthracene and of 3.61 μg L- 1 and 4.29 μg L- 1 for pyrene, respectively. Also, the N-PLS models were developed for quantitative analysis of anthracene and pyrene using EEM fluorescence spectra. The RMSEC and RMSEP were 3.97 μg L- 1 and 4.63 μg L- 1 for anthracene, 4.46 μg L- 1 and 4.52 μg L- 1 for pyrene, respectively. It was found that the N-PLS model using 2D fluorescence correlation spectra could provide better results comparing with EEM fluorescence spectra because of its low RMSEC and RMSEP. The methodology proposed has the potential to be an alternative method for detection of PAHs in environment.

  7. Highly fluorescent carbon dots as nanoprobes for sensitive and selective determination of 4-nitrophenol in surface waters

    International Nuclear Information System (INIS)

    Ahmed, Gaber Hashem Gaber; Laíño, Rosana Badía; Calzón, Josefa Angela García; García, Marta Elena Díaz

    2015-01-01

    We report on the synthesis of carbon dots (C-dots) by thermal carbonization of a mixture of ethyleneglycol bis-(2-aminoethyl ether)-N,N,N’,N’-tetraacetic acid (EGTA) and tris(hydroxymethyl)aminomethane (Tris). The resulting C-dots were characterized by X-ray diffraction, proton and carbon nuclear magnetic resonance, FTIR and fluorescence spectroscopy, and high-resolution TEM. The data reveal that the C-dots are mainly capped with hydroxy and carbonyl groups and are highly fluorescent with an emission peak that shifts from 427 to 438 nm if the excitation wavelength is increased from 310 to 360–370 nm. Fluorescence is quenched by 4-nitrophenol (4-NP), and this effect was exploited to design a simple and rapid protocol for the determination of 4-NP. The detection limit is 28 nM and the linear range extends from 0.1 to 50 μM. The method was successfully applied to the determination of 4-NP in spiked river and sea waters. (author)

  8. Simultaneous determination of arsenic and antimony by hydride generation atomic fluorescence spectrometry with dielectric barrier discharge atomizer

    Energy Technology Data Exchange (ETDEWEB)

    Xing Zhi [Department of Chemistry, Key Laboratory for Atomic and Molecular Nanosciences of the Education Ministry, Tsinghua University, Beijing 100084 (China); Kuermaiti, Biekesailike [Department of Chemistry, Key Laboratory for Atomic and Molecular Nanosciences of the Education Ministry, Tsinghua University, Beijing 100084 (China); Products Quality Inspection Institute, Yili, Xinjiang 835000 (China); Wang Juan; Han Guojun; Zhang Sichun [Department of Chemistry, Key Laboratory for Atomic and Molecular Nanosciences of the Education Ministry, Tsinghua University, Beijing 100084 (China); Zhang Xinrong, E-mail: xrzhang@mail.tsinghua.edu.cn [Department of Chemistry, Key Laboratory for Atomic and Molecular Nanosciences of the Education Ministry, Tsinghua University, Beijing 100084 (China)

    2010-12-15

    Simultaneous determination of As and Sb by hydride generation atomic fluorescence spectrometry was developed with the dielectric barrier discharge plasma as the hydride atomizer. The low-temperature and atmospheric-pressure micro-plasma was generated in a quartz cylindrical configuration device, which was constructed by an axial internal electrode and an outer electrode surrounding outside of the tube. The optimization of the atomizer construction and parameters for hydride generation and fluorescence detection systems were carried out. Under the optimized conditions, the detection limits for As and Sb were 0.04 and 0.05 {mu}g L{sup -1}, respectively. In addition, the applicability of the present method was confirmed by the detection of As and Sb in reference materials of quartz sandstone (GBW07106) and argillaceous limestone (GBW07108). The present work provided a new approach to exploit the miniaturized hydride generation dielectric barrier discharge atomic fluorescence spectrometry system for simultaneous multi-element determination.

  9. Simultaneous determination of arsenic and antimony by hydride generation atomic fluorescence spectrometry with dielectric barrier discharge atomizer

    International Nuclear Information System (INIS)

    Xing Zhi; Kuermaiti, Biekesailike; Wang Juan; Han Guojun; Zhang Sichun; Zhang Xinrong

    2010-01-01

    Simultaneous determination of As and Sb by hydride generation atomic fluorescence spectrometry was developed with the dielectric barrier discharge plasma as the hydride atomizer. The low-temperature and atmospheric-pressure micro-plasma was generated in a quartz cylindrical configuration device, which was constructed by an axial internal electrode and an outer electrode surrounding outside of the tube. The optimization of the atomizer construction and parameters for hydride generation and fluorescence detection systems were carried out. Under the optimized conditions, the detection limits for As and Sb were 0.04 and 0.05 μg L -1 , respectively. In addition, the applicability of the present method was confirmed by the detection of As and Sb in reference materials of quartz sandstone (GBW07106) and argillaceous limestone (GBW07108). The present work provided a new approach to exploit the miniaturized hydride generation dielectric barrier discharge atomic fluorescence spectrometry system for simultaneous multi-element determination.

  10. Fluorescence of sanguinarine: fundamental characteristics and analysis of interconversion between various forms.

    Science.gov (United States)

    Janovská, Marika; Kubala, Martin; Simánek, Vilím; Ulrichová, Jitka

    2009-09-01

    The quaternary isoquinoline alkaloid, sanguinarine (SG) plays an important role in both traditional and modern medicine, exhibiting a wide range of biological activities. Under physiological conditions, there is an equilibrium between the quaternary cation (SG+) and a pseudobase (SGOH) forms of SG. In the gastrointestinal tract, SG is converted to dihydrosanguinarine (DHSG). All forms exhibit bright fluorescence. However, their spectra overlap, which limited the use of powerful techniques based on fluorescence spectroscopy/microscopy. Our experiments using a combination of steady-state and time-resolved techniques enabled the separation of individual components. The results revealed that (a) the equilibrium constant between SG+ and SGOH is pKa = 8.06, while fluorescence of DHSG exhibited no changes in the pH range 5-12, (b) the SGOH has excitation/emission spectra with maxima at 327/418 nm and excited-state lifetime 3.2 ns, the spectra of the SG+ have maxima at 475/590 nm and excited-state lifetime 2.4 ns. The DHSG spectra have maxima at 327/446 nm and 2-exponential decay with components 4.2 and 2.0 ns, (c) NADH is able to convert SG to DHSG, while there is no apparent interaction between NADH and DHSG. These techniques are applicable for monitoring the SG to DHSG conversion in hepatocytes.

  11. Laser fluorescence on radioactive isotopes produced in very low yield

    International Nuclear Information System (INIS)

    Wells, S.A.; Evans, D.E.; Griffith, J.A.R.; Eastham, D.A.; Groves, J.; Tolfree, D.W.L.; Warner, D.D.; Dancy, M.P.; Billowes, J.; Grant, I.S.; Walker, P.M.

    1990-01-01

    Heavy ion accelerators such as the NSF at Daresbury Laboratory are capable of producing a wide variety of radio-active beams. The intensities of the beams of atoms or ions are always modest, and ultra-sensitive methods are needed to observe laser-induced fluorescence. The fast ion-photon coincidence technique has been applied to neutron-deficient barium ions down to 120 Ba. Nuclear moments and changes in charge radii have been determined from the measured hyperfine splittings and isotope shifts. An abrupt increase in the mean square radius is observed at 121 Ba, large enough to disrupt the systematic staggering seen for the isotopic series. The hyperfine structure has also been observed for an isomeric state of 127 Ba which has a lifetime of about 2 seconds. The measurements lead to an unambiguous assignment of the spin of the isomer. Another technique has been tested with stable krypton atoms. Fluorescent photons in the VUV wavelength region are detected with a high efficiency using a channel plate detector. The background is small enough that it should be possible to measure hyperfine spectra on beams with fewer than 10 3 atoms per second

  12. Detection of Colorectal Cancer by a Quantitative Fluorescence Determination of DNA Amplification in Stool

    Directory of Open Access Journals (Sweden)

    Daniele Calistri

    2004-09-01

    Full Text Available DNA amplification of exfoliated cells in stool repre sents an inexpensive and rapid test, but has only 50% to 60% sensitivity. A new quantitative method, calle( fluorescence long DNA, was developed and validate( in our laboratory on stool obtained from 86 patient., with primary colorectal cancer and from 62 health individuals. It consists of the amplification of stoo DNA with fluorescence primers and the quantification of the amplification using a standard curve. Results are arbitrarily expressed in nanograms. The potential of thi new method compared to the conventional approact was analyzed in a subgroup of 94 individuals (51 patients and 38 healthy volunteers. In the presen series, DNA amplification analysis showed a specific ity of 97% and a sensitivity of only 50%. Conversely fluorescence DNA evaluation, using the best cutoff o 25 ng, showed a sensitivity of about 76% and a spec ificity of 93%. Similar sensitivity was observed regard less of Dukes stage, tumor location, and size, thu., also permitting the detection of early-stage tumors The present study seems to indicate that quantitative fluorescence DNA determination in stool successfully identifies colorectal cancer patients with a sensitivity comparable, if not superior, to that of multiple gene analysis but at a lower cost and in a shorter time.

  13. Fluorescence enhancement effect for the determination of curcumin with yttrium(III)-curcumin-sodium dodecyl benzene sulfonate system

    International Nuclear Information System (INIS)

    Wang Feng; Huang Wei; Wang Yanwei

    2008-01-01

    It is found that the fluorescence of curcumin is greatly enhanced by yttrium(III) (Y 3+ ) in the presence of sodium dodecyl benzene sulfonate. Based on this, a sensitive fluorimetric method for the determination of curcumin in aqueous solution is proposed. In the potassium hydrogen phthalate (KHP) buffer, the fluorescence intensity of curcumin is proportional to the concentration of curcumin in the range of 7.37x10 -4 -0.18, 0.18-2.95 μg mL -1 and the detection limit is 0.1583 ng mL -1 . The actual samples are satisfactorily determined. In addition, the interaction mechanism is also studied

  14. Multipumping flow system for improving hydride generation atomic fluorescence spectrometric determinations

    International Nuclear Information System (INIS)

    Lopez-Garcia, Ignacio; Ruiz-Alcaraz, Irene; Hernandez-Cordoba, Manuel

    2006-01-01

    The advantages of using membrane micropumps rather than peristaltic pumps to introduce both sample and reagent solutions for hydride generation atomic fluorescence spectrometry are discussed. Arsenic was used as a test analyte to check the performance of the proposed manifold. Sample and reagent consumption was reduced 8-9 fold compared with continuous mode measurements made with peristaltic pumps, with no deterioration in sensitivity. The calibration graph was linear in the 0.05 to 2.5 μg l -1 As range using peak area as the analytical signal and maximum gain in the detector setting. A limit of detection (3σ) of 0.02 μg l -1 and relative standard deviation values close to 2% for 10 independent measurements of a 1 μg l -1 As solution were obtained. The sampling frequency increased from 45 to 102 h -1 with the subsequent saving in carrier gas used and reduction in wastes generated. The instrumental modification, which could be used for other elements currently determined by atomic fluorescence spectrometry, will permit hydride generators of more reduced dimensions to be constructed

  15. A new measurement of the Ξc+ lifetime

    Science.gov (United States)

    FOCUS Collaboration; Link, J. M.; Reyes, M.; Yager, P. M.; Anjos, J. C.; Bediaga, I.; Göbel, C.; Magnin, J.; Massafferri, A.; de Miranda, J. M.; Pepe, I. M.; Dos Reis, A. C.; Carrillo, S.; Casimiro, E.; Sánchez-Hernández, A.; Uribe, C.; Vázquez, F.; Cinquini, L.; Cumalat, J. P.; O'Reilly, B.; Ramirez, J. E.; Vaandering, E. W.; Butler, J. N.; Cheung, H. W. K.; Gaines, I.; Garbincius, P. H.; Garren, L. A.; Gottschalk, E.; Kasper, P. H.; Kreymer, A. E.; Kutschke, R.; Bianco, S.; Fabbri, F. L.; Zallo, A.; Cawlfield, C.; Kim, D. Y.; Rahimi, A.; Wiss, J.; Gardner, R.; Kryemadhi, A.; Chung, Y. S.; Kang, J. S.; Ko, B. R.; Kwak, J. W.; Lee, K. B.; Park, H.; Alimonti, G.; Boschini, M.; Chiodini, G.; D'Angelo, P.; DiCorato, M.; Dini, P.; Giammarchi, M.; Inzani, P.; Leveraro, F.; Malvezzi, S.; Menasce, D.; Mezzadri, M.; Milazzo, L.; Moroni, L.; Pedrini, D.; Pontoglio, C.; Prelz, F.; Rovere, M.; Sala, S.; Davenport, T. F.; Agostino, L.; Arena, V.; Boca, G.; Bonomi, G.; Gianini, G.; Liguori, G.; Merlo, M. M.; Pantea, D.; Ratti, S. P.; Riccardi, C.; Segoni, I.; Vitulo, P.; Hernandez, H.; Lopez, A. M.; Mendez, H.; Mendez, L.; Mirles, A.; Montiel, E.; Olaya, D.; Paris, A.; Quinones, J.; Rivera, C.; Xiong, W.; Zhang, Y.; Wilson, J. R.; Cho, K.; Handler, T.; Mitchell, R.; Engh, D.; Hosack, M.; Johns, W. E.; Nehring, M.; Sheldon, P. D.; Stenson, K.; Webster, M.; Sheaff, M.

    2001-12-01

    A precise determination of the charm-strange baryon Ξc+ lifetime is presented. The data were accumulated by the Fermilab high-energy photoproduction experiment FOCUS. The measurement is made with 300Ξc+-->Ξ- π+π+ decays, 130Ξc+-->Σ+K- π+ decays, 45Ξc+-->pK-π+ decays and 58Ξc+-->Λ0K- π+π+ decays. The Ξc+ lifetime is measured to be 0.439+/-0.022+/-0.009 ps.

  16. Statistical Analysis of Bending Rigidity Coefficient Determined Using Fluorescence-Based Flicker-Noise Spectroscopy.

    Science.gov (United States)

    Doskocz, Joanna; Drabik, Dominik; Chodaczek, Grzegorz; Przybyło, Magdalena; Langner, Marek

    2018-06-01

    Bending rigidity coefficient describes propensity of a lipid bilayer to deform. In order to measure the parameter experimentally using flickering noise spectroscopy, the microscopic imaging is required, which necessitates the application of giant unilamellar vesicles (GUV) lipid bilayer model. The major difficulty associated with the application of the model is the statistical character of GUV population with respect to their size and the homogeneity of lipid bilayer composition, if a mixture of lipids is used. In the paper, the bending rigidity coefficient was measured using the fluorescence-enhanced flicker-noise spectroscopy. In the paper, the bending rigidity coefficient was determined for large populations of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine and 1,2-dioleoyl-sn-glycero-3-phosphocholine vesicles. The quantity of obtained experimental data allows to perform statistical analysis aiming at the identification of the distribution, which is the most appropriate for the calculation of the value of the membrane bending rigidity coefficient. It has been demonstrated that the bending rigidity coefficient is characterized by an asymmetrical distribution, which is well approximated with the gamma distribution. Since there are no biophysical reasons for that we propose to use the difference between normal and gamma fits as a measure of the homogeneity of vesicle population. In addition, the effect of a fluorescent label and types of instrumental setups on determined values has been tested. Obtained results show that the value of the bending rigidity coefficient does not depend on the type of a fluorescent label nor on the type of microscope used.

  17. Prediction of the lifetime productive and reproductive performance of Holstein cows managed for different lactation durations, using a model of lifetime nutrient partitioning

    DEFF Research Database (Denmark)

    Gaillard, Charlotte; Martin, O; Blavy, P

    2016-01-01

    The GARUNS model is a lifetime performance model taking into account the changing physiological priorities of an animal during its life and through repeated reproduction cycles. This dynamic and stochastic model has been previously used to predict the productive and reproductive performance...... of various genotypes of cows across feeding systems. In the present paper, we used this model to predict the lifetime productive and reproductive performance of Holstein cows for different lactation durations, with the aim of determining the lifetime scenario that optimizes cows' performance defined...... by lifetime efficiency (ratio of total milk energy yield to total energy intake) and pregnancy rate. To evaluate the model, data from a 16-mo extended lactation experiment on Holstein cows were used. Generally, the model could consistently fit body weight, milk yield, and milk components of these cows...

  18. Measurement of the lifetime difference between Bs mass eigenstates

    International Nuclear Information System (INIS)

    Acosta, D.; The CDF Collaboration

    2005-01-01

    We present measurements of the lifetimes and polarization amplitudes for B s 0 → J/ψφ and B d 0 → J/ψ K* 0 decays. Lifetimes of the heavy (H) and light (L) mass eigenstates in the B s 0 system are separately measured for the first time by determining the relative contributions of amplitudes with definite CP as a function of the decay time

  19. Determination of small amounts of aluminum in iron and steel by x-ray fluorescence analysis

    International Nuclear Information System (INIS)

    Ito, Minao; Sato, Shoki; Narita, Masanao

    1981-01-01

    The accuracy of X-ray fluorescence analysis for determining aluminum in steel is generally poor when the aluminum content is below 0.1%. In this study, it is confirmed that the existence of inhomogeneous acid-insoluble aluminum causes the poor accuracy of X-ray fluorescence analysis. The accuracy can be improved by adding granular ferro-zirconium to molten steel when sampling. By using this method, the accuracy of determining aluminum was 0.0024% for low alloy steels and 0.0022% for stainless steels as the standard deviation obtained for a series of differences between an analytical result and a standard value. Corrections for overlapping of the chromium spectrum, and for absorption and enhancement effects of co-existent elements are not necessary for the analysis of low alloy steels, whereas these corrections are necessary for the analysis of stainless steels. (author)

  20. Determination of glutaredoxin enzyme activity and protein S-glutathionylation using fluorescent eosin-glutathione.

    Science.gov (United States)

    Coppo, Lucia; Montano, Sergio J; Padilla, Alicia C; Holmgren, Arne

    2016-04-15

    Glutaredoxins catalyze glutathione-dependent disulfide oxidoreductions, particularly reduction of glutathione (GSH)-protein mixed disulfides. Mammalian glutaredoxins are present in the cytosol/nucleus as Grx1 or in mitochondria as Grx2a. Here we describe di-eosin-glutathione disulfide (Di-E-GSSG) as a new tool to study glutaredoxin (Grx) activity. Di-E-GSSG has almost no fluorescence in its disulfide form due to self-quenching, whereas the reduced form (E-GSH) has a large fluorescence emission at 545 nm after excitation at 520 nm. Di-E-GSSG was a very poor substrate for glutathione reductase, but we discovered that the molecule was an excellent substrate for glutaredoxin in a coupled assay system with GSH, nicotinamide adenine dinucleotide phosphate (NADPH), and glutathione reductase or with lipoamide, NADH, and lipoamide dehydrogenase. In addition, Di-E-GSSG was used to glutathionylate the free SH group of bovine serum albumin (BSA), yielding eosin-glutathionylated BSA (E-GS-BSA) readily observed in ultraviolet (UV) light. E-GS-BSA also displayed a quenched fluorescence, and its Grx-catalyzed reduction could be followed by the formation of E-GSH by fluorescence emission using microtiter plates. This way of measuring Grx activity provided an ultrasensitive method that detected Grx1 and Grx2 at picomolar levels. Human Grx1 was readily quantified in 40 μl of plasma and determined to be 680 ± 208 pM in healthy controls. Copyright © 2016 Elsevier Inc. All rights reserved.

  1. Bias in the absorption coefficient determination of a fluorescent dye, standard reference material 1932 fluorescein solution

    International Nuclear Information System (INIS)

    DeRose, Paul C.; Kramer, Gary W.

    2005-01-01

    The absorption coefficient of standard reference material[registered] (SRM[registered]) 1932, fluorescein in a borate buffer solution (pH=9.5) has been determined at λ=488.0, 490.0, 490.5 and 491.0 nm using the US national reference UV/visible spectrophotometer. The purity of the fluorescein was determined to be 97.6% as part of the certification of SRM 1932. The solution measured was prepared gravimetrically by diluting SRM 1932 with additional borate buffer. The value of the absorption coefficient was corrected for bias due to fluorescence that reaches the detector and for dye purity. Bias due to fluorescence was found to be on the order of -1% for both monochromatic and polychromatic (e.g., diode-array based) spectrophotometers

  2. Hadronization, spin and lifetimes

    International Nuclear Information System (INIS)

    Grossman, Yuval; Nachshon, Itay

    2008-01-01

    Measurements of lifetimes can be done in two ways. For very short lived particles, the width can be measured. For long lived ones, the lifetime can be directly measured, for example, using a displaced vertex. Practically, the lifetime cannot be extracted for particles with intermediate lifetimes. We show that for such cases information about the lifetime can be extracted for heavy colored particles that can be produced with known polarization. For example, a t-like particle with intermediate lifetime hadronizes into a superposition of the lowest two hadronic states, T* and T (the equivalent of B* and B). Depolarization effects are governed by time scales that are much longer than the hadronization time scale, Λ QCD -1 . After a time of order 1/Δm, with Δm≡m(T*)-m(T), half of the initial polarization is lost. The polarization is totally lost after a time of order 1/Γ γ , with Γ γ = Γ(T* → Tγ). Thus, by comparing the initial and final polarization, we get information on the particle's lifetime.

  3. 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.

  4. 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...

  5. Instantaneous imaging of ozone in a gliding arc discharge using photofragmentation laser-induced fluorescence

    Science.gov (United States)

    Larsson, Kajsa; Hot, Dina; Gao, Jinlong; Kong, Chengdong; Li, Zhongshan; Aldén, Marcus; Bood, Joakim; Ehn, Andreas

    2018-04-01

    Ozone vapor, O3, is here visualized in a gliding arc discharge using photofragmentation laser-induced fluorescence. Ozone is imaged by first photodissociating the O3 molecule into an O radical and a vibrationally hot O2 fragment by a pump photon. Thereafter, the vibrationally excited O2 molecule absorbs a second (probe) photon that further transits the O2-molecule to an excited electronic state, and hence, fluorescence from the deexcitation process in the molecule can be detected. Both the photodissociation and excitation processes are achieved within one 248 nm KrF excimer laser pulse that is formed into a laser sheet and the fluorescence is imaged using an intensified CCD camera. The laser-induced signal in the vicinity of the plasma column formed by the gliding arc is confirmed to stem from O3 rather than plasma produced vibrationally hot O2. While both these products can be produced in plasmas a second laser pulse at 266 nm was utilized to separate the pump- from the probe-processes. Such arrangement allowed lifetime studies of vibrationally hot O2, which under these conditions were several orders of magnitude shorter than the lifetime of plasma-produced ozone.

  6. Determination of total iron in iron ore by x-ray fluorescence analysis using the Compton effect: comparison with others analytical techniques

    International Nuclear Information System (INIS)

    Castilho, M.V. de; Oliveira, R.C.

    1991-01-01

    Total iron in iron ores is determines by X-ray fluorescence analysis method using the compton effect. The Bragg angle is determined for compton no-coherent scattering related to K alpha of Rhodium. This measurement procedure can be used for best fitting of analytical results in X-ray fluorescence, when compared with others methods used for results corrections. (M.V.M.)

  7. Determination of human albumin in serum and urine samples by constant-energy synchronous fluorescence method.

    Science.gov (United States)

    Madrakian, Tayyebeh; Bagheri, Habibollah; Afkhami, Abbas

    2015-08-01

    A sensitive spectrofluorimetric method using constant-energy synchronous fluorescence technique is proposed for the determination of human albumin without separation. In this method, no reagent was used for enhancement of the fluorescence signal of albumin in the solution. Effects of some parameters, such as energy difference between excitation and emission monochromators (ΔE), emission and excitation slit widths and scan rate of wavelength were studied and the optimum conditions were established. For this purpose factorial design and response surface method were employed for optimization of the effective parameters on the fluorescence signal. The results showed that the scan rate of the wavelength has no significant effect on the analytical signal. The calibration curve was linear in the range 0.1-220.0 µg mL(-1) of albumin with a detection limit of 7.0 × 10(-3)  µg mL(-1). The relative standard deviations (RSD) for six replicate measurements of albumin were calculated as 2.2%, 1.7% and 1.3% for 0.5, 10.0 and 100.0 µg mL(-1) albumin, respectively. Furthermore the proposed method has been employed for the determination of albumin in human serum and urine samples. Copyright © 2014 John Wiley & Sons, Ltd.

  8. Light extraction efficiency enhancement for fluorescent SiC based white light-emitting diodes

    DEFF Research Database (Denmark)

    Ou, Haiyan; Ou, Yiyu; Argyraki, Aikaterini

    Fluorescent SiC based white light-emitting diodes(LEDs) light source, as an innovative energy-efficient light source, would even have longer lifetime, better light quality and eliminated blue-tone effect, compared to the current phosphor based white LED light source. In this paper, the yellow...

  9. Lifetime measurements and oscillator strengths in singly ionized scandium and the solar abundance of scandium

    Science.gov (United States)

    Pehlivan Rhodin, A.; Belmonte, M. T.; Engström, L.; Lundberg, H.; Nilsson, H.; Hartman, H.; Pickering, J. C.; Clear, C.; Quinet, P.; Fivet, V.; Palmeri, P.

    2017-12-01

    The lifetimes of 17 even-parity levels (3d5s, 3d4d, 3d6s and 4p2) in the region 57 743-77 837 cm-1 of singly ionized scandium (Sc II) were measured by two-step time-resolved laser induced fluorescence spectroscopy. Oscillator strengths of 57 lines from these highly excited upper levels were derived using a hollow cathode discharge lamp and a Fourier transform spectrometer. In addition, Hartree-Fock calculations where both the main relativistic and core-polarization effects were taken into account were carried out for both low- and high-excitation levels. There is a good agreement for most of the lines between our calculated branching fractions and the measurements of Lawler & Dakin in the region 9000-45 000 cm-1 for low excitation levels and with our measurements for high excitation levels in the region 23 500-63 100 cm-1. This, in turn, allowed us to combine the calculated branching fractions with the available experimental lifetimes to determine semi-empirical oscillator strengths for a set of 380 E1 transitions in Sc II. These oscillator strengths include the weak lines that were used previously to derive the solar abundance of scandium. The solar abundance of scandium is now estimated to logε⊙ = 3.04 ± 0.13 using these semi-empirical oscillator strengths to shift the values determined by Scott et al. The new estimated abundance value is in agreement with the meteoritic value (logεmet = 3.05 ± 0.02) of Lodders, Palme & Gail.

  10. Lifetime-management and lifetime-extension at PAKS nuclear power plant

    International Nuclear Information System (INIS)

    Katona, Tamas; Ratkai, Sandor; Janosi, Agnes Biro

    2002-01-01

    Paks Nuclear Power Plant provides 38-40% of domestic generation at lowest price. It has an important energy-policy role in Hungary. NPP Paks shall be a decisive and perspectively permanent element of the domestic electricity generation during the next two decades, which shall be ensured by plant safe operation, the lifetime extension and power uprating. Paks Nuclear Power Plant investigated the nuclear power plant's lifetime extension possibilities and alternatives, as well as technical and business feasibility of such alternatives. The feasibility study is based on the evaluation of a representative set of systems, structures and components, operational, test, in-service inspection and maintenance practice, experience and findings of the Periodic Safety Review. The most important results of this study showing the feasibility of 20 years lifetime extension is summarised in the paper. It was found that there are no technical or safety issues or limits, which may inhibit the operation of the Nuclear Power Plant Paks up to 50 years. In case of most systems and equipment the recent monitoring, maintenance and regular reconstruction practice of the NPP Paks allows the lifetime extension without outstanding cost. Replacement or reconstruction of a few equipment and systems requires significant investment costs. Material of reactor vessels of VVER/213 incorporated at Paks, compared to vessels of the similar units, is less sensitive to the embrittlement. At units 3-4 reactor vessels do not require any measure, consequently, any additional cost, even in case of a lifetime of 50 years. At unit 2 to extend the lifetime of the reactor vessel, only heating-up of emergency core cooling tanks is needed in order to decrease thermal stress levels caused by pressure thermal shock (PST) transients. For this purpose cost-effective technical solutions are available. At unit 1, beside the heating-up of the emergency core cooling tanks annealing of the welded joint No. 5/6 close to the

  11. The total lifetime health cost savings of smoking cessation to society

    DEFF Research Database (Denmark)

    Rasmussen, Gitte Susanne; Prescott, Eva; Sørensen, Thorkild I A

    2005-01-01

    Smoking cessation has major immediate and long-term health benefits. However, ex-smokers' total lifetime health costs and continuing smokers' costs remain uncompared, and hence the economic savings of smoking cessation to society have not been determined.......Smoking cessation has major immediate and long-term health benefits. However, ex-smokers' total lifetime health costs and continuing smokers' costs remain uncompared, and hence the economic savings of smoking cessation to society have not been determined....

  12. Silver content determination in mining and ore dressing using radionuclide-excited X-ray fluorescence analysis

    International Nuclear Information System (INIS)

    Koerner, B.; George, R.; Ratnikow, W.

    1989-01-01

    A laborsaving method based on radionuclide-excited X-ray fluorescence is described for the determination of silver in mining and ore dressing. Detection limits reached under operation conditions and measuring errors are discussed and compared with results from chemical analyses

  13. Determination of calcium and iron in limestone by X-ray fluorescence method

    International Nuclear Information System (INIS)

    Sovtsova, M.K.

    1977-01-01

    The results of determining calcium and iron content in limestone by X-ray fluorescence method are described. The 109 Cd isotape was chosen as a source for excitation, as it permited to reduce the concentration degeneration in the range of large Ca contents due to the larger energy of the primary radiation. The root-mean-square deviation from the data of chemical analysis was +-0.02%FeO and +-0.22%CaO

  14. Fluorescent carbon dots nanosensor for label-free determination of vitamin B12 based on inner filter effect

    Science.gov (United States)

    Ding, Longhua; Yang, Hongmei; Ge, Shenguang; Yu, Jinghua

    2018-03-01

    A simple and effective fluorescent assay for the determination of vitamin B12 was developed. In this study, carbon dots (CDs) were prepared by one-pot hydrothermal method and directly used as a fluorophore in the inner filter effect (IFE). Both of the maximum absorption peak of vitamin B12 and excitation maxima of CDs are located at 360 nm, hence, the excited light of CDs can be absorbed by vitamin B12, resulting in the fluorescence reduction of CDs. And the fluorescence intensity of CDs decreases with the increasing concentration of vitamin B12. This IFE-based sensing strategy shows a good linear relationship between the normalized fluorescence intensity and the concentration of vitamin B12 ranging from 0 to 60 μM, with a limit of detection (LOD) of 0.1 μM at a signal-to-noise ratio of 3. Furthermore, this proposed approach was successfully applied to vitamin B12 sensing in injections. This IFE sensing platform based on various fluorescent nanomaterials has a high promise for the detection of other biomolecules due to its inherent convenience.

  15. Positron lifetime studies of electron irradiated copper

    International Nuclear Information System (INIS)

    Hadnagy, T.D.

    1976-01-01

    Single-crystal copper was irradiated with 4.5-MeV electrons producing simple Frenkel defects as well as a significant concentration of divacancies. Mean positron lifetime characteristics, which are sensitive to the presence of vacancies and multivacancies in copper, was monitored after isochronal anneals between 80 and 800 0 K to determine the relative change of characteristic mean lifetimes and their associated intensities. Also a study of the dependence of the mean positron lifetime on the total electron fluence was made and compared with existing theories relating these lifetimes to vacancy or multivacancy concentrations. Numerical data from curve fitting procedures using a conventional trapping model for defect-induced changes in positron lifetimes indicate that upon irradiation with 4.5-MeV electrons at 80 0 K, about 8 percent of the defects produced are divacancy units. Divacancy units appear to be several times more effective in trapping positrons than are monovacancies. Further, the experimental data suggest that the stage III annealing processes in electron-irradiated copper most probably involve the motion and removal of both monovacancies and divacancies. A conglomerate (multivacancy) unit appears to exist as a stable entity even after annealing procedures are carried out at temperatures slightly above the stage III region. Such a stable unit could serve as a nucleation center for the appearance of voids

  16. Fluorescence Determination of Warfarin Using TGA-capped CdTe Quantum Dots in Human Plasma Samples.

    Science.gov (United States)

    Dehbozorgi, A; Tashkhourian, J; Zare, S

    2015-11-01

    In this study, some effort has been performed to provide low temperature, less time consuming and facile routes for the synthesis of CdTe quantum dots using ultrasound and water soluble capping agent thioglycolic acid. TGA-capped CdTe quantum dots were characterized through x-ray diffraction, transmission electron microscopy, Fourier transform infrared, ultraviolet-visible and fluorescence spectroscopy. The prepared quantum dots were used for warfarin determination based on the quenching of the fluorescence intensity in aqueous solution. Under the optimized conditions, the linear range of quantum dots fluorescence intensity versus the concentration of warfarin was 0.1-160.0 μM, with the correlation coefficient of 0.9996 and a limit of detection of 77.5 nM. There was no interference to coexisting foreign substances. The selectivity of the sensor was also tested and the results show that the developed method possesses a high selectivity for warfarin.

  17. A study of lifetime within the -12 sec range

    International Nuclear Information System (INIS)

    Dorobantu, V.

    1977-01-01

    Lifetimes of excited nuclear states are studied with a particular emphasis on the determination of nuclear lifetimes by means of the Doppler shift decomposition attenuation method - DSAM - which is based on the mathematical theory of stochastic processes; by its own nature, the emission of recoil nuclei, their motion through an amorphous medium and the detection of gamma rays resulting from these nuclei are all stochastic processes as well. Measurements of lifetimes together with other measurements can supply nuclear structure data, as well as information on the slowing-down process of an energetic ion through a certain material. The experimental tests have been carried out on the Tandem accelerator - IFIN, Bucharest. (author)

  18. A convenient method for determination of quizalofop-p-ethyl based on the fluorescence quenching of eosin Y in the presence of Pd(II)

    Science.gov (United States)

    Wu, Huan; Zhao, Yanmei; Tan, Xuanping; Zeng, Xiaoqing; Guo, Yuan; Yang, Jidong

    2017-03-01

    A convenient fluorescence quenching method for determination of Quizalofop-p-ethyl(Qpe) was proposed in this paper. Eosin Y(EY) is a red dye with strong green fluorescence (λex/λem = 519/540 nm). The interaction between EY, Pd(II) and Qpe was investigated by fluorescence spectroscopy, resonance Rayleigh scattering(RRS) and UV-Vis absorption. Based on changes in spectrum, Pd(II) associated with Qpe giving a positively charged chelate firstly, then reacted with EY through electrostatic and hydrophobic interaction formed ternary chelate could be demonstrated. Under optimum conditions, the fluorescence intensity of EY could be quenched by Qpe in the presence of Pd(II) and the RRS intensity had a remarkable enhancement, which was directly proportional to the Qpe concentration within a certain concentration range, respectively. Based on the fluorescence quenching of EY-Pd(II) system by Qpe, a novel, convenient and specific method for Qpe determination was developed. To our knowledge, this is the first fluorescence method for determination of Qpe was reported. The detection limit for Qpe was 20.3 ng/mL and the quantitative determination range was 0.04-1.0 μg/mL. The method was highly sensitive and had larger detection range compared to other methods. The influence of coexisting substances was investigated with good anti-interference ability. The new analytical method has been applied to determine of Qpe in real samples with satisfactory results.

  19. Measurement of the Lifetime of the $\\tau$ Lepton

    CERN Document Server

    Acciarri, M; Aguilar-Benítez, M; Ahlen, S P; Alpat, B; Alcaraz, J; Alemanni, G; Allaby, James V; Aloisio, A; Alverson, G; Alviggi, M G; Ambrosi, G; Anderhub, H; Andreev, V P; Angelescu, T; Anselmo, F; Antreasyan, D; Arefev, A; Azemoon, T; Aziz, T; Bagnaia, P; Baksay, L; Ball, R C; Banerjee, S; Banicz, K; Barillère, R; Barone, L; Bartalini, P; Baschirotto, A; Basile, M; Battiston, R; Bay, A; Becattini, F; Becker, U; Behner, F; Berdugo, J; Berges, P; Bertucci, B; Betev, B L; Bhattacharya, S; Biasini, M; Biland, A; Bilei, G M; Blaising, J J; Blyth, S C; Bobbink, Gerjan J; Böck, R K; Böhm, A; Borgia, B; Boucham, A; Bourilkov, D; Bourquin, Maurice; Boutigny, D; Branson, J G; Brigljevic, V; Brock, I C; Buffini, A; Buijs, A; Burger, J D; Burger, W J; Busenitz, J K; Buytenhuijs, A O; Cai, X D; Campanelli, M; Capell, M; Cara Romeo, G; Caria, M; Carlino, G; Cartacci, A M; Casaus, J; Castellini, G; Cavallari, F; Cavallo, N; Cecchi, C; Cerrada-Canales, M; Cesaroni, F; Chamizo-Llatas, M; Chan, A; Chang, Y H; Chaturvedi, U K; Chemarin, M; Chen, A; Chen, G; Chen, G M; Chen, H F; Chen, H S; Chen, M; Chiefari, G; Chien, C Y; Choi, M T; Cifarelli, Luisa; Cindolo, F; Civinini, C; Clare, I; Clare, R; Cohn, H O; Coignet, G; Colijn, A P; Colino, N; Costantini, S; Cotorobai, F; de la Cruz, B; Csilling, Akos; Dai, T S; D'Alessandro, R; De Asmundis, R; De Boeck, H; Degré, A; Deiters, K; Denes, P; De Notaristefani, F; DiBitonto, Daryl; Diemoz, M; Van Dierendonck, D N; Di Lodovico, F; Dionisi, C; Dittmar, Michael; Dominguez, A; Doria, A; Dorne, I; Dova, M T; Drago, E; Duchesneau, D; Duinker, P; Durán, I; Dutta, S; Easo, S; Efremenko, Yu V; El-Mamouni, H; Engler, A; Eppling, F J; Erné, F C; Ernenwein, J P; Extermann, Pierre; Fabre, M; Faccini, R; Falciano, S; Favara, A; Fay, J; Fedin, O; Felcini, Marta; Fenyi, B; Ferguson, T; Fernández, D; Ferroni, F; Fesefeldt, H S; Fiandrini, E; Field, J H; Filthaut, Frank; Fisher, P H; Forconi, G; Fredj, L; Freudenreich, Klaus; Furetta, C; Galaktionov, Yu; Ganguli, S N; García-Abia, P; Gau, S S; Gentile, S; Gerald, J; Gheordanescu, N; Giagu, S; Goldfarb, S; Goldstein, J; Gong, Z F; Gougas, Andreas; Gratta, Giorgio; Grünewald, M W; Gupta, V K; Gurtu, A; Gutay, L J; Hangarter, K; Hartmann, B; Hasan, A; Hatzifotiadou, D; Hebbeker, T; Hervé, A; Van Hoek, W C; Hofer, H; Hoorani, H; Hou, S R; Hu, G; Innocente, Vincenzo; Janssen, H; Jin, B N; Jones, L W; de Jong, P; Josa-Mutuberria, I; Kasser, A; Khan, R A; Kamyshkov, Yu A; Kapinos, P; Kapustinsky, J S; Karyotakis, Yu; Kaur, M; Kienzle-Focacci, M N; Kim, D; Kim, J K; Kim, S C; Kim, Y G; Kinnison, W W; Kirkby, A; Kirkby, D; Kirkby, Jasper; Kiss, D; Kittel, E W; Klimentov, A; König, A C; Korolko, I; Koutsenko, V F; Krämer, R W; Krenz, W; Kuijten, H; Kunin, A; Ladrón de Guevara, P; Landi, G; Lapoint, C; Lassila-Perini, K M; Laurikainen, P; Lebeau, M; Lebedev, A; Lebrun, P; Lecomte, P; Lecoq, P; Le Coultre, P; Lee Jae Sik; Lee, K Y; Leggett, C; Le Goff, J M; Leiste, R; Leonardi, E; Levchenko, P M; Li Chuan; Lieb, E H; Lin, W T; Linde, Frank L; Lista, L; Liu, Z A; Lohmann, W; Longo, E; Lu, W; Lü, Y S; Lübelsmeyer, K; Luci, C; Luckey, D; Luminari, L; Lustermann, W; Ma Wen Gan; Maity, M; Majumder, G; Malgeri, L; Malinin, A; Maña, C; Mangla, S; Marchesini, P A; Marin, A; Martin, J P; Marzano, F; Massaro, G G G; McNally, D; Mele, S; Merola, L; Meschini, M; Metzger, W J; Von der Mey, M; Mi, Y; Mihul, A; Van Mil, A J W; Mirabelli, G; Mnich, J; Molnár, P; Monteleoni, B; Moore, R; Morganti, S; Moulik, T; Mount, R; Müller, S; Muheim, F; Nagy, E; Nahn, S; Napolitano, M; Nessi-Tedaldi, F; Newman, H; Nippe, A; Nowak, H; Organtini, G; Ostonen, R; Pandoulas, D; Paoletti, S; Paolucci, P; Park, H K; Pascale, G; Passaleva, G; Patricelli, S; Paul, T; Pauluzzi, M; Paus, C; Pauss, Felicitas; Peach, D; Pei, Y J; Pensotti, S; Perret-Gallix, D; Petrak, S; Pevsner, A; Piccolo, D; Pieri, M; Pinto, J C; Piroué, P A; Pistolesi, E; Plyaskin, V; Pohl, M; Pozhidaev, V; Postema, H; Produit, N; Prokofev, D; Prokofiev, D O; 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    1996-01-01

    The lifetime of the tau lepton is measured using data collected in 1994 by the L3 detector at LEP. The precise track position information of the Silicon Microvertex Detector is exploited. The tau lepton lifetime is determined from the signed impact parameter distribution for 30 322 tau decays into one charged particle and from the decay length distribution for 3891 tau decays into three charged particles. Combining the two methods we obtain $\\tau_{\\tau}$ = 290.1 $\\pm$ 4.0 fs.

  20. The role of academic motivation in high school students' current and lifetime alcohol consumption: adopting a self-determination theory perspective.

    Science.gov (United States)

    Wormington, Stephanie V; Anderson, Kristen G; Corpus, Jennifer Henderlong

    2011-11-01

    The current study investigated the relationship between different types of academic motives-specifically, intrinsic motivation, introjected regulation, and external regulation-and high school students' current and lifetime alcohol consumption. One thousand sixty-seven high school students completed measures of academic motivation, other school-related factors, and lifetime and current alcohol consumption. Using structural equation modeling, different types of motivation and school-related factors were differentially related to student drinking. Specifically, intrinsic motivation was negatively related to lifetime and current alcohol consumption. External regulation, on the other hand, was positively associated with current drinking. Grade point average was the only school-related factor related to student alcohol use. These findings suggest that motivation is an important construct to consider in predicting students' alcohol use, even when other more commonly studied educational variables are considered. In addition, it supports the adoption of a motivation framework that considers different types of motivation in understanding the relationship between academic motivation and alcohol use. Suggestions for incorporating the self-determination model of motivation into studies of alcohol and substance use, as well as potential impacts on intervention efforts, are discussed. In particular, it may be important to foster only certain types of motivation, rather than all types of academically-focused motives, in efforts to deter alcohol use.