Science.gov

Sample records for fluorometers

  1. Pulse amplitude modulated chlorophyll fluorometer

    SciTech Connect

    Greenbaum, Elias; Wu, Jie

    2015-12-29

    Chlorophyll fluorometry may be used for detecting toxins in a sample because of changes in micro algae. A portable lab on a chip ("LOAC") based chlorophyll fluorometer may be used for toxin detection and environmental monitoring. In particular, the system may include a microfluidic pulse amplitude modulated ("PAM") chlorophyll fluorometer. The LOAC PAM chlorophyll fluorometer may analyze microalgae and cyanobacteria that grow naturally in source drinking water.

  2. Expedition automated flow fluorometer

    NASA Astrophysics Data System (ADS)

    Krikun, V. A.; Salyuk, P. A.

    2015-11-01

    This paper describes an apparatus and operation of automated flow-through dual-channel fluorometer for studying the fluorescence of dissolved organic matter, and the fluorescence of phytoplankton cells with open and closed reaction centers in sea areas with oligotrophic and eutrophic water type. The step-by step excitation by two semiconductor lasers or two light-emitting diodes is realized in the current device. The excitation wavelengths are 405nm and 532nm in the default configuration. Excitation radiation of each light source can be changed with different durations, intensities and repetition rate. Registration of the fluorescence signal carried out by two photo-multipliers with different optical filters of 580-600 nm and 680-700 nm band pass diapasons. The configuration of excitation sources and spectral diapasons of registered radiation can be changed due to decided tasks.

  3. Multiple protocol fluorometer and method

    DOEpatents

    Kolber, Zbigniew S.; Falkowski, Paul G.

    2000-09-19

    A multiple protocol fluorometer measures photosynthetic parameters of phytoplankton and higher plants using actively stimulated fluorescence protocols. The measured parameters include spectrally-resolved functional and optical absorption cross sections of PSII, extent of energy transfer between reaction centers of PSII, F.sub.0 (minimal), F.sub.m (maximal) and F.sub.v (variable) components of PSII fluorescence, photochemical and non-photochemical quenching, size of the plastoquinone (PQ) pool, and the kinetics of electron transport between Q.sub.a and PQ pool and between PQ pool and PSI. The multiple protocol fluorometer, in one embodiment, is equipped with an excitation source having a controlled spectral output range between 420 nm and 555 nm and capable of generating flashlets having a duration of 0.125-32 .mu.s, an interval between 0.5 .mu.s and 2 seconds, and peak optical power of up to 2 W/cm.sup.2. The excitation source is also capable of generating, simultaneous with the flashlets, a controlled continuous, background illumination.

  4. 21 CFR 862.2560 - Fluorometer for clinical use.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...) MEDICAL DEVICES CLINICAL CHEMISTRY AND CLINICAL TOXICOLOGY DEVICES Clinical Laboratory Instruments § 862.2560 Fluorometer for clinical use. (a) Identification. A fluorometer for clinical use is a device... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Fluorometer for clinical use. 862.2560 Section...

  5. 21 CFR 862.2560 - Fluorometer for clinical use.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...) MEDICAL DEVICES CLINICAL CHEMISTRY AND CLINICAL TOXICOLOGY DEVICES Clinical Laboratory Instruments § 862.2560 Fluorometer for clinical use. (a) Identification. A fluorometer for clinical use is a device... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Fluorometer for clinical use. 862.2560 Section...

  6. The Fraunhofer line discriminator: An airborne fluorometer

    NASA Technical Reports Server (NTRS)

    Stoertz, G. E.

    1969-01-01

    An experimental Fraunhofer Line Discriminator (FLD) can differentiate and measure solar-stimulated luminescence when viewed against a background of reflected light. Key elements are two extremely sensitive photomultipliers, two glass-spaced Fabry-Perot filters having a bandwidth less than 1 A, and an analog computer. As in conventional fluorometers, concentration of a fluorescent substance is measured by comparison with standards. Quantitative use is probably accurate only at low altitudes but detection of luminescent substances should be possible from any altitude. Applications of the present FLD include remote sensing of fluorescent dyes used in studies of current dynamics. The basic technique is applicable to detection of oil spills, monitoring of pollutants, and sensing over land areas.

  7. Portable and modularized fluorometer based on optical fiber

    NASA Astrophysics Data System (ADS)

    Yue, WeiWei; Zhang, Lei; Guo, ZhenYa; Jiang, ShouZhen; Bai, ChengJie

    2015-02-01

    A portable and modularized fluorometer based on optical fiber was proposed in this work. The fluorometer included a light emitter diode (LED) light source module (LSM), a sample cell module (SCM), an optical-electrical converter module (OCM) and a signal process module (SAM). The LEDs in LSM were driven by a constant current source to provide stable exciting light with different wavelength. The OCM included a modularized optical filter and used a photomultiplier tube (PMT) to detect fluorescence signal. The SCM was used to locate sample cuvette and could be connected by optical fibers with the LSM and OCM. Via modularized design, the LSM and OCM could both selected and replaced based on different fluorescence dyes. In order to improve the detecting dynamic range of the fluorometer, the SAM could control the light intensity of LED source in LSM, to control the gain of PMT in OCM, and particularly, four channel signal acquisition circuits with different gain were constructed to collect fluorescence signal simultaneously. Fluorescein isothiocyanate (FITC) was selected as sample to test the fluorometer. The fluorometer has shown a high sensitivity with FITC concentration of 10ng/mL and presented a good linearity from 10 ng/mL to 10 μg/mL.

  8. Three color laser fluorometer for studies of phytoplankton fluorescence

    NASA Technical Reports Server (NTRS)

    Phinney, David A.; Yentsch, C. S.; Rohrer, J.

    1988-01-01

    A three-color laser fluorometer has been developed for field work operations. Using two tunable dye lasers (excitation wavelengths at 440 nm and 530 nm), broadband wavelength optical filters were selected to obtain maximum fluorescence sensitivity at wavelengths greater than 675 nm (chlorophyll) and 575 + or - 15 nm (phycoerythrin). The laser fluorometer permits the measurement of phytoplankton pigments under static or flowing conditions and more closely resembles the time scales (ns) and energy levels (mW) of other laser-induced fluorescence instruments.

  9. 21 CFR 862.2560 - Fluorometer for clinical use.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Fluorometer for clinical use. 862.2560 Section 862.2560 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES CLINICAL CHEMISTRY AND CLINICAL TOXICOLOGY DEVICES Clinical Laboratory Instruments §...

  10. 21 CFR 862.2560 - Fluorometer for clinical use.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Fluorometer for clinical use. 862.2560 Section 862.2560 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED... radiating, when illuminated, a light of a different wavelength. This device is used in conjunction...

  11. High-Resolution Fluorometer for Mapping Microscale Phytoplankton Distributions

    PubMed Central

    Doubell, Mark J.; Seuront, Laurent; Seymour, Justin R.; Patten, Nicole L.; Mitchell, James G.

    2006-01-01

    A new high-resolution, in situ profiling fluorometer maps fluorescence distributions with a spatial resolution of 0.5 to 1.5 mm to a depth of 70 m in the open ocean. We report centimeter-scale patterns for phytoplankton distributions associated with gradients exhibiting 10- to 30-fold changes in fluorescence in contrasting marine ecosystems. PMID:16751572

  12. High-resolution fluorometer for mapping microscale phytoplankton distributions.

    PubMed

    Doubell, Mark J; Seuront, Laurent; Seymour, Justin R; Patten, Nicole L; Mitchell, James G

    2006-06-01

    A new high-resolution, in situ profiling fluorometer maps fluorescence distributions with a spatial resolution of 0.5 to 1.5 mm to a depth of 70 m in the open ocean. We report centimeter-scale patterns for phytoplankton distributions associated with gradients exhibiting 10- to 30-fold changes in fluorescence in contrasting marine ecosystems. PMID:16751572

  13. Improved Underwater Excitation-Emission Matrix Fluorometer

    NASA Technical Reports Server (NTRS)

    Moore, Casey; daCunha, John; Rhoades, Bruce; Twardowski, Michael

    2007-01-01

    A compact, high-resolution, two-dimensional excitation-emission matrix fluorometer (EEMF) has been designed and built specifically for use in identifying and measuring the concentrations of organic compounds, including polluting hydrocarbons, in natural underwater settings. Heretofore, most EEMFs have been designed and built for installation in laboratories, where they are used to analyze the contents of samples collected in the field and brought to the laboratories. Because the present EEMF can be operated in the field, it is better suited to measurement of spatially and temporally varying concentrations of substances of interest. In excitation-emission matrix (EEM) fluorometry, fluorescence is excited by irradiating a sample at one or more wavelengths, and the fluorescent emission from the sample is measured at multiple wavelengths. When excitation is provided at only one wavelength, the technique is termed one-dimensional (1D) EEM fluorometry because the resulting matrix of fluorescence emission data (the EEM) contains only one row or column. When excitation is provided at multiple wavelengths, the technique is termed two-dimensional (2D) EEM fluorometry because the resulting EEM contains multiple rows and columns. EEM fluorometry - especially the 2D variety - is well established as a means of simultaneously detecting numerous dissolved and particulate compounds in water. Each compound or pool of compounds has a unique spectral fluorescence signature, and each EEM is rich in information content, in that it can contain multiple fluorescence signatures. By use of deconvolution and/or other mixture-analyses techniques, it is often possible to isolate the spectral signature of compounds of interest, even when their fluorescence spectra overlap. What distinguishes the present 2D EEMF over prior laboratory-type 2D EEMFs are several improvements in packaging (including a sealed housing) and other aspects of design that render it suitable for use in natural underwater

  14. Photon-counting phase-modulation fluorometer for lifetime measurements

    NASA Astrophysics Data System (ADS)

    Iwata, Tetsuo; Hori, Akio; Kamada, Takeshi

    2001-05-01

    We propose a phase-modulation fluorometer that is applicable to a very weak fluorescence intensity level. In order to counter the single-photon event situation, we have introduced a combination of a time-to-amplitude converter (TAC) and a pulse height analyzer (PHA) to the phase- modulation fluorometer, the combination of which is usually used in the single-photon correlation method to measure fluorescence decay waveforms by pulsed excitation. In the proposed fluorometer, a sinusoidal response waveform that is shifted in phase over the reference one is obtained statistically as a histogram in the PHA memory and then the fluorescence lifetime can be calculated by the same procedure as the conventional analog phase-modulation method. The excitation light source used was a current- modulated ultraviolet light-emitting diode (UV LED), whose center wavelength was 370 nm and its spectral bandwidth was 10 nm. Fluorescence lifetimes of 17.6 ns and 5.7 ns obtained for 10 ppb quinine sulfate in 0.1 N H2SO4 and for 10 ppb rhodamine 6G in ethanol, respectively, agreed well with those reported in the literature.

  15. FORTRAN PROCESSING OF FLUOROMETRIC DATA LOGGED BY A TURNER DESIGNS FIELD FLUOROMETER

    EPA Science Inventory

    Continuous recording of dye fluorescence using field fluorometers at selected sampling sites facilitate acquisition of real-time dye-tracing data. The Turner Designs Model 10-AU-005 Field Fluorometer allows for frequent fluorescence readings, data logging, and easy downloading t...

  16. Computer controlled fluorometer device and method of operating same

    DOEpatents

    Kolber, Z.; Falkowski, P.

    1990-07-17

    A computer controlled fluorometer device and method of operating same, said device being made to include a pump flash source and a probe flash source and one or more sample chambers in combination with a light condenser lens system and associated filters and reflectors and collimators, as well as signal conditioning and monitoring means and a programmable computer means and a software programmable source of background irradiance that is operable according to the method of the invention to rapidly, efficiently and accurately measure photosynthetic activity by precisely monitoring and recording changes in fluorescence yield produced by a controlled series of predetermined cycles of probe and pump flashes from the respective probe and pump sources that are controlled by the computer means. 13 figs.

  17. Computer controlled fluorometer device and method of operating same

    DOEpatents

    Kolber, Zbigniew; Falkowski, Paul

    1990-01-01

    A computer controlled fluorometer device and method of operating same, said device being made to include a pump flash source and a probe flash source and one or more sample chambers in combination with a light condenser lens system and associated filters and reflectors and collimators, as well as signal conditioning and monitoring means and a programmable computer means and a software programmable source of background irradiance that is operable according to the method of the invention to rapidly, efficiently and accurately measure photosynthetic activity by precisely monitoring and recording changes in fluorescence yield produced by a controlled series of predetermined cycles of probe and pump flashes from the respective probe and pump sources that are controlled by the computer means.

  18. Construction of a Fourier-transform phase-modulation fluorometer

    NASA Astrophysics Data System (ADS)

    Shibata, Hironobu; Iwata, Tetsuo

    2005-12-01

    We have constructed a Fourier-transform phase-modulation fluorometer (FT-PMF) by which a fluorescence decay waveform can be obtained. In the FT-PMF, the modulation frequency of the excitation light source is swept continuously from a direct current (dc) to a high frequency f max with a time duration T. The resultant fluorescence signal waveform is Fourier-transformed to obtain its amplitude and phase spectra. The ratio of the amplitude spectrum and the difference of the phase spectrum over those of the reference spectra that are obtained from a non-fluorescent material are calculated, respectively, and the pair of both spectral data is inverse-Fourier-transformed again to obtain the fluorescence decay waveform. The light source used was an ultraviolet light emitting- diode (UV LED) whose typical operating condition was f max = 100 MHz and T = 10 μs. To demonstrate the performance of the FT-PMF, we carried out (1) measurement of a fluorescent decay waveform of YAG materials packed in a white LED, and (2) determination of fluorescence lifetime of 10 ppm quinine sulfate in 0.1N H IISO 4.

  19. Construction of a Fourier-transform phase-modulation fluorometer

    NASA Astrophysics Data System (ADS)

    Iwata, Tetsuo; Shibata, Hironobu; Araki, Tsutomu

    2005-11-01

    We have constructed a Fourier-transform phase-modulation fluorometer (FT-PMF) by which a fluorescence decay waveform can be obtained. In the FT-PMF, the modulation frequency of the excitation light source is swept continuously from a direct current (dc) to a high frequency fmax with a time duration T. The resultant fluorescence signal waveform is Fourier transformed to obtain its amplitude and phase spectra. The ratio of the amplitude spectrum and the difference of the phase spectrum over those of the reference spectra from an excitation waveform are calculated, respectively, and the pair of both spectral data is inverse-Fourier-transformed again to obtain the fluorescence decay waveform. The light source used was an ultraviolet light-emitting diode (UV LED) whose operating condition was fmax = 50-120 MHz and T = 10 µs. To demonstrate the performance of the FT-PMF, we carried out (1) the measurement of a fluorescent decay waveform of YAG materials enclosed in a white LED and (2) determinations of fluorescence lifetimes of 10 ppm quinine sulfate in 0.1 N H2SO4 and 10 ppm rhodamine 6G in ethanol.

  20. Versatile portable fluorometer for time-resolved luminescence analysis

    NASA Astrophysics Data System (ADS)

    Chen, Guoying

    2005-06-01

    A robust, filter-based portable fluorometer was designed, prototyped, and tested for time-resolved luminescence (TRL) analysis. Its flexible optical design allows interchangeable configurations to support three measurement modes: liquid-phase TRL using a sample cuvette, solid-matrix TRL using a sorbent strip, and evanescent-field TRL using a quartz-rod waveguide. A xenon flashlamp is used as the light source and a photomultiplier tube (PMT) as the photodetector. A gating technique was implemented to overcome PMT saturation by the intense xenon lamp flash, therefore higher gains can be set to measure weak luminescence signals. The TRL signal is digitized at a 4μs time resolution and a 12bit amplitude resolution. Individual flashes were monitored by a photodiode and its current was integrated to compensate for source light fluctuation. Using tetracycline as a model analyte, a 0.025ppb limit of detection (LOD) with a typical 2% relative standard deviation, and a 3 orders of magnitude (0.5-300ppb) linear dynamic range (r2=0.9996) were achieved.

  1. A vertical-axis transmission-type filter-fluorometer for solutions

    USGS Publications Warehouse

    Fletcher, M.H.

    1963-01-01

    A vertical-axis transmission-type filter-fluorometer for solutions is described and scale drawings are presented. Data from studies of several chemical-fluorescence systems show that filters effectively isolate fluorescence from exciting energy. Other data illustrate the relationships between fluorescence intensity and solution depth.

  2. Fiber-Optic Fluorometer for Microscale Mapping of Photosynthetic Pigments in Microbial Communities

    PubMed Central

    Thar, Roland; Kühl, Michael; Holst, Gerhard

    2001-01-01

    Microscale fluorescence measurements were performed in photosynthetic biofilms at a spatial resolution of 100 to 200 μm with a new fiber-optic fluorometer which allowed four different excitation and emission wavelengths and was configured for measuring phycobiliproteins, chlorophylls, and bacteriochlorophylls. We present details of the measuring system and describe examples of applications in different microbial communities. PMID:11375200

  3. A compact frequency domain fluorometer with a directly modulated deuterium light source

    NASA Astrophysics Data System (ADS)

    Morgan, C. G.; Hua, Y.; Mitchell, A. K.; Murray, J. G.; Boardman, A. D.

    1996-01-01

    A phase fluorometer based on a low-cost and versatile high-frequency modulated light source and a fast gain-modulated photomultiplier is described. The apparatus is particularly well-suited to high-sensitivity frequency-domain fluorescence measurements requiring ultraviolet excitation. The system is very compact since it features a directly modulated light source, a miniature photomultiplier tube, and an rf synthesizer on a PC board. Equipped with a suitable fiber optic probe sensor, the device has potential as a portable unit for a wide range of remote sensing applications. The lamp can be modulated at frequencies up to 120 MHz and the phase fluorometer has been tested at up to 70 MHz with a range of fluorescent lifetime standards containing quinine sulfate quenched with sodium chloride.

  4. Linking fluorescence spectroscopy to the scale of spectral sensitivity: the BAM reference fluorometer

    NASA Astrophysics Data System (ADS)

    Monte, Christian; Pilz, Walter; Resch-Genger, Ute

    2005-08-01

    Providing fluorescence and fluorescence excitation spectra traceable to the scale of spectral sensitivity (responsivity) and spectral radiance at minimized uncertainty is currently limited by two factors: The uncertainty of the available transfer standards and the uncertainty of the measurement process itself. Here the requirements on a reference fluorometer enabling measurements at minimized uncertainty, its design, the simulation and the realization are presented. The fluorometer is designed with minimized chromatic and geometrical aberrations. To realize an efficient reduction of stray light and subtractive dispersion a double monochromator design was necessary. The basic element is a so-called U-type Czerny-Turner single monochromator featuring off-axis parabolas and an entrance and exit slit virtually at the same place. Thereby spherical aberration, coma and astigmatism are effectively minimized. The here employed special double monochromator design further cancels the remaining aberrations of the single monochromator. The design of the whole spectrometer was optimized with a ray tracing program. To minimize uncertainties due to the transfer standards, the reference fluorometer is exclusively traceable to the spectral sensitivity (responsivity) scale. This enables the use of transfer standards with much smaller uncertainty. Here trap detectors are employed of common design but specially calibrated for a divergent light bundle. Based on this instrument with its achromatic design and precisely known numerical apertures the determination of absolute fluorescence spectra will be addressed.

  5. Fast repetition rate (FRR) fluorometer and method for measuring fluorescence and photosynthetic parameters

    DOEpatents

    Kolber, Z.; Falkowski, P.

    1995-06-20

    A fast repetition rate fluorometer device and method for measuring in vivo fluorescence of phytoplankton or higher plants chlorophyll and photosynthetic parameters of phytoplankton or higher plants is revealed. The phytoplankton or higher plants are illuminated with a series of fast repetition rate excitation flashes effective to bring about and measure resultant changes in fluorescence yield of their Photosystem II. The series of fast repetition rate excitation flashes has a predetermined energy per flash and a rate greater than 10,000 Hz. Also, disclosed is a flasher circuit for producing the series of fast repetition rate flashes. 14 figs.

  6. Fast repetition rate (FRR) fluorometer and method for measuring fluorescence and photosynthetic parameters

    DOEpatents

    Kolber, Zbigniew; Falkowski, Paul

    1995-06-20

    A fast repetition rate fluorometer device and method for measuring in vivo fluorescence of phytoplankton or higher plants chlorophyll and photosynthetic parameters of phytoplankton or higher plants by illuminating the phytoplankton or higher plants with a series of fast repetition rate excitation flashes effective to bring about and measure resultant changes in fluorescence yield of their Photosystem II. The series of fast repetition rate excitation flashes has a predetermined energy per flash and a rate greater than 10,000 Hz. Also, disclosed is a flasher circuit for producing the series of fast repetition rate flashes.

  7. Detection of water quality parameters in Hangzhou Bay using a portable laser fluorometer.

    PubMed

    Chen, Peng; Pan, Delu; Mao, Zhihua; Tao, Bangyi

    2015-04-15

    A field, light-weight laser fluorometer based on the method of laser induced fluorescence was developed for water quality monitoring. The basic instrument configuration uses a high pulse repetition frequency microchip laser, a confocal reflective fluorescent probe and a broadband hyperspectral micro spectrometer; it weights only about 1.7 kg. Simultaneous estimates of three important water quality parameters, namely, chlorophyll a (chl-a), colored dissolved organic matter (CDOM), and total suspended matter (TSM) measured by the laser fluorometer were observed to agree well with those measured by traditional methods (0.27-0.84 μg L(-3) chl-a, R(2)=0.88; 0.104-0.295 m(-)(1) CDOM absorption, R(2)=0.90; and 59.8-994.9 mg L(-)(3) TSM, R(2)=0.86) in Hangzhou Bay water. Subsequently, distribution and characteristics of CDOM and chl-a laser fluorescence in Hangzhou Bay were analyzed, which will enhance our understanding of biogeochemical processes in this complex estuarine system at high-resolution, high-frequency and long-term scale. PMID:25697817

  8. Methods and Best Practice to Intercompare Dissolved Oxygen Sensors and Fluorometers/Turbidimeters for Oceanographic Applications

    PubMed Central

    Pensieri, Sara; Bozzano, Roberto; Schiano, M. Elisabetta; Ntoumas, Manolis; Potiris, Emmanouil; Frangoulis, Constantin; Podaras, Dimitrios; Petihakis, George

    2016-01-01

    In European seas, ocean monitoring strategies in terms of key parameters, space and time scale vary widely for a range of technical and economic reasons. Nonetheless, the growing interest in the ocean interior promotes the investigation of processes such as oxygen consumption, primary productivity and ocean acidity requiring that close attention is paid to the instruments in terms of measurement setup, configuration, calibration, maintenance procedures and quality assessment. To this aim, two separate hardware and software tools were developed in order to test and simultaneously intercompare several oxygen probes and fluorometers/turbidimeters, respectively in the same environmental conditions, with a configuration as close as possible to real in-situ deployment. The chamber designed to perform chlorophyll-a and turbidity tests allowed for the simultaneous acquisition of analogue and digital signals of several sensors at the same time, so it was sufficiently compact to be used in both laboratory and onboard vessels. Methodologies and best practice committed to the intercomparison of dissolved oxygen sensors and fluorometers/turbidimeters have been used, which aid in the promotion of interoperability to access key infrastructures, such as ocean observatories and calibration facilities. Results from laboratory tests as well as field tests in the Mediterranean Sea are presented. PMID:27196908

  9. Development of a multi-sensor in situ fiber optic fluorometer

    SciTech Connect

    Lohrenz, S.E.; Asper, V.L. . Center for Marine Sciences); Morris, M.J.; Walters, R.A. )

    1992-12-01

    Objective is to develop and evaluate a multi-sensor in situ fiber optic fluorometer. The instrument is designed to sample and store in vivo strobe-stimulated fluorescence data at multiple depths and high frequencies (1 Hz). This information may be used for estimating the distribution and abundance of particulate pigment biomass, for supporting models of water column primary production and as a complement to remotely sensed ocean color estimates of pigment biomass. The instrument is unique in that it uses fiber optic technology to increase vertical resolution. While it is theoretically possible to accomplish this task using a large number of commercially available fluorometers, our proposed design would provide a less expensive approach. A laboratory prototype has been built and is being tested. Preliminary results indicate that the instrument meets all the project goals and that low cost, high frequency, high spatial resolution chlorophyll data are obtainable with the current design. Further work is required to develop the seagoing version, and optimize the configuration of the fiber sensors.

  10. Methods and Best Practice to Intercompare Dissolved Oxygen Sensors and Fluorometers/Turbidimeters for Oceanographic Applications.

    PubMed

    Pensieri, Sara; Bozzano, Roberto; Schiano, M Elisabetta; Ntoumas, Manolis; Potiris, Emmanouil; Frangoulis, Constantin; Podaras, Dimitrios; Petihakis, George

    2016-01-01

    In European seas, ocean monitoring strategies in terms of key parameters, space and time scale vary widely for a range of technical and economic reasons. Nonetheless, the growing interest in the ocean interior promotes the investigation of processes such as oxygen consumption, primary productivity and ocean acidity requiring that close attention is paid to the instruments in terms of measurement setup, configuration, calibration, maintenance procedures and quality assessment. To this aim, two separate hardware and software tools were developed in order to test and simultaneously intercompare several oxygen probes and fluorometers/turbidimeters, respectively in the same environmental conditions, with a configuration as close as possible to real in-situ deployment. The chamber designed to perform chlorophyll-a and turbidity tests allowed for the simultaneous acquisition of analogue and digital signals of several sensors at the same time, so it was sufficiently compact to be used in both laboratory and onboard vessels. Methodologies and best practice committed to the intercomparison of dissolved oxygen sensors and fluorometers/turbidimeters have been used, which aid in the promotion of interoperability to access key infrastructures, such as ocean observatories and calibration facilities. Results from laboratory tests as well as field tests in the Mediterranean Sea are presented. PMID:27196908

  11. Development of miniaturized submersible fluorometers for the detection of aromatic hydrocarbons in marine waters

    NASA Astrophysics Data System (ADS)

    Tedetti, Marc; Bachet, Caroline; Joffre, Pascal; Ferretto, Nicolas; Guigue, Catherine; Goutx, Madeleine

    2014-05-01

    Polycyclic aromatic hydrocarbons (PAHs) are among the most widespread organic contaminants in aquatic environments. Due to their physico-chemical properties, PAHs are persistent and mobile, can strongly bioaccumulate in food chains and are harmful to living organisms. They are thus recognized by various international organizations as priority contaminants and are included in the list of 45 priority regulated substances by the European Union. Because of their aromatic structure, PAHs are "optically active" and have inherent fluorescence properties in the ultraviolet (UV) spectral domain (200-400 nm). Therefore, UV fluorescence spectroscopy has been successfully used to develop PAH sensors (i.e. UV fluorometers). Currently, five UV submersible fluorometers are commercially available for in situ measurements of PAHs: EnviroFlu-HC (TriOS Optical Sensors, Germany), Hydrocarbon Fluorometer (Sea & Sun Technology, Germany), HydroC ™ / PAH (CONTROS, Germany), UviLux AquaTracka (Chelsea Technology Group, UK) and Cyclops-7 (Turner Designs, US). These UV fluorometers are all dedicated to the measurement of phenanthrene (λEx /λEm: 255/360 nm), one of the most abundant and fluorescent PAHs found in the aquatic environment. In this study, we developed original, miniaturized submersible fluorometers based on deep UV light-emitting diodes (LEDs) for simultaneous measurements of two PAHs of interest: the MiniFluo-UV 1 for the detection of phenanthrene (PHE, at λEx /λEm: 255/360 nm) and naphthalene (NAP, at λEx /λEm: 270/340 nm), and the MiniFluo-UV 2 for the detection of fluorene (FLU, at λEx /λEm: 255/315 nm) and pyrene (PYR, at λEx /λEm: 270/380 nm). The MiniFluo-UV sensors have several features: measurements of two PAHs at the same time, small size (puck format, 80 x 60 mm), very low energy consumption (500 mW at 12V), LED monitoring, analog and numerical communication modes. The two MiniFluo-UV sensors were first tested in the laboratory: 1) on standard solutions of

  12. Fluorometer with a quartz-rod waveguide-integrating sphere configuration to measure evanescent-field luminescence

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A fluorometer was designed to measure evanescent-field luminescence. A quartz-rod waveguide (d = 2 mm) was installed coaxally inside a cylindrical flow-through cell (id = 2.3 mm, od = 6.3 mm, l = 116 mm). An excitation beam from a UV LED or a miniature xenon flashlamp was focused by a ball lens and ...

  13. Multi-angle fluorometer technique for the determination of absorption and scattering coefficients of subwavelength nanoparticles.

    PubMed

    Shortell, Matthew P; Hewins, Rodney A; Fernando, Joseph F S; Walden, Sarah L; Waclawik, Eric R; Jaatinen, Esa A

    2016-07-25

    A thorough analysis of the resonance light scattering (RLS) technique for quantitative scattering measurements of subwavelength nanoparticles is reported. The systematic error associated with using a measurement at a single angle to represent all of the scattered light is investigated. In-depth analysis of the reference material was performed to identify and minimize the error associated with the reference material. Semiconductor ZnO nanobullets and spherical Au nanoparticles of various sizes were used to verify the approach. A simple and inexpensive modification to standard fluorometers is demonstrated using a glass prism allowing scattering measurements in the slightly forward and backwards directions. This allows quantification of the systematic error associated with RLS which is consistently overlooked. PMID:27464160

  14. Novel blue LED-based handheld fluorometer for detection of terrestrial algae on solid surfaces

    NASA Astrophysics Data System (ADS)

    Brechet, Eric; McStay, Daniel; Wakefield, Rachael D.; Campbell, Ian

    1998-09-01

    The application of an hand-held fluorometer used to monitor algal growth on stone surfaces is reported. The system is based on a modulated ultra-bright blue LED, used to induce chlorophyll-a fluorescence, as well as that of accessory pigments. With the addition of an encoding wheel and when linked to computer this system can produce real time map of the algae population on solid surface. The system has been shown to have a linear response to algal concentration, making it a viable tool for algal monitoring. The hand-held system is relatively immune to ambient light allowing it to be used on-site in various daylight conditions. Results from field and laboratory tests of the system on historically important sites and test samples are presented.

  15. Inexpensive, pocket-sized LED-based fluorometer for undergraduate teaching laboratories and in-the-field chemical detection

    NASA Astrophysics Data System (ADS)

    Tiber, Gage; Basu, Partha; Corcovilos, Theodore A.

    2015-05-01

    Fluorometry is a standard experimental technique for the detection of chemical compounds in solution. Excitation light is absorbed by a sample and then longer-wavelength light is emitted. Typical laboratory fluorometers are large and expensive, making them poorly suited for field work and teaching laboratories. We present a simple battery-powered fluorometer built with off-the-shelf components and a 3D-printed body. The light sources are user-replaceable light emitting diodes (LEDs). Two independent light sources of different wavelengths allow ratiometric measurements of the sample. The detectors are photodiodes with interchangeable dielectric Fabry-Perot stack spectral filters. The light gathering optics are designed using non-imaging optics principles to maximize the amount of detected fluorescence light. We present the design of the device and demonstrate the sensitivity using a molecular detector of Pb2+ ions in solution. The absorption and emission wavelengths of the detector molecule change from 415 nm and 465 nm, resp., in the absence of Pb2+ to 389 nm and 423 nm, resp., in the presence of Pb2+. The estimated sensitivity of the fluorometer with this molecular detector is a few p.p.b.

  16. Photon-counting 1.0 GHz-phase-modulation fluorometer

    SciTech Connect

    Mizuno, T.; Nakao, S.; Mizutani, Y.; Iwata, T.

    2015-04-15

    We have constructed an improved version of a photon-counting phase-modulation fluorometer (PC-PMF) with a maximum modulation frequency of 1.0 GHz, where a phase domain measurement is conducted with a time-correlated single-photon-counting electronics. While the basic concept of the PC-PMF has been reported previously by one of the authors, little attention has been paid to its significance, other than its weak fluorescence measurement capability. Recently, we have recognized the importance of the PC-PMF and its potential for fluorescence lifetime measurements. One important aspect of the PC-PMF is that it enables us to perform high-speed measurements that exceed the frequency bandwidths of the photomultiplier tubes that are commonly used as fluorescence detectors. We describe the advantages of the PC-PMF and demonstrate its usefulness based on fundamental performance tests. In our new version of the PC-PMF, we have used a laser diode (LD) as an excitation light source rather than the light-emitting diode that was used in the primary version. We have also designed a simple and stable LD driver to modulate the device. Additionally, we have obtained a sinusoidal histogram waveform that has multiple cycles within a time span to be measured, which is indispensable for precise phase measurements. With focus on the fluorescence intensity and the resolution time, we have compared the performance of the PC-PMF with that of a conventional PMF using the analogue light detection method.

  17. Modified cytotoxic T lymphocyte precursor frequency assay by measuring released europium in a time resolved fluorometer.

    PubMed

    Haque, K; Truman, C; Dittmer, I; Laundy, G; Denning-Kendall, P; Hows, J; Feest, T; Bradley, B

    1997-01-01

    The frequency of cytotoxic T lymphocyte precursors (CTLpf) can be quantified by using the principle of limiting dilution analysis (LDA). Chromium 51 (51Cr) and europium (Eu) release assays are based on the measurement of marker release after lysis of targets by the effector cells. Although, 51Cr release has been widely used to quantify cell lysis since its introduction, it has several disadvantages such as handling and disposal of radioisotopes as well as health risk to personnel involved performing the assay. This situation has led us to adopt a non-radioactive cytotoxicity assay. After 7 days culture the PHA-stimulated targets are labeled with europium DTPA chelate. Lysis of labeled targets by effectors releases the Eu-DTPA complex in culture medium--a highly fluorescent substance. The amount of fluorescence can be measured in a time resolved fluorometer. We describe here some modifications of the original protocol which include optimising IL-2 requirements, reduction of incubation times, addition of an extra spin before 37 degrees C incubation, readjustment of target cells per volume of labeling buffer and other crucial parameters increasing the specificity and sensitivity of CTLpf assay. We are in agreement with others that the Eu-release assay is specific and reproducible. It can be used for the CTLpf estimation as well as other T cell and non-T cell cytotoxicity assays. PMID:9090438

  18. Coaxial fiber-optic chemical-sensing excitation-emission matrix fluorometer.

    PubMed

    Kim, Yoon-Chang; Jordan, James A; Chávez, Diana; Booksh, Karl S

    2011-02-01

    Great reductions in the overall size and complexity of high throughput multichannel UV-visible fluorometers were achieved by coupling a compact optical fiber array to compact dispersive transmission optics. The coaxial configuration centers on the insertion of a silica/silica optical fiber into the hollow region of a UV-fused silica capillary waveguide. The outer core delivers the maximum power of the narrow wavelength region of the excitation spectrum created by coupling a xenon arc discharge lamp to a compact spectrometer. The molecular fluorescence resulting from the interaction of light emitted at the distal end of the hollow waveguide and the sample matrix is received and transmitted to a CCD via a compact dispersive grating-prism (grism) optical assembly. A linear array of the coaxial optical fibers permits a full excitation-emission matrix spectrum of the analyte matrix to be projected onto the face of the CCD. The in situ identification and monitoring of polycyclic aromatic hydrocarbons was carried out for the initial application testing for this prototype. PMID:21283188

  19. Design of a portable fluorometer capable of time-resolved luminescence measurements

    NASA Astrophysics Data System (ADS)

    Chen, Guoying

    2004-11-01

    A rugged, filter-based fluorometer capable of time-resolved luminescence (TRL) measurements was designed, prototyped and tested for field applications. The instrument operation and data processing were controlled by a laptop computer running a custom LabVIEW program. A xenon flashlamp was used as the light source and a photomultiplier tube (PMT) as the photodetector. A gating technique was implemented to effectively overcome PMT saturation by intense xenon lamp flash so signal integrity was maintained even at very high gains, leading to improved sensitivity and reproducibility. The instrument was tested by TRL using tetracycline as a model analyte; and the signal was digitized at a 2-μs time resolution and a 12-bit amplitude resolution. Its performance was similar to or slightly better than that of a commercial fluorescence spectrophotometer. A 0-300 ppb linear dynamic range (r2 = 0.9996) and a 0.025-ppb limit of detection (LOD) were achieved with a <=5% relative standard deviation.

  20. Enhanced optical fiber fluorometer using a periodic perturbation in the fiber core

    NASA Astrophysics Data System (ADS)

    Chiniforooshan, Yasser; Bock, Wojtek J.; Ma, Jianjun

    2013-10-01

    Tracing of the specific chemicals and biological agents in a solution is becoming a vital interest in health, security and safety industries. Although a number of standard laboratory-based testing systems exists for detecting such targets, but the fast, real-time and on-site methods could be more efficient and cost-effective. One of the most common ways to detect a target in the solution is to use the fluorophore molecules which will be selectively attached to the targets and will emit or quench the fluorescence in presence of the target. The fiber-optic fluorometers are developed for inexpensive and portable detection. In this paper, we explain a novel multi-segment fiber structure which uses the periodic perturbation on the side-wall of a highly multi-mode fiber to enhance collecting the fluorescent light. This periodic perturbation is fabricated and optimized on the core of the fiber using a CO2 laser. The theoretical explanation to show the physical principle of the structure is followed by the experimental evidence of its functioning.

  1. Utilization of a submersible UV fluorometer for monitoring anthropogenic inputs in the Mediterranean coastal waters.

    PubMed

    Tedetti, Marc; Guigue, Catherine; Goutx, Madeleine

    2010-03-01

    We evaluated the performances of a submersible ultraviolet fluorometer (EnviroFlu-HC, TriOS Optical Sensors) dedicated to the real time measurement of polycyclic aromatic hydrocarbons (PAHs) in the aquatic media. We conducted calibration experiments and in situ measurements in the coastal Mediterranean Sea. We found that the EnviroFlu-HC was not strictly specific to PAHs, even though it exhibited the highest sensitivity for phenanthrene, but could response to tryptophan-like material as well, and in a much less extent, to humic substances. The sensor signal showed great spatial and temporal variations in clean and polluted sites, with likely a high contribution of PAHs in the harbors, and a high contribution of tryptophan-like and humic-like materials in the sewage effluent. We conclude that the EnviroFlu-HC is a good tool for monitoring anthropogenic inputs in the coastal waters, although its utilization should be combined to other fluorescence measurements to improve the information about the nature of the aromatic compounds detected. PMID:19948348

  2. In situ tryptophan-like fluorometers: assessing turbidity and temperature effects for freshwater applications.

    PubMed

    Khamis, K; Sorensen, J P R; Bradley, C; Hannah, D M; Lapworth, D J; Stevens, R

    2015-04-01

    Tryptophan-like fluorescence (TLF) is an indicator of human influence on water quality as TLF peaks are associated with the input of labile organic carbon (e.g. sewage or farm waste) and its microbial breakdown. Hence, real-time measurement of TLF could be particularly useful for monitoring water quality at a higher temporal resolution than available hitherto. However, current understanding of TLF quenching/interference is limited for field deployable sensors. We present results from a rigorous test of two commercially available submersible tryptophan fluorometers (ex ∼ 285, em ∼ 350). Temperature quenching and turbidity interference were quantified in the laboratory and compensation algorithms developed. Field trials were then undertaken involving: (i) an extended deployment (28 days) in a small urban stream; and, (ii) depth profiling of an urban multi-level borehole. TLF was inversely related to water temperature (regression slope range: -1.57 to -2.50). Sediment particle size was identified as an important control on the turbidity specific TLF response, with signal amplification apparent <150 NTU for clay particles and <650 NTU for silt particles. Signal attenuation was only observed >200 NTU for clay particles. Compensation algorithms significantly improved agreement between in situ and laboratory readings for baseflow and storm conditions in the stream. For the groundwater trial, there was an excellent agreement between laboratory and raw in situ TLF; temperature compensation provided only a marginal improvement, and turbidity corrections were unnecessary. These findings highlight the potential utility of real time TLF monitoring for a range of environmental applications (e.g. tracing polluting sources and monitoring groundwater contamination). However, in situations where high/variable suspended sediment loads or rapid changes in temperature are anticipated concurrent monitoring of turbidity and temperature is required and site specific calibration is

  3. Preliminary evaluation of an in vivo fluorometer to quantify algal periphyton biomass and community composition

    USGS Publications Warehouse

    Harris, Theodore D.; Graham, Jennifer

    2015-01-01

    The bbe-Moldaenke BenthoTorch (BT) is an in vivo fluorometer designed to quantify algal biomass and community composition in benthic environments. The BT quantifies total algal biomass via chlorophyll a (Chl-a) concentration and may differentiate among cyanobacteria, green algae, and diatoms based on pigment fluorescence. To evaluate how BT measurements of periphytic algal biomass (as Chl-a) compared with an ethanol extraction laboratory analysis, we collected BT- and laboratory-measured Chl-a data from 6 stream sites in the Indian Creek basin, Johnson County, Kansas, during August and September 2012. BT-measured Chl-a concentrations were positively related to laboratory-measured concentrations (R2 = 0.47); sites with abundant filamentous algae had weaker relations (R2 = 0.27). Additionally, on a single sample date, we used the BT to determine periphyton biomass and community composition upstream and downstream from 2 wastewater treatment facilities (WWTF) that discharge into Indian Creek. We found that algal biomass increased immediately downstream from the WWTF discharge then slowly decreased as distance from the WWTF increased. Changes in periphyton community structure also occurred; however, there were discrepancies between BT- and laboratory-measured community composition data. Most notably, cyanobacteria were present at all sites based on BT measurements but were present at only one site based on laboratory-analyzed samples. Overall, we found that the BT compared reasonably well with laboratory methods for relative patterns in Chl-a but not as well with absolute Chl-aconcentrations. Future studies need to test the BT over a wider range of Chl-aconcentrations, in colored waters, and across various periphyton assemblages.

  4. Optics and experimental resolution of the Heidelberg slit-scan flow fluorometer

    NASA Astrophysics Data System (ADS)

    Hausmann, Michael; Wickert, Burkhard; Vogel, Michael; Schurwanz, Michael; Doelle, Juergen; Wolf, Dietmar; Aldinger, Klaus; Cremer, Christoph G.

    1996-01-01

    Slit-scan flow fluorometry is a laser-technological approach for accelerated screening and sorting of fluorescence labelled metaphase chromosomes. Details of the optics of the Heidelberg slit-scan sorter are presented. In a fluid stream the fluorescence labelled chromosomes rapidly pass one at a time by a scanning laser beam. The laser can be focused by a less complex optic consisting of only a few commercially available lenses. The laser intensity distribution around the focus was measured for 488 nm for two lens configurations. Although the light distribution obtained by such an optic is normally not aberration free, the requirements of a 'ribbonlike' shape in the center of the fluid stream can be fulfilled. Since the chromosomes are oriented perpendicularly to the laser beam by hydrodynamic focusing of the fluid stream, the fluorescence intensity along the chromosome axis can be measured time (equals spatially) resolved. According to their intensity profiles the chromosomes can be classified. Signal processing of the profiles can be performed in less than 600 microseconds, so that in the order of hundred chromosomes per second can be sorted out by a computer controlled electro-acoustic sorting unit. The final spatial resolution of a slit-scan flow sorter is not only affected by the focusing optics of the laser but also by the fluid stream, the detection optics and electronics, as well as by the computer analysis algorithm. Calculations often consider only the optics under ideal conditions. Here, a method is shown how to estimate the overall resolution of a slit-scan flow fluorometer experimentally. According to this criterion the resolution of the Heidelberg slit-scan sorter for 488 nm fluorescence excitation was estimated to be 2.4 micrometer in its basic optical configuration and 1.7 micrometer with additional correction of chromatic aberration effects.

  5. Analysis of data obtained from a frequency-multiplexed phase-modulation fluorometer using an autoregressive model.

    PubMed

    Iwata, Tetsuo; Muneshige, Akitaka; Araki, Tsutomu

    2007-09-01

    In order to derive plural values of fluorescence lifetimes simultaneously from a multi-component sample, we formulate a mathematical method for analyzing data obtained from a frequency-multiplexed phase-modulation fluorometer (FM-PMF) using an autoregressive (AR) model. Various parameter settings necessary for performing accurate data analysis based on the AR model are studied through numerical simulations. Measurement results of fluorescence lifetimes of real samples, 10 ppm quinine sulfate in 0.1 N H(2)SO(4), 10 ppm rhodamine 6G in ethanol, and their mixture with a volume ratio of 1:1, demonstrate that the proposed method works quite well. PMID:17910791

  6. Mapping Bronchial Carcinoma In Situ Lung Cancer Lesions By Combined Imaging Fluorescence Bronchoscopy And Ratioing Fluorometer Probe

    NASA Astrophysics Data System (ADS)

    Balchum, Oscar J.; Profio, A. Edward; Razum, Nickolas J.

    1988-06-01

    A two-component system of instrumentation and methods, IFB and RFP, when used in combination employing hematoporphyrin derivates, DHE, had highly satisfactory sensitivity (95%) and specificity (100%) for localizing carcinoma in situ lesions in the bronchi of individuals with early stage lung cancer, having a normal chest x-ray and detected by a positive sputum cytology test. The more detailed mapping of additional subjects may increase the specificity of RFP (Ratioing Fluorometer Probe) by itself to an adequate level. Digital computer subs traction of background antofluorescence may increase contrast to enhance the specificity of IFB (Imaging Fluorescence Bronchoscopy).

  7. Electrets and plant fluorometers used in field studies to measure hydrogen chloride produced during Space Shuttle launches

    NASA Technical Reports Server (NTRS)

    Milligan, J. E.; Swoboda, G. D.; Susko, M.

    1985-01-01

    The results of the field tests of two monitoring device techniques, electrets and plant fluorometers are analyzed in order to determine the environmental effects of launch by-products and the extent of these effects. The STS launches are used because the Shuttle emits 2 1/2 times more HCl than any previous systems, it produces a voluminous ground cloud and, most important, it produces near field HCl deposition and revolatilization, far-field acid washout/rainout, and gaseous HCl diffusion. Field evaluations of electrets at STS-5, STS-6, and STS-8 have shown that qualitative assessments can be made for areas lightly or moderately impacted by gaseous and aerosol HCl. Field evaluation of the plant productivity fluorometer at STS-8 has shown that this system is also useful for qualitative assessment in areas lightly, moderately, or heavily affected by gaseous and aerosol HCl. Quantitative prediction of HCl may be possible in lightly and moderately affected areas, given deposition rates correlation.

  8. SEEP II, Shelf Edge Exchange Processes-II: Chlorophyll a fluorescence, temperature, and beam attenuation measurements from moored fluorometers

    SciTech Connect

    Medeiros, W.H.; Wirick, C.D.

    1992-02-01

    The Shelf Edge Exchange Processes (SEEP) program sponsored by the United States Department of Energy is a multi-institutional effort designed to investigate the flux of suspended material from the continental shelf to the waters of the upper slope, and then possibly into the slope sediments. The first SEEP experiment (SEEP I) was across the outer continental shelf of New England during 1983--1984 and consisted of a series of nine cruises and a mooring array. The second experiment (SEEP II) focused specifically of the shelf/slope frontal region of the mid-Atlantic Bight off the Delmarva peninsula. This report presents data collected during SEEP II. The SEEP II experiment consisted of a series of ten cruises and mooring arrays as well as over-flights by NASA aircraft. The cruises were consecutively designated SEEP2-01 to SEEP2-10. Hydrographic data were collected on all cruises except SEEP2-04 and SEEP2-07 during which benthic processes were investigated. Mooring arrays were deployed during three cruises in the Spring, Summer and Winter of 1988. Brookhaven National Laboratory deployed sixteen fluorometer instrument packages on their moorings with sensors to measure: the in vivo fluorescence of phytoplankton, temperature, subsurface light, dissolved oxygen, and water transparency. Data from the fluorometer, temperature, and transmissometer sensors are reported herein.

  9. Development of a multi-sensor in situ fiber optic fluorometer. Progress report, June 1, 1992--October 31, 1993

    SciTech Connect

    Lohrenz, S.E.; Asper, V.L.; Morris, M.J.; Walters, R.A.

    1993-11-01

    Our objective is to develop and evaluate a multi-sensor in situ fiber optic fluorometer. The instruments is designed to sample and store in vivo strobe-stimulated fluorescence data at multiple depths and high frequencies (1 Hz). This information may be used for estimating the distribution and abundance of particulate pigment biomass, for supporting models of water column primary production and as a complement to remotely sensed ocean color estimates of pigment biomass. The instrument is unique in that it uses fiber optic technology to increase vertical resolution. While it is theoretically possible to accomplish this task using a large number of commercially available fluorometers, our proposed design would provide a less expensive approach. Two prototype instruments have been built and are being tested. The first, a single sensor instrument interfaced with a 486 personal computer, has been used to optimize hardware and sensor design and to evaluate fiber performance an instrument detection limits. The second instrument, containing 8 sensors and capable of autonomous operation with time-series data acquisition and storage, was recently deployed in a cruise in the Gulf of Mexico. Preliminary results indicate that the instrument meets all the project goals and that low cost, high frequency, high spatial resolution fluorescence data are obtainable with the current design. Additional work will focus on further optimization of hardware design and software algorithms, and construction of an additional instrument specifically designed for deployement in the benthic boundary layer.

  10. Development of a multi-sensor in situ fiber optic fluorometer. Progress report, June 1, 1992--December 31, 1992

    SciTech Connect

    Lohrenz, S.E.; Asper, V.L.; Morris, M.J.; Walters, R.A.

    1992-12-01

    Objective is to develop and evaluate a multi-sensor in situ fiber optic fluorometer. The instrument is designed to sample and store in vivo strobe-stimulated fluorescence data at multiple depths and high frequencies (1 Hz). This information may be used for estimating the distribution and abundance of particulate pigment biomass, for supporting models of water column primary production and as a complement to remotely sensed ocean color estimates of pigment biomass. The instrument is unique in that it uses fiber optic technology to increase vertical resolution. While it is theoretically possible to accomplish this task using a large number of commercially available fluorometers, our proposed design would provide a less expensive approach. A laboratory prototype has been built and is being tested. Preliminary results indicate that the instrument meets all the project goals and that low cost, high frequency, high spatial resolution chlorophyll data are obtainable with the current design. Further work is required to develop the seagoing version, and optimize the configuration of the fiber sensors.

  11. Low-cost phase-modulation fluorometer for measuring nanosecond lifetimes using a lock-in amplifier

    NASA Astrophysics Data System (ADS)

    Harms, Peter D.; Rao, Govind

    1999-05-01

    A Stanford Research Systems SR844 lock-in amplifier was used to build a sub $10,000 phase-modulation fluorometer capable of measuring nanosecond fluorescence lifetimes. The lock-in directly provided both the DC bias and the AC signal used to modulate the intensity of a blue LED excitation source. A photomultiplier tube measured the emission, and the resulting signal was sent back through a DC block to the lock-in with no external signal processing or heterodyning required. A simple computer program was developed to automate the measuring process and correct for the most common sources of error, namely coherent pickup and stray ambient light. Several standard fluorophores were measured, and the results compare favorably with those from a research grade cross-correlation phase fluorometer up to frequencies of 100 MHz. This system can operate in several configurations, each with benefits and limitations. The system is particularly well suited for fluorescence lifetime based sensing applications, demonstrated by measuring dissolved carbon dioxide online in a bacterial fermentation.

  12. A fiber optic, ultraviolet light-emitting diode-based, two wavelength fluorometer for monitoring reactive adsorption

    NASA Astrophysics Data System (ADS)

    Granz, Christopher D.; Schindler, Bryan J.; Peterson, Gregory W.; Whitten, James E.

    2016-03-01

    Construction and use of an ultraviolet light-emitting diode-based fluorometer for measuring photoluminescence (PL) from powder samples with a fiber optic probe is described. Fluorescence at two wavelengths is detected by miniature photomultiplier tubes, each equipped with a different band pass filter, whose outputs are analyzed by a microprocessor. Photoluminescent metal oxides and hydroxides, and other semiconducting nanoparticles, often undergo changes in their emission spectra upon exposure to reactive gases, and the ratio of the PL intensities at two wavelengths is diagnostic of adsorption. Use of this instrument for reactive gas sensing and gas filtration applications is illustrated by measuring changes in the PL ratio for zirconium hydroxide and zinc oxide particles upon exposure to air containing low concentrations of sulfur dioxide.

  13. A phase-shift fluorometer using a laser and a transverse electrooptic modulator for subnanosecond lifetime measurements.

    PubMed Central

    Salmeen, I; Rimai, L

    1977-01-01

    We described a simple phase-shift fluorometer using continuous laser excitation. The laser enables the use of a transverse mode electrooptic modulator with a half-wave retardation voltage of about 200 V (in contrast to many kilovolts of longitudinal modulators) at frequencies up to 100 MHz. The modulated fluorescence signal is detected, after passing through a double monochromator, by a photomultiplier tube feeding a radio frequency (RF) tuned amplifier. THE RF phase is then determined by phase-sensitive detection using a double balanced mixer with the reference obtained from a PIN photodiode-turned amplifier combination which detects light split off from the main exciting beam. The laser and double monochromator allow the observation of modulated Raman solvent and Rayleigh scatterin, which are convenient for determining the zero reference phase. PMID:922124

  14. Study of multicomponent mixtures in solution with a vertical-axis transmission-type filter-fluorometer

    USGS Publications Warehouse

    Fletcher, M.H.

    1963-01-01

    Fluorescence intensity, sensitivity, and the effect of diverse ions are discussed in relation to chemical equilibrium and the general equation for fluorescence. High sensitivity is the common denominator in eliminating or reducing all types of interference and the general equation is the key for quickly selecting conditions that give maximum sensitivity. With a transmission-type fluorometer, experimental fluorescence intensities adhere to the general equation over a very wide range of light absorption. If the instrument has a vertical axis, solution depth can be adjusted so that the data fall on that portion of the theoretical curve which is essentially a straight line. When the fluorescence wavelength selected for measurement is not absorbed by the solution, the general equation reduces to a much simpler expression, and then, under proper circumstances, fluorescence values can be used for calculations as confidently and as easily as absorbance values.

  15. A rapid excitation-emission matrix fluorometer utilizing supercontinuum white light and acousto-optic tunable filters

    NASA Astrophysics Data System (ADS)

    Wang, Wenbo; Wu, Zhenguo; Zhao, Jianhua; Lui, Harvey; Zeng, Haishan

    2016-06-01

    Scanning speed and coupling efficiency of excitation light to optic fibres are two major technical challenges that limit the potential of fluorescence excitation-emission matrix (EEM) spectrometer for on-line applications and in vivo studies. In this paper, a novel EEM system, utilizing a supercontinuum white light source and acousto-optic tunable filters (AOTFs), was introduced and evaluated. The supercontinuum white light, generated by pumping a nonlinear photonic crystal fiber with an 800 nm femtosecond laser, was efficiently coupled into a bifurcated optic fiber bundle. High speed EEM spectral scanning was achieved using AOTFs both for selecting excitation wavelength and scanning emission spectra. Using calibration lamps (neon and mercury argon), wavelength deviations were determined to vary from 0.18 nm to -0.70 nm within the spectral range of 500-850 nm. Spectral bandwidth for filtered excitation light broadened by twofold compared to that measured with monochromatic light between 650 nm and 750 nm. The EEM spectra for methanol solutions of laser dyes were successfully acquired with this rapid fluorometer using an integration time of 5 s.

  16. A rapid excitation-emission matrix fluorometer utilizing supercontinuum white light and acousto-optic tunable filters.

    PubMed

    Wang, Wenbo; Wu, Zhenguo; Zhao, Jianhua; Lui, Harvey; Zeng, Haishan

    2016-06-01

    Scanning speed and coupling efficiency of excitation light to optic fibres are two major technical challenges that limit the potential of fluorescence excitation-emission matrix (EEM) spectrometer for on-line applications and in vivo studies. In this paper, a novel EEM system, utilizing a supercontinuum white light source and acousto-optic tunable filters (AOTFs), was introduced and evaluated. The supercontinuum white light, generated by pumping a nonlinear photonic crystal fiber with an 800 nm femtosecond laser, was efficiently coupled into a bifurcated optic fiber bundle. High speed EEM spectral scanning was achieved using AOTFs both for selecting excitation wavelength and scanning emission spectra. Using calibration lamps (neon and mercury argon), wavelength deviations were determined to vary from 0.18 nm to -0.70 nm within the spectral range of 500-850 nm. Spectral bandwidth for filtered excitation light broadened by twofold compared to that measured with monochromatic light between 650 nm and 750 nm. The EEM spectra for methanol solutions of laser dyes were successfully acquired with this rapid fluorometer using an integration time of 5 s. PMID:27370436

  17. Extension of transverse relaxation-optimized spectroscopy techniques to allosteric proteins: CO- and paramagnetic fluoromet-hemoglobin [beta (15N-valine)].

    PubMed

    Nocek, J M; Huang, K; Hoffman, B M

    2000-03-14

    We present the first steps in applying transverse relaxation-optimized spectroscopy (TROSY) techniques to the study of allosterism. Each beta-chain of the hemoglobin (Hb) tetramer has 17 valine residues. We have (15)N-labeled the beta-chain Val residues and detected 16 of the 17 (1)H-(15)N correlation peaks for beta-chain Val of the R state CO-Hb structure by using the TROSY technique. Sequence-specific assignments are suggested, based mainly on analysis of the (1)H pseudocontact-shift increments produced by oxidizing the diamagnetic R state HbCO to the paramagnetic R state fluoromet form. When possible, we support these assignments with sequential nuclear Overhauser effect (NOE) information obtained from a two-dimensional [(1)H,(1)H]-NOESY-TROSY experiment (NOESY, NOE spectroscopy). We have induced further the R-T conformational change by adding the allosteric effector, inositol hexaphosphate, to the fluoromet-Hb sample. This change induces substantial increments in the (1)H and (15)N chemical shifts, and we discuss the implication of these findings in the context of the tentative sequence assignments. These preliminary results suggest that amide nitrogen and amide proton chemical shifts in a selectively labeled sample are site-specific probes for monitoring the allosteric response of the ensemble-averaged solution structure of Hb. More important, the chemical-shift dispersion obtained is adequate to permit a complete assignment of the backbone (15)N/(13)C resonances upon nonselective labeling. PMID:10716987

  18. Calibration and performance of a new in situ multi-channel fluorometer for measurement of colored dissolved organic matter in the ocean

    NASA Astrophysics Data System (ADS)

    Conmy, Robyn N.; Coble, Paula G.; Castillo, Carlos E. Del

    2004-02-01

    The development of multispectral in situ fluorescence instruments greatly enhances the study of the optical properties of Colored Organic Matter (COM). Here, we tested the inter-instrument variability of three WetLabs, Inc. SAFIres using quinine sulfate standards. As with any fluorometer, intensity and spectral biases in fluorescence output due to properties of the SAFIre's optical components necessitate corrections. Low response of the instrument to quinine sulfate and lack of an excitation/emission channel at the fluorescence maximum of this standard precluded direct spectral calibration. Calibrations conducted using seawater as a secondary standard provided an acceptable alternative. The field performance of the SAFIre from two experiments is presented here. Time series contour plots show that the instrument has the ability to detect small differences in COM optical properties, and observed fluorescence emission ratios are indicative of changes in sources of the material over the course of the study. The SAFIre was found to extend multispectral measurements to include high spatial and high temporal resolution.

  19. Development of a fluorescent enzyme-linked DNA aptamer-magnetic bead sandwich assay and portable fluorometer for sensitive and rapid leishmania detection in sandflies.

    PubMed

    Bruno, John G; Richarte, Alicia M; Phillips, Taylor; Savage, Alissa A; Sivils, Jeffrey C; Greis, Alex; Mayo, Michael W

    2014-01-01

    A fluorescent peroxidase-linked DNA aptamer-magnetic bead sandwich assay is described which detects as little as 100 ng of soluble protein extracted from Leishmania major promastigotes with a high molarity chaotropic salt. Lessons learned during development of the assay are described and elucidate the pros and cons of using fluorescent dyes or nanoparticles and quantum dots versus a more consistent peroxidase-linked Amplex Ultra Red (AUR; similar to resazurin) fluorescence version of the assay. While all versions of the assays were highly sensitive, the AUR-based version exhibited lower variability between tests. We hypothesize that the AUR version of this assay is more consistent, especially at low analyte levels, because the fluorescent product of AUR is liberated into bulk solution and readily detectable while fluorophores attached to the reporter aptamer might occasionally be hidden behind magnetic beads near the detection limit. Conversely, fluorophores could be quenched by nearby beads or other proximal fluorophores on the high end of analyte concentration, if packed into a small area after magnetic collection when an enzyme-linked system is not used. A highly portable and rechargeable battery-operated fluorometer with on board computer and color touchscreen is also described which can be used for rapid (<1 h) and sensitive detection of Leishmania promastigote protein extracts (∼ 100 ng per sample) in buffer or sandfly homogenates for mapping of L. major parasite geographic distributions in wild sandfly populations. PMID:24222436

  20. Development of a fluorescent enzyme-linked DNA aptamer-magnetic bead sandwich assay and portable fluorometer for sensitive and rapid listeria detection.

    PubMed

    Bruno, John G; Phillips, Taylor; Montez, Tiffany; Garcia, Adrian; Sivils, Jeffrey C; Mayo, Michael W; Greis, Alex

    2015-01-01

    A fluorescent DNA aptamer-magnetic bead sandwich assay was developed to detect listeriolysin O (LLO) protein from pathogenic Listeria bacteria using a peroxidase-linked system, Amplex Ultra Red (AUR; derivatized resazurin) substrate, and a custom-designed handheld fluorometer. The assay is highly sensitive with demonstrated limits of detection (LODs) in the range of 4 to 61 L. monocytogenes cells or the equivalent LLO produced by 4 to 61 cells on average in separate titration trials. Total assay processing and analysis time was approximately 30 mins. The assay has demonstrated the ability to detect 6 species of Listeria as desired by the USDA's Food Safety Inspection Service (FSIS). The portable system was designed to be used primarily with surface swab samples from fomites, but it can also be used to assess enrichment cultures. The minimal time to detect a positive enrichment culture in our hands from an initial 10 cell inoculum in 200 ml of broth has been 8 h post-incubation at 37 °C in shaker flask cultures. An optional automated magnetic bead assay processing and wash device capable of simultaneously processing 6 samples with low and consistent fluorescence background for higher volume central laboratories is also described. PMID:25511112

  1. Application of a laser fluorometer for discriminating phytoplankton species

    NASA Astrophysics Data System (ADS)

    Chen, Peng; Pan, Delu; Mao, Zhihua

    2015-04-01

    A portable laser-induced fluorescence system for discriminating phytoplankton species has been developed. It consists of a high pulsed repetition frequency (10-kHz) microchip laser at 405 nm, a reflective fluorescent probe and a broadband micro spectrometer. The measured fluorescent spectra were overlapped by various fluorescent components, and were then decomposed by a bi-Gaussian mixture model. A spectral shape description index was designed to characterize fluorescent spectral shapes for descriminating the phytoplankton species cultured in our laboratory. Using clustering analysis, the samples of eight phytoplankton species belonging to two divisions of Bacillariophyta and Dinophyta were divided into six categories: 1) Chaetoceros debilis, Thalassiosira rotula; 2) Prorocentrum donghaiense, Prorocentrum dentatum; 3) Gymnodinium simplex; 4) Alexandrium tamarense; 5) Karenia mikimotoi; and 6) Akashiwo sanguinea. The phytoplankton species belonging to Bacillariophyta were well separated from those belonging to Dinophyta. In addition, the phytoplankton species belonging to Dinophyta were successfully distinguished from each other at genus level. The portable system is expected to be used both in vivo and in the field.

  2. Optical fiber underwater fluorometer for measuring chlorophyll-a concentration

    NASA Astrophysics Data System (ADS)

    Zheng, Longjiang; Hou, Peiguo; Wang, Yutian

    2000-10-01

    This paper describes an efficient method for in-situ measurement of chlorophyll-a concentration in the seawater with fluorescence method and optical fiber techniques. The instrument uses the pulsed xenon lamp as the excited light resources. Both the exciting light and the fluorescence from algae chlorophyll-a are transmitted along two fiber bundles. The fluorescent signal is detected by using the relevant pulsed detecting technology. The minimal detecting concentration of chlorophyll-a in the ocean can reach 1x10-5mg/cm3. The system has advantages of simple structure, passive sensor head and high sensitivity. The experimental results show that this measurement method is realizable.

  3. 21 CFR 862.2560 - Fluorometer for clinical use.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... intended to measure by fluorescence certain analytes. Fluorescence is the property of certain substances of... certain materials to measure a variety of analytes. (b) Classification. Class I (general controls)....

  4. Airborne fluorometer applicable to marine and estuarine studies

    USGS Publications Warehouse

    Stoertz, George E.; Hemphill, William R.; Markle, David A.

    1969-01-01

    An experimental Fraunhofer line discriminator detected solar-stimulated yellow fluorescence (5890 A) emitted by Rhodamine WT dye in aqueous solutions. Concentration of 1 part per billion was detected in tap water 1/2-meter deep. In extremely turbid San Francisco Bay, dye was monitored in concentrations of less than 5 parts per billion from helicopter and ship. Applications include studies of current dynamics and dispersion. Potential applications of the technique could include sensing oil spills, fish oils, lignin sulfonates, other fluorescent pollutants, and chlorophyll fluorescence.

  5. Exploring Photosynthesis and Plant Stress Using Inexpensive Chlorophyll Fluorometers

    ERIC Educational Resources Information Center

    Cessna, Stephen; Demmig-Adams, Barbara; Adams, William W., III

    2010-01-01

    Mastering the concept of photosynthesis is of critical importance to learning plant physiology and its applications, but seems to be one of the more challenging concepts in biology. This teaching challenge is no doubt compounded by the complexity by which plants alter photosynthesis in different environments. Here we suggest the use of chlorophyll…

  6. Integrating fluorometer for the measurement of chlorophyll fluorescence induction in intact plants

    NASA Astrophysics Data System (ADS)

    Toivonen, Peter; Vidaver, William

    1984-10-01

    An economical device for monitoring the integrated chlorophyll fluorescence emission of plant material is described. The system, which uniquely incorporates an integrating sphere, light source, photographic shutter, optical filters, and a photodetector is applicable to intact plants, whole leaves, or other materials. It is noninvasive and a single sample may be repeatedly tested over time intervals of minutes to months. Data obtained provide information about the sample size (i.e., leaf area, total chlorophyll content), and the photosynthetic activity of the sample. Samples of from a few mm2 to several cm2 can be accommodated depending on the diameter of the integrating sphere and excitation light source intensity. The device should be of interest to workers in plant breeding, genetic engineering, tissue culture, horticulture, herbicides, pollution, pathology, and environmental stress.

  7. Confocal fluorometer for diffusion tracking in 3D engineered tissue constructs

    NASA Astrophysics Data System (ADS)

    Daly, D.; Zilioli, A.; Tan, N.; Buttenschoen, K.; Chikkanna, B.; Reynolds, J.; Marsden, B.; Hughes, C.

    2016-03-01

    We present results of the development of a non-contacting instrument, called fScan, based on scanning confocal fluorometry for assessing the diffusion of materials through a tissue matrix. There are many areas in healthcare diagnostics and screening where it is now widely accepted that the need for new quantitative monitoring technologies is a major pinch point in patient diagnostics and in vitro testing. With the increasing need to interpret 3D responses this commonly involves the need to track the diffusion of compounds, pharma-active species and cells through a 3D matrix of tissue. Methods are available but to support the advances that are currently only promised, this monitoring needs to be real-time, non-invasive, and economical. At the moment commercial meters tend to be invasive and usually require a sample of the medium to be removed and processed prior to testing. This methodology clearly has a number of significant disadvantages. fScan combines a fiber based optical arrangement with a compact, free space optical front end that has been integrated so that the sample's diffusion can be measured without interference. This architecture is particularly important due to the "wet" nature of the samples. fScan is designed to measure constructs located within standard well plates and a 2-D motion stage locates the required sample with respect to the measurement system. Results are presented that show how the meter has been used to evaluate movements of samples through collagen constructs in situ without disturbing their kinetic characteristics. These kinetics were little understood prior to these measurements.

  8. Fast repetition rate (FRR) fluorometer for making in situ measurements of primary productivity

    SciTech Connect

    Kolber, Z.S.; Falkowski, P.G.

    1992-01-01

    Understanding the ocean carbon cycle and predicting how climate-induced changes in ocean circulation will affect ocean productivity requires that (a) primary productivity be measured with high spatial and temporal resolution, and (b) natural variability in primary productivity be parameterized with regardto environmental factors such as nutrient availabuity, irradiance, and temperature. Instrumentation to measure primary productivity from the stimulated in vivo fluoresence of phytoplankton chlorophyll is currendy being developed at Brookhaven National Laboratory. The instrumentation is based on fast repetition rate (FRR) fluorometry, and provides a robust technique for deriving the photosynthetic rates in situ. Moreover, the FRR methodology directly measures several photosynthetic parameters such as effective absorption cross- section, photo-conversion efficiency, and turnover time of photosynthesis, and relate them to primary productivity. Since photosynthetic parameters are affected by environmental factors such as fight and nutrient availability, the relationship between these parameters and primary productivity can be established. By understanding such relationships, prognostic models of primary productivity can be developed and parameterized.

  9. A remote sensing laser fluorometer. [for detecting oil, ligninsulfonates, and chlorophyll in water

    NASA Technical Reports Server (NTRS)

    Oneill, R. A.; Davis, A. R.; Gross, H. G.; Kruus, J.

    1975-01-01

    A sensor is reported which is able to identify certain specific substances in water by means of their fluorescence spectra. In particular, the sensor detects oil, ligninsulfonates and chlorophyll. The device is able to measure the fluorescence spectra of water at ranges up to 75 m and to detect oil spills on water at altitudes up to 300 m. Blue light from a laser is used to excite the fluorescence of the target. Any light from the ambient background illumination, from the reflected laser light or from the induced fluorescence is gathered by a small telescope focused on the target. Optical filters are used to block the reflected laser light and to select the wavelengths of interest in the fluorescence spectrum of the target. The remaining light is detected with a photomultiplier tube. The amplitude of the laser induced fluorescence in the wavelength interval selected by the optical filters is displayed on a meter or strip chart recorder.

  10. Fast repetition rate (FRR) fluorometer for making in situ measurements of primary productivity

    SciTech Connect

    Kolber, Z.S.; Falkowski, P.G.

    1992-10-01

    Understanding the ocean carbon cycle and predicting how climate-induced changes in ocean circulation will affect ocean productivity requires that (a) primary productivity be measured with high spatial and temporal resolution, and (b) natural variability in primary productivity be parameterized with regardto environmental factors such as nutrient availabuity, irradiance, and temperature. Instrumentation to measure primary productivity from the stimulated in vivo fluoresence of phytoplankton chlorophyll is currendy being developed at Brookhaven National Laboratory. The instrumentation is based on fast repetition rate (FRR) fluorometry, and provides a robust technique for deriving the photosynthetic rates in situ. Moreover, the FRR methodology directly measures several photosynthetic parameters such as effective absorption cross- section, photo-conversion efficiency, and turnover time of photosynthesis, and relate them to primary productivity. Since photosynthetic parameters are affected by environmental factors such as fight and nutrient availability, the relationship between these parameters and primary productivity can be established. By understanding such relationships, prognostic models of primary productivity can be developed and parameterized.

  11. Imaging and quantitation of a tissue-selective lanthanide chelate using an endoscopic fluorometer

    NASA Astrophysics Data System (ADS)

    Houlne, Michael P.; Hubbard, Darren S.; Kiefer, Garry; Bornhop, Darryl J.

    1998-04-01

    Tissue spectroscopy and endoscopy are combined with a tissue site-selective fluorescent probes molecule to demonstrate in vitro, spatial, remote, quantitative imaging of the rat small intestine. The probe molecule employed, Tb-3,6,9- tris(methylene phosphonic acid n-butyl ester)-3,6,9,15- tetraaza-bicyclco(9.3.1)pentadeca-1(15),11,13-triene(Tb- PTCMB), is shown to bind with the small intestine and provide improved image contrast. High sensitivity is possible due to the absorption-emission Stoke's shift exhibited by the Tb-PTCMB complex. Excitation is centered near 270 nm and multifeatured emission is observed at 490, 550, 590, and 625 nm. Sprague-Dawley rats were dosed with the Tb-PTCMB complex, which shows biodistribution, leading to preferential binding to the inner surface of the small intestine. It is shown that the fluorescent image, taken at 550 nm, can be used to quantify the amount of Tb-PCTMB present in an excised tissue sample. The 3(sigma) detection limits are found to be in the femtomole range. An optical mass balance for Tb-PCTMB-dosed small intestine is performed and along with radiotracer biodistribution, demonstrates that approximately 40 percent of the marker probe resides in the endothelial tissue of the small intestine inner lumen. This result is of particular interest since most adult colon cancers develop in this region. These result demonstrate the ability to perform spatial, quantitative, in vitro, endoscopic imaging of a complex biological sample using a probe marker.

  12. Use of a portable time-resolved fluorometer to determine oxytetracycline residue in four fruit crops

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Worldwide, oxytetracycline (OTC) is used in fruit and vegetable crops to prevent and treat bacteria diseases. In the U.S., the Environmental Protection Agency approved its use in apple, pear, peach, and nectarine, and set tolerance at 350 ng/g. OTC residues in 12 varieties of these fruits are determ...

  13. Simulation analysis of moored fluorometer time series from the Mid-Atlantic Bight during 1987--1990

    SciTech Connect

    Walsh, J.J.

    1990-01-01

    The goal of the previous research during 1987-1990 within the DOE (Department of Energy) Shelf Edge Exchange Processes (SEEP) program in the Mid-Atlantic Bight was to understand the physical and biogeochemical processes effecting the diffusive exchange of the proxies of energy-related, by-products associated with particulate matter between estuarine, shelf, and slope waters on this continental margin. As originally envisioned in the SEEP program plan, SEEP-III would take place at Cape Hatteras to study the advective exchange of materials by a major boundary current. One problem of continuing interest is the determination of the local assimilative capacity of slope waters and sediments off the eastern seaboard of the US to lengthen the pathway between potentially harmful energy by-products and man. At basin scales, realistic specification of the lateral transport by western boundary currents of particulate matter is a necessary input to global models of carbon/nitrogen cycling. Finally, at these global scales, the generic role of continental margins in cycling greenhouse gases, e.g. CO{sub 2}, CH{sub 4}, and N{sub 2}O, is now of equal interest. This continuing research of model construction and evaluation within the SEEP program focuses on all three questions at local, regional, and basin scales. Results from SEEP-I and II are discussed as well as plans for SEEP-III. 14 figs., 3 tabs.

  14. Rapid Processing of Turner Designs Model 10-Au-005 Internally Logged Fluorescence Data

    EPA Science Inventory

    Continuous recording of dye fluorescence using field fluorometers at selected sampling sites facilitates acquisition of real-time dye tracing data. The Turner Designs Model 10-AU-005 field fluorometer allows for frequent fluorescence readings, data logging, and easy downloading t...

  15. Ultraviolet fluorescence monitor

    SciTech Connect

    Hargis, P.J. Jr.; Preppernau, B.L.; Aragon, B.P.

    1997-05-01

    A multispectral ultraviolet (UV) fluorescence imaging fluorometer and a pulsed molecular beam laser fluorometer were developed to detect volatile organic compounds of interest in environmental monitoring and drug interdiction applications. The UV fluorescence imaging fluorometer is a relatively simple instrument which uses multiple excitation wavelengths to measure the excitation/emission matrix for irradiated samples. Detection limits in the high part-per-million to low part-per-million range were measured for a number of volatile organic vapors in the atmosphere. Detection limits in the low part-per-million range were obtained using cryogenic cooling to pre-concentrate unknown samples before introducing them into the imaging fluorometer. A multivariate analysis algorithm was developed to analyze the excitation/emission matrix and used to determine the relative concentrations of species in computer synthesized mixtures containing up to five organic compounds. Analysis results demonstrated the utility of multispectral UV fluorescence in analytical measurements. A transportable UV fluorescence imaging fluorometer was used in two field tests. Field test results demonstrated that detection limits in the part-per-billion range were needed to reliably identify volatile organic compounds in realistic field test measurements. The molecular beam laser fluorometer, a more complex instrument with detection limits in the part-per-billion to part-per-trillion range, was therefore developed to satisfy detection sensitivity requirements for field test measurements. High-resolution spectroscopic measurements made with the molecular beam laser fluorometer demonstrated its utility in identifying volatile organic compounds in the atmosphere.

  16. A 5-kg time-resolved luminescence photometer with multiple excitation sources

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A portable fluorometer was developed to detect food contaminants and environmental pollutants including, in particular, two classes of antibiotics: tetracyclines and fluoroquinolones. Time resolution was implemented to take advantage of lanthanide-sensitized luminescence. Excitation sources included...

  17. Oceanographic Instrument

    NASA Technical Reports Server (NTRS)

    1994-01-01

    Developed under NASA contract, the Fast Repetition Rate (FRR) fluorometer is a computer-controlled instrument for measuring the fluorescence of phytoplankton, microscopic plant forms that provide sustenance for animal life in the oceans. The fluorometer sensor is towed by ship through the water and the resulting printouts are compared with satellite data. The instrument is non-destructive and can be used in situ, providing scientific information on ocean activity and productivity.

  18. In situ Measurements of Phytoplankton Fluorescence Using Low Cost Electronics

    PubMed Central

    Leeuw, Thomas; Boss, Emmanuel S.; Wright, Dana L.

    2013-01-01

    Chlorophyll a fluorometry has long been used as a method to study phytoplankton in the ocean. In situ fluorometry is used frequently in oceanography to provide depth-resolved estimates of phytoplankton biomass. However, the high price of commercially manufactured in situ fluorometers has made them unavailable to some individuals and institutions. Presented here is an investigation into building an in situ fluorometer using low cost electronics. The goal was to construct an easily reproducible in situ fluorometer from simple and widely available electronic components. The simplicity and modest cost of the sensor makes it valuable to students and professionals alike. Open source sharing of architecture and software will allow students to reconstruct and customize the sensor on a small budget. Research applications that require numerous in situ fluorometers or expendable fluorometers can also benefit from this study. The sensor costs US$150.00 and can be constructed with little to no previous experience. The sensor uses a blue LED to excite chlorophyll a and measures fluorescence using a silicon photodiode. The sensor is controlled by an Arduino microcontroller that also serves as a data logger. PMID:23783738

  19. Tissue-based water quality biosensors for detecting chemical warfare agents

    DOEpatents

    Greenbaum, Elias; Sanders, Charlene A.

    2003-05-27

    A water quality sensor for detecting the presence of at least one chemical or biological warfare agent includes: a cell; apparatus for introducing water into the cell and discharging water from the cell adapted for analyzing photosynthetic activity of naturally occurring, free-living, indigenous photosynthetic organisms in water; a fluorometer for measuring photosynthetic activity of naturally occurring, free-living, indigenous photosynthetic organisms drawn into the cell; and an electronics package that analyzes raw data from the fluorometer and emits a signal indicating the presence of at least one chemical or biological warfare agent in the water.

  20. FLUORESCENCE ASSESSMENT OF THE MAXIMUM QUANTUM EFFICIENCY PHOTOSYNTHESIS IN THE WESTERN NORTH ATLANTIC

    EPA Science Inventory

    The maximum quantum efficiency of phytoplankton photosystem II photochemistry was assessed using a pump and probe fluorometer on an offshore-onshore transect from the oligotrophic blue waters of the western Sargasso Sea to the eutrophic waters of lower Delaware Bay. ow values of ...

  1. Efficient Chlorophyll Fluorescence Measurements of Sugarcane

    Technology Transfer Automated Retrieval System (TEKTRAN)

    As with many crops, chlorophyll fluorescence emission is a promising tool for measuring responses of sugarcane (Saccharum spp.) to biotic and abiotic stresses. Chlorophyll fluorescence can be easily measured using portable fluorometers. However, several factors should be considered in order to op...

  2. "Open-Box" Approach to Measuring Fluorescence Quenching Using an iPad Screen and Digital SLR Camera

    ERIC Educational Resources Information Center

    Koenig, Michael H.; Yi, Eun P.; Sandridge, Matthew J.; Mathew, Alexander S.; Demas, James N.

    2015-01-01

    Fluorescence quenching is an analytical technique and a common undergraduate laboratory exercise. Unfortunately, a typical quenching experiment requires the use of an expensive fluorometer that measures the relative fluorescence intensity of a single sample in a closed compartment unseen by the experimenter. To overcome these shortcomings, we…

  3. Detection of plant injury from application of non-selective herbicide by measurement of chlorophyll reflectance and fluorescene

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Subtle changes in canopy reflectance could present useful information to detect the onset of crop stress. This study was conducted in a greenhouse to evaluate a portable spectroradiometer and a portable chlorophyll fluorometer for the detection of crop injury caused by glyphosate spray. In this stud...

  4. Smart oxygen cuvette for optical monitoring of dissolved oxygen in biological blood samples

    NASA Astrophysics Data System (ADS)

    Dabhi, Harish; Alla, Suresh Kumar; Shahriari, Mahmoud R.

    2010-02-01

    A smart Oxygen Cuvette is developed by coating the inner surface of a cuvette with oxygen sensitive thin film material. The coating is glass like sol-gel based sensor that has an embedded ruthenium compound in the glass film. The fluorescence of the ruthenium is quenched depending on the oxygen level. Ocean Optics phase fluorometer, NeoFox is used to measure this rate of fluorescence quenching and computes it for the amount of oxygen present. Multimode optical fibers are used for transportation of light from an LED source to cuvette and from cuvette to phase fluorometer. This new oxygen sensing system yields an inexpensive solution for monitoring the dissolved oxygen in samples for biological and medical applications. In addition to desktop fluorometers, smart oxygen cuvettes can be used with the Ocean Optics handheld Fluorometers, NeoFox Sport. The Smart Oxygen Cuvettes provide a resolution of 4PPB units, an accuracy of less than 5% of the reading, and 90% response in less than 10 seconds.

  5. Water Quality and Plankton in the United States Nearshore Waters of Lake Huron

    EPA Science Inventory

    We conducted an intensive survey for the US nearshore of Lake Huron along a continuous segment (523 km) from Port Huron Michigan to Detour Passage. A depth contour of 20 m was towed with a CTD, fluorometer, transmissometer, and laser optical plankton counter (LOPC). The continu...

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

    ERIC Educational Resources Information Center

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

    2007-01-01

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

  7. 40 CFR 53.63 - Test procedure: Wind tunnel inlet aspiration test.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    .... Relative aspiration is the ratio (expressed as a percentage) of the aerosol mass concentration measured by... using a calibrated fluorometer. Calculate and record the mass concentration as: Equation 15 ER18jy97.108 where: i = replicate number; Mref = mass of material collected with the reference method sampler;...

  8. Fluorescence Spectroscopy in a Shoebox

    ERIC Educational Resources Information Center

    Wahab, M. Farooq

    2007-01-01

    The construction of a fluorometer and a spectrofluorometer using flashlight or a sunlight excitation source in a shoebox and the eye as a detector is being described. The assembly helps in understanding several fundamental ideas related to the subject very easily.

  9. Measuring indigenous photosynthetic organisms to detect chemical warefare agents in water

    DOEpatents

    Greenbaum, Elias; Sanders, Charlene A.

    2005-11-15

    A method of testing water to detect the presence of a chemical or biological warfare agent is disclosed. The method is carried out by establishing control data by providing control water containing indigenous organisms but substantially free of a chemical and a biological warfare agent. Then measuring photosynthetic activity of the control water with a fluorometer to obtain control data to compare with test data to detect the presence of the chemical or agent. The test data is gathered by providing test water comprising the same indigenous organisms as contained in the control water. Further, the test water is suspected of containing the chemical or agent to be tested for. Photosynthetic activity is also measured by fluorescence induction in the test water using a fluorometer.

  10. Use of a Fluorometric Imaging Plate Reader in high-throughput screening

    NASA Astrophysics Data System (ADS)

    Groebe, Duncan R.; Gopalakrishnan, Sujatha; Hahn, Holly; Warrior, Usha; Traphagen, Linda; Burns, David J.

    1999-04-01

    High-throughput screening (HTS) efforts at Abbott Laboratories have been greatly facilitated by the use of a Fluorometric Imaging Plate Reader. The FLIPR consists of an incubated cabinet with integrated 96-channel pipettor and fluorometer. An argon laser is used to excite fluorophores in a 96-well microtiter plate and the emitted fluorometer. An argon laser is used to excite fluorophores in a 96-well microtiter plate and the emitted fluorescence is imaged by a cooled CCD camera. The image data is downloaded from the camera and processed to average the signal form each well of the microtiter pate for each time point. The data is presented in real time on the computer screen, facilitating interpretation and trouble-shooting. In addition to fluorescence, the camera can also detect luminescence form firefly luciferase.

  11. An optimised method for correcting quenched fluorescence yield

    NASA Astrophysics Data System (ADS)

    Biermann, L.; Guinet, C.; Bester, M.; Brierley, A.; Boehme, L.

    2014-05-01

    Under high light intensity, phytoplankton protect their photosystems from bleaching through non-photochemical quenching processes. The consequence of this is suppression of fluorescence emission, which must be corrected when measuring in situ yield with fluorometers. Previously, this has been done using the limit of the mixed layer, assuming that phytoplankton are uniformly mixed from the surface to this depth. However, the assumption of homogeneity is not robust in oceanic regimes that support deep chlorophyll maxima. To account for these features, we correct from the limit of the euphotic zone, defined as the depth at which light is at ~1% of the surface value. This method was applied to fluorescence data collected by eleven animal-borne fluorometers deployed in the Southern Ocean over four austral summers. Six tags returned data showing evidence of deep chlorophyll features. Using the depth of the euphotic layer, quenching was corrected without masking subsurface fluorescence signals.

  12. Validation of Ferrybox Observations and Comparison between Ferrybox Ferries

    NASA Astrophysics Data System (ADS)

    Kaitala, Seppo; Seppälä, Jukka; Maunula, Petri; Kallio, Kari Y.; Ruohola, Jani

    2016-04-01

    The annual Alg@line ferrybox instrument calibration is carried out annually in January in Finnish Environment institute (SYKE). The chlorophyll and phycocyanin fluorometers are calibrated with algae cultures. All sensors are compared also with each other. Fluorometer readings are validated against water sample analysis in laboratory biweekly during the year. This validation step using water samples is necessary due to different optical properties of phytoplankton species. When calibration and validation are carried out with similar protocols, there is a possibility to compare the ferrybox observations in the same area by different ferries. The Ferryboxes operating in the Central Baltic occasionally occur in the same area within the 24 hours. Spatiotemporal comparisons of chlorophyll fluorescence, temperature and salinity observations are demonstrated.

  13. Tissue-based standoff biosensors for detecting chemical warfare agents

    DOEpatents

    Greenbaum, Elias; Sanders, Charlene A.

    2003-11-18

    A tissue-based, deployable, standoff air quality sensor for detecting the presence of at least one chemical or biological warfare agent, includes: a cell containing entrapped photosynthetic tissue, the cell adapted for analyzing photosynthetic activity of the entrapped photosynthetic tissue; means for introducing an air sample into the cell and contacting the air sample with the entrapped photosynthetic tissue; a fluorometer in operable relationship with the cell for measuring photosynthetic activity of the entrapped photosynthetic tissue; and transmitting means for transmitting analytical data generated by the fluorometer relating to the presence of at least one chemical or biological warfare agent in the air sample, the sensor adapted for deployment into a selected area.

  14. Fluorometric procedures for dye tracing

    USGS Publications Warehouse

    Wilson, James F.; Cobb, Ernest D.; Kilpatrick, F.A.

    1986-01-01

    This manual describes the current fluorometric procedures used by the U.S. Geological Survey in dye tracer studies such as time of travel, dispersion, reaeration, and dilution-type discharge measurements. The advantages of dye tracing are (1) low detection and measurement limits and (2) simplicity and accuracy in measuring dye tracer concentrations using fluorometric techniques. The manual contains necessary background information about fluorescence, dyes, and fluorometers and a description of fluorometric operation and calibration procedures as a guide for laboratory and field use. The background information should be useful to anyone wishing to experiment with dyes, fluorometer components, or procedures different from those described. In addition, a brief section on aerial photography is included because of its possible use to supplement ground-level fluorometry.

  15. Fluorometric procedures for dye tracing

    USGS Publications Warehouse

    Wilson, James E., Jr.; Cobb, E.D.; Kilpatrick, F.A.

    1984-01-01

    This manual describes the current fluorometric procedures used by the U.S. Geological Survey in dye tracer studies such as time of travel, dispersion, reaeration, and dilution-type discharge measurements. The outstanding characteristics of dye tracing are: (1) the low detection and measurement limits, and (2) the simplicity and accuracy of measuring dye tracer concentrations using fluorometric techniques. The manual contains necessary background information about fluorescence, dyes, and fluorometers and a description of fluorometric operation and calibration procedures as a general guide for laboratory and field use. The background information should be useful to anyone wishing to experiment with dyes, fluorometer components, or procedures different from those described. In addition, a brief section is included on aerial photography because of its possible use to supplement ground-level fluorometry. (USGS)

  16. Fluorometric procedures for dye tracing

    USGS Publications Warehouse

    Wilson, James F.

    1968-01-01

    This manual describes the current fluorometric procedures used by the U.S. Geological Survey in dye tracer studies such as time of travel, dispersion, reaeration, and dilution-type discharge measurements. The advantages of dye tracing are (1) low detection and measurement limits and (2) simplicity and accuracy in measuring dye tracer concentrations using fluorometric techniques. The manual contains necessary background information about fluorescence, dyes, and fluorometers and a description of fluorometric operation and calibration procedures as a guide for laboratory and field use. The background information should be useful to anyone wishing to experiment with dyes, fluorometer components, or procedures different from those described. In addition, a brief section on aerial photography is included because of its possible use to supplement ground-level fluorometry.

  17. Continuous environmental monitoring for aqueous effluents

    SciTech Connect

    Pitt, Jr., W. W.; Jones, G. Jr.

    1980-05-01

    An aquatic environmental monitor has been developed that will continuously monitor aqueous waste streams from coal processing plants. The monitor contains three different instruments: a continuous chemical oxygen demand monitor and two continuous-flow fluorometers with different excitation-emission characteristics. A prototype instrument was fabricated and evaluated for several different applications. The details of the instrument design and results of its evaluation are presented in this report.

  18. Bio-optical profile data report coastal transition zone program, R/V Point Sur, June 15-28, 1987

    NASA Technical Reports Server (NTRS)

    Davis, Curtiss O.; Rhea, W. Joseph

    1990-01-01

    Twenty vertical profiles of the bio-optical properties of the ocean were made during a research cruise on the R/V Point Sur, June 15 to 28, 1987, as part of the Coastal Transition Zone Program off Point Arena, California. Extracted chlorophyll values were also measured at some stations to provide calibration data for the in situ fluorometer. This summary provides investigators with an overview of the data collected. The entire data set is available in digital form.

  19. Fluorometric analysis for neoplasm diagnostics and localization

    NASA Astrophysics Data System (ADS)

    Kwasny, Miroslaw; Mierczyk, Zygmunt

    1997-10-01

    The paper presents methods of laser-induced fluorescence with the use of endo- and exogenous dyes for diagnosis of early tumors of aerodigestive tracts, colons, bladder, GYN, and skin, as well as a review of equipment developed during laboratory examination, construction of diagnostic instruments and clinical use of fluorometric methods with application of various devices, from simple fluorometers to sophisticated endoscopic spectra-analyzers.

  20. Laser-induced fluorescence over optical fibers for real time in situ measurement of petroleum hydrocarbons in seawater

    NASA Astrophysics Data System (ADS)

    Lieberman, S. H.; Inman, S. M.; Theriault, G. A.

    1993-04-01

    A fiber optic-based fluorometer system is described that uses a pulsed laser to induce fluorescence and a time-gated linear photodiode array coupled to a spectrograph for rapid measurement of fluorescence emission spectra and fluorescence decay times. Data is presented from studies conducted in San Diego Bay where the system has been used for real-time in situ measurements of temporal and spatial variability of petroleum hydrocarbons in seawater. Results show that the optical fiber fluorometer system is capable of direct quantification of low level (parts-per-billion diesel fuel marine equivalent) concentrations of petroleum hydrocarbons. Analysis times for measurement of complete fluorescence emission spectra are fast (tens of milliseconds) and compare with the temporal response characteristics for temperature and conductivity sensors that are used for measuring standard physical hydrographic parameters. Results obtained with the fiber optic fluorometer system during a mapping study in San Diego Bay show good correlation with GC-MS analysis of total polycyclic aromatic hydrocarbons (PAHs) measured on discrete samples collected during the study.

  1. Swallowable fluorometric capsule for wireless triage of gastrointestinal bleeding.

    PubMed

    Nemiroski, A; Ryou, M; Thompson, C C; Westervelt, R M

    2015-12-01

    Real-time detection of gastrointestinal bleeding remains a major challenge because there does not yet exist a minimally invasive technology that can both i) monitor for blood from an active hemorrhage and ii) uniquely distinguish it from blood left over from an inactive hemorrhage. Such a device would be an important tool for clinical triage. One promising solution, which we have proposed previously, is to inject a fluorescent dye into the blood stream and to use it as a distinctive marker of active bleeding by monitoring leakage into the gastrointestinal tract with a wireless fluorometer. This paper reports, for the first time to our knowledge, the development of a swallowable, wireless capsule with a built-in fluorometer capable of detecting fluorescein in blood, and intended for monitoring gastrointestinal bleeding in the stomach. The embedded, compact fluorometer uses pinholes to define a microliter sensing volume and to eliminate bulky optical components. The proof-of-concept capsule integrates optics, low-noise analog sensing electronics, a microcontroller, battery, and low power Zigbee radio, all into a cylindrical package measuring 11 mm × 27 mm and weighing 10 g. Bench-top experiments demonstrate wireless fluorometry with a limit-of-detection of 20 nM aqueous fluorescein. This device represents a major step towards a technology that would enable simple, rapid detection of active gastrointestinal bleeding, a capability that would save precious time and resources and, ultimately, reduce complications in patients. PMID:26490455

  2. Surfzone Tracer Transport and Dispersion during the IB09 Field Experiment

    NASA Astrophysics Data System (ADS)

    Hally-Rosendahl, K.; Feddersen, F.; Clark, D. B.; Guza, R. T.

    2010-12-01

    Transport and dilution of pollutants discharged into the surfzone are not well understood, limiting water quality forecast accuracy and exposing beachgoers to health risks. Surfzone dispersion in an alongshore current has been characterized using observations of the downstream evolution of Rhodamine WT fluorescent dye tracer. Continuously released near the shoreline, dye was advected by alongshore mean currents while spreading cross-shore, forming shore-parallel plumes. Initial observations (HB06) with a single jetski sampling near-surface dye concentrations in relatively short (400 m) plumes have been used to estimate bulk eddy diffusivities within the surfzone [Clark et al., JGR, in press 2010]. However, questions about the seaward extent of surfzone mixing, exchange with offshore waters, vertical tracer structure, and mixing over longer distances and times were unresolved. Results from more comprehensive observations during the fall 2009 IB09 experiment at Imperial Beach, California will be discussed. The near-surface dye field was mapped with two GPS- and fluorometer-equipped jetskis. Vertical dye concentration profiles were measured with a boat-towed vertical fluorometer array as dye leaked offshore from the surfzone. Additionally, near-shoreline fluorometers were deployed at various downstream distances, and a cross-shore array of six bottom-mounted instrument packages (1-4 m depth) measured dye, waves, and currents. Aerial photographs of the dye field were also acquired. Continuous near-shoreline dye releases spanned a range of wave and current conditions. Dye was measured up to 3 km downstream of the source and over 500 m from shore. Fluorescent Rhodamine WT dye tracer plume during the IB09 experiment at Imperial Beach, California, fall 2009.

  3. ALOHA Cabled Observatory: On-going results and new instruments

    NASA Astrophysics Data System (ADS)

    Howe, B. M.; Lukas, R.

    2013-12-01

    Since June 2011, the ALOHA Cabled Observatory (ACO) has been collecting abyssal oceanographic data. The ACO is at Station ALOHA 100 km north of Oahu, the field site of the Hawaii Ocean Time-series (HOT) program that has collected biological, physical, and chemical oceanographic data since 1988. At 4728 m water depth, it is the world's deepest operating cabled observatory. On-going results will be presented along with results from two new instrument packages to be deployed: a basic sensor package (CTDO2, fluorometer, acoustic modem, ADCP), and a video/light/hydrophone combination. Plans for future research will be discussed. [Work supported by the National Science Foundation.

  4. An assay for measurement of protein adsorption to glass vials.

    PubMed

    Varmette, Elizabeth; Strony, Brianne; Haines, Daniel; Redkar, Rajendra

    2010-01-01

    Protein adsorption to primary packaging is one of the problems faced by biopharmaceutical drug companies. An assay was developed to quantify loss of proteins to glass vial surfaces. The assay involves the labeling of protein with a fluorescent dye, incubation of the labeled protein with the vial surface, elution of the adsorbed protein using a stripping buffer, and determination of fluorescence of the adsorbed protein using a fluorometer. The assay is simple to set up, accurate, sensitive, and flexible. The assay can be modified for indirect measurement of protein adsorption and offers an attractive alternative for researchers to quantify protein adsorption to glass vials and syringes. PMID:21502031

  5. Fluorescence lifetime resolution with phase fluorometry

    NASA Astrophysics Data System (ADS)

    Ide, Geert; Engelborghs, Yves; Persoons, Andre

    1983-07-01

    A phase fluorometer for the measurement of fluorescence lifetimes was constructed from commercially available components. The instrument was tested by using optical delays of 4 and 2 cm, showing an accuracy of 10 ps. Lifetimes, as short as 0.3 ns, obtained by the quenching of fluorescein by KI, were analyzed with a standard deviation of 3 ps. The lifetime resolving power was checked using mixtures of acridine and quinine-sulphate and least-squares fitting procedures. Accurate amplitude ratios were obtained with the technique of phase-sensitive detection [J. R. Lakowicz and H. Cherek, J. Biochem. Biophys. Methods 5, 19 (1981)].

  6. Progress toward implantable fluorescence-based sensors for monitoring glucose levels in interstitial fluid

    NASA Astrophysics Data System (ADS)

    McShane, Michael J.; O'Neal, Dennis P.; Russell, Ryan J.; Pishko, Michael V.; Cote, Gerard L.

    2000-05-01

    A painless monitoring procedure for diabetics has proven to be an elusive goal. While completely noninvasive measurements are the desired technique, minimally invasive procedures using implanted fluorescence sensor chemistry offer significant advantages in specificity over current noninvasive approaches. The goal was to evaluate the potential for transdermal glucose sensing using intensity measurements from implanted microspheres. A fiber-optic probe and spectrometer were custom-built for collection of in vivo data. Comparisons with commercial fluorometers show the constructed device is adequate for this project.

  7. Fluorometric functional assay for ion channel proteins in lipid nanovesicle membranes

    NASA Astrophysics Data System (ADS)

    Patti, J. T.; Montemagno, C. D.

    2007-08-01

    Voltage-gated membrane proteins function as biomolecular transistors, making them attractive components for biologically based nanodevices. A functional assay for purified channel proteins is described and demonstrated with sodium selective, voltage-gated NaChBac ion channels. Purified NaChBac proteins were incorporated into a nanovesicle system utilizing oxonol VI, a fluorescent indicator of trans-membrane voltage. The ionophore valinomycin was used to trigger a change in membrane potential, allowing the observation of sodium permeability using a fluorometer. This method is suitable for concurrently testing a large population of purified proteins prior to incorporation in nanodevices.

  8. Surface fluorescence studies of tissue mitochondrial redox state in isolated perfused rat lungs.

    PubMed

    Staniszewski, Kevin; Audi, Said H; Sepehr, Reyhaneh; Jacobs, Elizabeth R; Ranji, Mahsa

    2013-04-01

    We designed a fiber-optic-based optoelectronic fluorometer to measure emitted fluorescence from the auto-fluorescent electron carriers NADH and FAD of the mitochondrial electron transport chain (ETC). The ratio of NADH to FAD is called the redox ratio (RR = NADH/FAD) and is an indicator of the oxidoreductive state of tissue. We evaluated the fluorometer by measuring the fluorescence intensities of NADH and FAD at the surface of isolated, perfused rat lungs. Alterations of lung mitochondrial metabolic state were achieved by the addition of rotenone (complex I inhibitor), potassium cyanide (KCN, complex IV inhibitor) and/or pentachlorophenol (PCP, uncoupler) into the perfusate recirculating through the lung. Rotenone- or KCN-containing perfusate increased RR by 21 and 30%, respectively. In contrast, PCP-containing perfusate decreased RR by 27%. These changes are consistent with the established effects of rotenone, KCN, and PCP on the redox status of the ETC. Addition of blood to perfusate quenched NADH and FAD signal, but had no effect on RR. This study demonstrates the capacity of fluorometry to detect a change in mitochondrial redox state in isolated perfused lungs, and suggests the potential of fluorometry for use in in vivo experiments to extract a sensitive measure of lung tissue health in real-time. PMID:23238793

  9. Improvement of urinary delta-aminolevulinic acid determination by HPLC and fluorescence detection using condensing reaction with acetylacetone and formaldehyde.

    PubMed

    Endo, Y; Okayama, A; Endo, G; Ueda, T; Nakazono, N; Horiguchi, S

    1994-03-01

    We improved the method for determining urinary delta-aminolevulinic acid (ALA) by HPLC-fluorometer after pre-column derivatization with acetylacetone and formaldehyde, and a stable ALA derivative was obtained without any effect from various urinary components as demonstrated by the complete recovery of ALA (100.9 +/- 5.5%, n = 85) from the urine samples. The modified procedure was as follows: Twenty microliters of urine sample, 5 ml of acetylacetone solution (acetylacetone/ethanol/distilled water containing 4 milligrams of NaCl; 15/10/75), and 0.45 ml of 9.3% formaldehyde solution were mixed and boiled for 15 min. The fluorescent derivative of ALA was separated and analyzed by HPLC with the fluorometer at Ex 246 nm and Em 458 nm. Using a gradient program, the retention time of the ALA derivative was 7.3 min and the analysis could be repeated at 13 min intervals. Concentrations of ALA in urine samples measured by this method were significantly correlated with those measured by the Mauzerall-Granick (M-G) method (n = 85, r = 0.993, p < 0.001). The values obtained by our method were, however, lower than those obtained by the M-G method. Urinary ALA concentrations of 40 non-lead workers ranged from 0.1 to 2.3 mg/g creatinine with the mean +/- SD of 1.1 +/- 0.4 mg/g creatinine as measured by the present method. PMID:8007435

  10. Fluorometry of ischemia reperfusion injury in rat lungs in vivo

    NASA Astrophysics Data System (ADS)

    Sepehr, R.; Staniszewski, K.; Jacobs, E. R.; Audi, S.; Ranji, Mahsa

    2013-02-01

    Previously we demonstrated the utility of optical fluorometry to evaluate lung tissue mitochondrial redox state in isolated perfused rats lungs under various chemically-induced respiratory states. The objective of this study was to evaluate the effect of acute ischemia on lung tissue mitochondrial redox state in vivo using optical fluorometry. Under ischemic conditions, insufficient oxygen supply to the mitochondrial chain should reduce the mitochondrial redox state calculated from the ratio of the auto-fluorescent mitochondrial metabolic coenzymes NADH (Nicotinamide Adenine Dinucleotide) and FAD (Flavoprotein Adenine Dinucleotide). The chest of anesthetized, and mechanically ventilated Sprague-Dawley rat was opened to induce acute ischemia by clamping the left hilum to block both blood flow and ventilation to one lung for approximately 10 minutes. NADH and FAD fluorescent signals were recorded continuously in a dark room via a fluorometer probe placed on the pleural surface of the left lung. Acute ischemia caused a decrease in FAD and an increase in NADH, which resulted in an increase in the mitochondrial redox ratio (RR=NADH/FAD). Restoration of blood flow and ventilation by unclamping the left hilum returned the RR back to its baseline. These results (increase in RR under ischemia) show promise for the fluorometer to be used in a clinical setting for evaluating the effect of pulmonary ischemia-reperfusion on lung tissue mitochondrial redox state in real time.

  11. Lightweight Fiber Optic Gas Sensor for Monitoring Regenerative Food Production

    NASA Technical Reports Server (NTRS)

    Schmidlin, Edward; Goswami, Kisholoy

    1995-01-01

    In this final report, Physical Optics Corporation (POC) describes its development of sensors for oxygen, carbon dioxide, and relative humidity. POC has constructed a phase fluorometer that can detect oxygen over the full concentration range from 0 percent to 100 percent. Phase-based measurements offer distinct advantages, such as immunity to source fluctuation, photobleaching, and leaching. All optics, optoelectronics, power supply, and the printed circuit board are included in a single box; the only external connections to the fluorometer are the optical fiber sensor and a power cord. The indicator-based carbon dioxide sensor is also suitable for short-term and discrete measurements over the concentration range from 0 percent to 100 percent. The optical fiber-based humidity sensor contains a porous core for direct interaction of the light beam with water vapor within fiber pores; the detection range for the humidity sensor is 10 percent to 100 percent, and response time is under five minutes. POC is currently pursuing the commercialization of these oxygen and carbon dioxide sensors for environmental applications.

  12. Portable system for the detection of micromolar concentrations of glucose

    PubMed Central

    Kostov, Yordan; Ge, Xudong; Rao, Govind; Tolosa, Leah

    2014-01-01

    Glucose in non-invasively collected biofluids is generally in the micromolar range and thus, requires sensing methodologies capable of measuring glucose at these levels. Here, we present a small fluorometer system that can quantify glucose in the range of 0–5 μM with resolution of ~0.07 μM. It relies on the glucose binding protein (GBP) fluorescently labeled with two fluorophores. Fluorescence signals from the dual-labeled GBP are utilized in a ratiometric mode, making the measurements insensitive to variations in protein concentration and other systematic errors. Fluorescence is quantified by a miniature, dedicated ratiometric fluorometer that is powered via USB. Concentration is calculated using an ultra-mobile personal computer (UMPC). The whole system is designed to be pocket sized suitable for point-of-care or bedside applications. Test results suggest that the system is a promising tool for accurate measurements of low glucose concentrations (0.1–10 μM) in biological samples. PMID:24587594

  13. Relationship between Chlorophyll a Concentration, Light Attenuation and Diving Depth of the Southern Elephant Seal Mirounga leonina

    PubMed Central

    Jaud, Thomas; Dragon, Anne-Cécile; Garcia, Jade Vacquie; Guinet, Christophe

    2012-01-01

    Recently, a number of Antarctic marine environmental studies have used oceanographic parameters collected from instrumented top predators for ecological and physical information. Phytoplankton concentration is generally quantified through active measurement of chlorophyll fluorescence. In this study, light absorption coefficient (K0.75) was used as an indicator of phytoplankton concentration. This measurement, easy to obtain and requiring low electric power, allows for assessing of the fine scale horizontal structuring of phytoplankton. As part of this study, Southern elephant seals (SES) were simultaneously equipped with a fluorometer and a light logger. Along the SES tracks, variations in K0.75 were strongly correlated with chlorophyll, a concentration measured by the fluorometer within the euphotic layer. With regards to SES foraging behaviour, bottom depth of the seal’s dive was highly dependent on light intensity at 150 m, indicating that the vertical distribution of SES’s prey such as myctophids is tightly related to light level. Therefore, change in phytoplankton concentration may not only have a direct effect on SES’s prey abundance but may also determine their vertical accessibility with likely consequences on SES foraging efficiency. PMID:23082166

  14. A hand-held electronic tongue based on fluorometry for taste assessment of tea.

    PubMed

    Chang, Kuang-Hua; Chen, Richie L C; Hsieh, Bo-Chuan; Chen, Po-Chung; Hsiao, Hsien-Yi; Nieh, Chi-Hua; Cheng, Tzong-Jih

    2010-12-15

    A hand-held electronic tongue was developed for determining taste levels of astringency and umami in tea infusions. The sensing principles are based on quenching the fluorescence of 3-aminophthalate by tannin, and the fluorogenic reaction of o-phthalaldehyde (OPA) with amino acids to determine astringency and umami levels, respectively. Both reactions were measured by a single fluorescence sensing system with same excitation and emission wavelengths (340/425 nm). This work describes in detail the design, fabrication, and performance evaluation of a hand-held fluorometer with an ultra-violet light emitted diode (UVLED) and a photo-detector with a filter built-in. The dimension and the weight of proposed electronic tongue prototype are only 120×60×65 mm(3) and 150 g, respectively. The detection limits of this prototype for theanine and tannic acid were 0.2 μg/ml and 1 μg/ml, respectively. Correlation coefficients of this prototype compared with a commercial fluorescence instrument are both higher than 0.995 in determinations of tannin acid and theanine. Linear detection ranges of the hand-held fluorometer for tannic acid and theanine are 1-20 μg/ml and 0.2-10 μg/ml (CV<5%, n=3), respectively. A specified taste indicator for tea, defined as ratio of umami to astringency, was adopted here to effectively distinguish flavour quality of partially fermented Oolong teas. PMID:20728331

  15. Spatial extent and dissipation of the deep chlorophyll layer in Lake Ontario during the Lake Ontario lower foodweb assessment, 2003 and 2008

    USGS Publications Warehouse

    Watkins, J. M.; Weidel, Brian M.; Rudstam, L. G.; Holek, K. T.

    2014-01-01

    Increasing water clarity in Lake Ontario has led to a vertical redistribution of phytoplankton and an increased importance of the deep chlorophyll layer in overall primary productivity. We used in situ fluorometer profiles collected in lakewide surveys of Lake Ontario in 2008 to assess the spatial extent and intensity of the deep chlorophyll layer. In situ fluorometer data were corrected with extracted chlorophyll data using paired samples from Lake Ontario collected in August 2008. The deep chlorophyll layer was present offshore during the stratified conditions of late July 2008 with maximum values from 4–20 μg l−1 corrected chlorophyll a at 10 to 17 m depth within the metalimnion. Deep chlorophyll layer was closely associated with the base of the thermocline and a subsurface maximum of dissolved oxygen, indicating the feature's importance as a growth and productivity maximum. Crucial to the deep chlorophyll layer formation, the photic zone extended deeper than the surface mixed layer in mid-summer. The layer extended through most of the offshore in July 2008, but was not present in the easternmost transect that had a deeper surface mixed layer. By early September 2008, the lakewide deep chlorophyll layer had dissipated. A similar formation and dissipation was observed in the lakewide survey of Lake Ontario in 2003.

  16. Quantitating Fluorescence Intensity From Fluorophore: Assignment of MESF Values

    PubMed Central

    Gaigalas, A. K.; Wang, Lili; Schwartz, Abe; Marti, Gerald E.; Vogt, Robert F.

    2005-01-01

    A procedure is presented to convert the comparison of measured fluorescence signals into a comparison of fluorescence yields (FY). The fluorescence yield, which is a property of a solution or a suspension, is defined as the product of the fluorophore concentration and the molecular quantum yield. The paper revises the measurement model which relates the measured fluorescence signal to the FY. The equality of FY of two solutions provides an equivalence between the concentrations of fluorophore in the two solutions. The equivalence is the basis for quantitation in terms of molecules of equivalent soluble fluorophore (MESF). The quantitation procedure starts with the measurement of fluorescence signals from a serial dilution of fluorescein solutions to obtain a calibration of a fluorometer. The fluorometer is used to measure the fluorescence signal of a suspension of microspheres with immobilized fluorescein isothiocyanate (FITC). The calibration is used to obtain the concentration of soluble fluorophores which gives the same fluorescence signal as the microsphere suspension. The number concentration of microspheres is measured and the equality of fluorescence yields is used to obtain the number of soluble fluorescein molecules equivalent to a single microsphere. PMID:27308107

  17. Making Sense of Plant Health

    NASA Technical Reports Server (NTRS)

    2001-01-01

    Ciencia, Inc. created a new device, known as a Portable Photosynthesis Analyzer, or Phase Fluorometer, that provides real-time data about the photochemical efficiency of phytoplankton and other plant forms. The commercial version of this technology is used for photosynthesis research and offers major benefits to the field of life science. This new instrument is the first portable instrument of its kind. Through a license agreement with Ciencia, Oriel Instruments, of Stratford, Connecticut, manufactures and markets the commercial version of the instrument under the name LifeSense.TMLifeSense is a 70 MHz single-frequency fluorometer that offers unrivaled capabilities for fluorescence lifetime sensing and analysis. LifeSense provides information about all varieties of photosynthetic systems. Photosynthesis research contributes important health assessments about the plant, be it phytoplankton or a higher form of plant life. With its unique sensing capabilities, LifeSense furnishes data regarding the yield of a plant's photochemistry, as well as its levels of photosynthetic activity. The user can then gain an extremely accurate estimate of the plant's chlorophyll biomass, primary production rates, and a general overview of the plant's physiological condition.

  18. Effects of habitat light conditions on the excitation quenching pathways in desiccating Haberlea rhodopensis leaves: an Intelligent FluoroSensor study.

    PubMed

    Solti, Ádám; Lenk, Sándor; Mihailova, Gergana; Mayer, Péter; Barócsi, Attila; Georgieva, Katya

    2014-01-01

    Resurrection plants can survive dehydration to air-dry state, thus they are excellent models of understanding drought and dehydration tolerance mechanisms. Haberlea rhodopensis, a chlorophyll-retaining resurrection plant, can survive desiccation to relative water content below 10%. Leaves, detached from plants of sun and shade habitats, were moderately (∼50%) dehydrated in darkness. During desiccation, chlorophyll a fluorescence was detected by the recently innovated wireless Intelligent FluoroSensor (IFS) chlorophyll fluorometer, working with three different detectors: a pulse-amplitude-modulated (PAM) broadband channel and two channels to measure non-modulated red and far-red fluorescence. No change in area-based chlorophyll content of leaves was observed. The maximal quantum efficiency of photosystem II decreased gradually in both shade and sun leaves. Shade leaves could not increase antennae-based quenching, thus inactivated photosystem II took part in quenching of excess irradiation. Sun leaves seemed to be pre-adapted to quench excess light as they established an intensive increase in antennae-based non-photochemical quenching parallel to desiccation. The higher far-red to red antennae-based quenching may sign light-harvesting complex reorganization. Thus, compared to PAM, IFS chlorophyll fluorometer has additional benefits including (i) parallel estimation of changes in the Chl content and (ii) prediction of underlying processes of excitation energy quenching. PMID:24345600

  19. Satellite Remote Sensing Studies of Biological and Biogeochemical Processing in the Ocean

    NASA Technical Reports Server (NTRS)

    Vernet, Maria

    2001-01-01

    The remote sensing of phycoerythrin-containing phytoplankton by ocean color was evaluated. Phycoerythrin (PE) can be remotely sensed by three methods: surface reflectance (Sathyendranath et al. 1994), by laser-activated fluorescence (Hoge and Swift 1986) and by passive fluorescence (Letelier et al. 1996). In collaboration with Dr. Frank Hoge and Robert Swift during Dr. Maria Vernet's tenure as Senior Visiting Scientist at Wallops Island, the active and passive methods were studied, in particular the detection of PE fluorescence and spectral reflectance from airborne LIDAR (AOL). Airborne instrumentation allows for more detailed and flexible sampling of the ocean surface than satellites thus providing the ideal platform to test model and develop algorithms than can later be applied to ocean color by satellites such as TERRA and AQUA. Dr. Vernet's contribution to the Wallops team included determination of PE in the water column, in conjunction with AOL flights in the North Atlantic Bight. In addition, a new flow-through fluorometer for PE determination by fluorescence was tested and calibrated. Results: several goals were achieved during this period. Cruises to the California Current, North Atlantic Bight, Gulf of Maine and Chesapeake Bay provided sampling under different oceanographic and optical conditions. The ships carried the flow-through fluorometer and samples for the determination of PE were obtained from the flow-through flow. The AOL was flown over the ship's track, usually several flights during the cruise, weather permitting.

  20. Fluorometric Determination of Deoxyribonucleic Acid in Bacteria with Ethidium Bromide

    PubMed Central

    Donkersloot, J. A.; Robrish, S. A.; Krichevsky, M. I.

    1972-01-01

    A simple, sensitive, and rapid method is presented for the determination of deoxyribonucleic acid (DNA) in both gram-positive and gram-negative bacteria. It is based upon the fluorometric determination of DNA with ethidium bromide after alkaline digestion of the bacteria to hydrolyze the interfering ribonucleic acid. The assay takes less than 2 hr. Its sensitivity is at least 0.2 μg of DNA in a final solution of 4 ml and it uses commonly available filter or double monochromator fluorometers. Judicious choice of light source and filters allows an additional 10-fold increase in sensitivity with a filter fluorometer. Turbidity caused by bacteria or insoluble polysaccharides does not interfere with the fluorescence measurements. There was no significant difference between the results obtained with this method and those obtained with the indole and diphenylamine methods when these assays were applied to Escherichia coli and sucrose- or glucose-grown Streptococcus mutans. The method was also tested by determining the specific growth rate of E. coli. This new procedure should be especially useful for the determination of bacterial DNA in dilute suspensions and for the estimation of bacterial growth or DNA replication where more conventional methods are not applicable or sensitive enough. PMID:4561101

  1. An alternative method for correcting fluorescence quenching

    NASA Astrophysics Data System (ADS)

    Biermann, L.; Guinet, C.; Bester, M.; Brierley, A.; Boehme, L.

    2015-01-01

    Under high light intensity, phytoplankton protect their photosystems from bleaching through non-photochemical quenching processes. The consequence of this is suppression of fluorescence emission, which must be corrected when measuring in situ yield with fluorometers. We present data from the Southern Ocean, collected over five austral summers by 19 southern elephant seals tagged with fluorometers. Conventionally, fluorescence data collected during the day (quenched) were corrected using the limit of the mixed layer, assuming that phytoplankton are uniformly mixed from the surface to this depth. However, distinct deep fluorescence maxima were measured in approximately 30% of the night (unquenched) data. To account for the evidence that chlorophyll is not uniformly mixed in the upper layer, we propose correcting from the limit of the euphotic zone, defined as the depth at which photosynthetically available radiation is ~ 1% of the surface value. Mixed layer depth exceeded euphotic depth over 80% of the time. Under these conditions, quenching was corrected from the depth of the remotely derived euphotic zone Zeu, and compared with fluorescence corrected from the depth of the density-derived mixed layer. Deep fluorescence maxima were evident in only 10% of the day data when correcting from mixed layer depth. This was doubled to 21% when correcting from Zeu, more closely matching the unquenched (night) data. Furthermore, correcting from Zeu served to conserve non-uniform chlorophyll features found between the 1% light level and mixed layer depth.

  2. Relationship between chlorophyll a concentration, light attenuation and diving depth of the Southern elephant seal Mirounga leonina.

    PubMed

    Jaud, Thomas; Dragon, Anne-Cécile; Garcia, Jade Vacquie; Guinet, Christophe

    2012-01-01

    Recently, a number of Antarctic marine environmental studies have used oceanographic parameters collected from instrumented top predators for ecological and physical information. Phytoplankton concentration is generally quantified through active measurement of chlorophyll fluorescence. In this study, light absorption coefficient (K(0.75)) was used as an indicator of phytoplankton concentration. This measurement, easy to obtain and requiring low electric power, allows for assessing of the fine scale horizontal structuring of phytoplankton. As part of this study, Southern elephant seals (SES) were simultaneously equipped with a fluorometer and a light logger. Along the SES tracks, variations in K(0.75) were strongly correlated with chlorophyll, a concentration measured by the fluorometer within the euphotic layer. With regards to SES foraging behaviour, bottom depth of the seal's dive was highly dependent on light intensity at 150 m, indicating that the vertical distribution of SES's prey such as myctophids is tightly related to light level. Therefore, change in phytoplankton concentration may not only have a direct effect on SES's prey abundance but may also determine their vertical accessibility with likely consequences on SES foraging efficiency. PMID:23082166

  3. Fluorescence Spectroscopy in a Shoebox

    NASA Astrophysics Data System (ADS)

    Farooq Wahab, M.

    2007-08-01

    This article describes construction of a simple, inexpensive fluorometer. It utilizes a flashlight or sunlight source, highlighter marker ink, bowl of water with mirror as dispersing element, and colored cellophane sheets as filters. The human eye is used as a detector. This apparatus is used to demonstrate important concepts related to fluorescence spectroscopy. Using ink from a highlighter marker, one can demonstrate the difference between light scattering and fluorescence emission, the need for an intense light source, phenomenon of the Stokes shift, the choice of filters, the preferred geometry of excitation source and emission detector, and the low detection limits that can be achieved by fluorescence measurements. By reflecting the fluorescence emission from a compact disk, it can be seen that the light emitted by molecules is not monochromatic. Furthermore, a spectrofluorometer is constructed using gratings made from a DVD or a CD. The shoebox fluorometer and spectrofluorometer can serve as useful teaching aids in places where commercial instruments are not available, and it avoids the black box problem of modern instruments.

  4. An Automated Method for the Optical Characterization of Dissolved Organic Matter in a Rapidly Suburbanizing Watershed, Southeastern New Hampshire

    NASA Astrophysics Data System (ADS)

    Gettel, G. M.; McDowell, W.; Pisani, O.

    2006-12-01

    Dissolved organic matter (DOM) is exported from watersheds to downstream ecosystems where it can contribute to eutrophication problems by enhancing microbial respiration and lowering oxygen levels. DOM quality affects microbial respiration; however, little is known about how watershed processes affect the quality of DOM export. In order to document temporal and spatial variability in DOM quality at the watershed scale, we are developing a method to automate the optical characterization of DOM in the Lamprey River watershed in southeastern New Hampshire. This method employs a refrigerated autosampler and column heater associated with a Shimadzu high-pressure liquid chromatograph (HPLC) with a photo-diode array (PDA) UV absorbance detector and an in-line Horiba Jobin Yvon Fluoromax 3 fluorometer capable of 3D excitation- emission scans (EEM). One advantage of this method is that the HPLC flow-through cell in the fluorometer reduces inner-filter effects due to its small volume. Furthermore, specific UV absorbance or SUVA can also be calculated because an in-line UV PDA is used. We found that a number of fluorescence indices are related to DOC, DON, or NO3 concentrations throughout the Lamprey River watershed. For example, Fluorescence Index (F.I.), an indicator of autochthonous sources of DOM, is positively correlated with nitrate and negatively correlated with DOC concentrations (R2=0.95; p<0.01; R2=0.86; p<0.05 respectively). The highest F.I. occurred in the highest-population density sub-basin with the highest nitrate concentrations, while the lowest F.I. occurred in the lowest-population density sub-basin with highest DOC concentrations. These results indicate that nitrate may increase within-stream generation of DOC at high- population sites while DOC from low-population, low-nitrate sites is predominately allochthonous. This allows DOM characterization to be performed in conjunction with weekly and monthly monitoring of many water quality parameters and to be

  5. Detection of specific DNA sequences by fluorescence amplification: a color complementation assay.

    PubMed Central

    Chehab, F F; Kan, Y W

    1989-01-01

    We have developed a color complementation assay that allows rapid screening of specific genomic DNA sequences. It is based on the simultaneous amplification of two or more DNA segments with fluorescent oligonucleotide primers such that the generation of a color, or combination of colors, can be visualized and used for diagnosis. Color complementation assay obviates the need for gel electrophoresis and has been applied to the detection of a large and small gene deletion, a chromosomal translocation, an infectious agent, and a single-base substitution. DNA amplification with fluorescent oligonucleotide primers has also been used to multiplex and discriminate five different amplified DNA loci simultaneously. Each primer set is conjugated to a different dye, and the fluorescence of each dye respective to its amplified DNA locus is scored on a fluorometer. This method is valuable for DNA diagnostics of genetic, acquired, and infectious diseases, as well as in DNA forensics. It also lends itself to complete automation. Images PMID:2594760

  6. A Low Cost, Customizable Turbidostat for Use in Synthetic Circuit Characterization

    PubMed Central

    2015-01-01

    Engineered biological circuits are often disturbed by a variety of environmental factors. In batch culture, where the majority of synthetic circuit characterization occurs, environmental conditions vary as the culture matures. Turbidostats are powerful characterization tools that provide static culture environments; however, they are often expensive, especially when purchased in custom configurations, and are difficult to design and construct in a lab. Here, we present a low cost, open source multiplexed turbidostat that can be manufactured and used with minimal experience in electrical or software engineering. We demonstrate the utility of this system to profile synthetic circuit behavior in S. cerevisiae. We also demonstrate the flexibility of the design by showing that a fluorometer can be easily integrated. PMID:25036317

  7. Portable flow immunosensor for detecting drugs and explosives

    NASA Astrophysics Data System (ADS)

    Kusterbeck, Anne W.; Gauger, Paul R.; Charles, Paul T.

    1997-02-01

    To assist in airport surveillance efforts, a biosensor based on antibody recognition of individual explosives and drugs has been developed at the Naval Research Laboratory. Analysis of samples containing ng/mL levels of the material are completed in under one minute. Immunoassays for the explosives and the five major drugs of abuse are currently available. The intrinsic nature of antigen-antibody binding also provides the unit with an inherently high degree of selectivity. A portable version of the biosensor that can be run by non-technical personnel is also being engineered. The device, including pumps and fluorometer, will be housed on a modified PCMCIA cartridge fitted into a laptop computer. To run assays, a disposable coupon containing the antibody/fluorescent-antigen complex is inserted into the unit and samples are introduced via a sampling port. Results can be viewed in real time or stored on the computer for later data retrieval and analysis.

  8. Laboratory studies of in vivo fluorescence of phytoplankton

    NASA Technical Reports Server (NTRS)

    Brown, C. A., Jr.; Farmer, F. H.; Jarrett, O., Jr.; Staton, W. L.

    1978-01-01

    A lidar system is developed that uses four selected excitation wavelengths to induce chlorophyll 'a' fluorescence which is indicative of both the concentration and diversity of phytoplankton. The operating principles of the system and the results of measurements of phytoplankton fluorescence in a controlled laboratory environment are presented. A comparative study of results from lidar fluorosensor laboratory tank tests using representative species of phytoplankton in single and multispecies cultures from each of four color groups reveals that (1) there is good correlation between the fluorescence of chlorophyll 'a' remotely simulated and detected by the lidar system and in-situ measurements using four similar excitation wavelengths in a flow-through fluorometer; (2) good correlation exists between the total chlorophyll 'a' calculated from lidar-fluorosensor data and measurements obtained by the Strickland-Parsons method; and (3) the lidar fluorosensor can provide an index of population diversity.

  9. Spectral and Temporal Laser Fluorescence Analysis Such as for Natural Aquatic Environments

    NASA Technical Reports Server (NTRS)

    Chekalyuk, Alexander (Inventor)

    2015-01-01

    An Advanced Laser Fluorometer (ALF) can combine spectrally and temporally resolved measurements of laser-stimulated emission (LSE) for characterization of dissolved and particulate matter, including fluorescence constituents, in liquids. Spectral deconvolution (SDC) analysis of LSE spectral measurements can accurately retrieve information about individual fluorescent bands, such as can be attributed to chlorophyll-a (Chl-a), phycobiliprotein (PBP) pigments, or chromophoric dissolved organic matter (CDOM), among others. Improved physiological assessments of photosynthesizing organisms can use SDC analysis and temporal LSE measurements to assess variable fluorescence corrected for SDC-retrieved background fluorescence. Fluorescence assessments of Chl-a concentration based on LSE spectral measurements can be improved using photo-physiological information from temporal measurements. Quantitative assessments of PBP pigments, CDOM, and other fluorescent constituents, as well as basic structural characterizations of photosynthesizing populations, can be performed using SDC analysis of LSE spectral measurements.

  10. Adiabatic compressibility of myoglobin. Effect of axial ligand and denaturation.

    PubMed

    Leung, W P; Cho, K C; Lo, Y M; Choy, C L

    1986-03-01

    An ultrasonic technique has been employed to study the adiabatic compressibility of three metmyoglobin derivatives (aquomet-, fluoromet- and azidometmyoglobin) at neutral pH, and aquometmyoglobin as a function of pH in the frequency range of 1-10 MHz at 20 degrees C. No difference was observed in the adiabatic compressibility of the various derivatives. This indicates that the binding of different axial ligands to myoglobin does not affect significantly the conformational fluctuations of the protein. The finding is consistent with the results of the hydrogen exchange rate experiment, indicating that both types of measurements are useful for the study of protein dynamics. Upon acid-induced denaturation, the adiabatic compressibility of myoglobin drops from 5.3 X 10(-12) cm2/dyn to 0.5 X 10(-12) cm2/dyn. Plausible reasons for such a decrease are discussed. PMID:3947645

  11. Fraunhofer line-dept sensing applied to water

    NASA Technical Reports Server (NTRS)

    Stoertz, G. E.

    1969-01-01

    An experimental Fraunhofer line discriminator is basically an airborne fluorometer, capable of quantitatively measuring the concentration of fluorescent substances dissolved in water. It must be calibrated against standards and supplemented by ground-truth data on turbidity and on approximate vertical distribution of the fluorescent substance. Quantitative use requires that it be known in advance what substance is the source of the luminescence emission; qualitative sensing, or detection of luminescence is also possible. The two approaches are fundamentally different, having different purposes, different applications, and different instruments. When used for sensing of Rhodamine WT dye in coastal waters and estuaries, the FLD is sensing in the spectral region permitting nearly maximum depth of light penetration.

  12. An ingestible, NIR-fluorometric, cancer-screening capsule.

    PubMed

    Demosthenous, Panayiota; Georgiou, Julius

    2015-08-01

    Asymptomatic, early-stage, cancer detection is a problem in the small intestine, that is largely inaccessible. This paper presents a cost-effective screening capsule prototype, which is able to detect infrared (IR) fluorescence emitted by indocyanine green (ICG) fluorophore dye. The presented mixed-signal system has a small size, consumes little power and works as a high-sensitivity fluorometer that records fluorescence levels throughout the small intestine, rather than collecting images that need labour intensive video analysis. Ex-vivo experiments, on ICG-impregnated swine intestine, have shown that the prototype system is able to detect low concentrations of ICG in the nanomolar and micromolar region, which is required to detect early cancer in the small intestine. PMID:26736713

  13. Novel imaging techniques for diabetic macular edema.

    PubMed

    Lobo, C; Bernardes, R; Faria de Abreu, J R; Cunha-Vaz, J G

    1999-01-01

    Retinal edema should be defined as any increase of water of the retinal tissue resulting in an increase in its volume. It may be of cytotoxic or vasogenic origin. Development of vasogenic macular edema is dependent on a series of factors such as blood pressure, blood-retinal barrier permeability, retinal cell damage, retinal tissue osmotic pressure and retinal tissue compliance. Objective measurements of retinal thickness are now possible using the Retinal Thickness Analyser. Localised measurements of blood-retinal barrier permeability may also be obtained using the Retinal Leakage Analyser, a modified confocal scanning laser fluorometer, while obtaining simultaneously angiographic images of the choroid and retina. These new imaging techniques show that cytotoxic and vasogenic retinal edema may occur independently in the early stages of diabetic retinopathy. These findings offer new perspectives for designing novel therapeutic strategies. PMID:10896349

  14. Fluorescence dynamics of biological systems using synchrotron radiation

    SciTech Connect

    Gratton, E.; Mantulin, W.W.; Weber, G.; Royer, C.A.; Jameson, D.M.; Reininger, R.; Hansen, R.

    1996-09-01

    A beamline for time-resolved fluorescence spectroscopy of biological systems is under construction at the Synchrotron Radiation Center. The fluorometer, operating in the frequency domain, will take advantage of the time structure of the synchrotron radiation light pulses to determine fluorescence lifetimes. Using frequency-domain techniques, the instrument can achieve an ultimate time resolution on the order of picoseconds. Preliminary experiments have shown that reducing the intensity of one of the fifteen electron bunches in the storage ring allows measurement of harmonic frequencies equivalent to the single-bunch mode. This mode of operation of the synchrotron significantly extends the range of lifetimes that can be measured. The wavelength range (encompassing the visible and ultraviolet), the range of measurable lifetimes, and the stability and reproducibility of the storage ring pulses should make this beamline a versatile tool for the investigation of the complex fluorescence decay of biological systems. {copyright} {ital 1996 American Institute of Physics.}

  15. A microfluidic nano-biosensor for the detection of pathogenic Salmonella.

    PubMed

    Kim, Giyoung; Moon, Ji-Hea; Moh, Chang-Yeon; Lim, Jong-guk

    2015-05-15

    Rapid detection of pathogenic Salmonella in food products is extremely important for protecting the public from salmonellosis. The objective of the present study was to explore the feasibility of using a microfluidic nano-biosensor to rapidly detect pathogenic Salmonella. Quantum dot nanoparticles were used to detect Salmonella cells. For selective detection of Salmonella, anti-Salmonella polyclonal antibodies were covalently immobilized onto the quantum dot surface. To separate and concentrate the cells from the sample, superparamagnetic particles and a microfluidic chip were used. A portable fluorometer was developed to measure the fluorescence signal from the quantum dot nanoparticles attached to Salmonella in the samples. The sensitivity for detection of pathogenic Salmonella was evaluated using serially diluted Salmonella Typhimurium in borate buffer and chicken extract. The fluorescence response of the nano-biosensor increased with increasing cell concentration. The detection limit of the sensor was 10(3) CFU/mL Salmonella in both borate buffer and food extract. PMID:25172028

  16. Hadamard-transform fluorescence-lifetime imaging.

    PubMed

    Mizuno, Takahiko; Iwata, Tetsuo

    2016-04-18

    We discuss a Hadamard-transform-based fluorescence-lifetime-imaging (HT-FLI) technique for fluorescence-lifetime-imaging microscopy (FLIM). The HT-FLI uses a Fourier-transform phase-modulation fluorometer (FT-PMF) for fluorescence-lifetime measurements, where the modulation frequency of the excitation light is swept linearly in frequency from zero to a specific maximum during a fixed duration of time. Thereafter, fluorescence lifetimes are derived through Fourier transforms for the fluorescence and reference waveforms. The FT-PMF enables the analysis of multi-component samples simultaneously. HT imaging uses electronic exchange of HT illumination mask patterns, and a high-speed, high-sensitivity photomultiplier, to eliminate frame-rate issues that accompany two-dimensional image detectors. PMID:27137259

  17. Submersible optical sensors exposed to chemically dispersed crude oil: wave tank simulations for improved oil spill monitoring.

    PubMed

    Conmy, Robyn N; Coble, Paula G; Farr, James; Wood, A Michelle; Lee, Kenneth; Pegau, W Scott; Walsh, Ian D; Koch, Corey R; Abercrombie, Mary I; Miles, M Scott; Lewis, Marlon R; Ryan, Scott A; Robinson, Brian J; King, Thomas L; Kelble, Christopher R; Lacoste, Jordanna

    2014-01-01

    In situ fluorometers were deployed during the Deepwater Horizon (DWH) Gulf of Mexico oil spill to track the subsea oil plume. Uncertainties regarding instrument specifications and capabilities necessitated performance testing of sensors exposed to simulated, dispersed oil plumes. Dynamic ranges of the Chelsea Technologies Group AQUAtracka, Turner Designs Cyclops, Satlantic SUNA and WET Labs, Inc. ECO, exposed to fresh and artificially weathered crude oil, were determined. Sensors were standardized against known oil volumes and total petroleum hydrocarbons and benzene-toluene-ethylbenzene-xylene measurements-both collected during spills, providing oil estimates during wave tank dilution experiments. All sensors estimated oil concentrations down to 300 ppb oil, refuting previous reports. Sensor performance results assist interpretation of DWH oil spill data and formulating future protocols. PMID:24377909

  18. [Photosynthetic fluorescence characteristics of floating-leaved and submersed macrophytes commonly found in Taihu Lake].

    PubMed

    Song, Yu-zhi; Cai, Wei; Qin, Bo-qiang

    2009-03-01

    Some aquatic macrophytes commonly found in Taihu Lake, including Trapa bispinosa, Nymphyoides peltatum, Vallisneria natans, and Hydrilla verticillata were collected, and their maximal quantum yield of photosystem II (Fv/Fm) as well as the rapid light curves (RLCs) under conditions of light adaptation and dark adaptation were measured in situ by using a submersible and pulse-amplitude modulated fluorometer (Diving-PAM). The results showed that floating-leaved plants T. bispinosa and N. peltatum had higher potential maximum photosynthetic capacity than submerged macrophytes V. natans and H. verticillata. The measured maximal quantum yield of T. bispinosa, N. peltatum, V. natans, and H. verticillata was 0.837, 0.831, 0.684, and 0.764, respectively. Both the maximal relative electron transport rate and the half saturation point of light intensity of T. bispinosa and N. peltatum were higher than those of V. natans and H. verticillata, especially under the condition of light adaptation. PMID:19637593

  19. Development of advanced methods for early detection of lung cancer in the uranium miner/worker population

    SciTech Connect

    Profio, A.E.; Balchum, O.J.; Saccomanno, G.; Huth, G.C.

    1983-01-01

    Fluorescence bronchoscopy with a violet laser and image intensifier has been developed for imaging the red fluorescence of a tumor-specific agent, hematoporphyrin derivative, that has been injected before the examination. The instrument was developed to localize carcinoma in situ and early, small bronchogenic tumors diagnosed by sputum cytology but invisible on chest x-ray and conventional bronchoscopy, in underground uranium miners and others at risk for lung cancer. In addition to the imaging devices, a video system including a processor and electronics for digital background image subtraction has been developed to enhance contrast. A ratio fluorometer and a rapid-scan spectrum analyzer have been designed for quantitative measurements of fluorescence intensity and dependence on dosage and time after injection of the fluorescent agent. Clinical trials demonstrate detection of carcinoma in situ, and the true positive rate should be improved by the new instrumentation and optimization of time delay and dosage. 14 references, 6 figures.

  20. Real-time Monitoring of Dissolved Organic Matter (DOM) Amount, Composition, Source and Reactivity Using Fluorescence Spectroscopy: Applications for Drinking Water Quality

    NASA Astrophysics Data System (ADS)

    Kraus, T. E.; Saraceno, J.; Downing, B. D.; Goldman, J. H.; Carpenter, K. D.; McGhee, G.; Bergamaschi, B. A.

    2010-12-01

    There is growing interest in the use of in situ, continuous fluorescence spectroscopy as a proxy for dissolved organic carbon (DOC) concentration. To date, in situ fluorometers designed to estimate DOC concentration are single wavelength sensors centered near the excitation/emission (ex/em) pair 370/460 nm. Additional information about dissolved organic matter (DOM) composition has only been obtainable from benchtop fluorometers that provide multi-spectral data. Changes in DOM composition are important as they provide insight into DOM source (e.g. terrestrial, algal, wastewater) and reactivity. Recent advances in sensor technology make it possible to build in situ instruments for measuring multiple fluorescence ex/em pairs, including pairs with excitations in the lower “deep UV” region (e.g. 270/340 nm) associated with fresher and more labile DOM pools. The deployment of multi-spectral sensors will provide real-time continuous data showing not only changes in DOM concentration, but also changes in composition. This information is particularly pertinent to drinking water utilities because a fraction of DOM reacts upon disinfection (e.g. chlorination and ozonation) to form toxic disinfection byproducts (DBPs) which are regulated by the EPA. To test this application, we designed a multi-wavelength sensor that will measure three ex/em pairs (370/470, 370/520 and 270/340 nm) for deployment near a drinking water intake on the Clackamas River in Oregon. Comparison of the continuous data with discrete sample data indicates these tools can track both quantitative and qualitative changes in the DOM pool. The availability of this type of continuous data in real time could enable utilities to minimize the formation of DBPs by continuously optimizing treatment plant operations in response to changes in source water. In addition, collection of high-frequency data will improve understanding of watershed DOM dynamics and help identify sources of DOM and DBP precursors, thereby

  1. Spatial and Temporal Dynamics of Hyporheic Respiration Under Variable Discharge Conditions

    NASA Astrophysics Data System (ADS)

    Kurz, M. J.; Schmidt, C.; Knapp, J.; Romeijn, P.; Blaen, P.; Klaar, M. J.; Keller, T.; Krause, S.; Ward, A. S.; Fleckenstein, J. H.; Larned, S.; Zarnetske, J. P.; Martí Roca, E.; Datry, T.

    2014-12-01

    The hyporheic zone is the site of intensive biogeochemical cycling in streams. However, the controls on spatio-temporal variability in hyporheic processing, and the impact of this hyporheic processing on reach-scale processing, are largely unknown. We aimed to evaluate spatial variability in hyporheic respiration along an upland river over the course of a flood event using the reactive tracer resazurin (Raz). Raz, a weakly fluorescent dye, irreversibly transforms to resorufin (Rru) under mildly reducing conditions, providing a proxy for aerobic respiration in the hyporheic zone. Eight conductivity loggers and in-situ fluorometers, measuring in-stream concentrations of Raz, Rru, fluorescein, and turbidity, were evenly spaced along a 1km reach of the Selke River, a gravelly, third-order river in north-central Germany. Sub-reaches between fluorometers differed in the number of streambed structures (ex. pool-riffle sequences and gravel bars) hypothesized to impact hyporheic exchange, residence time distributions, and the development of biogeochemical hotspots. Discharge over the 5 days of the experiment in the Selke River ranged from baseflow conditions of 0.3 m3/s to peak flows of 2.6 m3/s. Seven in-stream slug injections of Raz, NaCl and the conservative tracer fluorescein were conducted at discharge conditions of 0.3, 0.8, 2.5, 2.1, 1.3, 1.0, and 0.9 m3/s. Aerobic respiration rates and residence time distributions in the reach and sub-reaches are evaluated relative to the changing discharge conditions. Preliminary results indicate that although reach-scale tracer travel times decrease with increasing discharge, the reach-scale transformation of Raz to Rru is lowest at intermediate discharge and highest at during baseflow and peak flow conditions. This suggests that the highest transformation rates occur during high discharge.

  2. Reverse transcription genome exponential amplification reaction assay for rapid and universal detection of human rhinoviruses.

    PubMed

    Guan, Li; Zhao, Lin-Qing; Zhou, Hang-Yu; Nie, Kai; Li, Xin-Na; Zhang, Dan; Song, Juan; Qian, Yuan; Ma, Xue-Jun

    2016-07-01

    Human rhinoviruses (HRVs) have long been recognized as the cause of more than one-half of acute viral upper respiratory illnesses, and they are associated with more-serious diseases in children, such as asthma, acute otitis media and pneumonia. A rapid and universal test for of HRV infection is in high demand. In this study, a reverse transcription genome exponential amplification reaction (RT-GEAR) assay targeting the HRV 5' untranslated region (UTR) was developed for pan-HRV detection. The reaction was performed in a single tube in one step at 65 °C for 60 min using a real-time fluorometer (Genie(®)II; Optigene). The RT-GEAR assay showed no cross-reactivity with common human enteroviruses, including HEV71, CVA16, CVA6, CVA10, CVA24, CVB5, Echo30, and PV1-3 or with other common respiratory viruses including FluA H3, FluB, PIV1-4, ADV3, RSVA, RSVB and HMPV. With in vitro-transcribed RNA containing the amplified regions of HRV-A60, HRV-B06 and HRV-C07 as templates, the sensitivity of the RT-GEAR assay was 5, 50 and 5 copies/reaction, respectively. Experiments to evaluate the clinical performance of the RT-GEAR assay were also carried out with a panel of 143 previously verified samples, and the results were compared with those obtained using a published semi-nested PCR assay followed by sequencing. The tested panel comprised 91 HRV-negative samples and 52 HRV-positive samples (18 HRV-A-positive samples, 3 HRV-B-positive samples and 31 HRV-C-positive samples). The sensitivity and specificity of the pan-HRVs RT-GEAR assay was 98.08 % and 100 %, respectively. The kappa correlation between the two methods was 0.985. The RT-GEAR assay based on a portable Genie(®)II fluorometer is a sensitive, specific and rapid assay for the universal detection of HRV infection. PMID:27132014

  3. Modeling and design of micromachined optical Söller collimators for lensless CCD-based fluorometry.

    PubMed

    Balsam, Joshua; Ossandon, Miguel; Bruck, Hugh Alan; Rasooly, Avraham

    2012-11-01

    To address the needs of medical diagnostics in resource-poor settings, it is necessary to develop low cost, simple and portable Point of Care detectors for integrated medical diagnostics. Previously, we have described a simple lensless fluorometer with sensitivity in the range of current ELISA plate readers. The key to the lensfree fluorometer is the uniform spatial distribution of light, which we achieved using a simple optical collimator based on a "stack of pinholes" (a stack of black PMMA plates with arrays of pinholes machined via laser) enabling the light to be collimated from the LED light source through the necessary wavelength filters and the assay's microfluidics directly onto the CCD without a lens. In this paper, we describe the optical principle for designing these Söller collimators for lensfree CCD-based fluorometry. The illuminating surface was modeled as a collection of differential areas emitting uniformly and spherically, and the intensity contribution of each emitting area was summed over the detector surface. To compute the final light intensity distribution from such a differential model we derived an integral equation to sum the individual intensity contributions from the two-dimensional emitting surface. The equation is for a single-hole collimator. Light intensity measurements were taken by placing a collimator with a particular aspect ratio (the ratio of hole length to diameter (L/d)) over the CCD image sensor and capturing an image. The resulting image is the 2D light intensity profile generated by the collimator. As the aspect ratio is increased the slope of the light intensity profile increases, corresponding to an increased degree of collimation. To test the model, the measured maximum and mean light intensities were compared with the theoretical predictions generated from the model. There was an agreement between the variation of the mean (R(2) = 0.990) and maximum (R(2) = 0.938) values of light intensities with aspect ratios based

  4. Spatial distribution and seasonal variability of chlorophyll-a concentration in the Azov Sea turbid waters by means of remote sensing and continuous fluorescence measurements

    NASA Astrophysics Data System (ADS)

    Saprygin, V. V.

    2011-12-01

    The goal of this study was to apply continuous fluorometric and remote estimation of chlorophyll-a concentration (Cchl) techniques to complex turbid waters of Azov Sea and explore Cchl temporal variation and spatial pattern. Azov Sea is the shallowest sea in the world with maximum depth below 15 m. Its maximum salinity is about 14%; total suspended solids and chlorophyll-a concentrations reach 120 [tex]g m^{-3}[/tex] and 100 [tex]mg m^{-3}[/tex] respectively in Taganrog Bay, daily production varies up to 3.5 [tex]gC_{org} m^{-3}[/tex]. Chlorophyll-a concentrations were measured in 2008-2010 year-round spectrophotometrically, 446 water samples were taken to calibrate fluorometerical and remote sensing data. The highest recorded concentration was 149.3, the lowest - 0.3 [tex]mg m^{-3}[/tex]. Continuous-flow fluorometer was applied in the course of 3 expeditions to Taganrog Bay to measure chlorophyll-a fluorescence (Fchl) each 30 meters along the ship path. Two-cuvette fluorometer was used to discount the influence of dissolved organic matter. Fchl measurements were calibrated and Cchl profiles derieved to estimate Cchl spatial heterogeneity in close scale. Fchl measurements were also made during moorings each 6 seconds to estimate temporal Cchl variability. Recently published algorithm based on reflectance in the red and the near-infrared (NIR) spectral regions was applied to MERIS data for the remote estimation of Cchl. Taking in account fluorometric Cchl spatial heterogeneity estimation, the algorithm for culling the outliers in Cchl fields derived from satellite data was developed. 74 images were processed to Cchl maps and then averaged monthly. Consequently, Cchl spatial distribution and seasonal variability were studied. Spectrophotometric, flourumetric measurements and values obtained by NIR-red algorithm showed strong correlation in turbid Case II waters of Azov Sea. Fluorometric and remote measurements showed high Cchl variations in short and long terms

  5. Real time monitoring of urban surface water quality using a submersible, tryptophan-like fluorescence sensor

    NASA Astrophysics Data System (ADS)

    Khamis, Kieran; Bradley, Chris; Hannah, David; Stevens, Rob

    2014-05-01

    Due to the recent development of field-deployable optical sensor technology, continuous quantification and characterization of surface water dissolved organic matter (DOM) is possible now. Tryptophan-like (T1) fluorescence has the potential to be a particularly useful indicator of human influence on water quality as T1 peaks are associated with the input of labial organic carbon (e.g. sewage or farm waste) and its microbial breakdown. Hence, real-time recording of T1 fluorescence could be particular useful for monitoring waste water infrastructure, treatment efficiency and the identification of contamination events at higher temporal resolution than available hitherto. However, an understanding of sensor measurement repeatability/transferability and interaction with environmental parameters (e.g. turbidity) is required. Here, to address this practical knowledge gap, we present results from a rigorous test of a commercially available submersible tryptophan fluorometer (λex 285, λem 350). Sensor performance was first examined in the laboratory by incrementally increasing turbidity under controlled conditions. Further to this the sensor was integrated into a multi-parameter sonde and field tests were undertaken involving: (i) a spatial sampling campaign across a range of surface water sites in the West Midlands, UK; and (ii) collection of high resolution (sub-hourly) samples from an urban stream (Bournbrook, Birmingham, U.K). To determine the ability of the sensor to capture spatiotemporal dynamics of urban waters DOM was characterized for each site or discrete time step using Excitation Emission Matrix spectroscopy and PARAFAC. In both field and laboratory settings fluorescence intensity was attenuated at high turbidity due to suspended particles increasing absorption and light scattering. For the spatial survey, instrument readings were compared to those obtained by a laboratory grade fluorometer (Varian Cary Eclipse) and a strong, linear relationship was apparent

  6. Global spectral-kinetic analysis of room temperature chlorophyll a fluorescence from light-harvesting antenna mutants of barley.

    PubMed Central

    Gilmor, A M; Itoh, S; Govindjee

    2000-01-01

    This study presents a novel measurement, and simulation, of the time-resolved room temperature chlorophyll a fluorescence emission spectra from leaves of the barley wild-type and chlorophyll-b-deficient chlorina (clo) f2 and f104 mutants. The primary data were collected with a streak-camera-based picosecond-pulsed fluorometer that simultaneously records the spectral distribution and time dependence of the fluorescence decay. A new global spectral-kinetic analysis programme method, termed the double convolution integral (DCI) method, was developed to convolve the exciting laser pulse shape with a multimodal-distributed decay profile function that is again convolved with the spectral emission band amplitude functions. We report several key results obtained by the simultaneous spectral-kinetic acquisition and DCI methods. First, under conditions of dark-level fluorescence, when photosystem II (PS II) photochemistry is at a maximum at room temperature, both the clo f2 and clo f104 mutants exhibit very similar PS II spectral-decay contours as the wild-type (wt), with the main band centred around 685 nm. Second, dark-level fluorescence is strongly influenced beyond 700 nm by broad emission bands from PS I, and its associated antennae proteins, which exhibit much more rapid decay kinetics and strong integrated amplitudes. In particular a 705-720 nm band is present in all three samples, with a 710 nm band predominating in the clo f2 leaves. When the PS II photochemistry becomes inhibited, maximizing the fluorescence yield, both the clo f104 mutant and the wt exhibit lifetime increases for their major distribution modes from the minimal 205-500 ps range to the maximal 1500-2500 ps range for both the 685 nm and 740 nm bands. The clo f2 mutant, however, exhibits several unique spectral-kinetic properties, attributed to its unique PS I antennae and thylakoid structure, indicating changes in both PS II fluorescence reabsorption and PS II to PS I energy transfer pathways

  7. Image stacking approach to increase sensitivity of fluorescence detection using a low cost complementary metal-oxide-semiconductor (CMOS) webcam.

    PubMed

    Balsam, Joshua; Bruck, Hugh Alan; Kostov, Yordan; Rasooly, Avraham

    2012-01-01

    Optical technologies are important for biological analysis. Current biomedical optical analyses rely on high-cost, high-sensitivity optical detectors such as photomultipliers, avalanched photodiodes or cooled CCD cameras. In contrast, Webcams, mobile phones and other popular consumer electronics use lower-sensitivity, lower-cost optical components such as photodiodes or CMOS sensors. In order for consumer electronics devices, such as webcams, to be useful for biomedical analysis, they must have increased sensitivity. We combined two strategies to increase the sensitivity of CMOS-based fluorescence detector. We captured hundreds of low sensitivity images using a Webcam in video mode, instead of a single image typically used in cooled CCD devices.We then used a computational approach consisting of an image stacking algorithm to remove the noise by combining all of the images into a single image. While video mode is widely used for dynamic scene imaging (e.g. movies or time-lapse photography), it is not used to capture a single static image, which removes noise and increases sensitivity by more than thirty fold. The portable, battery-operated Webcam-based fluorometer system developed here consists of five modules: (1) a low cost CMOS Webcam to monitor light emission, (2) a plate to perform assays, (3) filters and multi-wavelength LED illuminator for fluorophore excitation, (4) a portable computer to acquire and analyze images, and (5) image stacking software for image enhancement. The samples consisted of various concentrations of fluorescein, ranging from 30 μM to 1000 μM, in a 36-well miniature plate. In the single frame mode, the fluorometer's limit-of-detection (LOD) for fluorescein is ∼1000 μM, which is relatively insensitive. However, when used in video mode combined with image stacking enhancement, the LOD is dramatically reduced to 30 μM, sensitivity which is similar to that of state-of-the-art ELISA plate photomultiplier-based readers. Numerous medical

  8. Calibration procedures and first dataset of Southern Ocean chlorophyll a profiles collected by elephant seals equipped with a newly developed CTD-fluorescence tags

    NASA Astrophysics Data System (ADS)

    Guinet, C.; Xing, X.; Walker, E.; Monestiez, P.; Marchand, S.; Picard, B.; Jaud, T.; Authier, M.; Cotté, C.; Dragon, A. C.; Diamond, E.; Antoine, D.; Lovell, P.; Blain, S.; D'Ortenzio, F.; Claustre, H.

    2013-02-01

    In situ observation of the marine environment has traditionally relied on ship-based platforms. The obvious consequence is that physical and biogeochemical properties have been dramatically undersampled, especially in the remote Southern Ocean (SO). The difficulty in obtaining in situ data represents the major limitations to our understanding, and interpretation of the coupling between physical forcing and the biogeochemical response. Southern elephant seals (Mirounga leonina) equipped with a new generation of oceanographic sensors can measure ocean structure in regions and seasons rarely observed with traditional oceanographic platforms. Over the last few years, seals have allowed for a considerable increase in temperature and salinity profiles from the SO, but we were still lacking information on the spatiotemporal variation of phytoplankton concentration. This information is critical to assess how the biological productivity of the SO, with direct consequences on the amount of CO2 "fixed'' by the biological pump, will respond to global warming. In this research programme, we use an innovative sampling fluorescence approach to quantify phytoplankton concentration at sea. For the first time, a low energy consumption fluorometer was added to Argos CTD-SRDL tags, and these novel instruments were deployed on 27 southern elephant seals between 25 December 2007 and the 4 February 2011. As many as 3388 fluorescence profiles associated with temperature and salinity measurements were thereby collected from a vast sector of the Southern Indian Ocean. This paper addresses the calibration issue of the fluorometer before being deployed on elephant seals and presents the first results obtained for the Indian sector of the Southern Ocean. This in situ system is implemented in synergy with satellite ocean colour radiometry. Satellite-derived data is limited to the surface layer and is restricted over the SO by extensive cloud cover. However, with the addition of these new tags

  9. Calibration procedures and first data set of Southern Ocean chlorophyll a profiles collected by elephant seal equipped with a newly developed CTD-fluorescence tags

    NASA Astrophysics Data System (ADS)

    Guinet, C.; Xing, X.; Walker, E.; Monestiez, P.; Marchand, S.; Picard, B.; Jaud, T.; Authier, M.; `Cotté, C.; Dragon, A. C.; Diamond, E.; Antoine, D.; Lovell, P.; Blain, S.; D'Ortenzio, F.; Claustre, H.

    2012-08-01

    In-situ observation of the marine environment has traditionally relied on ship-based platforms. The obvious consequence is that physical and biogeochemical properties have been dramatically undersampled, especially in the remote Southern Ocean (SO). The difficulty in obtaining in situ data represents the major limitations to our understanding, and interpretation of the coupling between physical forcing and the biogeochemical response. Southern elephant seals (Mirounga leonina) equipped with a new generation of oceanographic sensors can measure ocean structure in regions and seasons rarely observed with traditional oceanographic platforms. Over the last few years, seals have allowed for a considerable increase in temperature and salinity profiles from the SO. However we were still lacking information on the spatio-temporal variation of phytoplankton concentration. This information is critical to assess how the biological productivity of the SO, with direct consequences on the amount of CO2 "fixed" by the biological pump, will respond to global warming. In this research program, we use an innovative sampling fluorescence approach to quantify phytoplankton concentration at sea. For the first time, a low energy consumption fluorometer was added to Argos CTD-SRDL tags, and these novel instruments were deployed on 27 southern elephant seals between 25 December 2007 and the 4 February 2011. As many as 3388 fluorescence profiles associated with temperature and salinity measurements were thereby collected from a vast sector of the Southern Indian Ocean. This paper address the calibration issue of the fluorometer before being deployed on elephant seals and present the first results obtained for the Indian Sector of the Southern Ocean. This in situ system is implemented in synergy with satellite ocean colour radiometry. Satellite-derived data is limited to the surface layer and is restricted over the SO by extensive cloud cover. However, with the addition of these new tags

  10. FRET-Aptamer Assays for Bone Marker Assessment, C-Telopeptide, Creatinine, and Vitamin D

    NASA Technical Reports Server (NTRS)

    Bruno, John G.

    2013-01-01

    Astronauts lose 1.0 to 1.5% of their bone mass per month on long-duration spaceflights. NASA wishes to monitor the bone loss onboard spacecraft to develop nutritional and exercise countermeasures, and make adjustments during long space missions. On Earth, the same technology could be used to monitor osteoporosis and its therapy. Aptamers bind to targets against which they are developed, much like antibodies. However, aptamers do not require animal hosts or cell culture and are therefore easier, faster, and less expensive to produce. In addition, aptamers sometimes exhibit greater affinity and specificity vs. comparable antibodies. In this work, fluorescent dyes and quenchers were added to the aptamers to enable pushbutton, one-step, bind-and-detect fluorescence resonance energy transfer (FRET) assays or tests that can be freeze-dried, rehydrated with body fluids, and used to quantitate bone loss of vitamin D levels with a handheld fluorometer in the spacecraft environment. This work generated specific, rapid, one-step FRET assays for the bone loss marker C-telopeptide (CTx) when extracted from urine, creatinine from urine, and vitamin D congeners in diluted serum. The assays were quantified in nanograms/mL using a handheld fluorometer connected to a laptop computer to convert the raw fluorescence values into concentrations of each analyte according to linear standard curves. DNA aptamers were selected and amplified for several rounds against a 26- amino acid form of CTx, creatinine, and vitamin D. The commonalities between loop structures were studied, and several common loop structures were converted into aptamer beacons with a fluorophore and quencher on each end. In theory, when the aptamer beacon binds its cognate target (CTx bone peptide, creatinine, or vitamin D), it is forced open and no longer quenched, so it gives off fluorescent light (when excited) in proportion to the amount of target present in a sample. This proportional increase in fluorescence is

  11. InSitu-Eye: oceanological and atmospheric data processing and analyzing system

    NASA Astrophysics Data System (ADS)

    Stepochkin, Igor E.; Salyuk, Pavel A.; Shmirko, Konstantin A.; Golik, Irina A.; Burov, Denis V.

    2014-11-01

    In this study we introduce brief description and the main approaches used in system development. System is devising with a participation of Pacific Oceanological Institute (FEB RAS), Institute of Automation and Control Processes (FEB RAS) and also Maritime State University, n.a. G.I. Nevelskoy. For many years research team of these institutions carried out a lot of field measurements and collected a lot of remote sensing data, using spectrophotometers, LIDARs, fluorometers. The primary goal of this development - bring all this data together to integrated database and design user-friendly interface to work with. "InSitu-Eye" will perform standard routine operations, such as sampling data according to certain parameters; gridding and timing of data; filtering and quality check of data; visualization. After setting system up and testing it will provide a benefit. At first it gives 24/7 access to "clean", checked "in-situ" data, ready for further research. Also presence of such system gives "converse effect" - it will become necessary to develop strict protocols for measurements carrying out and increase their quality. In future, "InSitu-Eye" can become a platform, connecting research teams for data keeping and exchange.

  12. Role of the mannose receptor in phagocytosis of Enterococcus faecalis strain EC-12 by antigen-presenting cells

    PubMed Central

    Tsuruta, Takeshi; Inoue, Ryo; Nagino, Takayuki; Nishibayashi, Ryoichiro; Makioka, Yuko; Ushida, Kazunari

    2013-01-01

    Abstract The aim of this study was to clarify the phagocytic mechanisms of a heat-killed cell preparation of Enterococcus faecalis strain EC-12 (EC-12) by antigen-presenting cells (APCs). Fluorescein isothiocyanate (FITC)-labeled EC-12 was cocultured with peritoneal macrophage and the amount of EC-12 phagocytosed by peritoneal macrophages was measured using a microplate fluorometer. Peritoneal macrophages from toll-like receptor (TLR)2-, TLR7-, and MyD88-deficient knockout (KO) mice exhibited similar levels of EC-12 phagocytosis to those from wild-type mice. Similarly, dectin-1 neutralization of peritoneal macrophages had no effect on EC-12 phagocytosis. However, blockade of the mannose receptor (MR) significantly decreased the amount of EC-12 phagocytosed by peritoneal macrophages; the same effect was observed in bone marrow-derived macrophages and dendritic cells. Our findings suggest that MR plays a major role in EC-12 phagocytosis by the APCs. This aim of this study was to clarify the phagocytic mechanisms of a heat-killed cell preparation of Enterococcus faecalis strain EC-12 (EC-12) by antigen-presenting cells (APCs). Our findings suggest that mannose receptor (MR) plays a major role in EC-12 phagocytosis by the APCs. PMID:23801521

  13. An operational fluorescence system for crop assessment

    NASA Astrophysics Data System (ADS)

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

    2004-03-01

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

  14. Effects of ozone exposure or fungal pathogen on white lupin leaves as determined by imaging of chlorophyll a fluorescence.

    PubMed

    Guidi, Lucia; Mori, Sauro; Degl'Innocenti, Elena; Pecchia, Susanna

    2007-01-01

    Chlorophyll fluorescence has been used routinely to investigate photosynthetic activity in plants subjected to both biotic and abiotic stresses. The aim of this work was to compare the perturbations in photosynthesis induced by ozone and by a pathogen. By using a conventional fluorometer a similar response pattern was observed in inoculated and O(3)-fumigated leaves. The application of chlorophyll fluorescence imaging provided further detailed information on the spatial-temporal heterogeneity of the response of white lupin leaves to fungal pathogen or to ozone fumigation. In particular, 48 h after artificial inoculation with the necrotrophic fungal pathogen Pleiochaeta setosa, the leaves showed a remarkable alteration in PSII operating efficiency (Phi(PSII)), which affected the whole surface. Afterwards, the infection site was surrounded by a ring of increased photosynthetic activity. The response of ozonated leaves was quite different. The reduction in Phi(PSII) was already evident 24h after fumigation; moreover, a distinct heterogeneity of the fluorescence yield was observed and the major veins displayed a lowered Phi(PSII). PMID:17900916

  15. High frequency sampling of the 1984 spring bloom within the mid-Atlantic Bight: Synoptic shipboard, aircraft, and in situ perspectives of the SEEP-I experiment

    NASA Technical Reports Server (NTRS)

    Walsh, J. J.; Wirick, C. D.; Pietrafesa, L. J.; Whitledge, T. E.; Hoge, F. E.; Swift, R. N.

    1986-01-01

    Moorings of current meters, thermistors, transmissometers, and fluorometers on the mid-Atlantic shelf, south of Long Island, suggest a cumulative seaward export of perhaps 0.35 g C/sq m/day between the 80 and 120 m isobaths during February-April 1984. Such a horizontal loss of algal carbon over the lower third of the water column would be 23 to 78% of the March-April 1984 primary production. This physical carbon loss is similar to daily grazing losses from zooplankton of 32-40% of the algal fixation of carbon. Metabolic demands of the benthos could be met by just the estimated fecal pellet flux, without direct consumption of algal carbon, while bacterioplankton needs could be served by excretory release of dissolved organic matter during photosynthesis. Sediment traps tethered 10 m off the bottom at the 120 m isobath and 50 m above the 500 m isobath caught as much as 0.16 to 0.26 g C /sq m/day during March-April 1984, in reasonable agreement with the flux estimated from the other moored instruments.

  16. Horizontal dispersion in shelf seas: High resolution modelling as an aid to sparse sampling

    NASA Astrophysics Data System (ADS)

    Stashchuk, Nataliya; Vlasenko, Vasiliy; Inall, Mark E.; Aleynik, Dmitry

    2014-11-01

    The ability of a hydrodynamic model to reproduce the results of a dye release experiment conducted in a wide shelf sea environment was investigated with the help of the Massachusetts Institute of Technology general circulation model (MITgcm). In the field experiment a fluorescent tracer, Rhodamine WT, was injected into the seasonal pycnocline, and its evolution was tracked for two days using a towed undulating vehicle equipped with a fluorometer and a CTD. With a 50 m horizontal resolution grid, and with three different forcings initialized in the model (viz: tides, stationary current, and wind stress on the free surface), it was possible to replicate the dye patch evolution quite accurately. The mechanisms responsible for the enhancement of horizontal dispersion were investigated on the basis of the model results. It was found that enhancement of the dye dispersion was controlled by vertically sheared currents that, in combination with vertical diapycnal mixing, led to a substantial increase in the “effective” horizontal mixing. The values of “effective” horizontal mixing found from the model runs were in good agreement with those obtained from in-situ data, and the probable degree to which the observational techniques undersampled the dye patch was revealed.

  17. The Use of Chlorophyll Fluorescence Lifetime to Assess Phytoplankton Physiology within a River-Dominated Environment

    NASA Technical Reports Server (NTRS)

    Hall, Callie M.; Miller, Richard L.; Redalje, Donald G.; Fernandez, Salvador M.

    2002-01-01

    Chlorophyll a fluorescence lifetime was measured for phytoplankton populations inhabiting the three physical zones surrounding the Mississippi River's terminus in the Gulf of Mexico. Observations of river discharge volume, nitrate + nitrite, silicate, phosphate, PAR (Photosynthetically Active Radiation) diffuse attenuation within the water column, salinity, temperature, SPM, and chl a concentration were used to characterize the distribution of chl fluorescence lifetime within a given region within restricted periods of time. 33 stations extending from the Mississippi River plume to the shelf break of the Louisiana coast were surveyed for analysis of chlorophyll fluorescence lifetime during two cruises conducted March 31 - April 6, 2000, and October 24 - November 1, 2000. At each station, two to three depths were chosen for fluorescence lifetime measurement to represent the vertical characteristics of the water column. Where possible, samples were taken from just below the surface and from just above and below the pycnocline. All samples collected were within the 1% light level of the water column (the euphotic zone). Upon collection, samples were transferred to amber Nalgene bottles and left in the dark for at least 15 minutes to reduce the effects of non-photochemical quenching and to insure that photosynthetic reaction centers were open. Before measurements within the phase fluorometer were begun, the instrument was allowed to warm up for no less than one hour.

  18. Assessment of benthic flux of dissolved organic carbon in wetland and estuarine sediments using the eddy-correlation technique

    NASA Astrophysics Data System (ADS)

    Swett, M. P.; Amirbahman, A.; Boss, E.

    2009-12-01

    Wetland and estuarine sediments release significant amounts of dissolved organic carbon (DOC) due to high levels of microbial activity, particularly sulfate reduction. Changes in climate and hydrologic conditions have a potential to alter DOC release from these systems as well. This is a concern, as high levels of DOC can lead to mobilization of toxic metals and organics in natural waters. In addition, source waters high in DOC produce undesirable disinfection byproducts in water treatment. Various in situ methods, such as peepers and sediment core centrifugation, exist to quantify vertical benthic fluxes of DOC and other dissolved species from the sediment-water interface (SWI). These techniques, however, are intrusive and involve disturbance of the sediment environment. Eddy-correlation allows for real-time, non-intrusive, in situ flux measurement of important analytes, such as O2 and DOC. An Acoustic Doppler Velocimeter (ADV) is used to obtain three-dimensional fluid velocity measurements. The eddy-correlation technique employs the mathematical separation of fluid velocity into mean velocity and fluctuating velocity components, with the latter representing turbulent eddy velocity. DOC concentrations are measured using a colored dissolved organic matter (CDOM) fluorometer, and instantaneous vertical flux is determined from the correlated data. This study assesses DOC flux at three project sites: a beaver pond in the Lower Penobscot Watershed, Maine; a mudflat in Penobscot River, Maine; and a mudflat in Great Bay, New Hampshire. Eddy flux values are compared with results obtained using peepers and centrifugation, as well as vertical profiling.

  19. Comparison of breast milk vitamin A concentration measured in fresh milk by a rapid field assay (the iCheck FLUORO) with standard measurement of stored milk by HPLC.

    PubMed

    Engle-Stone, R; Haskell, M J; La Frano, M R; Ndjebayi, A O; Nankap, M; Brown, K H

    2014-08-01

    Availability of rapid, point-of-contact analytical methods would facilitate the use of breast milk vitamin A concentration (BMVA) to assess vitamin A (VA) status. We compared BMVA concentrations measured by high-performance liquid chromatography (HPLC) (the standard technique) with those by iCheck FLUORO, a new portable fluorometer that can rapidly quantify BMVA. Casual breast milk samples (n=154) were collected during a representative survey in Yaoundé and Douala, Cameroon. Milk fat and BMVA concentrations (by iCheck) were measured in fresh milk in the field. After storage at <-20 °C, BMVA concentrations were also measured by HPLC. BMVA values from the two methods were highly correlated (R(2)=0.72 for BMVA/l; R(2)=0.62 for BMVA/g fat, both P<0.0001). HPLC values were greater than iCheck values on average, and the difference increased with increasing BMVA. The iCheck FLUORO could be useful for monitoring fortification programs, but before-after surveys to assess change in BMVA concentrations should use one method consistently. PMID:24736678

  20. Optical studies of tissue mitochondrial redox in isolated perfused rat lungs

    NASA Astrophysics Data System (ADS)

    Sepehr, R.; Staniszewski, K.; Jacobs, E. R.; Audi, S.; Ranji, M.

    2012-02-01

    Through the monitoring of the auto-fluorescent mitochondrial metabolic coenzymes, NADH (Nicotinamide Adenine Dinucleotide) and FAD (Flavoprotein Adenine Dinucleotide), the redox state of metabolism can be probed in real time in many intact organs, but its use has not been fully developed in lungs. The ratio of these fluorophores, (NADH/FAD), referred to as the mitochondrial redox ratio (RR), can be used as a quantitative metabolic marker of tissue. We have designed a fluorometer that can be used to monitor lung surface NADH and FAD fluorescence in isolated perfused lungs. Surface fluorescence NADH and FAD signals were acquired in the absence (control) and presence of pentachlorophenol (PCP), rotenone, and potassium cyanide (KCN). Rotenone, an inhibitor of complex I, increased RR by 18%, predominantly due to an increase in NADH signal. KCN, an inhibitor of complex IV reduced the chain and resulted in an increase of 33% in RR, as a result of 23% increase in NADH and 8% in FAD . PCP, an uncoupler which oxidizes the respiratory chain, decreased RR by 18% as a result of 14% decrease in NADH signal and 4% increase in FAD signal. These results demonstrate the ability of surface fluorometry to detect changes in lung tissue mitochondrial redox state in isolated perfused lungs.

  1. Multimode evanescent wave-based sensors: enhancement strategies

    NASA Astrophysics Data System (ADS)

    Saaski, Elric W.; Bizak, Michael; Yeatts, Jennifer

    1995-04-01

    There is currently a need for new technologies that are designed specifically for the economical field monitoring of toxins, explosives, and chemical contaminants. The United States has, for example, implemented five regulatory acts to protect its ecologies and its citizens from environmental pollution, and these acts all mandate the monitoring of various chemical contaminants. It is generally accepted that the number of analyses that would be required to meet these new standards would exceed the capacity of all the certified testing labs in the country. New field-portable equipment is needed that can supplement lab-based diagnostic analytical instrumentation, but a continuing problem has been the development of field hardware that can identify and quantify with high specificity a particular species of interest. One of the most promising strategies for performing such narrowly targeted field assays is based on sensors that harness natural immune and protective responses of animals and humans to hone in on a specific compound. This paper discusses the design of a new solid-state portable fluorometer that can be used for the interrogation of a wide range of multimode fiber optic biosensors.

  2. Firefly Luciferase-Based Sequential Bioluminescence Resonance Energy Transfer (BRET)-Fluorescence Resonance Energy Transfer (FRET) Protease Assays.

    PubMed

    Branchini, Bruce

    2016-01-01

    We describe here the preparation of ratiometric luminescent probes that contain two well-separated emission peaks produced by a sequential bioluminescence resonance energy transfer (BRET)-fluorescence resonance energy transfer (FRET) process. The probes are single soluble fusion proteins consisting of a thermostable firefly luciferase variant that catalyzes yellow-green (560 nm maximum) bioluminescence and a red fluorescent protein covalently labeled with a near-Infrared fluorescent dye. The two proteins are connected by a decapeptide containing a protease recognition site specific for factor Xa, thrombin, or caspase 3. The rates of protease cleavage of the fusion protein substrates were monitored by recording emission spectra and plotting the change in peak ratios over time. Detection limits of 0.41 nM for caspase 3, 1.0 nM for thrombin, and 58 nM for factor Xa were realized with a scanning fluorometer. This method successfully employs an efficient sequential BRET-FRET energy transfer process based on firefly luciferase bioluminescence to assay physiologically important protease activities and should be generally applicable to the measurement of any endoprotease lacking accessible cysteine residues. PMID:27424898

  3. Remote monitoring of chlorophyll fluorescence in two reef corals during the 2005 bleaching event at Lee Stocking Island, Bahamas

    NASA Astrophysics Data System (ADS)

    Manzello, D.; Warner, M.; Stabenau, E.; Hendee, J.; Lesser, M.; Jankulak, M.

    2009-03-01

    Zooxanthellae fluorescence was measured in situ, remotely, and in near real-time with a pulse amplitude modulated (PAM) fluorometer for a colony of Siderastrea siderea and Agaricia tenuifolia at Lee Stocking Island, Bahamas during the Caribbean-wide 2005 bleaching event. These colonies displayed evidence of photosystem II (PS II) inactivation coincident with thermal stress and seasonally high doses of solar radiation. Hurricane-associated declines in temperature and light appear to have facilitated the recovery of maximum quantum yield of PS II within these two colonies, although both corals responded differently to individual storms. PAM fluorometry, coupled with long-term measurement of in situ light and temperature, provides much more detail of coral photobiology on a seasonal time scale and during possible bleaching conditions than sporadic, subjective, and qualitative observations. S. siderea displayed evidence of PS II inactivation over a month prior to the issuing of a satellite-based, sea surface temperature (SST) bleaching alert by the National Oceanic and Atmospheric Administration (NOAA). In fact, recovery had already begun in S. siderea when the bleaching alert was issued. Fluorescence data for A. tenuifolia were difficult to interpret because the shaded parts of a colony were monitored and thus did not perfectly coincide with thermal stress and seasonally high doses of solar radiation as in S. siderea. These results further emphasize the limitations of solely monitoring SST (satellite or in situ) as a bleaching indicator without considering the physiological status of coral-zooxanthellae symbioses.

  4. Cu(2+) inhibits photosystem II activities but enhances photosystem I quantum yield of Microcystis aeruginosa.

    PubMed

    Deng, Chunnuan; Pan, Xiangliang; Wang, Shuzhi; Zhang, Daoyong

    2014-08-01

    Responses of photosystem I and II activities of Microcystis aeruginosa to various concentrations of Cu(2+) were simultaneously examined using a Dual-PAM-100 fluorometer. Cell growth and contents of chlorophyll a were significantly inhibited by Cu(2+). Photosystem II activity [Y(II)] and electron transport [rETRmax(II)] were significantly altered by Cu(2+). The quantum yield of photosystem II [Y(II)] decreased by 29 % at 100 μg L(-1) Cu(2+) compared to control. On the contrary, photosystem I was stable under Cu(2+) stress and showed an obvious increase of quantum yield [Y(I)] and electron transport [rETRmax(I)] due to activation of cyclic electron flow (CEF). Yield of cyclic electron flow [Y(CEF)] was enhanced by 17 % at 100 μg L(-1) Cu(2+) compared to control. The contribution of linear electron flow to photosystem I [Y(II)/Y(I)] decreased with increasing Cu(2+) concentration. Yield of cyclic electron flow [Y(CEF)] was negatively correlated with the maximal photosystem II photochemical efficiency (F v/F m). In summary, photosystem II was the major target sites of toxicity of Cu(2+), while photosystem I activity was enhanced under Cu(2+) stress. PMID:24920130

  5. Optical sensor for rapid microbial detection

    NASA Astrophysics Data System (ADS)

    Al-Adhami, Mustafa; Tilahun, Dagmawi; Rao, Govind; Kostov, Yordan

    2016-05-01

    In biotechnology, the ability to instantly detect contaminants is key to running a reliable bioprocess. Bioprocesses are prone to be contaminated by cells that are abundant in our environment; detection and quantification of these cells would aid in the preservation of the bioprocess product. This paper discusses the design and development of a portable kinetics fluorometer which acts as a single-excitation, single-emission photometer that continuously measures fluorescence intensity of an indicator dye, and plots it. Resazurin is used as an indicator dye since the viable contaminant cells reduce Resazurin toResorufin, the latter being strongly fluorescent. A photodiode detects fluorescence change by generating current proportional to the intensity of the light that reached it, and a trans-impedance differential op-amp ensures amplification of the photodiodes' signal. A microfluidic chip was designed specifically for the device. It acts as a fully enclosed cuvette, which enhances the Resazurin reduction rate. E. coli in LB media, along with Resazurin were injected into the microfluidic chip. The optical sensor detected the presence of E. coli in the media based on the fluorescence change that occurred in the indicator dye in concentrations as low as 10 CFU/ml. A method was devised to detect and determine an approximate amount of contamination with this device. This paper discusses application of this method to detect and estimate sample contamination. This device provides fast, accurate, and inexpensive means to optically detect the presence of viable cells.

  6. In-situ optical and acoustical measurements of the buoyant cyanobacterium p. Rubescens: spatial and temporal distribution patterns.

    PubMed

    Hofmann, Hilmar; Peeters, Frank

    2013-01-01

    Optical (fluorescence) and acoustic in-situ techniques were tested in their ability to measure the spatial and temporal distribution of plankton in freshwater ecosystems with special emphasis on the harmful and buoyant cyanobacterium P. rubescens. Fluorescence was measured with the multi-spectral FluoroProbe (Moldaenke FluoroProbe, MFP) and a Seapoint Chlorophyll Fluorometer (SCF). In-situ measurements of the acoustic backscatter strength (ABS) were conducted with three different acoustic devices covering multiple acoustic frequencies (614 kHz ADCP, 2 MHz ADP, and 6 MHz ADV). The MFP provides a fast and reliable technique to measure fluorescence at different wavelengths in situ, which allows discriminating between P. rubescens and other phytoplankton species. All three acoustic devices are sensitive to P. rubescens even if other scatterers, e.g., zooplankton or suspended sediment, are present in the water column, because P. rubescens containing gas vesicles has a strong density difference and hence acoustic contrast to the ambient water and other scatterers. After calibration, the combination of optical and acoustical measurements not only allows qualitative and quantitative observation of P. rubescens, but also distinction between P. rubescens, other phytoplankton, and zooplankton. As the measuring devices can sample in situ at high rates they enable assessment of plankton distributions at high temporal (minutes) and spatial (decimeters) resolution or covering large temporal (seasonal) and spatial (basin scale) scales. PMID:24303028

  7. Self-adjustment of stream bed roughness and flow velocity in a steep mountain channel

    NASA Astrophysics Data System (ADS)

    Schneider, Johannes M.; Rickenmann, Dieter; Turowski, Jens M.; Kirchner, James W.

    2015-10-01

    Understanding how channel bed morphology affects flow conditions (and vice versa) is important for a wide range of fluvial processes and practical applications. We investigated interactions between bed roughness and flow velocity in a steep, glacier-fed mountain stream (Riedbach, Ct. Valais, Switzerland) with almost flume-like boundary conditions. Bed gradient increases along the 1 km study reach by roughly 1 order of magnitude (S = 3-41%), with a corresponding increase in streambed roughness, while flow discharge and width remain approximately constant due to the glacial runoff regime. Streambed roughness was characterized by semivariograms and standard deviations of point clouds derived from terrestrial laser scanning. Reach-averaged flow velocity was derived from dye tracer breakthrough curves measured by 10 fluorometers installed along the channel. Commonly used flow resistance approaches (Darcy-Weisbach equation and dimensionless hydraulic geometry) were used to relate the measured bulk velocity to bed characteristics. As a roughness measure, D84 yielded comparable results to more laborious measures derived from point clouds. Flow resistance behavior across this large range of steep slopes agreed with patterns established in previous studies for both lower-gradient and steep reaches, regardless of which roughness measures were used. We linked empirical critical shear stress approaches to the variable power equation for flow resistance to investigate the change of bed roughness with channel slope. The predicted increase in D84 with increasing channel slope was in good agreement with field observations.

  8. Impact of Emission Anisotropy on Fluorescence Spectroscopy and FRET Distance Measurements

    PubMed Central

    Ivanov, Vassili; Li, Min; Mizuuchi, Kiyoshi

    2009-01-01

    Abstract The objective of this report is to provide a practical and improved method for estimating Förster resonance energy transfer distance measurement error due to unknown angles in the dipole orientation factor based on emission anisotropy measurements. We improve on the method of Dale et al. (1979), which has minor mistakes and is frequently interpreted in overly optimistic ways in the literature. To facilitate proper fluorescence intensity measurements, we also evaluated instrument parameters that could impact the measurement. The apparent fluorescence intensity of isotropic samples depends on the sample emission anisotropy, fluorometer geometry, and optical apertures. We separate parameters of the sample, and those of the cylindrically symmetric illumination source and detector in the equations describing results of unpolarized and polarized fluorescence intensity measurements. This approach greatly simplifies calculations compared with the more universal method of Axelrod (1989). We provide a full computational method for calculating the Förster resonance energy transfer distance error and present a graph describing distance error in the simplest case. PMID:19651051

  9. Impairment of benthic diatom adhesion and photosynthetic activity by 2E,4E-decadienal.

    PubMed

    Leflaive, Joséphine; Ten-Hage, Loïc

    2011-11-01

    Within biofilms, microorganisms are exposed to a wide range of chemicals released by phototrophic organisms. Those chemicals are likely to influence the dynamics and functioning of biofilms. 2E,4E-decadienal (DD) is a polyunsaturated aldehyde produced by diatoms which is known to induce adverse effects in many aquatic organisms. It has been shown to inhibit the adhesion and motility of one benthic diatom. The aim of this article was to determine if the effects of DD on diatom adhesion were widespread and if it could affect biofilm formation and functioning. The adhesion of 5 of 10 benthic diatom strains was strongly inhibited at 2.5 μg ml(-1) DD. This indicates a high variability in diatom sensitivity to DD. Several experiments in microcosms showed that the presence of DD diffusing from a substrate decreased biofilm formation. This effect was dose-dependent and persisted for 72 h, though the molecule is highly volatile. Using a PHYTO-PAM fluorometer, we also showed that the effective quantum efficiency of charge separation of PSII of biofilms exposed to DD was negatively affected. This indicates a decrease in the efficiency of the photochemical processes. All these results suggest that the presence of DD-producing strains may have a significant impact on the composition and physiology of biofilms. PMID:21704156

  10. Fluorescence quantum yields of a series of red and near-infrared dyes emitting at 600-1000 nm.

    PubMed

    Rurack, Knut; Spieles, Monika

    2011-02-15

    The determination of the fluorescence quantum yields (QY, Φ(f)) of a series of fluorescent dyes that span the absorption/excitation and emission ranges of 520-900 and 600-1000 nm is reported. The dyes encompass commercially available rhodamine 101 (Rh-101, Φ(f) = 0.913), cresyl violet (0.578), oxazine 170 (0.579), oxazine 1 (0.141), cryptocyanine (0.012), HITCI (0.283), IR-125 (0.132), IR-140 (0.167), and four noncommercial cyanine dyes with specific spectroscopic features, all of them in dilute ethanol solution. The QYs have been measured relative to the National Institute of Standards and Technology's standard reference material (SRM) 936a (quinine sulfate, QS) on a traceably characterized fluorometer, employing a chain of transfer standard dyes that include coumarin 102 (Φ(f) = 0.764), coumarin 153 (0.544), and DCM (0.435) as links between QS and Rh-101. The QY of Rh-101 has also been verified in direct measurements against QS using two approaches that rely only on instrument correction. In addition, the effects of temperature and the presence of oxygen on the fluorescence quantum yield of Rh-101 have been assessed. PMID:21250654

  11. Optimized parameters for fluorescence-based verification of ballast water exchange by ships.

    PubMed

    Murphy, Kathleen R; Ruiz, Gregory M; Dunsmuir, William T M; Waite, T David

    2006-04-01

    Mid-ocean ballast water exchange is mandatory for ships discharging foreign ballast in US territorial waters in order to reduce the risk of biological invasions. However, a reliable tool for determining whether the procedure took place is lacking. We investigated chromophoric dissolved organic matter (CDOM) fluorescence as a tracer of mid-ocean exchange on nine research cruises out of Asia, Europe, and the USA, focusing on challenging source conditions (high salinity, low CDOM). Using parallel factor analysis, we identified nine independent fluorescent components present in varying concentrations in the ocean and in ballast water. One component was sufficient for predicting the coastal vs oceanic source of most ballast water samples. Across nine cruises, thresholds (1.7 and 0.7 ppb quinine sulfate equivalent units) at two fixed wavelength pairs (lambda(ex)/lambda(em) = 320/414 and 370/496 nm, respectively) discriminated coastal from oceanic ballast water in > 95% of samples (N = 514). Our results suggest that single- and dual-channel fluorometers could be optimized for verifying ballast water exchange. PMID:16646474

  12. Fluorescence Intercalibration Experiment: a Multi-laboratory Comparison of Correction Procedures for Fluorescence Analysis of Dissolved Organic Matter

    NASA Astrophysics Data System (ADS)

    Boehme, J.; Stedmon, C.; Boyd, T.; Chen, R.; Coble, P.; Cooper, W.; Mopper, K.; Wells, M.; Zepp, R.

    2006-12-01

    Measurement of the fluorescence properties of dissolved organic matter (DOM) provides a window into the biological, chemical and physical processes that affect this significant portion of the global carbon pool. Parameters such as fluorescence intensity, quantum yields, peak bandwidth and peak position provide the basis for interpretation of DOM chemical and environmental variability. Generating reliable parameters from fluorescence data requires both correction for instrument bias and standardized experimental methods. The development, publication and use of correction procedures across different fluorometer platforms has proceeded, however the level of variability among corrected fluorescence data in the general DOM community has not been assessed recently. To that end, an intercalibration study was undertaken to examine the current status of correction procedures with the excitation emission matrix spectroscopy technique (EEMS). Analyses of quinine sulfate standard reference material, Suwannee river fulvic acid, and unconcentrated seawater from the Hudson Canyon were performed by 8 participating laboratories. Statistical analysis of fluorescence variability among laboratories will be discussed, along with implications for future fluorescence analysis of DOM.

  13. Long-term changes in the chlorophyll fluorescence of bleached and recovering corals from Hawaii.

    PubMed

    Rodrigues, Lisa J; Grottoli, Andréa G; Lesser, Michael P

    2008-08-01

    Chlorophyll fluorescence has been used to predict and monitor coral bleaching over short timescales (hours to days), but long-term changes during recovery remain largely unknown. To evaluate changes in fluorescence during long-term bleaching and recovery, Porites compressa and Montipora capitata corals were experimentally bleached in tanks at 30 degrees C for 1 month, while control fragments were maintained at 27 degrees C. A pulse amplitude modulated fluorometer measured the quantum yield of photosystem II fluorescence (Fv/Fm) of the zooxanthellae each week during bleaching, and after 0, 1.5, 4 and 8 months recovery. M. capitata appeared bleached 6 days sooner than P. compressa, yet their fluorescence patterns during bleaching did not significantly differ. Changes in minimum (Fo), maximum (Fm) and variable (Fv) fluorescence throughout bleaching and recovery indicated periods of initial photoprotection followed by photodamage in both species, with P. compressa requiring less time for photosystem II (PS II) repair than M. capitata. Fv/Fm fully recovered 6.5 months earlier in P. compressa than M. capitata, suggesting that the zooxanthellae of P. compressa were more resilient to bleaching stress. PMID:18626085

  14. Fiber optic-based fluorescence detection system for in vivo studies of exogenous chromophore pharmacokinetics

    NASA Astrophysics Data System (ADS)

    Doiron, Daniel R.; Dunn, J. B.; Mitchell, W. L.; Dalton, Brian K.; Garbo, Greta M.; Warner, Jon A.

    1995-05-01

    The detection and quantification of the concentration of exogenous chromophores in-vivo by their fluorescence is complicated by many physical and geometrical parameters. Measurement of such signals is advantageous in determining the pharmacokinetics of photosensitizers such as those used in photodynamic therapy (PDT) or to assist in the diagnosis of tissue histological state. To overcome these difficulties a ratio based fiber optic contact fluorometer has been developed. This fluorescence detection system (FDS) uses the ratio of the fluorescence emission peak of the exogenous chromophore to that of endogenous chromophores, i.e. autofluorescence, to correct for a variety of parameters affecting the magnitude of the measured signals. By doing so it also minimizes the range of baseline measurements prior to exogenous drug injection, for various tissue types. Design of the FDS and results of its testing in animals and patients using the second generation photosensitizer Tin ethyletiopurpurin (SnET2) are presented. These results support the feasibility and usefulness of the Ratio FDS system.

  15. Assessing the poplar photochemical response to high zinc concentrations by image processing and statistical approach.

    PubMed

    Sighicelli, Maria; Guarneri, Massimiliano

    2014-12-01

    Exposure of plants to high-heavy metals concentration inhibits multiple metabolic processes in plants and leads to an oxidative stress commonly referred as heavy metal ion toxicity. Chlorophyll a fluorescence has enhanced understanding of heavy metal ion action on the photosynthetic system. A rapid and non-invasive technique involving imaging of chlorophyll fluorescence is a useful tool for early detection of plant responses to heavy metal ion toxicity. In this work chlorophyll fluorescence emission and photochemical parameters in plants of Populus x euramericana clone I-214 were investigated by the portable Imaging PAM fluorometer at different days after soil treatment with zinc. Custom software for analysis of the photochemical parameters images has been developed in order to gain a better assessing of the plant performance in response of metal stress. The imaging analysis allowed visualizing heterogeneity in plant response to high zinc concentrations. The heterogeneity of images suggests spatial differences in photochemical activity and changes in the antenna down-regulation. PMID:25086626

  16. Photosynthesis rates in cyanobacteria-dominated sub-tidal biofilms near Zanzibar, Tanzania

    NASA Astrophysics Data System (ADS)

    Lugomela, Charles; Söderbäck, Erik; Björk, Mats

    2005-05-01

    The surface cover (during 1998) and photosynthesis rates (measured intermittently between 1998 and 2000) of submerged cyanobacteria-dominated biofilms near a patchy reef in Zanzibar were determined using a line intercept transect method and pulse-amplitude modulated fluorometer (PAM), respectively. The biofilm surface cover ranged between 5 and 56% with an annual average value of 25%. Photosynthetic activity on deep-dwelling biofilms was low light adapted compared to shallow-dwelling biofilms. Biofilms also regulated their photosynthetic activity depending on the light regime over the day, manifesting high light utilization coefficient ( α), light saturation index ( Ek) and maximum electron transport rate (ETR max) values at around noon compared to morning and evening measurements. We calculated carbon fixation rates of 0.05, 0.3 and 0.5 kg C m -2 y -1 for thin (˜0.5 mm), medium (˜1 mm) and thick (˜2 mm) biofilms, respectively, and estimated an overall primary production rate of 0.14 kg C m -2 y -1 at depths of about 5 m. This study shows that biofilms in the area actively fix carbon and may contribute substantially to the primary productivity of coastal ecosystems. More experiments are required to precisely determine the absorption factor for robust determination of the ETR and to explain the significance of the biofilms on the overall productivity of coastal ecosystems.

  17. Biological membrane modeling with a liquid/liquid interface. Probing mobility and environment with total internal reflection excited fluorescence.

    PubMed Central

    Morrison, L E; Weber, G

    1987-01-01

    Total internal reflection of exciting light, in combination with fluorescence intensity and polarization measurements, was used to selectively study fluorescent compounds adsorbed to the interface region between two immiscible liquids. A fluorometer was constructed which provided excitation at variable angles of incidence and allowed sensitive detection of polarized fluorescence emitted from the interface. The compound 4,4'-bis-1-phenylamino-8-naphthalenesulfonate (bis-ANS) was examined at a decalin/water interface and was found to possess remarkable affinity for the interface region with the bulk of the adsorbed molecule residing in the decalin phase. The adsorbed fluorophore displayed an apparent hindered rotation in the plane of the interface with a rotational diffusion coefficient 3- to 12-fold lower than that expected for bis-ANS in solution. While other dyes examined were not found to be significantly surface active, the addition of cationic surfactant sufficed to induce adsorption of the anionic fluorophore 1-aminonaphthalene-3,6,8-trisulfonic acid. This fluoropore was found to reside in an aqueous environment when bound to the interface, and it also exhibited hindered rotation in the plane of the interface. As the concentrations of the dyes were increased, both adsorbed dyes exhibited polarization reductions consistent with excitation energy transfer. Adsorption of bis-ANS was reversed by addition of bovine serum albumin. The membrane protein cytochrome b5 was found not to bind at the decalin/water interface, indicating that interaction with lipid is required for its adherence to biological membranes. PMID:3651556

  18. Simultaneous measurement of thrombin generation and fibrin formation in whole blood under flow conditions.

    PubMed

    Kelchtermans, Hilde; Pelkmans, Leonie; Bouwhuis, Anne; Schurgers, Evelien; Lindhout, Theo; Huskens, Dana; Miszta, Adam; Hemker, H Coenraad; Lancé, Marcus D; de Laat, Bas

    2016-07-01

    Assays based on the formation of thrombin and fibrin are frequently used, and results are considered exchangeable in research/clinical settings. However, thrombin generation and fibrin formation do not always go hand in hand and flow profoundly influences thrombus formation. We describe the technical/clinical evaluation of an assay to simultaneously measure thrombin generation and fibrin formation under conditions of flow. Introduction of a fluorometer into a 'cone and base principle'-based rheometer allowed the measurement of thrombin generation (using a thrombin-sensitive substrate) and fibrin formation (changes in viscosity), while applying a linear shear flow. Increasing shear rates inversely related with thrombin generation and fibrin formation. Increasing fibrinogen concentrations in defibrinated plasma resulted in increased thrombin generation and fibrin formation. In pre-operative samples of 70 patients undergoing cardiothoracic surgery, fibrin formation and thrombin generation parameters correlated with fibrinogen content, rotational thromboelastometry (ROTEM) and whole blood Calibrated Automated Thrombinography (CAT) parameters, respectively. Upon dividing patients into two groups based on the median clot strength, a significant difference in perioperative/total blood loss was established. In conclusion, we clinically evaluated a method capable of simultaneously measuring thrombin generation and fibrin formation in plasma/whole blood under continuous flow, rendering our method one step closer to physiology. Importantly, our test proved to be indicative for the amount of blood loss during/after cardiothoracic surgery. PMID:27074907

  19. Development of a no-wash assay for mitochondrial membrane potential using the styryl dye DASPEI.

    PubMed

    Jensen, Kristian H R; Rekling, Jens C

    2010-10-01

    Mitochondrial dysfunction is a hallmark of several diseases and may also result from drugs with unwanted side effects on mitochondrial biochemistry. The mitochondrial membrane potential is a good indicator of mitochondrial function. Here, the authors have developed a no-wash mitochondrial membrane potential assay using 2-(4-(dimethylamino)styryl)-N-ethylpyridinium iodide (DASPEI), a rarely used mitochondrial potentiometric probe, in a 96-well format using a fluorescent plate reader. The assay was validated using 2 protonophores (CCCP, DNP), which are known uncouplers, and the neuroleptic thioridazine, which is a suspected mitochondrial toxicant. CCCP and DNP have short-term depolarizing effects, and thioridazine has long-term hyperpolarizing effects on the mitochondrial membrane potential of Chinese hamster ovary (CHO) cells. The assay also detected changes of the mitochondrial membrane potential in CHO cells exposed to cobalt (mimicking hypoxia) and in PC12 cells exposed to amyloid β, demonstrating that the assay can be used in cellular models of hypoxia and Alzheimer's disease. The assay needs no washing steps, has a Z' value >0.5, can be used on standard fluorometers, has good post liquid-handling stability, and thus is suitable for large-scale screening efforts. In summary, the DASPEI assay is simple and rapid and may be of use in toxicological testing, drug target discovery, and mechanistic models of diseases involving mitochondrial dysfunction. PMID:20713988

  20. Mid-summer mesozooplankton biomass, its size distribution, and estimated production within a glacial Arctic fjord (Hornsund, Svalbard)

    NASA Astrophysics Data System (ADS)

    Trudnowska, E.; Basedow, S. L.; Blachowiak-Samolyk, K.

    2014-09-01

    The estimation of secondary production constitutes an integrating proxy of pelagic ecosystem status, its functions as well as its responses to environmental stressors. The combination of high-resolution automatic measurements with a Laser Optical Plankton Counter (LOPC) and size spectrum analyses was utilized to estimate the secondary production of a high Arctic fjord during a summer post bloom situation in 2012. The dataset comprised 28 vertical and extensive horizontal hauls of a LOPC-CTD-fluorometer platform plus four zooplankton net sampling stations for taxonomic composition designation. A clear gradient in temperature, salinity, chlorophyll a concentrations as well as mesozooplankton abundance, biomass and production was demonstrated along Hornsund fjord axis. The outer fjord part was under the influence of water advection and had the highest chlorophyll a concentrations, numerous opaque mesozooplankton individuals and flat slopes of size spectra, pointing to long food chains in which biomass is recycled several times. The opposite state was found in the glacial bays, where the glacier meltwater discharge led to low chlorophyll a concentrations but high abundance of small and amorphous particles. It resulted in steep size spectra slopes and high intercepts implying higher potential productivity there. The model of mesozooplankton production demonstrated that Hornsund fjord is a highly productive ecosystem, particularly its upper water layer and its central parts. However, we would like to emphasize that a careful approach is needed before going deeper into ecological interpretations based on size spectra analysis, especially in reservoirs, where non-zooplankton particles contribute to the size spectra.

  1. Rapid measurements of intracellular calcium using a fluorescence plate reader.

    PubMed

    Lin, K; Sadée, W; Quillan, J M

    1999-02-01

    Intracellular calcium is a universal second messenger that can serve as a broad-based measure of receptor activity. Recent developments in multi-well plate fluorescence readers facilitate measurement of intracellular free-calcium levels and reduce reliance on slower, more cumbersome or expensive data collection methods. In this report, we describe a rapid and sensitive method to assay intracellular calcium ions in human embryonic kidney (HEK293) and Chinese hamster ovary (CHO) cells from multi-well plates using a fluorometer equipped with on-line injectors. We examine the compatibility of visible-light excitable dyes Calcium Green-1 and Oregon Green 488 BAPTA-1. Using this assay, we were able to detect and quantify activity from muscarinic and beta-adrenergic receptors endogenous to HEK293 cells and detect calcium signals generated by activation of Gi-coupled recombinant mu-opioid and dopamine D2L receptors, and the Gs-coupled melanocortin subtype 4 (MC4) receptor. Fluorescence signals, stable in HEK293 cells, required the use of Oregon Green 488 BAPTA-1 and an inhibitor of organic anion transport in CHO cells. Under appropriate conditions, both cell types can be used to collect complete concentration-response data for a variety of receptors (including a recombinant muscarinic M1 receptor expressed in CHO cells) from a single plate of dye-loaded cells. PMID:10023544

  2. Eat-by-light: fiber-optic and micro-optic devices for food safety and quality assessment

    NASA Astrophysics Data System (ADS)

    Mignani, A. G.; Ciaccheri, L.; Cucci, C.; Mencaglia, A. A.; Cimato, A.; Attilio, C.; Thienpont, H.; Ottevaere, H.; Paolesse, R.; Mastroianni, M.; Monti, D.; Buonocore, G.; Del Nobile, A.; Mentana, A.; Dall'Asta, C.; Faccini, A.; Galaverna, G.; Dossena, A.

    2007-07-01

    A selection of fiber-optic and micro-optic devices is presented designed and tested for monitoring the quality and safety of typical foods, namely the extra virgin olive oil, the beer, and the milk. Scattered colorimetry is used for the authentication of various types of extra virgin olive oil and beer, while a fiber-optic-based device for UV-VIS-NIR absorption spectroscopy is exploited in order to obtain the hyperspectral optical signature of olive oil. This is done not only for authentication purposes, but also so as to correlate the spectral data with the content of fatty acids that are important nutritional factors. A micro-optic sensor for the detection of olive oil aroma is presented. It is capable of distinguishing different ageing levels of extra virgin olive oil. It shows effective potential for acting as a smart cap of bottled olive oil in order to achieve a non-destructive olfactory perception of oil ageing. Lastly, a compact portable fluorometer is experimented for the rapid monitoring of the carcinogenic M1 aflatoxin in milk.

  3. Eat-by-light fiber-optic and micro-optic devices for food quality and safety assessment

    NASA Astrophysics Data System (ADS)

    Mignani, A. G.; Ciaccheri, L.; Cucci, C.; Mencaglia, A. A.; Cimato, A.; Attilio, C.; Thienpont, H.; Ottevaere, H.; Paolesse, R.; Mastroianni, M.; Monti, D.; Buonocore, G.; Del Nobile, A.; Mentana, A.; Grimaldi, M. F.; Dall'Asta, C.; Faccini, A.; Galaverna, G.; Dossena, A.

    2007-06-01

    A selection is presented of fiber-optic and micro-optic devices that have been designed and tested for guaranteeing the quality and safety of typical foods, such as extra virgin olive oil, beer, and milk. Scattered colorimetry is used to authenticate various types of extra virgin olive oil and beer, while a fiber-optic-based device for UV-VIS-NIR absorption spectroscopy is exploited in order to obtain the hyperspectral optical signature of olive oil. This is done not only for authentication purposes, but also so as to correlate the spectral data with the content of fatty acids, which are important nutritional factors. A micro-optic sensor for the detection of olive oil aroma that is capable of distinguishing different ageing levels of extra virgin olive oil is also presented. It shows effective potential for acting as a smart cap of bottled olive oil in order to achieve a non-destructive olfactory perception of oil ageing. Lastly, a compact portable fluorometer for the rapid monitoring of the carcinogenic M1 aflatoxin in milk, is experimented.

  4. A Practical Solution for 77 K Fluorescence Measurements Based on LED Excitation and CCD Array Detector.

    PubMed

    Lamb, Jacob; Forfang, Kristin; Hohmann-Marriott, Martin

    2015-01-01

    The fluorescence emission spectrum of photosynthetic microorganisms at liquid nitrogen temperature (77 K) provides important insights into the organization of the photosynthetic machinery of bacteria and eukaryotes, which cannot be observed at room temperature. Conventionally, to obtain such spectra, a large and costly table-top fluorometer is required. Recently portable, reliable, and largely maintenance-free instruments have become available that can be utilized to accomplish a wide variety of spectroscopy-based measurements in photosynthesis research. In this report, we show how to build such an instrument in order to record 77K fluorescence spectra. This instrument consists of a low power monochromatic light-emitting diode (LED), and a portable CCD array based spectrometer. The optical components are coupled together using a fiber optic cable, and a custom made housing that also supports a dewar flask. We demonstrate that this instrument facilitates the reliable determination of chlorophyll fluorescence emission spectra for the cyanobacterium Synechocystis sp. PCC 6803, and the green alga Chlamydomonas reinhardtii. PMID:26177548

  5. Satellite detection of phytoplankton export from the mid-Atlantic Bight during the 1979 spring bloom

    NASA Technical Reports Server (NTRS)

    Walsh, J. J.; Dieterle, D. A.; Esaias, W. E.

    1986-01-01

    Analysis of Coastal Zone Color Scanner (CZCS) imagery confirms shipboard and in situ moored fluorometer observations of resuspension of near-bottom chlorophyll within surface waters (1 to 10 m) by northwesterly wind events in the mid-Atlantic Bight. As much as 8 to 16 micrograms chl/l are found during these wind events from March to May, with a seasonal increase of algal biomass until onset of stratification of the water column. Rapid sinking or downwelling apparently occurs after subsequent wind events, however, such that the predominant surface chlorophyll pattern is approx. 0.5 to 1.5 micrograms/l over the continental shelf during most of the spring bloom. Perhaps half of the chlorophyll increase observed by satellite during a wind resuspension event represents in-situ production during the 4 to 5 day interval, with the remainder attributed to accumulation of algal biomass previously produced and temporarily stored within near-bottom water. Present calculations suggest that about 10% of the primary production of the spring bloom may be exported as ungrazed phytoplankton carbon from mid-Atlantic shelf waters to those of the continental slope.

  6. Investigating mechanisms for the variability in phytoplankton assemblages in the northeastern waters of the Philippines

    NASA Astrophysics Data System (ADS)

    Yniguez, A. T.; Camoying, M.; Bollozos, I.; Palermo, J. H.; Villanoy, C. L.

    2013-12-01

    The northeastern side of the Philippines, particularly off of Luzon Island, is strongly influenced by the Pacific Western Boundary current system. The North Equatorial Current bifurcates offshore feeding into the Mindanao current flowing south and the Kuroshio Current that begins to consolidate in the waters off of Luzon. Two cruises were conducted during May/June 2011 and April/May 2012 which showed distinctly different circulation patterns and strengths of water masses. Phytoplankton abundances and composition measurements were obtained using microscopy, as well as through an in situ fluorometer. Analysis of the phytoplankton assemblages from these two cruises showed variability in both space and time. Distinct assemblages could be observed amongst different water masses both in 2011 and 2012. There was also a significant difference in the phytoplankton abundances and composition seen between the two sampling years. Mechanisms such as circulation patterns and related physico-chemical characteristics leading to these spatio-temporal variabilities in phytoplankton concentrations and distributions are explored in this study. These shifts in primary producers and productivity have important implications in this region which is a key fishing area for both municipal and commercial fisheries.

  7. On the calibration of large-scale fluorometric chlorophyll measurements from towed undulating vehicles

    NASA Astrophysics Data System (ADS)

    Strass, Volker

    1990-03-01

    Calibrating in situ fluorescence profiles to give the concentration of chlorophyll a at high resolution on the oceanic gyre scale must take into account large variations of fluorescence yield R (the ratio of the fluorescence signal and the chlorophyll concentration within the fluorometer detection volume). The calibration method suggested in this paper has been developed to handle fluorescence measurements made from a towed undulating vehicle along a section about 2000 km long between the Azores and Greenland repeated during the years 1984-1986. The calibration is based on equally spaced samples with photometrically determined near-surface chlorophyll concentrations and incorporates profiles of solar irradiance as auxillary variable. Judging from the auto-correlation function of R, the large variations of fluorescence yield are well resolved by the time interval of 4 h (corresponding to a horizontal distance of about 65 km) between adjacent chlorophyll samples. Using a filter based on the auto-correlation function the variability of yield is smoothed; the smoothed yield function is employed to calibrate the fluoresence signal continuously, in which yield between supporting points is interpreted linearly. Significant correlations of R with solar irradiance appear to be negative; this ambient-light-dependent fluorescence quenching is eliminated during calibration. The relative error in the chlorophyll concentration derived in this way is estimated to be 4% near the surface and 15% deeper down. The suggested calibration method is as accurate as more expensive procedures involving bottle cast series.

  8. Fluorometric determination of zirconium in minerals

    USGS Publications Warehouse

    Alford, W.C.; Shapiro, L.; White, C.E.

    1951-01-01

    The increasing use of zirconium in alloys and in the ceramics industry has created renewed interest in methods for its determination. It is a common constituent of many minerals, but is usually present in very small amounts. Published methods tend to be tedious, time-consuming, and uncertain as to accuracy. A new fluorometric procedure, which overcomes these objections to a large extent, is based on the blue fluorescence given by zirconium and flavonol in sulfuric acid solution. Hafnium is the only element that interferes. The sample is fused with borax glass and sodium carbonate and extracted with water. The residue is dissolved in sulfuric acid, made alkaline with sodium hydroxide to separate aluminum, and filtered. The precipitate is dissolved in sulfuric acid and electrolysed in a Melaven cell to remove iron. Flavonol is then added and the fluorescence intensity is measured with a photo-fluorometer. Analysis of seven standard mineral samples shows excellent results. The method is especially useful for minerals containing less than 0.25% zirconium oxide.

  9. Effect of arsenic on reflectance spectra and chlorophyll fluorescence of aquatic plants.

    PubMed

    Iriel, Analia; Dundas, Gavin; Fernández Cirelli, Alicia; Lagorio, Maria G

    2015-01-01

    Arsenic pollution of groundwater is a serious problem in many regions of Latin America that causes severe risks to human health. As a consequence, non-destructive monitoring methodologies, sensitive to arsenic presence in the environment and able to perform a rapid screening of large polluted areas, are highly sought-after. Both chlorophyll - a fluorescence and reflectance of aquatic plants may be potential indicators to sense toxicity in water media. In this work, the effects of arsenic on the optical and photophysical properties of leaves of different aquatic plants (Vallisneria gigantea, Azolla filiculoides and Lemna minor) were evaluated. Reflectance spectra were recorded for the plant leaves from 300 to 2400 nm. The spectral distribution of the fluorescence was also studied and corrected for light re-absorption processes. Photosynthetic parameters (Fv/Fm and ΦPSII) were additionally calculated from the variable chlorophyll fluorescence recorded with a pulse amplitude modulated fluorometer. Fluorescence and reflectance properties for V. gigantea and A. filiculoides were sensitive to arsenic presence in contrast to the behaviour of L. minor. Observed changes in fluorescence spectra could be interpreted in terms of preferential damage in photosystem II. The quantum efficiency of photosystem II for the first two species was also affected, decreasing upon arsenic treatment. As a result of this research, V. gigantea and A. filiculoides were proposed as bioindicators of arsenic occurrence in aquatic media. PMID:25150973

  10. Carbon transport in the bottom boundary layer. Final report

    SciTech Connect

    Lohrenz, S.E.; Asper, V.L.

    1997-09-01

    The authors objective was to characterize distributions of chloropigment fluorescence in relation to physical processes in the benthic boundary layer in support of the Department of Energy (DOE) Ocean Margins Program`s (OMP) goal of quantifying carbon transport across the continental shelf. Their approach involved participation in the Ocean Margins Program (OMP) field experiment on the continental shelf off Cape Hatteras by conducting multi-sensor fluorescence measurements of photosynthetic pigments. Specific tasks included (1) pre- and post-deployment calibration of multiple fluorescence sensors in conjunction with Woods Hole personnel; (2) collection and analysis of photosynthetic pigment concentrations and total particulate carbon in water column samples to aid in interpretation of the fluorescence time-series during the field experiment; (3) collaboration in the analysis and interpretation of 1994 and 1996 time-series data in support of efforts to quantify pigment and particulate organic carbon transport on the continental shelf off Cape Hatteras. This third component included analysis of data obtained with a multi-sensor fiber-optic fluorometer in the benthic boundary layer of the inner shelf off Cape Hatteras during summer 1994.

  11. SeaWiFS technical report series. Volume 32: Level-3 SeaWiFS data products. Spatial and temporal binning algorithms

    NASA Technical Reports Server (NTRS)

    Hooker, Stanford B. (Editor); Firestone, Elaine R. (Editor); Acker, James G. (Editor); Campbell, Janet W.; Blaisdell, John M.; Darzi, Michael

    1995-01-01

    The level-3 data products from the Sea-viewing Wide Field-of-view Sensor (SeaWiFS) are statistical data sets derived from level-2 data. Each data set will be based on a fixed global grid of equal-area bins that are approximately 9 x 9 sq km. Statistics available for each bin include the sum and sum of squares of the natural logarithm of derived level-2 geophysical variables where sums are accumulated over a binning period. Operationally, products with binning periods of 1 day, 8 days, 1 month, and 1 year will be produced and archived. From these accumulated values and for each bin, estimates of the mean, standard deviation, median, and mode may be derived for each geophysical variable. This report contains two major parts: the first (Section 2) is intended as a users' guide for level-3 SeaWiFS data products. It contains an overview of level-0 to level-3 data processing, a discussion of important statistical considerations when using level-3 data, and details of how to use the level-3 data. The second part (Section 3) presents a comparative statistical study of several binning algorithms based on CZCS and moored fluorometer data. The operational binning algorithms were selected based on the results of this study.

  12. Chemical, biochemical, and environmental fiber sensors IV; Proceedings of the Meeting, Boston, MA, Sept. 8, 9, 1992

    NASA Astrophysics Data System (ADS)

    Lieberman, Robert A.

    Various paper on chemical, biochemical, and environmental fiber sensors are presented. Some of the individual topics addressed include: evanescent-wave fiber optic (FO) biosensor, refractive-index sensors based on coupling to high-index multimode overlays, advanced technique in FO sensors, design of luminescence-based temperature sensors, NIR fluorescence in FO applications, FO sensor based on microencapsulated reagents, emitters and detectors for optical gas and chemical sensing, tunable fiber laser source for methane detection at 1.68 micron, FO fluorometer based on a dual-wavelength laser excitation source, thin polymer films as active components of FO chemical sensors, submicron optical sources for single macromolecule detection, nanometer optical fiber pH sensor. Also discussed are: microfabrication of optical sensor array, luminescent FO sensor for the measurement of pH, time-domain fluorescence methods as applied to pH sensing, characterization of a sol-gel-entrapped artificial receptor, FO technology for nuclear waste cleanup, spectroscopic gas sensing with IR hollow waveguides, dissolved-oxygen quenching of in situ fluorescence measurements.

  13. Chemical, biochemical, and environmental fiber sensors IV; Proceedings of the Meeting, Boston, MA, Sept. 8, 9, 1992

    SciTech Connect

    Lieberman, R.A.

    1993-01-01

    Various paper on chemical, biochemical, and environmental fiber sensors are presented. Some of the individual topics addressed include: evanescent-wave fiber optic (FO) biosensor, refractive-index sensors based on coupling to high-index multimode overlays, advanced technique in FO sensors, design of luminescence-based temperature sensors, NIR fluorescence in FO applications, FO sensor based on microencapsulated reagents, emitters and detectors for optical gas and chemical sensing, tunable fiber laser source for methane detection at 1.68 micron, FO fluorometer based on a dual-wavelength laser excitation source, thin polymer films as active components of FO chemical sensors, submicron optical sources for single macromolecule detection, nanometer optical fiber pH sensor. Also discussed are: microfabrication of optical sensor array, luminescent FO sensor for the measurement of pH, time-domain fluorescence methods as applied to pH sensing, characterization of a sol-gel-entrapped artificial receptor, FO technology for nuclear waste cleanup, spectroscopic gas sensing with IR hollow waveguides, dissolved-oxygen quenching of in situ fluorescence measurements.

  14. Development of loop-mediated isothermal amplification (LAMP) assay for rapid and sensitive identification of ostrich meat.

    PubMed

    Abdulmawjood, Amir; Grabowski, Nils; Fohler, Svenja; Kittler, Sophie; Nagengast, Helga; Klein, Guenter

    2014-01-01

    Animal species identification is one of the primary duties of official food control. Since ostrich meat is difficult to be differentiated macroscopically from beef, therefore new analytical methods are needed. To enforce labeling regulations for the authentication of ostrich meat, it might be of importance to develop and evaluate a rapid and reliable assay. In the present study, a loop-mediated isothermal amplification (LAMP) assay based on the cytochrome b gene of the mitochondrial DNA of the species Struthio camelus was developed. The LAMP assay was used in combination with a real-time fluorometer. The developed system allowed the detection of 0.01% ostrich meat products. In parallel, a direct swab method without nucleic acid extraction using the HYPLEX LPTV buffer was also evaluated. This rapid processing method allowed detection of ostrich meat without major incubation steps. In summary, the LAMP assay had excellent sensitivity and specificity for detecting ostrich meat and could provide a sampling-to-result identification-time of 15 to 20 minutes. PMID:24963709

  15. Design for manufacturablility (DFM) in the life sciences: fluorescence spectroscopy product platform realized with TracePro suite of opto-mechanical design software tools

    NASA Astrophysics Data System (ADS)

    Freniere, Edward; Hassler, Richard; Heinz, Eric; Smith, Linda

    2007-02-01

    This paper describes the design-for-manufacture (DFM) process for a multi-channel fluorometer product platform. The multi-disciplinary team eliminated the cost of quality by design, using a formal design method, facilitated by Lambda Research Corporation's suite of TraceProTM suite of optical design software. Development of this platform presented rigorous design challenges - from identifying feasible design alternatives to minimizing the exponential cumulative effect of component quality and quantity to optimizing tolerances to thoroughly documenting the design. The design was highly constrained in terms of cost and the ability of the platform to accommodate a breadth of fluorescence-tagged media. Furthermore, the inherently interdisciplinary nature of developing medical devices required a high level of collaboration between scientists and engineers across the areas of optics, mechanics, materials, biology and clinical chemistry. While fluorescence tag technologies enable very sensitive detection of molecules, the anisotropic nature of fluorescence in both intensity and polarization severely complicate system design. TracePro's fluorescence modeling capability enabled adherence to a methodical design process of (1) testing system design alternatives, (2) evaluating off-the- shelf and custom optical component and fluorophore feasibility, and (3) tolerancing for robustness without the cost and time associated with iterative hardware prototyping.

  16. Artemisinin induces caspase-8/9-mediated and Bax/Bak-independent apoptosis in human lung adenocarcinoma (ASTC-a-1) cells.

    PubMed

    Xiao, Feng-Lian; Gao, Wei-Jie; Liu, Cheng-Yi; Wang, Xiao-Ping; Chen, Tong-Sheng

    2011-01-01

    Artemisinin (ARTE), an antimalarial phytochemical component from the sweet wormwood plant, has been shown a potential anticancer activity by inducing cell apoptosis. The aim of this report is to explore the mechanism of ARTE-induced human lung adenocarcinoma (ASTC-a-1) cell apoptosis. Cell counting kit (CCK-8) assay showed that ARTE induced cytotoxcity in a dose- and time-dependent manner. Confocal microscopy fluorescence imaging of cells stained with Hoechst 33258 and flow cytometry (FCM) analysis of cells stained with Annexin V-FITC/propidium iodide (PI) showed that ARTE induced reactive oxygen species (ROS)-dependent apoptosis. Confocal fluorescence resonance energy transfer (FRET) imaging of single living cells expressing SCAT3, SCAT9 or CFP-Bid-YFP and fluorometic substrate assay showed that ARTE induced the activation of caspase-3, -8 and -9. Moreover, inhibition of caspase-8 or -9 completely blocked ARTE-induced apoptosis which was only partially attenuated by caspase-3 inhibitor. Interestingly, silencing Bax and Bak by RNA interference (RNAi) did not attenuate ARTE-induced apoptosis. Collectively, ARTE induces caspase-dependent but Bax/Bak-independent apoptosis in ASTC-a-1 cells. PMID:25214386

  17. Classification of tissue pathological state using optical multiparametric monitoring approach

    NASA Astrophysics Data System (ADS)

    Kutai-Asis, Hofit; Kanter, Ido; Barbiro-Michaely, Efrat; Mayevsky, Avraham

    2008-12-01

    In order to diagnose the development of pathophysiological events in the brain, the evaluation of multiparametric data in real time is highly important. The current work presents a new approach of using cluster analysis for the evaluation of relationship between: mitochondrial NADH, tissue blood flow and hemoglobin oxygenation under various pathophysiological conditions. The Time-Sharing Fluorometer Reflectometer (TSFR) was used for monitoring of mitochondrial NADH, oxyhemoglobin (HbO2), and microcirculatory blood flow simultaneously at the same location from the rat or gerbils cortex. This allows a more accurate assessment of brain functions in real time and a better understanding of the relationship between tissue oxygen supply and demand. Moreover, in some pathophysiological cases, monitoring of only one or two parameters in the cerebral cortex may be misleading. The classification was based on the data collected in experiments where different pathophysiological conditions, such as anoxia, ischemia, and SD were used. These three parameters were plotted in three dimensions. The clustering approach results showed similar patterns in each type of treatment. The distribution of data points in space was used to define the spatial behavior of each treatment in order to produce an index for identifying different treatments. In conclusion, our present study offers a new approach of data analysis that can serve as a reliable tool for tissue pathophysiology.

  18. Simultaneous three-dimensional velocity and mixing measurements by use of laser Doppler velocimetry and fluorescence probes in a water tunnel

    NASA Technical Reports Server (NTRS)

    Neuhart, Dan H.; Wing, David J.; Henderson, Uleses C., Jr.

    1994-01-01

    A water tunnel investigation was conducted to demonstrate the capabilities of a laser-based instrument that can measure velocity and fluorescence intensity simultaneously. Fluorescence intensity of an excited fluorescent dye is directly related to concentration level and is used to indicate the extent of mixing in flow. This instrument is a three-dimensional laser Doppler velocimeter (LDV) in combination with a fluorometer for measuring fluorescence intensity variations. This capability allows simultaneous flow measurements of the three orthogonal velocity components and mixing within the same region. Two different flows which were generated by two models were studied: a generic nonaxisymmetric nozzle propulsion simulation model with an auxiliary internal water source that generated a jet flow and an axisymmetric forebody model with a circular sector strake that generated a vortex flow. The off-body flow fields around these models were investigated in the Langley 16- by 24-Inch Water Tunnel. The experimental results were used to calculate 17 quantities that included mean and fluctuating velocities, Reynolds stresses, mean and fluctuating dye fluorescence intensities (proportional to concentration), and fluctuating velocity and dye concentration correlations. An uncertainty analysis was performed to establish confidence levels in the experimental results. In general, uncertainties in mean velocities varied between 1 and 7 percent of free-stream velocity; uncertainties in fluctuating velocities varied between 1 and 5 percent of reference values. The results show characteristics that are unique to each type of flow.

  19. Moorable spectroradiometers in the BIOWATT experiment

    NASA Technical Reports Server (NTRS)

    Booth, C. R.; Smith, R. C.

    1988-01-01

    A new oceanographic instrument, the MER-2020, has been developed for the long term measurement of bio-optical properties. This instrument is a self-contained reflectance spectroradiometer with 40 megabytes of internal data storage and battery power permitting deployments for as long as 6 months. It can serve as the control and recording center for a variety of other sensors including fluorometers, transmissometers, temperature and conductivity probes. Instruments of this type were deployed at depths to 70 meters on a 5000 meter mooring during the BIOWATT experiment between February and November 1987. Measurements of the spectral diffuse attenuation coefficient and of the remote sensed reflectance from the ten month period in 1987 in the Northern Sargasso Sea are presented. In addition, the temperature, pressure, package orientation and sunlight stimulated natural fluorescence of phytoplankton are discussed. This instrument will be a key part of in situ optical calibration for future remote color sensors, and presently permits direct measurement of long term optical variability, important in visibility and communication problems.

  20. Parameters of photosynthetic energy partitioning.

    PubMed

    Lazár, Dušan

    2015-03-01

    Almost every laboratory dealing with plant physiology, photosynthesis research, remote sensing, and plant phenotyping possesses a fluorometer to measure a kind of chlorophyll (Chl) fluorescence induction (FLI). When the slow Chl FLI is measured with addition of saturating pulses and far-red illumination, the so-called quenching analysis followed by the so-called relaxation analysis in darkness can be realized. These measurements then serve for evaluation of the so-called energy partitioning, that is, calculation of quantum yields of photochemical and of different types of non-photochemical processes. Several theories have been suggested for photosynthetic energy partitioning. The current work aims to summarize all the existing theories, namely their equations for the quantum yields, their meaning and their assumptions. In the framework of these theories it is also found here that the well-known NPQ parameter ( [Formula: see text] ; Bilger and Björkman, 1990) equals the ratio of the quantum yield of regulatory light-induced non-photochemical quenching to the quantum yield of constitutive non-regulatory non-photochemical quenching (ΦNPQ/Φf,D). A similar relationship is also found here for the PQ parameter (ΦP/Φf,D). PMID:25569797

  1. Multi-Parametric Relationships between PAM Measurements and Carbon Incorporation, an In Situ Approach

    PubMed Central

    Napoléon, Camille; Claquin, Pascal

    2012-01-01

    Primary production (PP) in the English Channel was measured using 13C uptake and compared to the electron transport rate (ETR) measured using PAM (pulse amplitude modulated fluorometer). The relationship between carbon incorporation (Pobs) and ETR was not linear but logarithmic. This result can be explained by alternative electron sinks at high irradiance which protect the phytoplankton from photoinhibition. A multi-parametric model was developed to estimate PP by ETR. This approach highlighted the importance of taking physicochemical parameters like incident light and nutrient concentrations into account. The variation in the ETR/Pobs ratio as a function of the light revealed different trends which were characterized by three parameters (Rmax, the maximum value of ETR/Pobs; ERmax, the light intensity at which Rmax is measured; γ the initial slope of the curve). Based on the values of these three parameters, data were divided into six groups which were highly dependent on the seasons and on the physicochemical conditions. Using the multi-parametric model which we defined by Pobs and ETR measurements at low frequencies, the high frequency measurements of ETR enabled us to estimate the primary production capacity between November 2009 and December 2010 at high temporal and spatial scales. PMID:22911698

  2. Zeaxanthin and the Heat Dissipation of Excess Light Energy in Nerium oleander Exposed to a Combination of High Light and Water Stress 1

    PubMed Central

    Demmig, Barbara; Winter, Klaus; Krüger, Almuth; Czygan, Franz-Christian

    1988-01-01

    Upon termination of watering of plants of Nerium oleander exposed to high light, photochemical efficiency became reduced as leaf water content decreased. Evidence is presented that this type of photoinhibition reflects to a substantial degree radiationless dissipation of excitation energy, probably mediated by the carotenoid zeaxanthin. During the imposition of water stress, the zeaxanthin content of leaves increased at the expense of violaxanthin and β-carotene as a water deficit developed over a period of several days. The increase in zeaxanthin content was linearly related to an increase in the rate of radiationless energy dissipation in the antenna chlorophyll as calculated from the characteristics of chlorophyll a fluorescence measured with a pulse amplitude modulated fluorometer at room temperature. The increase in the rate of radiationless dissipation was also linearly related to a decrease in PSII photochemical efficiency as indicated by the ratio of variable to maximum fluorescence. Leaves of well-watered shade plants of N. oleander exposed to strong light showed a similar increase in zeaxanthin content as sun leaves of the same species subjected to drought in strong light. Shade leaves possessed the same capacity as sun leaves to form zeaxanthin at the expense of both violaxanthin and β-carotene. The resistance of this species to the destructive effects of excess light appears to be related to interconversions between β-carotene and the three carotenoids of the xanthophyll cycle. PMID:16666096

  3. Protocol Improvements for Low Concentration DNA-Based Bioaerosol Sampling and Analysis

    PubMed Central

    Ng, Chun Kiat; Miller, Dana; Cao, Bin

    2015-01-01

    Introduction As bioaerosol research attracts increasing attention, there is a need for additional efforts that focus on method development to deal with different environmental samples. Bioaerosol environmental samples typically have very low biomass concentrations in the air, which often leaves researchers with limited options in choosing the downstream analysis steps, especially when culture-independent methods are intended. Objectives This study investigates the impacts of three important factors that can influence the performance of culture-independent DNA-based analysis in dealing with bioaerosol environmental samples engaged in this study. The factors are: 1) enhanced high temperature sonication during DNA extraction; 2) effect of sampling duration on DNA recoverability; and 3) an alternative method for concentrating composite samples. In this study, DNA extracted from samples was analysed using the Qubit fluorometer (for direct total DNA measurement) and quantitative polymerase chain reaction (qPCR). Results and Findings The findings suggest that additional lysis from high temperature sonication is crucial: DNA yields from both high and low biomass samples increased up to 600% when the protocol included 30-min sonication at 65°C. Long air sampling duration on a filter media was shown to have a negative impact on DNA recoverability with up to 98% of DNA lost over a 20-h sampling period. Pooling DNA from separate samples during extraction was proven to be feasible with margins of error below 30%. PMID:26619279

  4. Cross-shore surfzone tracer dispersion in an alongshore current

    NASA Astrophysics Data System (ADS)

    Clark, David B.; Feddersen, Falk; Guza, R. T.

    2010-10-01

    Cross-shore surfzone tracer dispersion in a wave driven alongshore current is examined over a range of wave and current conditions with 6 continuous dye releases, each roughly 1-2 hours in duration, at Huntington Beach, California. Fluorescent dye tracer released near the shoreline formed shore parallel plumes that were sampled on repeated cross-shore transects with a jet ski mounted fluorometer. Ensemble averaged cross-shore tracer concentration profiles are generally shoreline attached (maximum at or near the shoreline), with increasing cross-shore widths and decreasing peak values with downstream distance. More than a few 100 m from the source, tracer is often well mixed across the surfzone (i.e., saturated) with decreasing tracer concentrations farther seaward. For each release, cross-shore surfzone absolute diffusivities are estimated using a simple Fickian diffusion solution with a no-flux boundary at the shoreline, and range from 0.5-2.5 m2 s-1. Surfzone diffusivity scalings based on cross-shore bore dispersion, surfzone eddy mixing length, and undertow driven shear dispersion are examined. The mixing-length scaling has correlation r2 = 0.59 and the expected best-fit slope <1, indicating that horizontal rotational motions are important for cross-shore tracer dispersion in the surfzone.

  5. Laser Induced Fluorescence For Measurement Of Lignin Concentrations In Pulping Liquors

    NASA Astrophysics Data System (ADS)

    Horvath, J. J.; Semerjian, H. G.; Biasca, K. L.; Attala, R.

    1988-11-01

    Laser excited fluorescence of pulping liquors was investigated for use in the pulp and paper industry for process measurement and control applications. Liquors from both mill and laboratory cooks were studied. A Nd-YAG pumped dye laser was used to generate the excitation wavelength of 280 nm; measurements were also performed using a commercially available fluorometer. Measurements on mill pulping liquors gave strong signals and showed changes in the fluorescence intensity during the cook. Absorption spectra of diluted mill liquor samples showed large changes during the cook. Samples from well controlled and characterized laboratory cooks showed fluorescence to be linear with concentration over two decades with an upper limit of approximately 1000 ppm dissolved lignin. At the end of these cooks a possible chemical change was indicated by an increase in the observed fluorescence intensity. Results indicate that lignin concentrations in pulping liquors can be accurately determined with fluorescence in the linear optical region over a greater dynamic range than absorption spectroscopy. Laser induced fluorescence may also provide an indication of chemical changes occurring in the lignin structure during a cook.

  6. Pulsed laser fluorometry for environmental monitoring

    NASA Astrophysics Data System (ADS)

    Saunders, G. C.; Martin, J. C.; Jett, J. H.; Wilder, M. E.; Martinez, A.; Bentley, B. F.; Lopez, J.; Hutson, L.

    A compact pulsed laser fluorometer has been incorporated into a continuous flow system developed to detect acetylcholinesterase (AChE) inhibitors and/or primary amine compounds in air and water. A pulsed nitrogen laser pumped dye laser excites fluorescent reactants which flow continuously through a quartz flow cell. Data are collected, analyzed, and displayed using a Macintosh II personal computer. For detection of cholinesterase inhibitors the fluorogenic substrate N methylindoxyl acetate is used to monitor the activity of immobilized enzyme. Presence of inhibitors results in a decrease of steady state fluorescence. Detection of compounds containing primary amines is based on their reaction with fluorescamine to rapidly produce intensely fluorescent products. Compounds of interest to our research were amino acids, peptides, and proteins. An increase in steady state fluorescence could be cause to evaluate the reasons for the change. The detection limit of the protein, bovine serum albumin (BSA) in water, is 10 ppT. Nebulized BSA concentrated by the LANL air sampler can be detected at sub ppT original air concentration.

  7. Practical determination of hyaluronan by a new noncompetitive fluorescence-based assay on serum of normal and cirrhotic patients.

    PubMed

    Martins, João R M; Passerotti, Carlo C; Maciel, Rui M B; Sampaio, Lucia O; Dietrich, Carl P; Nader, Helena B

    2003-08-01

    A practical fluorescence-based assay method for determination of hyaluronan (HA) was developed. Plates were coated with hyaluronan-binding proteins (HABP) obtained from bovine cartilage and successively incubated with samples containing standard solutions of hyaluronan or serum from normal and cyrrhotic patients, biotin-conjugated HABP, and europium-labeled streptavidin. After release of europium from streptavidin with enhancement solution the final fluorescence is measured in a fluorometer. The method is specific for HA even in the presence of substantial amounts of other glycosaminoglycans (chondroitin, dermatan sulfate, and heparan sulfate, and heparin) or proteins. It is possible to quantify HA between 0.2 and 500.0 microg/L. Analyses of HA concentration in 545 normal subjects and 40 cirrhotic patients gave average values of 14.5 and 542.0 microg/L, respectively. It was also shown that older subjects (> or =51 years old) have more HA (28.0 microg/L) than younger subjects (12.0 to 14.0 microg/L). This new sandwich technique has shown high precision and sensitivity similar to those of a recently described fluorescence-based assay method, being able to measure HA in amounts as small as 0.2 microg/L. In addition, this noncompetitive assay avoids preincubation, consumes less time (<5 h) than the previous competitive fluorescence-based assay (>72 h), and avoids the use of radioactive materials. PMID:12842108

  8. An improved europium release assay for complement-mediated cytolysis.

    PubMed

    Cui, J; Bystryn, J C

    1992-02-14

    An improved assay for complement-mediated cytolysis is described. The target cells are labeled with europium complexed to diethylenetriaminopentaacetate (Eu-DTPA). Cytolysis caused by antibody plus complement leads to the release of the Eu-DTPA complex which is then formed into a highly fluorescent chelate by the addition of 2-naphthoyltrifluoroacetone (2-NTA). The amount of europium chelate formed--a measurement of cell death--is then quantified with a time-resolved fluorometer. The results of the assay are reproducible. Complement-mediated cytolysis when measured by europium release was five times more sensitive than when measured by conventional 51Cr release and three times than when measured by trypan blue exclusion. Because europium does not decay, target cells can be labelled in batches and stored frozen until use, which speeds and simplifies the assay. Thus, europium release assay is a simple and quantitative method to measure complement-mediated cytolysis which is sensitive and more rapid than conventional assays. PMID:1541836

  9. Photo-physiological variability in phytoplankton chlorophyll fluorescence and assessment of chlorophyll concentration.

    PubMed

    Chekalyuk, Alexander; Hafez, Mark

    2011-11-01

    Photo-physiological variability of in vivo chlorophyll fluorescence (CF) per unit of chlorophyll concentration (CC) is analyzed using a biophysical model to improve the accuracy of CC assessments. Field measurements of CF and photosystem II (PSII) photochemical yield (PY) with the Advanced Laser Fluorometer (ALF) in the Delaware and Chesapeake Bays are analyzed vs. high-performance liquid chromatography (HPLC) CC retrievals. It is shown that isolation from ambient light, PSII saturating excitation, optimized phytoplankton exposure to excitation, and phytoplankton dark adaptation may provide accurate in vivo CC fluorescence measurements (R2 = 0.90-0.95 vs. HPLC retrievals). For in situ or flow-through measurements that do not allow for dark adaptation, concurrent PY measurements can be used to adjust for CF non-photochemical quenching (NPQ) and improve the accuracy of CC fluorescence assessments. Field evaluation has shown the NPQ-invariance of CF/PY and CF(PY-1-1) parameters and their high correlation with HPLC CC retrievals (R2 = 0.74-0.96), while the NPQ-affected CF measurements correlated poorly with CC (R2 = -0.22). PMID:22109145

  10. In-Situ Optical and Acoustical Measurements of the Buoyant Cyanobacterium P. Rubescens: Spatial and Temporal Distribution Patterns

    PubMed Central

    Hofmann, Hilmar; Peeters, Frank

    2013-01-01

    Optical (fluorescence) and acoustic in-situ techniques were tested in their ability to measure the spatial and temporal distribution of plankton in freshwater ecosystems with special emphasis on the harmful and buoyant cyanobacterium P. rubescens. Fluorescence was measured with the multi-spectral FluoroProbe (Moldaenke FluoroProbe, MFP) and a Seapoint Chlorophyll Fluorometer (SCF). In-situ measurements of the acoustic backscatter strength (ABS) were conducted with three different acoustic devices covering multiple acoustic frequencies (614 kHz ADCP, 2 MHz ADP, and 6 MHz ADV). The MFP provides a fast and reliable technique to measure fluorescence at different wavelengths in situ, which allows discriminating between P. rubescens and other phytoplankton species. All three acoustic devices are sensitive to P. rubescens even if other scatterers, e.g., zooplankton or suspended sediment, are present in the water column, because P. rubescens containing gas vesicles has a strong density difference and hence acoustic contrast to the ambient water and other scatterers. After calibration, the combination of optical and acoustical measurements not only allows qualitative and quantitative observation of P. rubescens, but also distinction between P. rubescens, other phytoplankton, and zooplankton. As the measuring devices can sample in situ at high rates they enable assessment of plankton distributions at high temporal (minutes) and spatial (decimeters) resolution or covering large temporal (seasonal) and spatial (basin scale) scales. PMID:24303028

  11. Homogeneous assay technology based on upconverting phosphors.

    PubMed

    Kuningas, Katri; Rantanen, Terhi; Ukonaho, Telle; Lövgren, Timo; Soukka, Tero

    2005-11-15

    Upconversion photoluminescence can eliminate problems associated with autofluorescence and scattered excitation light in homogeneous luminescence-based assays without need for temporal resolution. We have demonstrated a luminescence resonance energy-transfer-based assay utilizing inorganic upconverting (UPC) lanthanide phosphor as a donor and fluorescent protein as an acceptor. UPC phosphors are excited at near-infrared and they have narrow-banded anti-Stokes emission at visible wavelengths enabling measurement of the proximity-dependent sensitized emission with minimal background. The acceptor alone does not generate any direct emission at shorter wavelengths under near-infrared excitation. A competitive model assay for biotin was constructed using streptavidin-conjugated Er3+,Yb3+-doped UPC phosphor as a donor and biotinylated phycobiliprotein as an acceptor. UPC phosphor was excited at near-infrared (980 nm) and sensitized acceptor emission was measured at red wavelength (600 nm) by using a microtitration plate fluorometer equipped with an infrared laser diode and suitable excitation and emission filters. Lower limit of detection was in the subnanomolar concentration range. Compared to time-resolved fluorometry, the developed assay technology enabled simplified instrumentation. Excitation at near-infrared and emission at red wavelengths render the technology also suitable to analysis of strongly colored and fluorescent samples, which are often of concern in clinical immunoassays and in high-throughput screening. PMID:16285685

  12. Real-time imaging of hydrogen peroxide dynamics in vegetative and pathogenic hyphae of Fusarium graminearum

    PubMed Central

    Mentges, Michael; Bormann, Jörg

    2015-01-01

    Balanced dynamics of reactive oxygen species in the phytopathogenic fungus Fusarium graminearum play key roles for development and infection. To monitor those dynamics, ratiometric analysis using the novel hydrogen peroxide (H2O2) sensitive fluorescent indicator protein HyPer-2 was established for the first time in phytopathogenic fungi. H2O2 changes the excitation spectrum of HyPer-2 with an excitation maximum at 405 nm for the reduced and 488 nm for the oxidized state, facilitating ratiometric readouts with maximum emission at 516 nm. HyPer-2 analyses were performed using a microtiter fluorometer and confocal laser scanning microscopy (CLSM). Addition of external H2O2 to mycelia caused a steep and transient increase in fluorescence excited at 488 nm. This can be reversed by the addition of the reducing agent dithiothreitol. HyPer-2 in F. graminearum is highly sensitive and specific to H2O2 even in tiny amounts. Hyperosmotic treatment elicited a transient internal H2O2 burst. Hence, HyPer-2 is suitable to monitor the intracellular redox balance. Using CLSM, developmental processes like nuclear division, tip growth, septation, and infection structure development were analyzed. The latter two processes imply marked accumulations of intracellular H2O2. Taken together, HyPer-2 is a valuable and reliable tool for the analysis of environmental conditions, cellular development, and pathogenicity. PMID:26446493

  13. Liquid-chromatographic separation and on-line bioluminescence detection of creatine kinase isoenzymes

    SciTech Connect

    Bostick, W.D.; Denton, M.S.; Dinsmore, S.R.

    1980-01-01

    Isoenzymes of creatine kinase were separated by anion-exchange chromatography, with use of an elution gradient containing lithium acetate (0.1 to 0.6 mol/L). A stream splitter was used to divert a 5% side stream of column effluent, which was subsequently mixed with the reagents necessary for bioluminescence assay of the separated isoenzymes. The use of the stream splitter greatly decreased the rate of consumption of reagent and, when combined with a peristaltic pumping system, permitted independent control of the side-stream flow rate. Thus both the residence interval in a delay coil in which the ATP reaction product is formed and the bioluminescence emission was monitored in a flow-through fluorometer without use of an external light source or filters. Separation and detection of the isoenzymes of creatine kinase were rapid, sensitive, and highly selective. The incremental decrease of bioluminescence response owing to inhibition by the ions in the eluent was less than 31% across the entire gradient.

  14. Oil Droplet Size Distribution and Optical Properties During Wave Tank Simulated Oil Spills

    NASA Astrophysics Data System (ADS)

    Conmy, R. N.; Venosa, A.; Courtenay, S.; King, T.; Robinson, B.; Ryan, S.

    2013-12-01

    Fate and transport of spilled petroleum oils in aquatic environments is highly dependent upon oil droplet behavior which is a function of chemical composition, dispersibility (natural and chemically-enhanced) and droplet size distribution (DSD) of the oil. DSD is influenced by mixing energy, temperature, salinity, pressure, presence of dissolved and particulate materials, flow rate of release, and application of dispersants. To better understand DSD and droplet behavior under varying physical conditions, flask-scale experiments are often insufficient. Rather, wave tank simulations allow for scaling to field conditions. Presented here are experiment results from the Bedford Institute of Oceanography wave tank facility, where chemically-dispersed (Corexit 9500; DOR = 1:20) Louisiana Sweet crude, IFO-120 and ANS crude oil were exposed to mixing energies to achieve dispersant effectiveness observed in the field. Oil plumes were simulated, both surface and subsea releases with varying water temperature and flow rate. Fluorometers (Chelsea Technologies Group AQUATracka, Turner Designs Cyclops, WET Labs Inc ECO) and particle size analyzers (Sequoia LISST) were used to track the dispersed plumes in the tank and characterize oil droplets. Sensors were validated with known oil volumes (down to 300 ppb) and measured Total Petroleum Hydrocarbons (TPH) and Benzene-Toluene-Ethylbenzene-Xylene (BTEX) values. This work has large implications for tracking surface and deep sea oil plumes with fluorescence and particle size analyzers, improved weathering and biodegradation estimates, and understanding the fate and transport of spill oil.

  15. Synthesis, structure and photoluminescence properties of Ho3+ doped TTB-BaTa2O6 phosphors

    NASA Astrophysics Data System (ADS)

    İlhan, Mustafa

    2014-12-01

    Ho3+ doped TTB-BaTa2O6 phosphors were produced by the solid state reaction method. XRD analysis confirmed the formation of BaTa2O6 single phase which was accomplished by heat treatment at 1425 °C for 20 h. The crystal structure of TTB-BaTa2O6 allowed doping concentration of Ho3+ ions up to 10 mol%, maintaining a single phase composition. A second phase of HoTaO4 begins to appear at 15 mol%. The lattice structure and the crystallite sizes changed with the concentration of Ho3+. In SEM analysis, it was also shown that BaTa2O6 grain sizes changed with the concentration of Ho3+. EDS analysis revealed that the Ta/Ba ratio increased on the grains depending on Ho3+ concentration. Charge balance of the structure was formulated through the EDS results. In fluorometer analysis, a strong green emission (λem = 546.9 nm) was observed in the visible spectral region. The emission increased with the doping concentration of up to 2.5 mol%, and above this level decreased due to concentration quenching.

  16. Multiemission wavelength picosecond time-resolved fluorescence decay data obtained on the millisecond time scale: application to protein:DNA interactions and protein-folding reactions

    NASA Astrophysics Data System (ADS)

    Beechem, Joseph M.

    1992-04-01

    One of the major aspects of fluorescence spectroscopy which differentiates this technique from many other spectroscopic approaches is the inherent multidimensional nature of the data. For instance, the basic pulsed-laser fluorescence data set is characterized by fluorescence versus: emission wavelength, polarization state (parallel and perpendicular intensities), time of emission (picoseconds to nanoseconds), and time of biological reaction (milliseconds to minutes). Usually, this six-dimensional data set is obtained piecemeal, single dimension at a time; often complete data sets are not even collected. This is especially true of the biological time scale axis. Data acquisition times for picosecond decay data are typically seconds to minutes, and, therefore, it has not been generally possible to perform this experiment in a kinetic mode. What is described in this report is the construction of a parallel multichannel time-correlated single-photon counting (TCSPC) fluorometer which is capable of simultaneous collection of: fluorescence vs. picosecond to nanosecond time vs. emission wavelength vs. polarization state vs. millisecond to second time. Use is made of two multi-anode microchannel plate detectors, each obtaining data at two different polarization states, six different emission wavelengths, along 12 independent TCSPC channels. This instrument is interfaced to a three-syringe stepper motor controlled stop-flow apparatus, and picosecond decay data along all of these channels is stored and collected by two 33 MHz 80486 computers at rates approaching 1200 - 12000 data sets per second.

  17. Micro-segmented flow and multisensor-technology for microbial activity profiling.

    PubMed

    Kürsten, Dana; Kothe, Erika; Wetzel, Katharina; Bergmann, Katja; Köhler, J Michael

    2014-01-01

    The combination of micro-segmented flow with miniaturized flow-through multisensor-technology has been utilized for metabolite profiling of soil bacteria. Series of sub-μl segments were generated containing soil sample slurry from historic copper mining sites and exposed to heavy metal salts of copper and nickel. Segments were examined for bacterial growth and spectral properties as well as for the effect of heavy metal-treatment after different incubation times. In order to evaluate microbial growth, extinction was recorded with 4 different spectral channels. Fluorescence was measured using a microflow-through fluorometer to detect both growth and production of fluorescent dyes or metabolites. The incidence of single segments with enhanced absorption in one of the spectral channels or enhanced fluorescence was scored to detect soil microorganisms with interesting properties for further screening. The study could show that the number of vegetated segments, the density of microorganisms in the segments after cultivation and the spectral response are different for separate soil samples and different metals. Thus, the highly parallelized and miniaturized segmented flow method is a promising tool for profiling of soil samples with regard to identifying micro-organisms with interesting profiles for secondary metabolite-production. PMID:25119668

  18. Vertical variation of mixing within porous sediment beds below turbulent flows

    PubMed Central

    Chandler, I. D.; Pearson, J. M.; van Egmond, R.

    2016-01-01

    Abstract River ecosystems are influenced by contaminants in the water column, in the pore water and adsorbed to sediment particles. When exchange across the sediment‐water interface (hyporheic exchange) is included in modeling, the mixing coefficient is often assumed to be constant with depth below the interface. Novel fiber‐optic fluorometers have been developed and combined with a modified EROSIMESS system to quantify the vertical variation in mixing coefficient with depth below the sediment‐water interface. The study considered a range of particle diameters and bed shear velocities, with the permeability Péclet number, PeK between 1000 and 77,000 and the shear Reynolds number, Re*, between 5 and 600. Different parameterization of both an interface exchange coefficient and a spatially variable in‐sediment mixing coefficient are explored. The variation of in‐sediment mixing is described by an exponential function applicable over the full range of parameter combinations tested. The empirical relationship enables estimates of the depth to which concentrations of pollutants will penetrate into the bed sediment, allowing the region where exchange will occur faster than molecular diffusion to be determined.

  19. Spring bloom onset in the Nordic Seas

    NASA Astrophysics Data System (ADS)

    Mignot, Alexandre; Ferrari, Raffaele; Mork, Kjell Arne

    2016-06-01

    The North Atlantic spring bloom is a massive annual growth event of marine phytoplankton, tiny free-floating algae that form the base of the ocean's food web and generates a large fraction of the global primary production of organic matter. The conditions that trigger the onset of the spring bloom in the Nordic Seas, at the northern edge of the North Atlantic, are studied using in situ data from six bio-optical floats released north of the Arctic Circle. It is often assumed that spring blooms start as soon as phytoplankton cells daily irradiance is sufficiently abundant that division rates exceed losses. The bio-optical float data instead suggest the tantalizing hypothesis that Nordic Seas blooms start when the photoperiod, the number of daily light hours experienced by phytoplankton, exceeds a critical value, independently of division rates. The photoperiod trigger may have developed at high latitudes where photosynthesis is impossible during polar nights and phytoplankton enters into a dormant stage in winter. While the first accumulation of biomass recorded by the bio-optical floats is consistent with the photoperiod hypothesis, it is possible that some biomass accumulation started before the critical photoperiod but at levels too low to be detected by the fluorometers. More precise observations are needed to test the photoperiod hypothesis.

  20. Impact of emission anisotropy on fluorescence spectroscopy and FRET distance measurements.

    PubMed

    Ivanov, Vassili; Li, Min; Mizuuchi, Kiyoshi

    2009-08-01

    The objective of this report is to provide a practical and improved method for estimating Förster resonance energy transfer distance measurement error due to unknown angles in the dipole orientation factor based on emission anisotropy measurements. We improve on the method of Dale et al. (1979), which has minor mistakes and is frequently interpreted in overly optimistic ways in the literature. To facilitate proper fluorescence intensity measurements, we also evaluated instrument parameters that could impact the measurement. The apparent fluorescence intensity of isotropic samples depends on the sample emission anisotropy, fluorometer geometry, and optical apertures. We separate parameters of the sample, and those of the cylindrically symmetric illumination source and detector in the equations describing results of unpolarized and polarized fluorescence intensity measurements. This approach greatly simplifies calculations compared with the more universal method of Axelrod (1989). We provide a full computational method for calculating the Förster resonance energy transfer distance error and present a graph describing distance error in the simplest case. PMID:19651051

  1. Light-induced dynamic changes of NADPH fluorescence in Synechocystis PCC 6803 and its ndhB-defective mutant M55.

    PubMed

    Mi, H; Klughammer, C; Schreiber, U

    2000-10-01

    Blue-green fluorescence emission of intact cells of Synechocystis PCC6803 and of its ndhB-defective mutant M55 was measured with a standard pulse-amplitude-modulation chlorophyll fluorometer equipped with a new type of emitter-detector unit featuring pulse-modulated UV-A measuring light and a photomultiplier detector. A special illumination program of repetitive saturating light pulses with intermittent dark periods (10 s light, 40 s dark) was applied to elicit dynamic fluorescence changes under conditions of quasi-stationary illumination. The observed effects of artificial electron acceptors and inhibitors on the responses of wild-type and mutant M55 cells lead to the conclusion that changes of NAD(P)H fluorescence are measured. In control samples, a rapid phase of light-driven NADP reduction is overlapped by a somewhat slower phase of NADPH oxidation which is suppressed by iodoacetic acid and, hence, appears to reflect NADPH oxidation by the Calvin cycle. Mercury chloride transforms the light-driven positive response into a negative one, suggesting that inhibition of NADP reduction at the acceptor side of PSI leads to reduction of molecular oxygen, with the hydrogen peroxide formed (via superoxide) causing rapid oxidation of NADPH. The new fluorescence approach opens the way for new insights into the complex interactions between photosynthetic and respiratory pathways in cyanobacteria. PMID:11148271

  2. Molecular diagnostics in a teacup: Non-Instrumented Nucleic Acid Amplification (NINA) for rapid, low cost detection of Salmonella enterica

    PubMed Central

    KUBOTA, Ryo; LABARRE, Paul; WEIGL, Bernhard H; LI, Yong; HAYDOCK, Paul; JENKINS, Daniel M

    2014-01-01

    We report on the use of a novel non-instrumented platform to enable a Loop Mediated isothermal Amplification (LAMP) based assay for Salmonella enterica. Heat energy is provided by addition of a small amount (<150 g) of boiling water, and the reaction temperature is regulated by storing latent energy at the melting temperature of a lipid-based engineered phase change material. Endpoint classification of the reaction is achieved without opening the reaction tube by observing the fluorescence of sequence-specific FRET-based assimilating probes with a simple handheld fluorometer. At or above 22°C ambient temperature the non-instrumented devices could maintain reactions above a threshold temperature of 61°C for over 90 min—significantly longer than the 60 min reaction time. Using the simple format, detection limits were less than 20 genome copies for reactions run at ambient temperatures ranging from 8 to 36°C. When used with a pre-enrichment step and non-instrumented DNA extraction device, trace contaminations of Salmonella in milk close to 1 CFU/mL could be reliably detected. These findings illustrate that the non- instrumented amplification approach is a simple, viable, low-cost alternative for field-based food and agricultural diagnostics or clinical applications in developing countries. PMID:25477717

  3. Time of travel and dispersion of solutes in a 36.4-mile reach of the North Platte River downstream from Casper, Wyoming

    USGS Publications Warehouse

    Armentrout, G.W., Jr.; Larson, L.R.

    1984-01-01

    Time-of-travel and dispersion measurements made during a dye study November 7-8, 1978, are presented for a reach of the North Platte River from Casper, Wyo., to a bridge 2 miles downstream from below the Dave Johnston Power Plant. Rhodamine WT dye was injected into the river at Casper, and the resultant dye cloud was traced by sampling as it moved downstream. Samples were taken in three equal-flow sections of the river 's lateral transect at three sites, then analyzed in a fluorometer. The flow in the river was 940 cubic feet per second. The data consist of measured stream mileages and time, distance, and concentration graphs of the dye cloud. The peak concentration traveled through the reach in 24 hours, averaging 1.5 miles per hour; the leading edge took about 22 hours, averaging 1.7 miles per hour; and the trailing edge took 35 hours, averaging 1.0 mile per hour. Data from this study were compared with methods for estimating time of travel for a range of stream discharges.

  4. Time-of-travel and dispersion studies, Lehigh River, Francis E. Walter Lake to Easton, Pennsylvania

    USGS Publications Warehouse

    Kauffman, C.D.

    1983-01-01

    Results of time-of-travel and dispersion studies are presented for the 77.0 mile reach of the Lehigh River from Francis E. Walter Lake to Easton, Pennsylvania. Rhodamine WT dye was injected at several points for a variety of several common flow conditions and its downstream travel was monitored at a number of downstream points by means of a fluorometer. Time-of-travel data have been related to stream discharge, distance along the river channel and dispersion. If 2.205 pounds of a conservative water soluble contaminant were accidentally spilled into the Lehigh River at Penn Haven Junction at Black Creek 6.09 miles downstream from Rockport, Pennsylvania, when the discharge at Walnutport, Pennsylvania, was 600 cubic feet per second, the leading edge, peak, and trailing edge of the contaminant would arrive 31.6 miles downstream at the Northhampton, Pennsylvania, water intakes 45, 54, and 66 hours later, respectively. The maximum concentration expected at the intakes would be about 1.450 micrograms per liter. From data and relations presented, time-of-travel and maximum concentration estimates can be made for any two points within the reach. (USGS)

  5. Development of Loop-Mediated Isothermal Amplification (LAMP) Assay for Rapid and Sensitive Identification of Ostrich Meat

    PubMed Central

    Abdulmawjood, Amir; Grabowski, Nils; Fohler, Svenja; Kittler, Sophie; Nagengast, Helga; Klein, Guenter

    2014-01-01

    Animal species identification is one of the primary duties of official food control. Since ostrich meat is difficult to be differentiated macroscopically from beef, therefore new analytical methods are needed. To enforce labeling regulations for the authentication of ostrich meat, it might be of importance to develop and evaluate a rapid and reliable assay. In the present study, a loop-mediated isothermal amplification (LAMP) assay based on the cytochrome b gene of the mitochondrial DNA of the species Struthio camelus was developed. The LAMP assay was used in combination with a real-time fluorometer. The developed system allowed the detection of 0.01% ostrich meat products. In parallel, a direct swab method without nucleic acid extraction using the HYPLEX LPTV buffer was also evaluated. This rapid processing method allowed detection of ostrich meat without major incubation steps. In summary, the LAMP assay had excellent sensitivity and specificity for detecting ostrich meat and could provide a sampling-to-result identification-time of 15 to 20 minutes. PMID:24963709

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

    NASA Astrophysics Data System (ADS)

    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. Fluorogenic Cell-Based Biosensors for Monitoring Microbes

    NASA Technical Reports Server (NTRS)

    Curtis, Theresa; Salazar, Noe; Tabb, Joel; Chase, Chris

    2010-01-01

    Fluorogenic cell-based sensor systems for detecting microbes (especially pathogenic ones) and some toxins and allergens are undergoing development. These systems harness the natural signaltransduction and amplification cascades that occur in mast cells upon activation with antigens. These systems include (1) fluidic biochips for automated containment of samples, reagents, and wastes and (2) sensitive, compact fluorometers for monitoring the fluorescent responses of mast cells engineered to contain fluorescent dyes. It should be possible to observe responses within minutes of adding immune complexes. The systems have been shown to work when utilizing either immunoglobulin E (IgE) antibodies or traditionally generated rat antibodies - a promising result in that it indicates that the systems could be developed to detect many target microbes. Chimeric IgE antibodies and rat immunoglobulin G (IgG) antibodies could be genetically engineered for recognizing biological and chemical warfare agents and airborne and food-borne allergens. Genetic engineering efforts thus far have yielded (1) CD14 chimeric antibodies that recognize both Grampositive and Gram-negative bacteria and bind to the surfaces of mast cells, eliciting a degranulation response and (2) rat IgG2a antibodies that act similarly in response to low levels of canine parvovirus.

  8. Bottom boundary layer measurements in OMP. Final report

    SciTech Connect

    Gross, T.F.; Williams, A.J.

    1998-11-01

    The main role of the Benthic Acoustic Stress Sensor (BASS) tripods within the Ocean Margins Program experiments was to detect and quantify organic carbon rich particle transport off the shelf. This requires measures of the turbulent boundary layer flow and bed stress, the physical forcing of the particle transport, as well as the concentration and type of particles which are being transported. The BASS tripods were deployed at sites 17 and 26. Data from site 26 were recovered spanning three periods: Feb. 2--April 6, May 13--June 27, June 28--Aug. 18. Site 17 was occupied Feb. 12--april 11. The BASS tripods were arrayed with five BASS sensors measuring detailed velocity parameters within four meters of the seabed. Velocity time series indicate a usually weak tidal flow which produces small bed stress by itself. On the occasions when a strong flow, probably the Gulf Stream, crosses the area, the bed shear stress increases dramatically to as much as 10 dyne cm{sup {minus}2}. This is competent to move unconsolidated sediments in the area. Other instruments from the tripods include: two conductivity/temperature sensor pairs, five WetStar fluorometers, thermistors, transmissometer, optical backscatterence sensors and a pressure sensor.

  9. Production of volatile organic compounds by cyanobacteria Synechococcus sp.

    NASA Astrophysics Data System (ADS)

    Hiraiwa, M.; Abe, M.; Hashimoto, S.

    2014-12-01

    Phytoplankton are known to produce volatile organic compounds (VOCs), which contribute to environmental problems such as global warming and decomposition of stratospheric ozone. For example, picophytoplankton, such as Prochlorococcus and Synechococcus, are distributed in freshwater and oceans worldwide, accounting for a large proportion of biomass and primary production in the open ocean. However, to date, little is known about the production of VOCs by picophytoplankton. In this study, VOCs production by cyanobacteria Synechococcus sp. (NIES-981) was investigated. Synechococcus sp. was obtained from the National Institute for Environmental Studies (NIES), Japan, and cultured at 24°C in autoclaved f/2-Si medium under 54 ± 3 µE m-2 s-1 (1 E = 1 mol of photons) with a 12-h light and 12-h dark cycle. VOCs concentrations were determined using a purge-and-trap gas chromatograph-mass spectrometer (Agilent 5973). The concentrations of chlorophyll a (Chl a) were also determined using a fluorometer (Turner TD-700). Bromomethane (CH3Br) and isoprene were produced by Synechococcus sp. Isoprene production was similar to those of other phytoplankton species reported earlier. Isoprene was produced when Chl a was increasing in the early stage of the incubation period (5-15 days of incubation time, exponential phase), but CH3Br was produced when Chl a was reduced in the late stage of the incubation period (30-40 days of incubation time, death phase).

  10. Autofluorescence lifetime metrology for label-free detection of cartilage matrix degradation

    NASA Astrophysics Data System (ADS)

    Nickdel, Mohammad B.; Lagarto, João. L.; Kelly, Douglas J.; Manning, Hugh B.; Yamamoto, Kazuhiro; Talbot, Clifford B.; Dunsby, Christopher; French, Paul; Itoh, Yoshifumi

    2014-03-01

    Degradation of articular cartilage extracellular matrix (ECM) by proteolytic enzyme is the hallmark of arthritis that leads to joint destruction. Detection of early biochemical changes in cartilage before irreversible structural damages become apparent is highly desirable. Here we report that the autofluorescence decay profile of cartilage is significantly affected by proteolytic degradation of cartilage ECM and can be characterised by measurements of the autofluorescence lifetime (AFL). A multidimensional fluorometer utilizing ultraviolet excitation at 355 nm or 375 nm coupled to a fibreoptic probe was developed for single point time-resolved AFL measurements of porcine articular cartilage explants treated with different proteinases. Degradation of cartilage matrix components by treating with bacterial collagenase, matrix metalloproteinase 1, or trypsin resulted in significant reduction of AFL of the cartilage in both a dose and time dependent manner. Differences in cartilage AFL were also confirmed by fluorescence lifetime imaging microscopy (FLIM). Our data suggest that AFL of cartilage tissue is a potential non-invasive readout to monitor cartilage matrix integrity that may be utilized for diagnosis of arthritis as well as monitoring the efficacy of anti-arthritic therapeutic agents.

  11. Pulsed laser fluorometry for environmental monitoring

    SciTech Connect

    Saunders, G. C.; Martin, J. C.; Jett, J. H.; Wilder, M. E.; Martinez, A.; Bentley, B. F.; Lopez, J.; Hutson, L.

    1990-01-01

    A compact pulsed laser fluorometer has been incorporated into a continuous flow system developed to detect acetylcholinesterase (AChE) inhibitors and/or primary amine compounds in air and water. A pulsed nitrogen laser pumped dye laser excites fluorescent reactants which flow continuously through a quartz flow cell. Data are collected, analyzed, and displayed using a Macintosh II personal computer. For detection of cholinesterase inhibitors the fluorogenic substrate N methylindoxyl acetate is used to monitor the activity of immobilized enzyme. Presence of inhibitors results in a decrease of steady state fluorescence. Detection of compounds containing primary amines is based on their reaction with fluorescamine to rapidly produce intensely fluorescent products. Compounds of interest to our research were amino acids, peptides, and proteins. An increase in steady state fluorescence could be cause to evaluate the reasons for the change. The detection limit of the protein, bovine serum albumin (BSA) in water is 10 ppT. Nebulized BSA concentrated by the LANL air sampler can be detected at sub ppT original air concentration. 16 refs., 14 figs., 3 tabs.

  12. Arsenic toxicity in the water weed Wolffia arrhiza measured using Pulse Amplitude Modulation Fluorometry (PAM) measurements of photosynthesis.

    PubMed

    Ritchie, Raymond J; Mekjinda, Nutsara

    2016-10-01

    Accumulation of arsenic in plants is a serious South-east Asian environmental problem. Photosynthesis in the small aquatic angiosperm Wolffia arrhiza is very sensitive to arsenic toxicity, particularly in water below pH 7 where arsenite (As (OH)3) (AsIII) is the dominant form; at pH >7 AsO4(2-) (As(V) predominates). A blue-diode PAM (Pulse Amplitude Fluorometer) machine was used to monitor photosynthesis in Wolffia. Maximum gross photosynthesis (Pgmax) and not maximum yield (Ymax) is the most reliable indicator of arsenic toxicity. The toxicity of arsenite As(III) and arsenate (H2AsO4(2-)) As(V) vary with pH. As(V) was less toxic than As(III) at both pH 5 and pH 8 but both forms of arsenic were toxic (>90% inhibition) at below 0.1molm(-3) when incubated in arsenic for 24h. Arsenite toxicity was apparent after 1h based on Pgmax and gradually increased over 7h but there was no apparent effect on Ymax or photosynthetic efficiency (α0). PMID:27318559

  13. In-Field Implementation of a Recombinant Factor C Assay for the Detection of Lipopolysaccharide as a Biomarker of Extant Life within Glacial Environments.

    PubMed

    Barnett, Megan J; Wadham, Jemma L; Jackson, Miriam; Cullen, David C

    2012-01-01

    The discovery over the past two decades of viable microbial communities within glaciers has promoted interest in the role of glaciers and ice sheets (the cryosphere) as contributors to subglacial erosion, global biodiversity, and in regulating global biogeochemical cycles. In situ or in-field detection and characterisation of microbial communities is becoming recognised as an important approach to improve our understanding of such communities. Within this context we demonstrate, for the first time, the ability to detect Gram-negative bacteria in glacial field-environments (including subglacial environments) via the detection of lipopolysaccharide (LPS); an important component of Gram-negative bacterial cell walls. In-field measurements were performed using the recently commercialised PyroGene® recombinant Factor C (rFC) endotoxin detection system and used in conjunction with a handheld fluorometer to measure the fluorescent endpoint of the assay. Twenty-seven glacial samples were collected from the surface, bed and terminus of a low-biomass Arctic valley glacier (Engabreen, Northern Norway), and were analysed in a field laboratory using the rFC assay. Sixteen of these samples returned positive LPS detection. This work demonstrates that LPS detection via rFC assay is a viable in-field method and is expected to be a useful proxy for microbial cell concentrations in low biomass environments. PMID:25585634

  14. Determination of lithium in rocks: Fluorometric method

    USGS Publications Warehouse

    White, C.E.; Fletcher, M.H.; Parks, J.

    1951-01-01

    The gravimetric method in general use for the determination of lithium is tedious, and the final weighed product often contains other alkali metals. A fluorometric method was developed to shorten the time required for the analysis and to assure that the final determination is for lithium alone. This procedure is based on the complex formed between lithium and 8-hydroxyquinoline. The fluorescence is developed in a slightly alkaline solution of 95% alcohol and measurement is made on a photoelectric fluorometer. Separation from the ore is carried out by the wet method or by the distillation procedure. Sodium and potassium are removed by alcohol and ether, but complete separation is not necessary. Comparison of analyzed samples shows excellent agreement with spectrographic and gravimetric methods. The fluorometric method is more rapid than the gravimetric and produces more conclusive results. Another useful application is in the preparation of standard lithium solutions from reagent quality salts when a known standard is available. In this case no separations are necessary.

  15. Examining concentrations and molecular weights of thiols in microorganism cultures and in Churchill River (Manitoba) using a fluorescent-labeling method coupled to asymmetrical flow field-flow fractionation.

    PubMed

    Mangal, Vaughn; Guéguen, Celine

    2015-06-01

    In this study, molecular weights of thiols from four laboratory cultures (Scenedesmus obliquus, Chlorella vulgaris, Euglena gracilis, and Attheya septentrionalis) and the Churchill River (Manitoba) were assessed using a fluorescent-labeling method such as monobromotrimethylammoniobimane (qBBr) and asymmetrical flow field-flow fractionation (AF4) coupled to a fluorescence detector. Concentrations of thiols in extracellular fractions ranged from 6.39 ± 3.39 to 39.2 ± 7.43 μmol g(-1), and intracellular concentrations ranged from 11.5 ± 4.52 to 41.0 ± 4.1 μmol g(-1). In addition, molecular weights (MW) of intracellular thiol ranged from 493 ± 24 to 946 ± 12 Da whereas extracellular thiol MWs varied from 443 ± 36 to 810 ± 174 Da. The novel method of combining AF4 to an on-line fluorometer and the incorporation of the thiol tag provided information regarding thiol concentration and composition of controlled and natural systems. Furthermore, the proposed methods allow for the simultaneous measurement of thiol and DOM MWs produced by microorganisms. By assessing characteristics of naturally produced thiols and lab-grown thiols, information regarding heavy metal complexation can be determined. PMID:25772566

  16. A Practical Solution for 77 K Fluorescence Measurements Based on LED Excitation and CCD Array Detector

    PubMed Central

    Lamb, Jacob; Forfang, Kristin; Hohmann-Marriott, Martin

    2015-01-01

    The fluorescence emission spectrum of photosynthetic microorganisms at liquid nitrogen temperature (77 K) provides important insights into the organization of the photosynthetic machinery of bacteria and eukaryotes, which cannot be observed at room temperature. Conventionally, to obtain such spectra, a large and costly table-top fluorometer is required. Recently portable, reliable, and largely maintenance-free instruments have become available that can be utilized to accomplish a wide variety of spectroscopy-based measurements in photosynthesis research. In this report, we show how to build such an instrument in order to record 77K fluorescence spectra. This instrument consists of a low power monochromatic light-emitting diode (LED), and a portable CCD array based spectrometer. The optical components are coupled together using a fiber optic cable, and a custom made housing that also supports a dewar flask. We demonstrate that this instrument facilitates the reliable determination of chlorophyll fluorescence emission spectra for the cyanobacterium Synechocystis sp. PCC 6803, and the green alga Chlamydomonas reinhardtii. PMID:26177548

  17. A new class of nontoxic nanoparticle tags based on surface enhanced Raman scattering

    NASA Astrophysics Data System (ADS)

    Qian, X.-M.; Ansari, D.; Nie, Shuming

    2007-02-01

    The advance of nanotechnology has boosted the development of ultra-sensitive biosensors for biomedical applications. Most recently, optical detection based biosensors have been demonstrated in medical imaging and diagnosis employing nanocrystals such as fluorescent quantum dots (QDs) and plasmon resonant metal nanoparticles to achieve femto-molar detection. An intriguing but far less explored approach for biological diagnostics relies on an emerging ultrasensitive technology -- surface enhanced Raman scattering (SERS) spectroscopy. We have developed a stable SERS nano-tag by grafting hydrophilic polymer to gold nanoparticle-dye molecule complexes to preserve the spectral signature and fully control the aggregation states. The light-emitting power and scattered light of both QDs and SERS nano-tags have been recorded under the same experimental conditions using dark field microscope, fluorometer, and Raman instrument. A comparison in brightness, sensitivity level, and quantum efficiency between SERS nano-tags and near infrared (NIR) QDs has been assessed on both bulk colloidal solution and single particle measurements. Well-designed SERS nano-tags exhibit excellent advantages over NIR QDs.

  18. Primary production off Southern California relative to surface layer carbon budgets: A component of the California Basins Study, CaBS. Final report, [1 June 1989--14 November 1991

    SciTech Connect

    Trees, C.C.

    1994-04-22

    This study started on 1 June 1989 and ended 14 November 1991. Two moored in situ natural fluorometers were deployed in January 1990 to collect bio-optical data for one year, making ground truth measurements around the mooring during 4 cruises. This one-year time series would investigate how the short-term physical forcing aliases the long-term primary production record such that the apparent, larger interannual variability in the record is in reality ``noise`` due to short-term fluctuations in the rate of nutrient input to the euphotic zone. These continuous measurements from moored bio-optical instruments would also allow better estimates of the mean and variance in primary production in these waters than has previously been available from shipboard measurements, as well as, phytoplankton response to short-term physical events. Ancillary measurements that were made were: (1) characterization of the apparent and inherent optical properties, (2) photosynthetic pigment distributions using both HPLC and standard fluorometric methods, (3) carbon, hydrogen and nitrogen content of suspended particulate matter, (4) primary production using conventional {sup 14}C methods from simulated in situ experiments.

  19. Early lung cancer: detection, treatment outcome

    SciTech Connect

    Balchum, O.J.; Huth, G.C.; Saccomanno, G.

    1984-01-01

    The performance of a room temperature mercuric iodide x-ray detector was investigated as a function of detector bias, amplifier time constant, and detector temperature. A Mn K/sub ..cap alpha../ line of 200 eV FWHM was obtained by using low noise electronics developed for Si(Li) detectors, including a cooled input FET. Measurements of the detector's resolution at various x-ray energies result in a Fano factor of 0.20. Fluorescence bronchoscopy with a violet laser and image intensifier has been developed for imaging the red fluorescence of a tumor-specific agent, hematoporphyrin derivative, that has been injected before the examination. The instrument was developed to localize carcinoma in situ and early, small bronchogenic tumors diagnosed by sputum cytology but invisible on chest x-ray and conventional bronchoscopy, in underground uranium miners and others at risk for lung cancer. In addition to the imaging devices, a video system including a processor and electronics for digital background image subtraction has been developed to enhance contrast. A ratio fluorometer and a rapid-scan spectrum analyzer have been designed for quantitative measurements of fluorescence intensity and dependence on dosage and time after injection of the fluorescent agent. Clinical trials demonstrate detection of carcinoma in situ, and the true positive rate should be improved by the new instrumentation and optimization of time delay and dosage.

  20. Production of terpenes in the culture of Chlorophyceae and Rhodophyta

    NASA Astrophysics Data System (ADS)

    Abe, M.; Hashimoto, S.

    2014-12-01

    Terpenes show high reactivity in the troposphere, contributing to organic aerosol reactions with OH radicals. One of the main sources of terpenes in the atmosphere is terrestrial plants. It has been recently reported that marine phytoplankton also produce monoterpenes (Yassaa et al: 2008). Because aerosol production of natural origin affects the cloud cover over the open ocean, it is important to investigate the origin of aerosol generation in the open ocean. In this study, we investigated the production of terpenes and isoprene with a focus on Chlamydomonas (Chlorophyceae) and Rhodella maculata (Rhodophyta). Concentrations of terpenes and isoprene were measured using a dynamic headspace (GERSTEL DHS)—gas chromatograph (Agilent 6890N)—mass spectrometer (Agilent 5975C). In addition, chlorophyll a was measured using a fluorometer (Turner TD-700). The results showed that isoprene, α-pinene, and β-pinene were produced by Chlamydomonas sp. and that isoprene, limonene, and camphene were produced by Rhodella maculata. Chlamydomonas sp. produced α-pinene and β-pinene, similar to land plants. The ratio of the pinene/isoprene concentrations in the atmosphere over seawater where phytoplankton are blooming has been reported as approximately 0.7 (Yassaa et al: 2008). In this experiment, the pinene/isoprene concentration ratios in the cultures were approximately 0.1. This result indicates that marine phytoplankton may not be ignored in the marine atmosphere chemistry of terpenes.

  1. ISHTAR: Inner shelf transport and recycling in the Bering/Chukchi Seas

    SciTech Connect

    Medeiros, W.H.; Wirick, C.D.

    1991-02-01

    ISHTAR is a multi-disciplinary, multi-university ecosystem study designed to test the hypothesis that interannual changes of atmospheric forcing on water transport through the Bering Strait result in a two-fold to four-fold difference in: (1) the flux of nutrients from the shelf break of the northwestern Bering Sea; (2) the primary production north of St. Lawrence Island; (3) the burial of carbon in Chukchi Sea sediments; (4) the amount of energy passed up the food web; and finally (5) the chemical properties of the Arctic Ocean water transported south across the Greenland-Scotland ridge system. The northern Bering Sea extending from St. Lawrence Island to Bering Strait was chosen as the site for studying phytoplankton populations and their role in transporting organic carbon. The authors major scientific responsibilities in the ISHTAR program are to: (1) provide phytoplankton boundary conditions for models constructed by other components; (2) obtain time series suitable for identifying interannual variations in phytoplankton standing stocks; and (3) identify the dominant time and length scales of the phytoplankton distributions. In addition, they construct, maintain, calibrate, and deploy the moored fluorometers used to make the required measurements. 3 refs., 6 figs., 12 tabs.

  2. In vivo dynamics of enterovirus protease revealed by fluorescence resonance emission transfer (FRET) based on a novel FRET pair

    SciTech Connect

    Hsu, Y.-Y.; Liu, Y.-N.; Wang Wenyen; Kao, Fu-Jen; Kung, S.-H. . E-mail: szkung@ym.edu.tw

    2007-02-23

    An in vivo protease assay suitable for analysis by fluorescence resonance energy transfer (FRET) was developed on the basis of a novel FRET pair. The specifically designed fusion substrate consists of green fluorescent protein 2 (GFP{sup 2})-peptide-red fluorescent protein 2 (DsRed2), with a cleavage motif for the enterovirus 2A protease (2A{sup pro}) embedded within the peptide region. FRET can be readily visualized in real-time from cells expressing the fusion substrate until a proteolytic cleavage by 2A{sup pro} from the input virus. The level of FRET decay is a function of the amount and infection duration of the inoculated virus as measured by a fluorometer assay. The FRET biosensor also responded well to other related enteroviruses but not to a phylogenetically distant virus. Western blot analysis confirmed the physical cleavage of the fusion substrate upon the infections. The study provides proof of principle for applying the FRET technology to diagnostics, screening procedures, and cell biological research.

  3. Velocity, bathymetry, and transverse mixing characteristics of the Ohio River upstream from Cincinnati, Ohio, October 2004-March 2006

    USGS Publications Warehouse

    Koltun, G.F.; Ostheimer, Chad J.; Griffin, Michael S.

    2006-01-01

    Velocity, bathymetry, and transverse (cross-channel) mixing characteristics were studied in a 34-mile study reach of the Ohio River extending from the lower pool of the Captain Anthony Meldahl Lock and Dam, near Willow Grove, Ky, to just downstream from the confluence of the Licking and Ohio Rivers, near Newport, Ky. Information gathered in this study ultimately will be used to parameterize hydrodynamic and water-quality models that are being developed for the study reach. Velocity data were measured at an average cross-section spacing of about 2,200 feet by means of boat-mounted acoustic Doppler current profilers (ADCPs). ADCP data were postprocessed to create text files describing the three-dimensional velocity characteristics in each transect. Bathymetry data were measured at an average transect spacing of about 800 feet by means of a boat-mounted single-beam echosounder. Depth information obtained from the echosounder were postprocessed with water-surface slope and elevation information collected during the surveys to compute stream-bed elevations. The bathymetry data were written to text files formatted as a series of space-delimited x-, y-, and z-coordinates. Two separate dye-tracer studies were done on different days in overlapping stream segments in an 18.3-mile section of the study reach to assess transverse mixing characteristics in the Ohio River. Rhodamine WT dye was injected into the river at a constant rate, and concentrations were measured in downstream cross sections, generally spaced 1 to 2 miles apart. The dye was injected near the Kentucky shoreline during the first study and near the Ohio shoreline during the second study. Dye concentrations were measured along transects in the river by means of calibrated fluorometers equipped with flow-through chambers, automatic temperature compensation, and internal data loggers. The use of flow-through chambers permitted water to be pumped continuously out of the river from selected depths and through the

  4. Physiology and cryosensitivity of coral endosymbiotic algae (Symbiodinium).

    PubMed

    Hagedorn, M; Carter, V L; Leong, J C; Kleinhans, F W

    2010-04-01

    Coral throughout the world are under threat. To save coral via cryopreservation methods, the Symbiodinium algae that live within many coral cells must also be considered. Coral juvenile must often take up these important cells from their surrounding water and when adult coral bleach, they lose their endosymbiotic algae and will die if they are not regained. The focus of this paper was to understand some of the cryo-physiology of the endosymbiotic algae, Symbiodinium, living within three species of Hawaiian coral, Fungia scutaria, Porites compressa and Pocillopora damicornis in Kaneohe Bay, Hawaii. Although cryopreservation of algae is common, the successful cryopreservation of these important coral endosymbionts is not common, and these species are often maintained in live serial cultures within stock centers worldwide. Freshly-extracted Symbiodinium were exposed to cryobiologically appropriate physiological stresses and their viability assessed with a Pulse Amplitude Fluorometer. Stresses included sensitivity to chilling temperatures, osmotic stress, and toxic effects of various concentrations and types of cryoprotectants (i.e., dimethyl sulfoxide, propylene glycol, glycerol and methanol). To determine the water and cryoprotectant permeabilities of Symbiodinium, uptake of radio-labeled glycerol and heavy water (D(2)O) were measured. The three different Symbiodinium subtypes studied demonstrated remarkable similarities in their morphology, sensitivity to cryoprotectants and permeability characteristics; however, they differed greatly in their sensitivity to hypo- and hyposmotic challenges and sensitivity to chilling, suggesting that standard slow freezing cryopreservation may not work well for all Symbiodinium. An appendix describes our H(2)O:D(2)O water exchange experiments and compares the diffusionally determined permeability with the two parameter model osmotic permeability. PMID:19857482

  5. Diurnal changes in epidermal UV transmittance of plants in naturally high UV environments.

    PubMed

    Barnes, Paul W; Flint, Stephan D; Slusser, James R; Gao, Wei; Ryel, Ronald J

    2008-06-01

    Studies were conducted on three herbaceous plant species growing in naturally high solar UV environments in the subalpine of Mauna Kea, Hawaii, USA, to determine if diurnal changes in epidermal UV transmittance (T(UV)) occur in these species, and to test whether manipulation of the solar radiation regime could alter these diurnal patterns. Additional field studies were conducted at Logan, Utah, USA, to determine if solar UV was causing diurnal T(UV) changes and to evaluate the relationship between diurnal changes in T(UV) and UV-absorbing pigments. Under clear skies, T(UV), as measured with a UV-A-pulse amplitude modulation fluorometer for leaves of Verbascum thapsus and Oenothera stricta growing in native soils and Vicia faba growing in pots, was highest at predawn and sunset and lowest at midday. These patterns in T(UV) closely tracked diurnal changes in solar radiation and were the result of correlated changes in fluorescence induced by UV-A and blue radiation but not photochemical efficiency (F(v)/F(m)) or initial fluorescence yield (F(o)). The magnitude of the midday reduction in T(UV) was greater for young leaves than for older leaves of Verbascum. Imposition of artificial shade eliminated the diurnal changes in T(UV) in Verbascum, but reduction in solar UV had no effect on diurnal T(UV) changes in Vicia. In Vicia, the diurnal changes in T(UV) occurred without detectable changes in the concentration of whole-leaf UV-absorbing compounds. Results suggest that plants actively control diurnal changes in UV shielding, and these changes occur in response to signals other than solar UV; however, the underlying mechanisms responsible for rapid changes in T(UV) remain unclear. PMID:18346077

  6. PhotoSpec - Ground-based Remote Sensing of Solar-Induced Chlorophyll Fluorescence

    NASA Astrophysics Data System (ADS)

    Grossmann, K.; Frankenberg, C.; Seibt, U.; Hurlock, S. C.; Pivovaroff, A.; Stutz, J.

    2015-12-01

    Solar-Induced Chlorophyll Fluorescence (SIF) emitted from vegetation can be used as a constraint for photosynthetic activity and is now observable on a global scale from space. However, many issues on a leaf-to-canopy scale remain poorly understood, such as influences on the SIF signal of environmental conditions, water stress, or radiation. Here, we report on the development and characterization of a novel ground-based spectrometer system for measuring SIF from natural ecosystems (http://www.kiss.caltech.edu/study/photosynthesis/technology.html). The instrumental set-up, requirements, and measurement technique are based on decades of experience using Differential Optical Absorption Spectroscopy (DOAS), an established method to measure atmospheric trace gases. The instrument consists of three thermally stabilized commercial spectrometers that are linked to a 2D scanning telescope unit via optical fiber bundles. The spectrometers cover an SIF retrieval wavelength range at high spectral resolution (670 - 780 nm, 0.1 nm FWHM), but also provide moderate resolution spectra (400 - 800 nm, 1.5 nm FWHM) in order to retrieve vegetation indices and the photochemical reflectance index (PRI). In addition to the instrumental set-up, we will show initial results of test and field measurements with the new instrument that examine the diurnal cycle of the SIF signal of different California native and non-native plants and its correlation with CO2 fluxes. Observations were made under different environmental conditions, variable water and nutrient stress, and with different viewing geometries. We also used concurrent observations by a photosynthetically active radiation (PAR) sensor and a portable chlorophyll fluorometer (PAM) to link the SIF signal to plant metabolism and carbon cycling under a range of environmental conditions.

  7. Autonomous Underwater Vehicle Data Management and Metadata Interoperability for Coastal Ocean Studies

    NASA Astrophysics Data System (ADS)

    McCann, M. P.; Ryan, J. P.; Chavez, F. P.; Rienecker, E.

    2004-12-01

    Data from over 1000 km of Autonomous Underwater Vehicle (AUV) surveys of Monterey Bay have been collected and cataloged in an ocean observatory data management system. The Monterey Bay Aquarium Institute's AUV is equipped with a suite of instruments that include a conductivity, temperature, depth (CTD) instrument, transmissometers, a fluorometer, a nitrate sensor, and an inertial navigation system. Data are logged on the vehicle and upon completion of a survey XML descriptions of the data are submitted to the Shore Side Data System (SSDS). Instrument data are then processed on shore to apply calibrations and produce scientifically useful data products. The SSDS employs a data model that tracks data from the instrument that created it through all the consuming processes that generate derived products. SSDS employs OPeNDAP and netCDF to provide data set interoperability at the data level. The core of SSDS is the metadata that is the catalog of these data sets and their relation to all other relevant data. The metadata is managed in a relational database and governed by a Enterprise Java Bean (EJB) server application. Cross-platform Java applications have been written to manage and visualize these data. A Java Swing application - the Hierarchical Ocean Observatory Visualization and Editing System (HOOVES) - has been developed to provide visualization of data set pedigree and data set variables. Because the SSDS data model is generalized according to "Data Producers" and "Data Containers" many different types of data can be represented in SSDS allowing for interoperability at a metadata level. Comparisons of appropriate data sets, whether they are from an autonomous underwater vehicle or from a fixed mooring are easily made using SSDS. The authors will present the SSDS data model and show examples of how the model helps organize data set metadata allowing for data discovery and interoperability. With improved discovery and interoperability the system is helping us

  8. Observations of a phytoplankton spring bloom onset triggered by a density front in NW Mediterranean

    NASA Astrophysics Data System (ADS)

    Olita, A.; Sparnocchia, S.; Cusí, S.; Fazioli, L.; Sorgente, R.; Tintoré, J.; Ribotti, A.

    2014-07-01

    Phytoplankton blooms in the northwestern Mediterranean Sea are seasonal events that mainly occur in a specific area comprising the Gulf of Lion and the Provençal basin, where they are promoted by a general cyclonic circulation, strong wind-driven mixing and subsequent re-stratification of the water column. At the southern boundary of this area, a persistent density front known as the north Balearic front can be found. The front is presumed to cause an early phytoplankton bloom in its vicinity because (a) it enhances the transport of nutrients into the euphotic layer and (b) it promotes the speedy re-stratification of the water column (through frontal instabilities). In February and March 2013, a glider, equipped with a CTD (conductivity, temperature, and depth device) and a fluorometer, was deployed on a mission that took it from the Balearic Islands to Sardinia and back. The frontal zone was crossed twice, once during the outbound leg and the once on the return leg. The data provided by the glider clearly showed the onset of a bloom soon after a decrease in wind-driven turbulent convection and mixing. The in situ observations were supported and confirmed by satellite imagery. It is shown that frontal dynamics play a key role in the promotion and acceleration of re-stratification, which is a necessary pre-conditioning factor for the onset of blooms much like other relevant processes such as an enhanced biological pump. Swift re-stratification stimulates new production by inhibiting mixing. Finally, viewing the blooming phenomenon from a regional perspective, it seems that Sverdrup's critical depth model applies in the northern well-mixed area whereas, in the south, front-related re-stratification seems to be the principal cause.

  9. In situ fluorescence measurements of protein-, humic- and HAP-like materials in the Northwestern Mediterranean Sea

    NASA Astrophysics Data System (ADS)

    Tedetti, Marc; Bachet, Caroline; Germain, Chloé; Ferretto, Nicolas; Bhairy, Nagib; Guigue, Catherine; Besson, Florent; Beguery, Laurent; Goutx, Madeleine

    2015-04-01

    Understanding the biogeochemical functioning of the ocean requires high frequency measurements of dissolved organic matter (DOM) descriptors. For 10 years, the technological developments of fluorescence sensors try to cover this need. In this context, our laboratory developed the MiniFluo-UV sensor, a prototype of miniaturized submersible fluorometer for the detection of aromatic compounds that fluoresce in the UV spectral domain. The qualification of the sensor consisted in measurements of drift, linearity, repeatability, sensitivity to light, temperature and pressure, and detection limits of phenanthrene (HAP) and tryptophan (aromatic amino acid) in standard solutions. Measurements were also conducted in crude oil water soluble fractions (WSFs). The MiniFluo-UV sensor was then deployed in two distinct areas of the Northwestern Mediterranean Sea: 1) in the Gulf of Lion during the continuous monitoring of the surface water layer (DEWEX cruise, winter and spring 2013) and 2) in the Bay of Marseilles, heavily impacted by urban activities, where the sensor was mounted onto the SeaExplorer underwater glider and onto a CTD vertical profiler (July-December 2014). These platforms were also equipped with a humic-like fluorescence sensor and other sensors for hydrological and biogeochemical parameters (T, S, Chla, oxygen, turbidity). The patterns of fluorescence signatures enabled to distinguish interesting distributions of DOM in relation with hydrological features and spring biological production in the Gulf of Lion, and showed the accumulation of contaminants in marine areas under anthropogenic pressure. This work was conducted within the framework of the ANR-09-ECOT-009-01 "IBISCUS" in collaboration with ALSEAMAR-ALCEN (Aix-en-Provence) and MicroModule (Brest) companies. It is relevant to WP5 NEXOS objectives. The SACEUP team of the DEWEX-MERMEX experiment is warmly acknowledged.

  10. Estimating phytoplankton photosynthesis by active fluorescence

    SciTech Connect

    Falkowski, P.G.; Kolber, Z.

    1992-01-01

    Photosynthesis can be described by target theory, At low photon flux densities, photosynthesis is a linear function of irradiance (I), The number of reaction centers (n), their effective absorption capture cross section {sigma}, and a quantum yield {phi}. As photosynthesis becomes increasingly light saturated, an increased fraction of reaction centers close. At light saturation the maximum photosynthetic rate is given as the product of the number of reaction centers (n) and their maximum electron transport rate (I/{tau}). Using active fluorometry it is possible to measure non-destructively and in real time the fraction of open or closed reaction centers under ambient irradiance conditions in situ, as well as {sigma} and {phi} {tau} can be readily, calculated from knowledge of the light saturation parameter, I{sub k} (which can be deduced by in situ by active fluorescence measurements) and {sigma}. We built a pump and probe fluorometer, which is interfaced with a CTD. The instrument measures the fluorescence yield of a weak probe flash preceding (f{sub 0}) and succeeding (f{sub 0}) a saturating pump flash. Profiles of the these fluorescence yields are used to derive the instantaneous rate of gross photosynthesis in natural phytoplankton communities without any incubation. Correlations with short-term simulated in situ radiocarbon measurements are extremely high. The average slope between photosynthesis derived from fluorescence and that measured by radiocarbon is 1.15 and corresponds to the average photosynthetic quotient. The intercept is about 15% of the maximum radiocarbon uptake and corresponds to the average net community respiration. Profiles of photosynthesis and sections showing the variability in its composite parameters reveal a significant effect of nutrient availability on biomass specific rates of photosynthesis in the ocean.